Nature Ecrogrow

During the growing seasons, from April to September, the site received 23.2% of the total precipitation in 2020 and only 2.1% in 2021 . In addition, there was minimal precipitation during data collection of this study from June to September, where only 2 mm and 1.2 mm of precipitation were received in 2020 and 2021. As for the air temperature during the growing seasons, the average maximum air temperature was slightly higher in July, August, and September in 2020 compared to 2021, but lower in March and April. The average minimum air temperature was constantly higher in 2020 compared to 2021 from March until harvest in September, except July. Similarly, the average air temperature was generally higher in 2020 than 2021 except both Julys which had the same average air temperature. As for GDD accumulation , the two seasons were slightly different. In 2020, there was 1525.4°C GDD accumulated when the berries reached 23.9°Brix on average; in 2021, there was 1292.3°C GDD accumulated when the berries reached 22.6°Brix on average. Thus, 2020 was a slightly drier and hotter season than 2021.LAI and crown porosity were assessed in both seasons, and leaf areas were calculated based on the unit ground area and LAI . In 2020, VSP80 had the most leaf area among the six trellis systems, VSP60 and GY had similar leaf areas, 25 liter pot followed by VSP . SH and HQ had the lowest leaf areas as the canopies in these two trellises still had gaps.

This was also confirmed with the fact that SH and HQ had the highest crown porosities among the six trellis systems . The other trellis systems had similar lower crown porosities than SH and HQ. There was no difference in canopy architecture among the three irrigation regimes in the first season . In 2021, all the trellis systems had similar leaf areas . HQ had higher crown porosity than VSP60, but the other trellis systems had similar crown porosities to either HQ or VSP60 . These effects were not modified by the irrigation treatments and no significant interactions between factors were found. For applied water amounts, 50% ETc had higher leaf area than 25% ETc, but there was no difference between 100% ETc with either 25% or 50% . However, 50% ETc still had the highest crown porosity compared to 100% ETc, and 25% ETc did not show any difference with the other two irrigation treatments .Grapevine leaf gas exchange was monitored throughout both seasons, and their integrals were calculated to represent the season-long plant response of grapevines for net carbon assimilation rate , stomatal conductance , and intrinsic water use efficiency . In 2020, there were no differences in gs and An among the six trellis systems . However, HQ had the highest WUEi, whereas VSP, VSP60, and SH had lower WUEi .

Regarding the irrigation treatments, there was no difference in gs integrals . However, a linear response to water amounts were observed for An and WUEi, with 100% ETc having the highest values of both gas exchange variable monitored .In 2021, there were no differences in gs, An, and WUEi among the six trellis systems . Nevertheless, a linear response to water amounts was recorded, with 100% ETc showing the highest An and gs, followed by 50% ETc, and 25% ETc which accounted for a higher WUEi in 25% ETc with 50% treatments compared with 100% ETc . The analysis of the gas exchange recorded at each measurement day indicated that in 2020, despite starting with the highest gs, SH had lower gs over the season . Contrarily, HQ trellis system showed higher gs in July and August which was connected with higher An over the season . On the other hand, GY and VSP80 systems enhanced An during some periods over the season. Regarding WUEi, VSP60 and HQ had the highest values while SH decreased it in the early season and increased it in early August . However, all these differences tended to diminish at the end of the season. For irrigation treatments, a constant effect of water amount was observed with 100% ETc increasing gs and An and decreasing WUEi . In 2021, GY and VSP60 showed higher gs and An values in general . HQ showed lower gs values and VSP had lower An values compared to the other trellis systems throughout the season. HQ increased WUEi throughout the whole season . Although GY had higher WUEi in the early season, it showed constantly lower WUEi values after 23 June 2021.

Besides GY, VSP showed lower WUEi in July and August. A similar effect of irrigation treatments was observed over the second season, with a linear response for increased gs and An and decreased WUEi when the irrigation water amount was increased .Yield components and berry quality parameters were assessed at harvest in both seasons . SH and HQ had the smallest berries among the six trellis systems in the two seasons. In 2020, SH and VSP increased the cluster number, while VSP80 and GY decreased it whereas, in 2021, SH and HQ accounted for increased the cluster number. VSP, VSP60, VSP80, and GY increased the cluster weight compared to SH in 2020. In 2021, SH showed the lowest cluster weight and skin weight. Regarding yield, differences were only significant in 2020 where SH enhanced vine yield compared to the other trellis systems. On the other hand, 100% ETc enhanced berry weight, cluster weight, and yield over the two seasons with no difference on leaf area to fruit ratio. Regarding berry quality parameters, SH had the highest TSS among and the lowest pH in 2020, whereas in 2021, VSPs and GY enhanced the TSS and the pH. Results also showed that irrigation treatments had little effect on the berry quality parameters over the two seasons with only TSS being increased in the 25% ETc treatment in the harvest of 2020.Different trellis systems affected not only the total anthocyanin concentration but also modified the anthocyanin composition, leading to modifications in the profile stability. In both seasons, SH had the highest concentrations in all the anthocyanin derivatives besides di- and tri-hydroxylated anthocyanins among the six trellis systems. In 2021, HQ also notably increased most of the anthocyanin derivatives, tri-hydroxylated, di-hydroxylated, and total anthocyanins compared to the VSP trellis systems. On the other hand, VSP trellis systems tended to decrease the anthocyanin concentrations. Regarding the irrigation treatments, 25% ETc generally showed the higher concentrations in petunidins, di- and tri-hydroxylated anthocyanins, and total anthocyanins in 2020 compared to 100% ETc. In 2021, 25% ETc increase most of the anthocyanin concentration in berries. 50% ETc performed similarly in 2021 and showed higher concentrations in malvidins, trihydroxylated anthocyanins, and total anthocyanins. In parallel with anthocyanin assessments, berry skin flavonols were measured at harvest in both seasons . In 2020, SH showed the highest concentration in myricetins. SH and HQ showed the highest concentrations in quercetins, isorhamnetins and kaempferols in both seasons. SH and HQ also showed the highest concentration in tri- and di-hydroxylated as well as total flavonols in both seasons. In 2020, there were no differences among the six trellis systems in laricetins and syringetins. While in 2021, VSPs enhanced syringetin concentration. Regarding applied water amounts, raspberry cultivation pot little effects of irrigation treatments were shown in 2020. However, in 2021, 25% ETc increased most of the flavonol derivatives except laricetins and syringetins compared to the other two treatments.The relationships between berry skin flavonol concentrations and canopy architecture were investigated in both seasons. In 2020, crown porosity had positive and significant correlations with quercetin , total flavonol concentration , R2 = 0.248, p< 0.0001, Figure 5Aa-2), and total flavonol concentration . Leaf area was also correlated with these variables, but the correlations were negative with quercetin , total flavonol concentration . In 2021, the correlations were similar but not as significant as 2020. Crown porosity still had significant and positive relationships with quercetin and total flavonols concentration . However, the relationship between crown porosity and total flavonol concentration did not persist, as was observed in 2020. The relationships between leaf area and quercetin and total flavonol concentration were significant, although not as strong . Leaf areas were negatively correlated with these two variables . The significant correlation between leaf area and total flavonol concentration did not hold in 2020 as compared to 2020 .

It was evident that when crown porosity was greater, there was greater flavonol accumulation as well greater molar percentage of quercetin.A trellis system selected in grapevine vineyard is usually aimed at optimizing canopy architecture to further maximize canopy photosynthetic activity and improve canopy microclimate, which can yield desirable production and berry composition . In historically cooler regions according to Winkler’s Index, VSPtrellis system is widely used as it offers relatively higher compatibility with mechanization and is suitable for the regional production goals . However, with the warming trend in air temperature getting more pronounced, VSPs have been showing greater chances of getting cluster overexposure, resulting in sunburnt berries with yield loss and color degradation . Under our experimental conditions, HQ trellis system showed less leaf area and greater crown porosity than the other trellis systems in 2020 in accordance with previous studies, where split trellis designs might allow more solar radiation to penetrate the canopy interior . Conversely, SH had similar leaf area but lower crown porosity in 2020. However, the differences in leaf areas and crown porosities were not as noticeable in 2021. This could be attributed to the fact that the HQ and SH might have still been filling up spaces with new growth compared to the VSPs, which might have already had relatively more established canopy architectures. In addition, the differences in leaf areas and crown porosities could be minimized by arid growing season in 2021, despite the supplemental irrigation applied to them, accounting for diminished leaf areas as shown in 2021 than 2020. Furthermore, precipitation received at the vineyard prior to bud break , as well as precipitation received immediately prior to flowering in semi-arid regions were deemed key determinants of canopy response for latter parts of the growing season.In this study, yield per vine was not constantly determined by the trellis systems in both years, although similar bud densities at pruning were achieved. Furthermore, more leaf area did not account for more yield at harvest, despite it was well established that a sufficient leaf area would support fruit development , and in contrast with some previous studies . This could be attributed to not only the total amount leaf area but also how the leaves were distributed within the canopy. Commonly, HQ would have more open space to distribute more exposed and photosynthetically active leaves to the sunlight to optimize production . Previous studies have shown that greater leaf area can also contribute to higher TSS accumulation , which was not observed in this study. On the contrary, more leaf area resulted in less TSS accumulation. This might be explained by the fact that the leaf area to fruit ratio, which represented the source-sink balance within the grapevine, might have a greater influence on the berry TSS accumulation. In this study, even though no statistical differences were observed in leaf area to fruit ratio, SH in 2020 showed relatively higher values with higher TSS accumulated at harvest. A similar situation was observed during the second season, where VSP80 showed relatively higher leaf area to fruit ratio and subsequently higher TSS at harvest. When crown porosity was considered, higher porosity resulted in greater TSS accumulation in berries, which could be attributed to the higher potential of berry exposure to the hot environment, causing the berries to experience greater dehydration . This relationship was not observed in 2021, and it might be derived from the relatively higher crop load and lower leaf area to fruit ratio in 2021 compared to 2020, especially by SH and HQ . SH and HQ did not have a similar source-sink balance as 2020, which lowered their capacity to translocate photosynthates into the berries, this might have been the reason why they had a more reduced TSS at harvest compared to the other trellis systems. Regarding the applied water amounts, the results were clear and consistent, with increased water status in grapevines irrigated with higher water amounts, and consequently, greater berry weight, cluster weight, and yield.

In a multi-center, double-blind, randomized controlled trial of very-low-birth-weight infants, no difference was found in the incidence of ROP between those supplemented with daily oral L and Z or placebo. However, the progression rate of threshold ROP showed a lower trend in the supplemented group. No adverse events were noted with L and Z supplementation, suggesting that they were well-tolerated. Another study examining the effects of daily oral L and Z supplementation in preterm infants from the seventh day after birth until 40 weeks of age or until hospital discharge found no change in the rate or severity of ROP compared to placebo. Further, a meta-analysis of three randomized controlled trials also found no protective association between L and Z supplementation and the risk of ROP. Additional studies are needed to assess the role of prenatal L and Z supplementation in pregnant women at risk for premature delivery. During development, L and Z are not interchangeable. Serum Z in newborns and in their mothers is strongly correlated with the MPOD of the babies, but no relationship was noted for either maternal or infant levels of L. During delivery, a high maternal plasma Z, but not plasma L, was significantly associated with a lower risk of visual acuity problems in children at three years of age. Further investigations that can accurately distinguish and quantify dietary and plasma Z from L are needed to better understand the role of these two carotenoids in visual performance during development.

The L and Z in human milk is particularly important for infant eye and brain development, grow bucket and may provide long term benefits to vision and cognition. Since humans cannot synthesize carotenoids, the fetus and breastfed infants must obtain these compounds from the mother through the placenta and the breast milk. During gestation, maternal lipoprotein synthesis increases, which accelerates the transport of carotenoids to the fetus. This transfer may deplete maternal stores if the dietary intake of carotenoids in general, and L and Z specifically, is inadequate to maintain body stores. The prevalence of AMD is higher in women than in men, even though on a global basis more men smoke. At the same time MPOD levels are lower in females. The potential reasons for an increased lifetime risk for AMD in women are complex and multifactorial in nature and may include maternal depletion of L and Z during pregnancy and lactation . Importantly, the average dietary consumption of L and Z among females in the US is far below the amount of 10 mg/d known to increase MPOD. Therefore, either the intake of supplements containing L and Z, or increased intake of foods rich in these two carotenoids for the duration of pregnancy and lactation may be of value. The concentration of β-carotene, lycopene, L and Z, the main carotenoids in breast milk are associated with maternal dietary intake over the first six months of lactation. Daily maternal supplementation of either 6 mg of L with 96 µg of Z, or 12 mg of L with 192 µg of Z, over six weeks resulted in a dose-dependent increase in L and Z levels in the breastmilk and of the mothers and their infants when assessed three to four months postpartum. 

Another study reported that more carotenes were present than xanthophylls in maternal plasma, whereas more xanthophylls such as L and Z were presented in breast milk, in comparison to carotenes. These findings support the notion that maternal-infant transfer of carotenoids may occur, possibly at the expense of the mother. Future studies are needed to clarify if breastfeeding or L and Z intake may impact their AMD risk. The L-ZIP supplementation trial is currently exploring whether prenatal supplementation of 10 mg of L and 2 mg of Z will maintain maternal body stores, prevent potential macular pigment depletion during pregnancy, or enhance systemic and ocular carotenoid stores for both mothers and infants. Clinical trials on the long-term effects of perinatal L and Z intake on MPOD changes among mothers and infants are also warranted.100 Unfortunately, longitudinal studies on AMD in females often do not include breast-feeding history. A useful study design would be to investigate MPOD levels and relative risks of AMD between multi-parous and nulliparous women, and in mothers practicing breastfeeding compared to formula feeding. Dietary intake of L and Z would be important to assess. Recognizing that such a study would take decades, shorter term studies could be conducted in non-human primates. Another challenge in retrospective studies is that breastfeeding history may not be accurate. Therefore, studies on the maternal transfer of L and Z during pregnancy and lactation with MPOD changes in infants and throughout the lifespan, could be important but difficult to conduct.

Future research should also focus on the measurement of L and Z status and MPOD in mother-infant pairs of twins or short birth intervals. Last, when accessing AMD risk in women, reproductive hormone status may be a confounding factor. The pathogenesis of AMD involves oxidative stress and immune dysregulation.Estrogen has been shown to reduce oxidative stress and inflammation in RPE cells as well as systemically. Lifetime estrogen exposure such as the number of pregnancies, menopause, reproductive period, oral contraceptive use, and hormone replacement therapy may all influence the risk of developing AMD. Current evidence regarding estrogen exposure and risk of AMD is inconsistent. One study reported that postmenopausal hormone use decreased the risk of neovascular AMD but increased the risk of early AMD, while parous women showed a reduced risk of early AMD but not neovascular AMD. Two nationwide studies from South Korea among postmenopausal women noted that exogenous estrogen exposure was not a protective factor for AMD. A cohort study found that hormone replacement therapy and a longer reproductive period was associated with an increased risk of neovascular AMD. A cross sectional study showed that oral contraceptive use was associated with an increased risk of late AMD.106 In addition, a review summarizing the effect of estrogen exposure and the risk of all age-related eye diseases concluded that HRT, or the use of oral contraceptives, could be either positively or negatively associated with the risk of AMD.103 In contrast, some studies have reported that a longer duration of breastfeeding may be protective from late but not early AMD,106,107 even when the estrogen level was low during lactation. Future studies on the interaction of different reproductive and estrogen exposure histories and AMD risk are needed. Humans cannot synthesize carotenoids, and the best dietary sources are fruits, vegetables, egg yolks, and dairy products. Consuming a diet rich in green leafy vegetables and fish is recommended by the National Eye Institute for the high carotenoid and DHA and EPA contents. Nevertheless, in the carotenoid group, L and Z are not yet considered essential, or even conditionally essential, so no dietary reference intakes for these two compounds exists. The US intake of L and Z has been decreasing. According to the U.S. National Health and Nutrition Examination Survey , the average intakes of L plus Z were 2.15 mg/d in males and 2.21 mg/d in females in 1987, and 2.15 mg/d in males and 1.86 mg/d in females in 1992. In NHANES 2013-2014, dutch bucket for tomatoes the average intakes of L and Z in males and females was 1.58 mg/d and 1.76 mg/d, respectively. Moreover, based on data from NHANES 2003-2004, the reported intake of L was significantly higher than Z in all age groups and ethnicities. Importantly, the Z to L ratio was also lower in females than males older than 31 years of age, which may result in a higher risk of AMD in women than in men. However, due to difficulties in analyzing dietary L and Z separately, most studies analyze both carotenoids together. Since the amount of L in most foods is significantly greater than Z, precise quantification of Z has been a challenge. The amount of L and Z in foods and dietary supplements appears to be safe. No adverse events were found in clinical trials giving L at 30 mg/d for 120 days or 40 mg/d for 63 days. The only reported adverse effect after a daily supplementation of 15 mg L in a 20-week trial was a single case of self-reported carotenodermia, a reversible condition of orange skin color. Although a higher amount has been used in human studies, after assessing the potential risks, the observed upper safety level for L has been proposed as 20 mg/d. 

The European Food Safety Authority concluded safe upper limits for L and Z for use in dietary supplements were 1 mg/kg body weight/d and 0.75 mg/kg body weight/d, respectively. In primate models, rhesus monkeys fed a xanthophyll-free diet for 3 to 6.5 years developed extremely deficient or absent macular yellow pigment and drusen-like bodies. When 3.9 µmol/kg per day of L or Z for 24 to 101 weeks were supplemented, the rhesus monkeys showed significant increases in their corresponding serum, retinal, and adipose tissue concentrations. In their retina samples, L and meso-Z, but not Z, appeared in the L-supplemented group, while only Z was found in the group supplemented with Z. In humans, a study investigating the serum and macular responses of L, Z, and meso-Z from dietary supplements found that 13.13 mg/d for 12 weeks provided maximum MPOD improvement, whereas 7.44 mg/d was the amount that increased serum levels at the highest efficacy. These and other studies support the need for dietary recommendations for L and Z, particularly as conditionally essential nutrients due to their protective effects on eye health. Lutein and Z fulfill many criteria as essential nutrients, including high concentrations in select tissues, biological plausibility for eye health, depletion outcomes such as vision impairment in primates, and inverse associations with certain diseases. In addition to their role in eye health, L and Z are involved in cognitive function at all stages of life. A randomized controlled trial reported that L and Z supplementation improved neural efficiency and learning performance by increasing the interaction of numerous brain regions in older adults. Other reports have demonstrated an association between L intake or circulating levels and preserving age-related cognitive decline, reducing the risks of certain cancers, coronary heart disease, stroke, metabolic syndrome, and achieving higher levels of physical activity. A systematic review of in vivo, ex vivo, and in vitro studies concluded that L may benefit vascular health by improving endothelial function, reducing inflammation, regulating favorable lipid profiles, and maintaining glucose homeostasis. Systematic reviews summarizing the amount of L needed for cognitive functions and enhancement of gray matter volume estimated that at least 10 mg/d for 12 months could be beneficial. Age must be considered when creating DRI values, since MPOD values are lower in older compared to younger individuals. Whether the proposed intake of L and Z should be based on the amount that can reduce AMD risk, benefit visual maturation in newborns, protect cognitive health, or reduce the risk of other diseases requires further consideration. Sex differences in AMD prevalence must also be considered, especially in relation to pregnancy and lactation, as discussed above. Nevertheless, L and Z are not included for DRI consideration due to inadequate details from food databases, limited large-scale dietary intake studies, and insufficient knowledge regarding their metabolism and biological functions. Many continue to advocate for a DRI for L, since it satisfies all nine criteria for bio-active compounds. A rich dietary source of L and Z is goji berry, also called wolf berry or Gou Qi Zi. The bright orange-red colored oval fruit, has been used for millennia in traditional Chinese medicine for its role in visual health, to provide immunoregulatory, neuro-protective, and anti-inflammatory benefits, and to help regulate liver and kidney meridians . Commercially-available goji berries and their products come primarily from the Ningxia and Xinjiang autonomous regions in western China. Goji berry is known for its high amount of carotenoids, with the Z content higher than any other known food. In addition to carotenoids, other bioactive compounds found in goji berries include Lycium barbarum polysaccharides , flavonoids, vitamins, minerals, betaine, cerebrosides, phenolic acids, and certain amino acids which may also support the overall health of the eye, particularly when working synergistically. 

All the architectures they review are modular with the low level control functions deployed on a microcontroller and higher level tasks performed by various systems—either ROS or a custom-designed middleware. The real-time control in two cases use a HAL, the others do not. Within the Autonomous Systems Laboratory there is an important related work called the Santa Cruz Low-Cost UAV GNC Subsystem, which was an early autopilot intended for UAVs. It was programmed using the embedded code generation capability of Matlab and Simulink. This project, though successful, was never widely adopted. One reason is that Matlab and Simulink are expensive commercial software packages and come with a significant learning curve. Another is that the project wasn’t open source limiting access to vehicle developers or potential contributors. These limitations motivate this work which is open source, both hardware and firmware.This dissertation is organized as follows. Chapter 2 is a review of the requirements for the real-time controller. Chapter 3 details the hardware development process, architecture, electrical design, and the final controller. Chapter 4 details the software design, architecture, and development process. In Chapter 5 we propose a benchmark algorithm to quantify the performance of the real-time controllers. Chapter 6 shows the results of using the benchmark on four different processors, three real-time systems using micro-controllers and one SBC running Debian Linux. Chapter 7 details the development of an AGV from the chassis to autonomous navigation used as the main test platform for the controller. Chapter 8 covers three new cases for different autonomous systems that use this architecture or are in the process of deploying it. Chapter 9 demonstrates several areas where this work can be extended. Finally, Chapter 10 provides the conclusions.

There are two supplementary appendices.Appendix A is a derivation of the complementary filter, nft growing system the basic algorithm behind the benchmark. Appendix B details the application of the autoregression with external inputs method for system identification using a simple motor model as an example. The primary motivation for this work is to provide a means for an autonomous vehicle developer to rapidly prototype the real-time guidance, navigation and control system of a new vehicle. To enable this we decided at the inception of the project to make the design and firmware open source and named it the Open Source Autonomous Vehicle Controller, or OSAVC for short. The advantages provided by open source methodologies to a would-be vehicle developer are that the designs and algorithms are freely accessible, they are easily modifiable , and can be sourced from numerous vendors. From the project side open sourcing allows contributors from all over the globe to add new functionality, to test and provide feedback, and iterate on the designs. Indeed, during the development of this project we were able to participate in the Google Summer of Code program which hires student interns to contribute to open source projects. Over the course of two summer sessions, the OSAVC project received over 45 applications from students all over the globe, and was able to provide six paid internships. Additionally, the OSAVC prototype was used by two other graduate researchers for an ASV and a quadcopter which was an early demonstration of the capability and modularity of the project. The OSAVC provides real-time control of the vehicle actuators and measurement of the sensors. In the context of control systems, real-time refers to a fixed output period of the controller, i.e., the period between updating the vehicle actuators is deterministic and constant. All modern control systems are digital, that is they do not provide continuous outputs, instead they update the outputs on a fixed clock cycle. The systems under control, however, exist in the continuous real world.

If the controller clock cycle, τc, is short compared to the dynamics of the vehicle, then the digital controller approximates a continuous controller. In other words, if the rise time1 of a system in response to a step change in input is τv, then the digital control system approximates the continuous case when τc << τv. In the frequency domain a usual figure of merit is that the update rate is 1/20th-1/30th the bandwidth of the system. A finite clock cycle introduces a lag in the response of the controlled system. In addition to having as small a lag as is practical, it is also important to minimize the variation in the clock cycle. Small variations are inevitable but larger variations can lead to poor control or, in the worst case, instability. Not all tasks in an autonomous system require real-time control. For example, logging of telemetry data for post-mission use does not require a fixed clock—the system can buffer the data until it is convenient to store it. Thus, when designing a distributed control system it is natural to divide tasks into real-time and non-real-time categories. Real-time tasks are the domain of the real-time controller and all others belong to a processor with an OS. Note that some control systems attempt to handle all tasks within the same processor by using an RTOS. We discuss the tradeoffs of this type of OS in Section 2.6.1. Strictly real-time tasks are ones that support the control of the vehicle. These comprise the measurement of all the inputs and computation of the algorithms necessary to provide the actuator output. These include any sensor that is used to estimate the vehicle’s state . A non-comprehensive list of these sensors include wheel encoders, airspeed sensors, GPS, gyroscopes, accelerometers, magnetometers, barometers, etc. Accordingly, the navigation tasks, which include state estimation, are also real-time as they provide the input to the controller.

Although mapping of the environment isn’t a strictly real-time task, simultaneous localization and mapping is, as it provides vehicle state information . Communications generally do not have real-time requirements with the exception of remote control, when the system is operated manually using a radio. Finally, the controller itself must operate on a fixed cycle. All other tasks are non-real-time. Note that this doesn’t imply that they are not time-sensitive. For example, a range sensor might be used for obstacle avoidance, or a camera may be used to identify landmarks in the environment. In both cases, these data are needed to support guidance of the vehicle. Guidance is the process of determining the vehicle trajectory and therefore must be done quickly, but not necessarily on a fixed clock. Generally, guidance tasks are not real-time but are typically time sensitive. Other tasks, such as logging of data, measurement of sensors not used for state estimation, and general communications are non-real-time. A generic taxonomy of the tasks of an autonomous system is summarized in Table 2.1 showing the classification of real-time versus non-real-time as discussed above.From a high level, the OSAVC should support the main hardware and communication interfaces used by commercially-available components. The components needed for a vehicle will include sensors, radios , and actuators . Additionally, as a real-time subsystem, vertical hydroponic nft system the OSAVC module should communicate efficiently and quickly to the larger vehicle control system. For example, a vehicle may have an onboard guidance and navigation SBC that requires periodic communication of the vehicle state variables in order to plan a route. The module should support the real-time requirements of latency and speed for a vehicle. That is, it should be able to operate as quickly as needed by the vehicle and meet the deterministic latency requirement for the control algorithms. A final requirement is that the OSAVC provide power, signal conditioning, and power management for the sensors and processor, and optionally for any external peripherals or modules . The most common hardware interfaces are the inter-integrated circuit interface, the serial peripheral interface , the universal serial bus , and the universal asynchronous receiver-transmitter —commonly known as a serial port. Less common but used extensively in automobiles and robotics is the controller area network bus . Other peripherals needed for the OSAVC are hardware timers, input capture modules —used for decoding incoming digital signals, and output compare modules used for generating pulse-width modulated signals needed for motor and servo actuator control. Finally, general purpose input-output pins are useful for turning on LEDs, setting switches, and other requirements. The OSAVC should have most or all of these hardware peripherals in order to support the broadest range of applications; while tradeoffs can be made with respect to capability versus size, weight, and power, we typically fall on the side of greater capability in this work. The firmware is written entirely in the C programming language for speed and efficiency. The most important requirement of the firmware is modularity. Each sensor driver, estimation algorithm, and control algorithm is modular. Each module has a header file which provides the interfaces to public methods and a source file containing the code; each source file has a built-in test harness to allow for module troubleshooting outside of an application.

To maximize efficient utilization of the processor each sensor module is non-blocking. This requirement forces the use of vectored interrupts and each sensor has a unique interrupt priority assigned to it. The firmware library contains a selection of commonly used sensors needed for vehicle autonomy both to provide a minimum of functionality for a new developer as well as a template for new sensor drivers. The application itself should be as simple as possible to achieve the desired control. An example application for an AGV will be provided as a template for other applications. The application is a bare metal application consisting of an infinite loop with a hardware timer for task scheduling. This avoids the complexity and overhead of an RTOS. This requirement merit some discussion and is covered in Section 2.6.1. An RTOS is a piece of software that brings in real-time and non-real-time tasks into the same controller. It uses a HAL that allows for programmers with no expertisein embedded programming to communicate with various sensors without any understanding of the underlying hardware. The difficulty in using an RTOS lies in modifying the real-time tasks. Even with well-structured code, debugging the latency of a new realtime algorithm can be challenging. Autopilot firmware running on an RTOS typically implements the algorithms with tuneable parameters . This may work well for many cases, but modifying an algorithm for faster operation or for different parameters is challenging. Developing a new algorithm is even more difficult. Of course any OS increases the complexity of an application as well using more computational resources. A bare metal application, on the other hand, allows for strict control over the sensing and control elements of an autonomous system. Measuring the latency is straightforward using a hardware timer of the processor, as is managing processor bandwidth. Debugging the function is similarly simplified. The challenge in programming on bare metal is the need for an understanding of the hardware in order to configure it properly, although device manufacturers often offer configuration apps or software development kits to ease this process. Another benefit of bare metal programming is efficiency due to the much lower overhead. The main downside for bare metal applications is the challenge in duplicating the common features of a standard OS. For this reason, a bare metal application is suited better for a distributed architecture where the real-time and non-real-time tasks are strictly segregated between the real-time core and the SBC as discussed earlier. The design of the OSAVC PCB consisted of two development efforts. As we had no prior experience designing a PCB we decided to start with a ‘daughter board’—a simpler I/O PCB that could interface with a commercially available development board1 . The first daughter board was hand-wired to determine an initial layout and to kick start the development process. This allowed us to develop PCB layout and assembly skills and at the same time acted as a platform to test sensor driver modules. The I/O board—a simple two layer PCB—required two revisions before its performance was satisfactory. We also selected a vehicle platform for the AGV test bed on which to validate OSAVC. To complement the AGV we purchased a suite of sensors that are widely used for small autonomous vehicles and proceeded to develop a library of sensor drivers and integrated the sensors onto the vehicle. The discussion of the software design is found in Chapter 4.

Viruses disrupt the plant cell cycle, inhibit cell death pathways, restrict macromolecular trafficking, alter cell signaling, protein turnover, and transcriptional regulation, and suppress defense mechanisms. The interference with these processes in the host leads to a wide range of plant developmental and physiological defects . Cultivated grapevines are highly susceptible to a variety of viruses and viroids, which cause significant crop losses and shorten the productive life of vineyards. More than 65 different viral species classified in at least 15 families have been reported to infect grapevines, which represents the highest number of viruses so far detected in a single cultivated plant species . Although these viruses are generally transmitted by plant-feeding insects or soilborne nematodes, they can also be spread through infected propagation material . Grapevine red blotch is a viral disease discovered in northern California in 2008 that has become a major economic problem for the wine industry in the USA . This disease is caused by the Grapevine red blotch-associated virus , a circular ssDNA virus with resemblance to geminiviruses, which infects wine grape cultivars with significant detrimental effects on productivity . The incidence and severity of the red blotch symptoms vary depending on the grape cultivar, environmental conditions, and cultural practices . In red-skinned varieties, GRBaV infections result in the appearance of red patches on the leaf blades, veins, and petioles; in white-skinned varieties, hydroponic bucket they manifest as irregular chlorotic regions on the leaf blades.

GRBaV also affects berry physiology, causing uneven ripening, higher titratable acidity, and lower sugar and anthocyanin content, among others . Consequently, must and wine produced from infected berries present altered flavor and aroma. To date, there is limited information on how GRBaV infections affect grape metabolism. Comprehensive analyses to study specific cellular processes that GRBaV exploits to promote infections in berries are still needed, in particular those that relate to changes in berry chemical composition during fruit development. Grape berry development exhibits a double sigmoid growth pattern with three distinct phases: early fruit development, lag phase, and berry ripening. Most metabolic pathways that promote desired quality traits in grape berries are induced during ripening. The onset of ripening is accompanied by significant changes in berry physiology and metabolism, including softening, sugar accumulation, decrease in organic acids, and synthesis of anthocyanins and other secondary metabolites that define the sensory properties of the fruit . Berry ripening is controlled by multiple regulatory pathways, and occurs in an organized and developmentally timed manner. Interactions between transcriptional regulators and plant hormones regulate the initiation and progression of ripening processes . Like other non-climacteric fruit, grape berries do not display a strong induction of ethylene production and respiration rate at véraison, and the activation of ripening events does not depend primarily on ethylene signaling.

Even though the hormonal control of grape berry development is not completely understood, it is established that abscisic acid , brassinosteroids, and ethylene are positive regulators of ripening processes, while auxin delays the initiation of ripening . In the context of virus–grape berry interactions, dissecting the mechanisms that regulate ripening and plant defenses may provide new opportunities to develop vineyard management strategies to control viral diseases and ameliorate the negative effects on berry quality. In this study, we integrated genome-wide transcriptional profiling, targeted chemical and biochemical analyses, and demonstrated that grapevine red blotch disrupts ripening and metabolism of red-skinned berries. We sampled berries at different ripening stages from vines infected with GRBaV and healthy vines in two vineyards. We identified grape metabolic pathways that were altered in ripening berries because of the viral infection. We determined that GRBaV-induced pathways that are normally associated with early fruit development in berries at late stages of ripening, and suppressed secondary metabolic pathways that occur during normal berry ripening and/ or in response to stress. Using targeted metabolite profiling and enzyme activity analyses, we confirmed the impact of GRBaV on phenylpropanoid metabolism. We identified specific ripening-related processes that were disturbed in GRBaV-infected berries. Remarkably, these processes included alterations in ripening regulatory networks mediated by transcriptional factors, post-transcriptional control, and plant hormones, which lead to berry developmental defects caused by red blotch.

To determine the impact of grapevine red blotch on berry physiology, we studied naturally occurring GRBaV infections in distinct wine grape-growing regions in northern California . We sampled red-skinned grape berries from two different vineyards, one in Oakville and one in Healdsburg . We used multiple vineyard sites to focus on observations consistently made across environments and, thus, to exclude factors associated with specific environmental or cultural conditions. Prior to sampling, vines were screened for the presence of GRBaV and other common grapevine viruses. The appearance of red blotch symptoms on leaves of GRBaV-positive vines and not on those of healthy controls confirmed the initial viral testing. We sampled grape berries from vines that tested positive for GRBaV and negative for other common grapevine viruses. At the same time, we also collected berries from vines that tested negative for all viruses and included them in the study as healthy controls. In order to determine the impact of the disease on berry development and metabolism, we collected GRBaV-positive and control berries at comparable developmental stages: pre-véraison , véraison , post-véraison , and harvest . This sampling strategy also aimed to limit confounding effects due to differences in the progression of ripening between berry clusters of GRBaV-positive and healthy vines. In some cases, we observed that GRBaV-positive vines presented grape clusters with evident uneven ripening . Comparisons between berries from GRBaV-positive vines and healthy controls indicated that, at equivalent stages of development, berries affected by red blotch had reduced soluble solids and total anthocyanins in agreement with previous reports on red-skinned wine grapes . Sampled berries were used for genome-wide transcriptional profiling of viral and grape genes. RNAseq was performed using 3–4 biological replicates of each ripening stage, infection status, and vineyard. We first confirmed the presence of the virus in the berries of GRBaV-positive vines by qPCR amplification of viral DNA . Viral activity in the berries was also assessed by quantifying plant-derived mRNA transcripts of GRBaV genes in the RNAseq data. Plant expression of five out of the six predicted genes in the GRBaV genome was detected in all berry samples obtained from GRBaV-positive vines but not in berries collected from the control vines . The most expressed GRBaV genes in the berries corresponded to V1, which encodes a coat protein, and V3 with unknown function. Expression levels of the GRBaV genes appeared to change as berries ripened. However, we could not determine to what extent the progression of ripening or other environmental factors influenced the plant’s transcription of viral genes because their pattern of variation between ripening stages differed in the two vineyards . Expression of 25 994 grape genes was detected by RNAseq across all berry samples. Principal component analysis was carried out with the normalized read counts of all detected genes. The two major PCs, which together accounted for 42.97% of the total variability, clearly separated the samples based on ripening stage, regardless of the vineyard of origin or their infection status . These results indicated that the inter vineyard variation was smaller than the ripening effect, stackable planters and the overall progression of ripening was similar between berries from GRBaV-positive and control vines. Therefore, we hypothesized that GRBaV infections in berries have altered the expression of particular grape genes and/or molecular pathways, which could subsequently have led to developmental and metabolic defects.While the PCA described above indicated that overall transcriptome dynamics associated with berry ripening were not perturbed by the infection, the lower levels of soluble solids and anthocyanins in GRBaV-positive berries, particularly later in development, suggested that red blotch may affect specific primary and secondary metabolic processes. We therefore focused the RNAseq analyses to identify grape molecular pathways that were differentially regulated as a result of GRBaV infections. We identified grape genes with significant differential expression due to red blotch by comparing GRBaV-positive and GRBaV-negative berries at each ripening stage and independently for each vineyard.

We then looked at the intersection of differentially expressed genes between the two vineyards to identify common responses to red blotch. A total of 932 grape DE genes were found to be consistently down- or up-regulated in infected berries in both vineyards at a given ripening stage, and were classified as GRBaV-responsive genes . On average these GRBaV-responsive genes showed 0.49 ± 0.22-fold changes compared with the healthy controls. Comparing berries at similar ripening stages may have contributed to exclude more dramatic changes in gene expression associated with more pronounced ripening delay due to GRBaV. Key metabolic processes that were suppressed or induced as a consequence of red blotch in ripening berries were identified by enrichment analyses of the functional categories defined by Grimplet et al. in the set of GRBaV-responsive genes . Amino acid biosynthetic pathways were repressed in GRBaV-positive berries, while amino acid catabolic pathways were induced. Changes in carbohydrate metabolism were also observed; in particular, genes involved in glycolysis/gluconeogenesis and starch metabolism had reduced expression in GRBaV-infected berries. Most enzymes involved in phenylpropanoid metabolism are encoded by large gene families. There is also high redundancy among these genes, which ensures the functional integrity and plasticity of the phenylpropanoid-related pathways . Therefore, to test the hypothesis that the red blotch-induced transcriptional changes had an actual impact on phenylpropanoid metabolism, we measured the activity of key enzymes and the abundance of compounds involved in these pathways . We detected significant reductions in activity of seven enzymes that catalyze important steps in the core phenylpropanoid, stilbene, flavonoid, and anthocyanin biosynthetic pathways due to GRBaV infections of berries at three ripening stages . In addition, the first enzyme committed to flavone and flavonol biosynthesis, flavonol synthase , had significantly lower activity at post-véraison and harvest stages . Red blotch altered the accumulation of 17 compounds that result from the phenylpropanoid metabolism and two compounds upstream of this pathway, shikimic acid and gallic acid . Most of these compounds showed significantly lower abundance in the GRBaV-positive berries compared with the controls at later stages of ripening. The main anthocyanins present in grape berries: malvidin-3-O-glucoside, petunidin-3-O-glucoside, delphinidin-3-O-glucoside, pelargodin-3-O-glucoside, and cyanidin-3-O-glucoside, were significantly reduced by red blotch at harvest. Gallic acid, sinapic acid, and quercetin also showed lower abundance in infected berries. Few exceptions to this general suppression of phenolic accumulation were the accumulation of the precursor shikimic acid, which significantly increased in infected berries at harvest, and of resveratrol that showed significantly greater accumulation at véraison and postvéraison. Additional experiments are necessary to understand the accumulation of these two metabolites in the presence of GRBaV: preliminarily, we can hypothesize that the higher abundance of resveratrol is due either to a restriction of subsequent enzymatic steps in stilbene metabolism for which this compound is a substrate, or to the enzymatic hydrolysis of resveratrol glycosides or stilbenoid dimers previously synthesized. The integrated analysis of transcriptomic, metabolite, and enzyme activity data supported a general repression of the core and peripheral phenylpropanoid pathways, which are normally triggered in red-skinned berries throughout ripening and in response to stress . These results suggest that GRBaV infections disrupt secondary metabolic pathways by altering the regulation of berry ripening processes and/or signaling mechanisms related to plant defense. Interestingly, GRBaV infections seemed to have a more a pronounced impact on enzymatic activities and metabolite accumulation than on the expression levels of the genes in the pathway, which in general displayed small fold change differences between healthy and infected samples. This observation further confirms the importance of evaluating metabolic perturbations at multiple regulatory levels.Understanding how plants respond to external stimuli in the field is crucial to improve agricultural traits under naturally fluctuating conditions. Most studies on plant–pathogen interactions are performed with model organisms in the greenhouse or laboratory, which reduce the confounding effects of the environment, but also challenge the reproducibility of the results in the field . Compatible plant–virus interactions in perennial woody crops are complex due to the presence of multiple and systemic infections, tissue and developmental stage-specific responses, differences between species and cultivars, and the combination of biotic and abiotic factors during the crop season . The application of a system biology approach to study red blotch under multiple vineyard conditions allowed us to explore grapevine responses to GRBaV infections in real agronomic settings and to characterize the influence of viral activity on berry physiology.

Building on research that has used empirical data to compare soil and above ground C stocks in vineyards and adjacent oak woodlands in California, this study sought to estimate the C composition of a vine, including the relative contributions of its component parts . By identifying the allometric relationships among trunk diameter, plant height, and other vine dimensions, growers could utilize a reliable mechanism for translating vine architecture and biomass into C estimates. In both natural and agricultural ecosystems, several studies have been performed using allometric equations in order to estimate above ground biomass to assess potential for C sequestration. For example, functional relationships between the ground-measured Lorey’s height and above ground biomass were derived from allometric equations in forests throughout the tropics. Similarly, functional relationships have been found in tropical agriculture for above ground, below ground, and field margin biomass and C. In the vineyard setting, however, horticultural intervention and annual pruning constrain the size and shape of vines making existing allometric relationships less meaningful, though it is likely that simple physical measurements could readily estimate above ground biomass. To date, most studies on C sequestration in vineyards have been focused on soil C as sinks and some attempts to quantify biomass C stocks have been carried out in both agricultural and natural systems. In vineyards, studies in California in the late 1990s have reported net primary productivity or total biomass values between 550 g C m−2 and 1100 g C m−2.

In terms of spatial distribution, hydroponic nft system some data of standing biomass collected by Kroodsma et al. from companies that remove trees and vines in California yielded values of 1.0–1.3 Mg C ha−1 year−1 woody C for nuts and stone fruit species, and 0.2–0.4 Mg C ha−1 year−1 for vineyards. It has been reported that mature California orchard crops allocate, on average, one third of their NPP to the harvested portion and mature vines 35–50% of the current year’s production to grape clusters. Pruning weight has also been quantified by two direct measurements which estimated 2.5 Mg of pruned biomass per ha for both almonds and vineyards. The incorporation of trees or shrubs in agroforestry systems can increase the amount of carbon sequestered compared to a monoculture field of crop plants or pasture. Additional forest planting would be needed to offset current net annual loss of above ground C, representing an opportunity for viticulture to incorporate the surrounding woodlands into the system. A study assessing C storage in California vineyards found that on average, surrounding forested wildlands had 12 times more above ground woody C than vineyards and even the largest vines had only about one-fourth of the woody biomass per ha of the adjacent wooded wildlands.The objectives of this study were to: measure standing vine biomass and calculate C stocks in Cabernet Sauvignon vines by field sampling the major biomass fractions ; calculate C fractions in berry clusters to assess C mass that could be returned to the vineyard from the winery in the form of rachis and pomace; determine proportion of perennially sequestered and annually produced C stocks using easy to measure physical vine properties ; and develop allometric relationships to provide growers and land managers with a method to rapidly assess vineyard C stocks. Lastly, we validate block level estimates of C with volumetric measurements of vine biomass generated during vineyard removal.

The study site is located in southern Sacramento County, California, USA , and the vineyard is part of a property annexed into a seasonal floodplain restoration program, which has since removed the levee preventing seasonal flooding. The ensuing vineyard removal allowed destructive sampling for biomass measurements and subsequent C quantification. The vineyard is considered part of the Cosumnes River appellation within the Lodi American Viticultural Area, a region characterized by its Mediterranean climate— cool wet winters and warm dry summers—and by nearby Sacramento-San Joaquin Delta breezes that moderate peak summer temperatures compared to areas north and south of this location. The study site is characterized by a mean summer maximum air temperature of 32 °C, has an annual average precipitation of 90 mm, typically all received as rain from November to April . During summer time, the daily high air temperatures average 24 °C, and daily lows average 10 °C. Winter temperatures range from an average low 5 °C to average high 15 °C . Total heating degree days for the site are approximately 3420 and the frost-free season is approximately 360 days annuall. Similar to other vineyards in the Lodi region, the site is situated on an extensive alluvial terrace landform formed by Sierra Nevada out wash with a San Joaquin Series soil . This soil-landform relationship is extensive, covering approximately 160,000 ha across the eastern Central Valley and it is used extensively for winegrape production. The dominant soil texture is clay loam with some sandy clay loam sectors; mean soil C content, based on three characteristic grab samples processed by the UC Davis Analytical Lab, in the upper 8 cm was 1.35% and in the lower 8–15 cm was 1.1% . The vineyard plot consisted of 7.5 ha of Cabernet Sauvignon vines, planted in 1996 at a density of 1631 plants ha−1 with flood irrigation during spring and summer seasons. The vines were trained using a quadrilateral trellis system with two parallel cordons and a modified Double Geneva Curtain structure attached to T-posts . Atypically, these vines were not grafted to rootstock, which is used often in the region to modify vigor or limit disease .In Sept.–Oct. of 2011, above ground biomass was measured from 72 vines. The vineyard was divided equally in twelve randomly assigned blocks, and six individual vines from each block were processed into major biomass categories of leaf, fruit, cane and trunk plus cordon . Grape berry clusters were collected in buckets, with fruit separated and weighed fresh in the field. Leaves and canes were collected separately in burlap sacks, and the trunks and cordons were tagged. Biomass was transported off site to partially air dry on wire racks and then fully dried in large ventilated ovens.

Plant tissues were dried at 60 °C for 48 h and then ground to pass through a 250 μm mesh sieve using a Thomas Wiley® Mini-Mill . Total C in plant tissues was analyzed using a PDZ Europa ANCA-GSL elemental analyzer at the UC Davis Stable Isotope Facility. For cluster and berry C estimations, grape clusters were randomly selected from all repetitions. Berries were removed from cluster rachis. While the berries were frozen, the seeds and skins were separated from the fruit flesh or “pulp”, and combined with the juice . The rachis, nft channel skins and seeds were dried in oven and weighed. The pulp was separated from the juice + pulp with vacuum filtration using a pre-weighed Q2 filter paper . The filter paper with pulp was oven dried and weighed to get insoluble solid fraction . The largest portion of grape juice soluble solids are sugars. Sugars were measured at 25% using a Refractometer PAL-1 . The C content of sugar was calculated at 42% using the formula of sucrose. Below ground biomass was measured by pneumatically excavating the root system with compressed air applied at 0.7 Mpa for three of the 12 sampling blocks, exposing two vines each in 8 m3 pits. A root restricting duripan, common in this soil, provided an effective rooting depth of about 40 cm at this site with only 5–10 fine and small roots able to penetrate below this depth in each plot. Roots were washed, cut into smaller segments and separated into four size classes , oven-dried at 60 °C for 48 h and weighed. Larger roots were left in the oven for 4 days. Stumps were considered part of the root system for this analysis.In vineyard ecosystems, annual C is represented by fruit, leaves and canes, and is either removed from the system and/or incorporated into the soil C pools, which was not considered further. Structures whose tissues remain in the plant were considered perennial C. Woody biomass volumes were measured and used for perennial C estimates. Cordon and trunk diameters were measured using a digital caliper at four locations per piece and averaged, and lengths were measured with a calibrated tape. Sixty vines were used for the analysis; twelve vines were omitted due to missing values in one or more vine fractions. All statistical estimates were conducted in R.The present study provides results for an assessment of vineyard biomass that is comparable with data from previous studies, as well as estimates of below ground biomass that are more precise than previous reports. While most studies on C sequestration in vineyards have focused on soil C, some have quantified above ground biomass and C stocks. For example, a study of grapevines in California found net primary productivity values between 5.5 and 11 Mg C ha−1—figures that are comparable to our mean estimate of 12.4 Mg C ha−1 . For pruned biomass, our estimate of 1.1 Mg C ha−1 were comparable to two assessments that estimated 2.5 Mg of pruned biomass ha−1 for both almonds and vineyards.

Researchers reported that mature orchard crops in California allocated, on average, one third of their NPP to harvestable biomass, and mature vines allocated 35–50% of that year’s production to grape clusters. Our estimate of 50% of annual biomass C allocated to harvested clusters represent the fraction of the structures grown during the season . Furthermore, if woody annual increments were considered this proportion would be even lower. Likewise the observed 1.7 Mg ha−1 in fruit represents ~14% of total biomass , which is within 10% of other studies in the region at similar vine densities. More importantly, this study reports the fraction of C that could be recovered from wine making and returned to the soil for potential long term storage. However, this study is restricted to the agronomic and environmental conditions of the site, and the methodology would require validation and potential adjustment in other locations and conditions. Few studies have conducted a thorough evaluation of below ground vine biomass in vineyards, although Elder field did estimate that fine roots contributed 20–30% of total NPP and that C was responsible for 45% of that dry matter. More recently, Brunori et al. studied the capability of grapevines to efficiently store C throughout the growing season and found that root systems contributed to between 9 and 26% of the total vine C fixation in a model Vitis vinifera sativa L. cv Merlot/berlandieri rupestris vineyard. The results of our study provide a utilitarian analysis of C storage in mature wine grape vines, including above and below ground fractions and annual vs. perennial allocations. Such information constitutes the basic unit of measurement from which one can then estimate the contribution of wine grapes to C budgets at multiple scales— fruit, plant or vineyard level—and by region, sector, or in mixed crop analyses. Our study builds on earlier research that focused on the basic physiology, development and allocation of biomass in vines. Previous research has also examined vineyard-level carbon at the landscape level with coarser estimates of the absolute C storage capacity of vines of different ages, as well as the relative contribution of vines and woody biomass in natural vegetation in mixed vineyard-wildland landscapes. The combination of findings from those studies, together with the more precise and complete carbon-by-vine structure assessment provided here, mean that managers now have access to methods and analytical tools that allow precise and detailed C estimates from the individual vine to whole-farm scales. As carbon accounting in vineyard landscapes becomes more sophisticated, widespread and economically relevant, such vineyard-level analyses will become increasingly important for informing management decisions. The greater vine-level measuring precision that this study affords should also translate into improved scaled-up C assessments . In California alone, for example, there are more than 230,000 ha are planted in vines. Given that for many, if not most of those hectares, the exact number of individual vines is known, it is easy to see how improvements in vine-level measuring accuracy can have benefits from the individual farmer to the entire sector.

Increased wind speed resulted in higher sensible heat losses and, therefore, berry temperatures closer to air temperature, specifically in sparse canopies . The wider row spacing in open canopies , besides providing less wind resistance, also allowed more heating of the ground and air which resulted in higher sensible heat transfer. The greater sensible heat losses in Unilateral compared to Goblet could be explained by the proximity of the Goblet clusters to the adjacent terrace slope, which has the potential for very large temperature variation that greatly affected the sensible heat fluxes. As expected, the cluster heat storage tended to be negative in the morning as the berries began to rapidly warm, and positive in the afternoon while generally cooling and releasing heat .The time series of measured and simulated berry temperature were compared graphically for east- and west-facing clusters in order to qualitatively assess model performance . Graphical results indicated good qualitative agreement between measured and modeled diurnal berry temperature variation at each site. At night time or when berries were in the shade, modeled and measured temperatures closely matched the air temperature. During intermittent periods of solar exposure, round plant pot the model was able to accurately replicate both the magnitude and duration of temperature increases over ambient.

There were a few brief periods, such as in the Unilateral vineyard, in which the timing between the measured and modeled transition from sunlit to shaded conditions lagged by about an hour. It is likely that the observed discrepancies could be explained by slight inaccuracies in the exact position of each berry and leaf. Small errors in the geometric model can translate into large errors in absorbed radiation and berry temperature during periods of sun-shade transition. The statistical error measures describing agreement between measured and modeled time series shown in Fig. 3.6 are summarized in Table 3.4. Quantitative agreement between measured and modeled berry temperature was excellent, with R2 between 0.94 and 0.97, index of agreement between 0.98 and 0.99, and NRMSE between 4.6% and 8.5%. Thus, while there could be brief periods of large error in predicted temperature at any instant when there were rapid transitions between sun and shade, their effect on daily averaged errors were relatively small.The different plant geometries and row spacing also influenced the berry heat storage and, therefore, the variability in berry temperature . Overall, including the heat storage term in the energy balance equation reduced the error of the temperature fluctuations. With no heat storage, the temperature increased or decreased too quickly, typically leading to over- or under-shooting of the berry temperature, which tended to increase model errors .The experimental data collected in this study corresponded to four field sites with different climatic and geographic conditions, and vineyard designs. The average within-canopy ambient microclimate is driven both by the local weather/climate at the site and by the canopy architecture.While it is difficult to directly compare the two Davis experiments because they were conducted during different years, the two Napa plots experienced virtually the same weather conditions and thus differences are likely to be dominated by canopy architecture.

The primary architectural differences between the Goblet and Unilateral plots were that the row spacing in Goblet was much smaller, and berries were in general much closer to the ground. This tighter row spacing and overall denser canopy led to an expected trend of lower wind speeds, more humid air, and cooler air temperatures on average. The ambient microclimate conditions have important implications for average berry temperatures because when there is minimal or no solar heating, such as at night or when the berries are in the shade, the berry temperature is nearly equal to the air temperature. Since berries spend the majority of their time with temperature near the air temperature, this period will dominate the overall average temperature of the berry. Accordingly, prior work measuring apple fruit temperature found that the long-term average fruit temperature was very close to the air temperature, although there could be large deviations from the air temperature at any instant. The differences in vineyard design not only created variability in ambient microclimate, but also introduced considerable temporal and spatial variability in berry temperature. Due to the predominantly north-south row orientation, there were often large differences between the exposed east- and west-facing clusters at any instant in time. Exposure for west-facing clusters coincided with warmer ambient afternoon temperatures, which meant that the average and maximum temperatures of west-facing clusters was typically higher than for east-facing clusters. The substantial asymmetry in temperature accumulation in north-south oriented vineyards is well-known, and has given rise to strategies based on oblique row angles aimed at achieving more even heating between both sides of the row.

Of the different variables explored, the berry temperature was also likely to be influenced by differences in wind speed created by the different vineyard geometries. The wider row spacing in open canopies provided less wind resistance and higher wind speeds that increased sensible heat losses. As observed in Unilateral compared to Goblet, the higher sensible heat losses resulted in fully-exposed berry temperatures closer to air temperature. The proximity of the clusters to the adjacent terrace slope in Goblet may have also increased berry temperatures since the ground has the potential to have high deviations in surface temperature relative to air temperature. The duration and temporal pattern of berry exposure could vary considerably depending on specifics of the berry position. Except for in the Wye vineyard, there were rarely periods in which all berries on a given side of the vine were at a similar temperature . Thus, depending on the horizontal or vertical position of the berry, and random positioning of neighboring leaves, there could be significant variability in berry temperature even on the same side of the vine.Helios simulates complex interactions between the environment and different parts of the vine from ambient weather variables that are not particularly difficult to measure, thereby making it possible to evaluate the applicability of the model at other spatial and temporal scales. Because of the spatially-explicit nature of the plant microclimate model used in this work, it was possible to resolve average differences in berry temperature due to vineyard geometry. Previous models have assumed that the berry temperature is equal to the air temperature, or used simple heuristic rules to represent the mean effect of the vines. The ability to represent berry temperature deviations from the ambient air temperature is likely important for processes that are sensitive to intermittent periods of high temperature, such as chemical composition or berry burn. Simple heuristic models are unlikely to be useful in evaluating the effects of different trellis systems, particularly systems with complex geometries such as Wye. The results of the present study showed significant differences in spatial and temporal patterns in berry temperature between trellis systems, which were well-replicated by the spatially explicit model used for prediction. The model formulation used in this work explicitly represented berry heat storage, and also compared the modeled result when berry heat storage was completely removed. As expected, removing berry heat storage resulted in much larger temporal fluctuations in response to high variability in ambient microclimate. Although inclusion of berry heat storage had a modest impact on average error metrics , it did provide a noticeable reduction in temperature variability when berries were exposed to the sun. The stabilizing effect of berry heat storage also decreased maximum berry temperatures significantly.A limitation of the proposed model is that it does not include the effect of rainfall or a wet canopy on berry temperature. However, round garden pot most quality wines are produced in regions with little average rainfall during the period of berry development. The model also did not explicitly represent the 3D variation of temperature and associated heat within the fruit as has been done in the model developed by Saudreau et al. and Saudreau et al for apple. However, validation results indicated that such detail was not necessary to achieve excellent agreement with measured temperatures, but rather an average exponential dampening of temperature fluctuations with appropriate time constant was sufficient. Another limitation of this study was that currently the model does not predict 3D spatial variations in wind speed, air temperature, or relative humidity, rather, these quantities were measured near the fruit clusters and used to drive the model.

If within-canopy microclimate was not available, it could be necessary to implement a canopy-scale energy and momentum transport model. Previous studies have shown that grape berry metabolism is sensitive to changes in both daily average temperatures and the magnitude of diurnal temperature fluctuations, therefore, advances in our understanding of the berry temperature fluctuations might help develop novel strategies to obtain the desired grape quality. The validation exercise in this work focused specifically on berry temperature from post-veraison to harvest. In future studies, the model could be modified to include latent heat fluxes and be validated to estimate berry temperature during ripening when berry evaporation may be significant and radiative properties of the berries likely differs. In addition to evaluating strategies for mitigating excessive berry temperatures, the model developed in this work could be used on the macro-scale to predict daily berry temperature fluctuations in different regions, such as hilly or mountainous areas and/or areas with arid continental weather subject to dramatic temperature fluctuations. The model could also be coupled with epidemiological and physiological models to study the effect of the spatial and temporal temperature variations on disease incidence or on physiological processes that determine grape yield and quality. Recent increases in average air temperatures and heat wave intensity can present challenges in maintaining grape productivity and quality. As a result, growers are exploring approaches to protect berries from excessive temperatures, however, they can be costly and time-consuming to evaluate experimentally and results may not be generalizable. In this work, we developed and evaluated a new 3D model that can predict metrics related to berry temperature and light interception in response to varying vineyard architecture, topography, and shade cloth density. The resulting modeling tool was applied to better understand and evaluate a range of potential vineyard design and management practices for mitigation of elevated berry temperatures in vertically-trained grapevines. Model validation showed close agreement between predicted and measured temperature dynamics, which responded appropriately to the application of shade cloth. In a simulation experiment, row spacing, row orientation, slope grade and aspect, and shade cloth density were varied in order to evaluate their effect on berry and canopy light interception, berry temperature spikes, and integrated berry heat accumulation. On flat terrain, NE-SW row orientation provided the best compromise of berry light and temperature balance between opposing vine faces while avoiding excessive berry temperatures, while N-S rows provided good daily symmetry but had risk of high afternoon berry temperatures. The efficacy of shade cloth in mitigating excessive temperatures depended strongly on all variables considered. Slopes with southern or western exposure increased imbalance and risk of high berry temperatures, which in some cases could not be well-managed by shade cloth. Overall, the modeling tool appears capable of providing quantitative guidance for vineyard design and management where excessive berry temperatures are a concern.Climate models predict that greenhouse gases will increase global average ambient temperatures by approximately 1 and 3◦C by 2030 and 2100, respectively, in addition to an increase in the frequency, duration and severity of heat waves, particularly in many wine grape growing regions. Elevated temperatures differentially affect rates of grape berry sugar accumulation and phenolic compound development, which can lead to trade-offs in harvest timing that can ultimately result in a reduction in overall grape quality. In Oakville, CA, Mart´ınez-Luscher et al. ¨ reported that elevated temperatures for grape clusters result in unbalanced wines with higher pH and lower levels of anthocyanins. Other research conducted in Murrumbidgee, Australia reported that temperatures exceeding 40◦C resulted in delayed ripening and caused berry sunburn. High ambient air temperatures can exacerbate problems created by excessive berry solar exposure due to the reduction of convective cooling. These high temperatures due to direct sunlight can result in berry cellular damage within a few minutes, while moderately high temperatures can result in injuries or death after long exposure. Grape growers have started to implement practices that modify vineyard microclimate in the short and long term to cope with elevated temperatures.

Proanthocyanidins are found in both the skin and seeds of grape berries . Tannin synthesis in the grape berry starts at flowering, continues in the early stages of grape development, and reaches its maximum around veraison . It then decreases throughout ripening due to a reduction in the extractability of tannins related to their interactions with cell wall components, rather than a decrease related to degradation . The effect of high temperatures on skin and seed tannins have shown inconsistent results across different studies. Cohen et al., found a linear relationship between the increase of skin tannin content and heat accumulation during phase I of grape development. However, Gouot et al., found an increase of total skin tannin concentration in vines exposed to short periods of heat stress before veraison, but found no differences in skin tannin concentration and content at maturity. Studies where heat was applied after veraison have shown no effect on tannin levels, which could be associated with the stop of tannin synthesis after this point in development . Lastly, Bonada et al., conducted a field experiment where vines were exposed to elevated temperatures throughout the entire growing season and found a decrease in total tannin content and concentration at maturity. This was mainly related to a decrease of seed tannins while skin tannins remained unresponsive to elevated temperatures. The different experimental conditions as well as differences in heat treatment design and time of application could be possible explanations for the discrepancies of these findings, as well as whether concentration or content of tannins was studied. Flavonols, anthocyanins, large plastic planting pots and flavan-3-ols are derived from the same precursors and are produced from the flavonoid biosynthetic pathway .

These precursors are derived from the Shikimate and phenylpropanoid pathways. Each step of these pathways are catalyzed by specific enzyme families. Flavonoid enzymes include flavone synthase , flavanone-3ß-hydroxylase , and flavonol synthase . Two enzymes, flavonoid-3’-hydroxylase and flavonoid-3’-5’-hydroxylase , are particularly important because they catalyze the reactions that form di- and tri-hydroxylated flavonoids from monohydroxylated precursors. The genes that code for F3’H and F3’5’H are expressed in the berry skin and contribute to the production of di- and tri-hydroxylated flavonols, anthocyanins, cyanidins, and delphinidin derivatives . The main enzymes for flavan-3-ol production are anthocyanidin reductase and leucoanthocyanidin reductase . However, the enzymes involved in producing proanthocyanidins are unknown. The enzyme that converts anthocyanidins to anthocyanins is UDP-glucose flavonoid-3-O-glucosyltransferase. It is also important to note the MYB transcription factors that are involved in the regulation of the phenylpropanoid pathway. MYBF1 regulates FLS for flavonol production. MYBA1&2 and MYB4 regulate UFGT for anthocyanin synthesis, with MYBA1&2 also being involved in several other steps in the pathway . The effects of high temperatures on the phenylpropanoid and flavonoid pathways is variable, and dependent upon the phenological stage during which heat events occur. Veraison is the most sensitive stage for anthocyanins, while flowering is the most sensitive stage for tannins . Although it has been widely reported that high temperatures impact anthocyanin synthesis, gene expression does not consistently reflect concentration.

A study done by Mori et al. on the variety Darkridge showed that high night temperature resulted in lower anthocyanin concentration due to a down regulation of genes including F3’H and UFGT. Similar results were seen in Kyoho grapes in that high day and night temperatures down-regulated UFGT . However, when the berries were exposed to high temperatures only at night, there was an initial decrease then sudden increase in UFGT activity during ripening. In contrast, a study done on Cabernet Sauvignon showed that UFGT was not strongly down-regulated in grapes that were exposed to high temperatures . Studies show that the effects of high temperatures on the expression of MYB transcription factors remains inconsistent, particularly with MYBA1. Some studies have shown that the expression of MYBA1 was unaffected by high temperatures , whereas other studies showed an extreme down-regulation of this transcription factor . Currently, there is a lack of published research that investigates the potential effects of HWs on berry temperature. While there have been studies published that look at how elevated temperatures affect berry development, primary metabolism, and secondary metabolism, they did not collect data measuring temperature of the berries themselves. For example, in a study done by Cohen et al. , individual cluster temperatures were manipulated to assess the impact of temperature on phenolic metabolism. While this study provided insight to the direct effects of high temperatures, it failed to show whether heat events result in higher berry temperature under different irrigation regimes, and if so, to what extent. The objective of this study was to evaluate the impact that different irrigation practices have on berry phenolics and gene expression carried out prior to and during HWs in a commercial vineyard of Cabernet Sauvignon. The results from this study provide a better understanding of how irrigation during heat events can help mitigate the effects of extreme temperatures and lead to more efficient water use.

In addition, by studying how HWs and irrigation practices influence the production of metabolites in the berry at the chemical and molecular levels provides the wine industry new knowledge that can be used to direct future efforts at preserving berry and wine quality under climate change conditions. In this experiment, a HW was defined as three consecutive days with maximum temperatures at or above 38 °C . Rather than creating artificial HWs with a heating system, this study relied on the occurrence of natural HWs. There were four HWs during the 2020 growing season, and there were two HWs during the 2021 growing season. Differential irrigation treatments were applied when HWs occurred and started one or two days before each HW and continued until the last day of the HW. There were three irrigation treatments: a baseline, which was exposed to deficit irrigation and held at 60% ET, a second treatment where irrigation was double the baseline , and a third treatment where irrigation was triple the baseline . In 2021, the irrigation treatments were adjusted in order to fine tune the amount of water used to see if less water could be used while maintaining the positive aspects of increased irrigation during HWs. The baseline treatment stayed the same at 60% ET, the second treatment was 1.5x baseline , and the third treatment was 2x baseline . Four pixels per treatment were randomly selected and distributed along the site using the VRDI system to provide differential irrigation to the individual plots. ET was estimated throughout the season using Landsat data, normalized difference vegetation index and crop coefficient. Variations in pre-treatment irrigation were implemented at the beginning of the growing season on the plots to even out vigor based on NDVI and thermal imaging due to the heterogeneous soil profile of the site. After, differences in irrigation schedules were based completely on implemented treatmentsBerry temperature was measured using 0.076 mm diameter type ‘E’ thermocouples . There were a total of 20 thermocouples placed in individual berries within three of the treatment blocks. The thermocouples were inserted into the center of the berries in exposed clusters facing the east and west side of the vine, and at each side of the vine 4 thermocouples were placed in different berries within the cluster. Because berries could develop necrosis from being punctured by the thermocouple, thermocouples were relocated to adjacent exposed berries at least every 12 days to maintain relatively fresh conditions . A subset of grape samples were stored at -20 °C for phenolic extraction. Three sets of sixty berries were weighed and volume occupied in water was recorded using a graduated cylinder.Skins were prepared for extraction by separating pulp and seed by hand. The skins were homogenized using a T25 digital ULTRA-TURRAX and S 25 N-18 G Dispersing tool. The skins were homogenized for one minute at 14 speed x 1000 rpm with 100 mL of a 66% acetone and transferred to an opaque polypropylene jar. The jar’s head space was filled with nitrogen gas, closed with a screw cap lid, black plastic planting pots and sealed with parafilm. The skins were extracted for 24 hours on a orbital shaker . The next day, samples were centrifuged for 10 minutes at 10,000 rpm. Samples were then poured through a Buchner funnel into a Buchner flask and filtered through Whatman filter paper with a pore size of 1.

Each filtered sample was transferred to a round-bottom flask and put onto the rotovap for 10 minutes with the water bath at 33 °C. Acetone was removed under reduced pressure. The extracts were brought back to 50 mL with milli-q water, and volume was recorded using a graduated cylinder. The samples were transferredto centrifuge tubes and stored at -20 °C until analysis. The next day, the samples were filtered, and acetone was separated from the solution using a rotovap. The samples stayed on the rotovap for 10 minutes with the water bath at 33 °C. The concentrated extracts were diluted with milli-q water, and volume was recorded using a graduated cylinder. The samples were transferred to falcon tubes and stored at -20 °C until analysis. Triplicate grape extracts were analyzed by methyl cellulose precipitation, which is the method used for analyzing tannin in extracts and wine, as previously published . Briefly, 25 µL of extraction sample was placed into 1200 µL deep well plates and combined with 300 µL of 0.04% methyl cellulose  solution or water , and mixed on an Thermomixer-C for 5 minutes at 1500 RPM and left to stand for 3 minutes. Following the mixing, 200 µL of saturated ammonium sulfate was added to the wells to prevent the re-release of proanthocyanidin material into solution following precipitation. Water was then added to the wells and again it was mixed for 5 minutes. The deep well plate was then allowed to stand for 10 minutes before being centrifuged for 5 minutes at 2,272 × g . Both the treated and control samples were taken and placed in a 96 half-area well plate . -Epicatechin was used as a quantitative standard. Proanthocyanidin quantification was conducted by calculating Δ280 nm with the linear regression from an -epicatechin standard curve. The samples were analyzed on the SpectraMax iD3 microplate reader. Endpoint analysis was at 280 nm, and a pathway correction made up for volume differences. Berry temperature was recorded across the August 2020 and September 2020 heat waves . Berry temperatures from 60% ET treatment were significantly different from the 120% ET and 180% ET treatments for the pre-HW date of August 13th . There were no significant differences in berry temperature between the three treatments for the August 2020 HW and post-HW dates. For the August 13th pre-HW date, peak berry temperature occurred at 13:00 hours for the 180% ET treatment. For the HW date, peak berry temperature was at 15:00 hours for the 120% ET treatment. For the post-HW date, peak berry temperature was at 17:00 hours for the 60% ET treatment. There were no significant differences among the three treatments during the September 2020 HW . For the pre-HW date, peak berry temperature was at 15:00 hours for the 60% ET treatment. Similarly, for the HW and post-HW dates, peak berry temperature was at 15:00 hours for the 180% ET treatment. Berries in the August 2020 HW consistently reached higher temperatures than berries in the September 2020 HW, which may be due to differences in the extremity of the respective HWs. Anthocyanins in both growing seasons were measured from pre-veraison until commercial harvest. As seen in Figure 4, in the 2020 growing season, the 60% ET treatment started out with the lowest total anthocyanin concentration and continued throughout the rest of the season. During HW3 and HW4, we see significant differences between the treatments. At harvest, there were no differences between the 120% ET and 180% ET treatments, but both treatments had significantly higher anthocyanin concentrations than the 60% ET treatment, suggesting that the additional water prior to the heat waves had mitigated the loss of anthocyanin material. Looking at Figure 4, again in 2021, the 60% ET treatment had consistently lower anthocyanin concentrations throughout the season. Unlike the 2020 growing season, the 60% ET treatment remained significantly different from the other two treatments starting at August 17th until close to harvest. However, at harvest, there were no significant differences between the three treatments.

What people say they prefer and what they actually do may, of course, be quite different. In order to get a sense of the extent to which people actually exercise the label preferences they indicated in the marketplace, we asked about their purchase patterns of foods with these labels. We asked specifically about fair trade, organic, and local, first defining the terms . Respondents were asked to check one of the following options regarding their purchasing habits: never, at least once a year, at least monthly, at least weekly, and don’t know. Respondents claimed to purchase these products quite often . At least 50% of the respondents reported purchasing products with one of these labels at least monthly or more often. Organic is the most frequently purchased item, with 42% buying such products at least weekly, and 68% at least monthly. Local products were the next most frequently purchased, with 63% purchasing them at least monthly. Almost 25% of people purchase fair trade at least weekly, and 52% at least monthly. A large number of respondents stated that they did not know whether they have purchased products with these labels. Almost a third of the respondents didn’t know whether they had purchased food that was fair trade and 27% didn’t know whether they had purchased local foods. And, even though the organic label is fairly well established, 16% of the people said they didn’t know whether they had purchased organic food. This information implies that either people don’t know enough about the labels, or that they just don’t look for them when they purchase their food. Considering that organic foods are fairly well known, plastic pot plant containers it is likely safe to assume that the high numbers of those who didn’t know whether they had purchased fair trade may be partially due to lack of information about the label. Thus, it would be useful to have more education about the Fair Trade label.

The large percentage of those who didn’t know whether they had purchased local is likely due to the fact that there isn’t a standard label for such products. For example, one of the factors that makes America’s relatively “cheap” food supply possible is low labor costs; the wages of workers in the food industry are often at poverty or below-poverty levels. To get at the criterion of social justice, we asked if people would be willing to pay more for their food if it meant better conditions for workers, and a living wage. In order to gauge this interest, survey respondents were asked whether they would be willing to pay more for a single product, strawberries. We chose strawberries because there had been a United Farm Workers campaign in the region to inform people about how much paying just five cents more for a pint of strawberries would do to improve wages and working conditions for strawberry farm workers. We asked survey respondents how much more they would be willing to pay for a pint of strawberries that guarantees a living wage and safe working conditions for farm workers. Survey respondents were randomly assigned 1 of 4 questions, asking if they would be willing to pay 5 cents, 25 cents, 50 cents or $1.50 more for strawberries that otherwise would cost $1.50 . Results indicate that nearly all respondents would be willing to pay at least 5 cents more and most people state they would pay significantly more than that ; 85% are willing to pay 25 cents more , 74% are willing to pay 50 cents more , and close to 50% claimed willingness to pay twice as much for a pint of strawberries. We posed a second “willingness to pay” question to meal-plan holders. We asked, “Would you be willing to pay more for your meal plan if the food had been produced in a socially just manner—for example, where workers have safe working conditions and receive a living wage ?” 

A third of meal-plan holders were willing to pay more for socially just food, a quarter were not, and the remainder were unsure . Why are there so few people willing to pay more in the dining commons than for a pint of strawberries? First, the lack of interest shown here is likely at least partially an artifact of the way the question was asked. Unlike the strawberry question, the meal plan holders were not asked a specific amount more that they might be willing to pay—which likely led to the large “unsure” numbers. We were not able to give a more specific amount because it is not clear how much a meal costs . Second, meal plan holders may just be less likely to pay more in general. From comparing how meal plan holders and all others answered the strawberry question, meal plan holders were somewhat less likely to pay more for the strawberries than the others . As noted, our survey results show that one-third of meal plan holders indicated a willingness to pay more for a meal plan for food produced in a socially just manner, and about one quarter were opposed to such a cost increase. Further research, with more specific scenarios, would be needed to get a better idea of the number of people that might really be willing to pay more. When considering these results, it is important to keep in mind that people don’t always do what they say they will—that they are more likely to respond positively on a survey than they may in practice. People often take many factors into account when making purchases, not just one. However, even given a likely inflated positive response, there appears to be significant support for paying more for socially just food.We asked respondents to indicate their preferences for how they would like to learn about the food system. Ten options were listed on the survey and respondents were asked to check four of them . As is shown in table 7, respondents’ preferred media for learning about their food were product labels and information available where they purchase or eat their food . Print media—e.g., newspapers, magazines, articles, and books—and web pages were the next most preferred at 48% and 47%, respectively. Other methods—tours, audiovisual media, talking to farmer/seller, and lectures—were preferred by less than 25% of respondents. Study groups garnered the least interest at 3%. Providing food system education in the dining halls, campus restaurants, and at coffee carts will likely be the most appreciated and most effective method for sharing information with a broad audience.

This study shows that food issues are important to a campus community, particularly regarding concerns for the environment and for people. Survey respondents showed high levels of interest in purchasing food produced in an environmentally sound and socially just manner. Key points from this study include – • There is significant interest in campus food that is nutritious, safe, supports workers, and is environmentally sound; interest in local food and GE-free food is lower. • People are interested in sustainably produced food and a majority of people already purchase food with labels based on these criteria. • Many people are willing to pay more for food that meets social justice criteria. • A campus community is likely to be receptive to education and discussion about food-system issues. • Since nutrition and food safety were of great importance to people, framing discussions of food-system issues in terms of health will meet people’s needs as well as capture their attention for education on other food-system issues, such as working conditions and environment. It would not be appropriate to extrapolate too much from a study of one campus; results from our 2007 national student survey will provide more comprehensive data. In the meantime, the results of the UC Santa Cruz study support the idea that colleges and universities are excellent choices for developing farm-to-institution programs and for popular education on food-system issues. In plants, a fruit is the seed-containing section, which is formed from the ovary after flowering. Fruits have their vivid colors due to the presence of phytochemicals as pigments, large plastic gardening pots which are natural compounds that protect against threats and insults such as insects and ultraviolet sunlight. Bright colors of fruits also attract animals and human beings for seed dispersing purposes.1 The belief of using fruits as traditional medicine exist in many cultures worldwide. For instance, cocoa beans and blueberries have been used traditionally as therapies among indigenous people in North America. The fruit, leaves, seed, and bark of the mango plant have been used as traditional medicine in Southeast Asia, Oceania, Africa, and Central America. Goji berries have been used as traditional Chinese medicine for two thousand years. Phytochemicals can be classified primarily as terpenoids, phenolics, alkaloids, nitrogencontaining plant constituents, and organosulfur compounds. Examples of major phytochemical groups that are abundant from dietary sources and related to human health include carotenoids and polyphenols. Carotenoids are a type of terpenoid. Carotenoids can be classified as carotenes and xanthophylls.4 Phenolics can be classified as phenolic acids and polyphenols.

Two primary subclasses of phenolic acids are hydroxybenzoic acid and hydroxycinnamic acid. Polyphenols include flavonoids, tannins, stilbenes, lignans, and xanthones. As one of the most studied categories of polyphenols, subclasses of flavonoids can be categorized to flavanones, flavones, anthocyanins, flavanols , chalcones, flavonols, and isoflavonoids. Among the thousands of phytochemicals that have been identified in plants, both health promoting and toxic compounds exist. For instance, some tannins decreased the activity of digestive enzymes or the bio-availability of protein or minerals and have been considered as anti-nutrients. Phytochemicals that exist in plant-based dietary sources and have value in human health maintenance and prevention of diseases are defined as phytonutrients. Fruits, vegetables, legumes, spices, nuts, wine, cocoa, tea, and olive oil are examples of foods rich in bio-active phytonutrients. The consumption of these dietary components has been related to decreased risk of developing chronic diseases, including cardiovascular diseases , age-related eye diseases, type II diabetes, cancers, and all-cause mortality. Observational studies also have reported that the total dietary polyphenol intake was inversely associated with the risk of hypertension, hypercholesterolemia, and cardiovascular events. Polyphenols under different categories may play various roles in reducing CVD risk. In the United States, the estimated flavonoid intake is 345 mg/day, with flavanols as the most abundant source. The three most consumed flavanols are catechin, epicatechin, and their polymers. Subanalyses of a cohort study indicated that dietary intakes of flavanols along with lignans, dihydrochalcones, and hydroxybenzoic acids showed a stronger inverse association with the risk of overall CVD events than other phenolic compounds.16 Another cohort study reported that the dietary intakes of anthocyanins, dihydrochalcones, dihydroflavonols, proanthocyanidins, catechins, flavonols, hydroxybenzoic acids, and stilbenes were significantly associated with decreased risks of total CVD. Blueberries and cranberries contain high amount of anthocyanin and proanthocyanidin, respectively, with moderate concentration of flavonoids. Cocoa is rich in flavanols, especially epicatechin and catechin. Mango, as the fourth leading fruit crop worldwide, is high in carotenoids, phenolic acids, and mangiferin, a polyphenol classified as a xanthonoid. Carotenes exist in dietary sources primarily as α-carotene, β-carotene, and lycopene. Major xanthophylls that exist in dietary sources include lutein , zeaxanthin , and β-cryptoxanthin. Epidemiological studies report inconsistent results on the relationship between dietary L and Z intakes and the risk of age-related macular degeneration. However, clinical studies have shown that the supplementation of L and Z was able to increase the level of these compounds in the retina, suggesting their protection against age-related macular degeneration . A major dietary source of L and particularly of Z is goji berry, which also have other in carotenoids, as well as phenolic acids, and flavonoids. While many examples of fruits used traditionally for health promotion exist, this literature review focuses on the evidence of mango, cocoa, blueberries, and cranberries in cardiovascular health, and goji berries in eye health. The application of modern scientific methods to assess traditional remedies is important because evidence-based data is necessary to transfer historical stories and ancient wisdom to contemporary life and advancement of health and human performance.Cocoa is the dried and fully fermented product obtained from the seeds of Theobroma cacao L. and is the main ingredient in chocolate products.

Each vessel in the body has different properties therefore when a stent is being designed, the geometry, structure, and mechanics of the target vessel should be considered. Several groups have examined the fluid dynamics governing blood flow through stented arteries. These studies concluded circular shape of the vessel lumen will optimize fluid flow characteristics at the blood/tissue interface, reducing regions of turbulence and/or stagnation in the immediate vicinity of stent struts. Laminar flow within the lumen of more circular stented arteries may decrease the platelet and inflammatory cell adhesion and activation, therefore, reducing neointimal hyperplasia. Although there was not a significant difference of stenosis between stents with five apices versus stents with six apices but Garasic et al., argued altering lumen shape by increasing the number of struts per cross section from 8 to 12 was associated with a 50% to 60% drop in mural thrombus burden after 3 days and a 2-fold reduction in neointimal thickening after 28 days in rabbit arteries. Therefore, more strut numbers and regularity of strut distribution provides a more circular vascular lumen which is associated with a smoother, more homogeneous arteriographic contour and more beneficial for the performance of a stent and needs to be considered for the future novel stent designs.Although this study was the first study to examine the effects of self-expanding stents on growth vessels, square plastic plant pots it was not enough for the assessment of complete aneurysm formation and full stent expansion in the arteries.

Longer studies are needed to assess for late aneurysm formation or erosion as well as stenosis. While this data set shows growth, it is not able to determine the extent of the stented vessel expansion. Will these stents ultimately expand to their full potential? If not, how close will they get. Future studies should consider more animals, longer implant times and should examine lower-radial force stents as well as various stent geometries and stent placement in different vessels. Stents that can grow with a small but rapidly growing artery can be designed and could be ideal for several pediatric applications. Despite obvious medial injury in some cases, all stents in this study grew with and beyond the native vessel’s growth without development of stenoses, aneurysms or dissections. Although this data set only includes 3- and 6-month pathology, it is likely to be useful in the design of the ideal set of pediatric growth stents. Agriculture is a key human activity in terms of food production, economic importance and impact on the global carbon cycle. As the human population heads toward 9 billion or beyond by 2050, there is an acute need to balance agricultural output with its impact on the environment, especially in terms of greenhouse gas production. An evolving set of tools, approaches and metrics are being employed under the term “climate smart agriculture” to help—from small and industrial scale growers to local and national policy setters—develop techniques at all levels and find solutions that strike that production-environment balance and promote various ecosystem services. California epitomizes the agriculture-climate challenge, as well as its opportunities.

As the United States’ largest agricultural producing state agriculture also accounted for approximately 8% of California’s greenhouse gas emissions statewide for the period 2000–2013. At the same time, California is at the forefront of innovative approaches to CSA . Given the state’s Mediterranean climate, part of an integrated CSA strategy will likely include perennial crops, such as winegrapes, that have a high market value and store C long term in woody biomass. Economically, wine production and retail represents an important contribution to California’s economy, generating $61.5 billion in annual economic impact. In terms of land use, 230,000 ha in California are managed for wine production, with 4.2 million tons of winegrapes harvested annually with an approximate $3.2 billion farm gate value. This high level of production has come with some environmental costs, however, with degradation of native habitats, impacts to wildlife, and over abstraction of water resources . Although many economic and environmental impacts of wine production systems are actively being quantified, and while there is increasing scientific interest in the carbon footprint of vineyard management activities [e.g., 11], efforts to quantify C capture and storage in annual and perennial biomass remain less well-examined. Studies from Mediterranean climates have focused mostly on C cycle processes in annual agroecosystems or natural systems. Related studies have investigated sources of GHGs , on-site energy balance, water use and potential impacts of climate change on productivity and the distribution of grape production. The perennial nature and extent of vineyard agroecosystems have brought increasing interest from growers and the public sector to reduce the GHG footprint associated with wine production. The ongoing development of carbon accounting protocols within the international wine industry reflects the increased attention that industry and consumers are putting on GHG emissions and offsets. In principle, an easy-to-use, wine industry specific, GHG protocol would measure the carbon footprints of winery and vineyard operations of all sizes.

However, such footprint assessment protocols remain poorly parameterized, especially those requiring time-consuming empirical methods. Data collected from the field, such as vine biomass, cover crop biomass, and soil carbon storage capacity are difficult to obtain and remain sparse, and thus limit the further development of carbon accounting in the wine sector. Simple yet accurate methods are needed to allow vineyard managers to measure C stocks in situ and thereby better parameterize carbon accounting protocols. Not only would removing this data bottleneck encourage broader participation in such activities, it would also provide a reliable means to reward climate smart agriculture.Building on research that has used empirical data to compare soil and above ground C stocks in vineyards and adjacent oak woodlands in California, this study sought to estimate the C composition of a vine, including the relative contributions of its component parts . By identifying the allometric relationships among trunk diameter, plant height, and other vine dimensions, growers could utilize a reliable mechanism for translating vine architecture and biomass into C estimates. In both natural and agricultural ecosystems, several studies have been performed using allometric equations in order to estimate above ground biomass to assess potential for C sequestration. For example, functional relationships between the ground-measured Lorey’s height and above ground biomass were derived from allometric equations in forests throughout the tropics. Similarly, functional relationships have been found in tropical agriculture for above ground, below ground, and field margin biomass and C. In the vineyard setting, however, horticultural intervention and annual pruning constrain the size and shape of vines making existing allometric relationships less meaningful, though it is likely that simple physical measurements could readily estimate above ground biomass. To date, most studies on C sequestration in vineyards have been focused on soil C as sinks and some attempts to quantify biomass C stocks have been carried out in both agricultural and natural systems. In vineyards, studies in California in the late 1990s have reported net primary productivity or total biomass values between 550 g C m−2 and 1100 g C m−2. In terms of spatial distribution, some data of standing biomass collected by Kroodsma et al. from companies that remove trees and vines in California yielded values of 1.0–1.3 Mg C ha−1 year−1 woody C for nuts and stone fruit species, square pot plastic and 0.2–0.4 Mg C ha−1 year−1 for vineyards. It has been reported that mature California orchard crops allocate, on average, one third of their NPP to the harvested portion and mature vines 35–50% of the current year’s production to grape clusters. Pruning weight has also been quantified by two direct measurements which estimated 2.5 Mg of pruned biomass per ha for both almonds and vineyards. The incorporation of trees or shrubs in agroforestry systems can increase the amount of carbon sequestered compared to a monoculture field of crop plants or pasture. Additional forest planting would be needed to offset current net annual loss of above ground C, representing an opportunity for viticulture to incorporate the surrounding woodlands into the system. A study assessing C storage in California vineyards found that on average, surrounding forested wild lands had 12 times more above ground woody C than vineyards and even the largest vines had only about one-fourth of the woody biomass per ha of the adjacent wooded wild lands.The objectives of this study were to: measure standing vine biomass and calculate C stocks in Cabernet Sauvignon vines by field sampling the major biomass fractions ; calculate C fractions in berry clusters to assess C mass that could be returned to the vineyard from the winery in the form of rachis and pomace; determine proportion of perennially sequestered and annually produced C stocks using easy to measure physical vine properties ; and develop allometric relationships to provide growers and land managers with a method to rapidly assess vineyard C stocks.

Lastly, we validate block level estimates of C with volumetric measurements of vine biomass generated during vineyard removal.The study site is located in southern Sacramento County, California, USA , and the vineyard is part of a property annexed into a seasonal floodplain restoration program, which has since removed the levee preventing seasonal flooding. The ensuing vineyard removal allowed destructive sampling for biomass measurements and subsequent C quantification. The vineyard is considered part of the Cosumnes River appellation within the Lodi American Viticultural Area, a region characterized by its Mediterranean climate— cool wet winters and warm dry summers—and by nearby Sacramento-San Joaquin Delta breezes that moderate peak summer temperatures compared to areas north and south of this location. The study site is characterized by a mean summer maximum air temperature of 32 °C, has an annual average precipitation of 90 mm, typically all received as rain from November to April. During summer time, the daily high air temperatures average 24 °C, and daily lows average 10 °C. Winter temperatures range from an average low 5 °C to average high 15 °C. Total heating degree days for the site are approximately 3420 and the frost-free season is approximately 360 days annually. Similar to other vineyards in the Lodi region, the site is situated on an extensive alluvial terrace landform formed by Sierra Nevada outwash with a San Joaquin Series soil . This soil-landform relationship is extensive, covering approximately 160,000 ha across the eastern Central Valley and it is used extensively for winegrape production. The dominant soil texture is clay loam with some sandy clay loam sectors; mean soil C content, based on three characteristic grab samples processed by the UC Davis Analytical Lab, in the upper 8 cm was 1.35% and in the lower 8–15 cm was 1.1% . The vineyard plot consisted of 7.5 ha of Cabernet Sauvignon vines, planted in 1996 at a density of 1631 plants ha−1 with flood irrigation during spring and summer seasons. The vines were trained using a quadrilateral trellis system with two parallel cordons and a modified Double Geneva Curtain structure attached to T-posts . Atypically, these vines were not grafted to root stock, which is used often in the region to modify vigor or limit disease .In Sept.–Oct. of 2011, above ground biomass was measured from 72 vines. The vineyard was divided equally in twelve randomly assigned blocks, and six individual vines from each block were processed into major biomass categories of leaf, fruit, cane and trunk plus cordon . Grape berry clusters were collected in buckets, with fruit separated and weighed fresh in the field. Biomass was transported off site to partially air dry on wire racks and then fully dried in large ventilated ovens. Plant tissues were dried at 60 °C for 48 h and then ground to pass through a 250 μm mesh sieve using a Thomas Wiley® Mini-Mill . Total C in plant tissues was analyzed using a PDZ Europa ANCA-GSL elemental analyzer at the UC Davis Stable Isotope Facility. For cluster and berry C estimations, grape clusters were randomly selected from all repetitions. Berries were removed from cluster rachis. While the berries were frozen, the seeds and skins were separated from the fruit flesh or “pulp”, and combined with the juice . The rachis, skins and seeds were dried in oven and weighed. The pulp was separated from the juice + pulp with vacuum filtration using a pre-weighed Q2 filter paper . The filter paper with pulp was oven dried and weighed to get insoluble solid fraction .

However,leaf petioles analysis of grapes from both vineyards showed considerable differences in nutrient levels, especially in the primary macro-nutrients . During both seasons, the amount of nitrogen in the form of nitrate in LP-V9 was roughly 2 to 3 times higher than the normal levels, in contrast to its counterpart in LP-V7, which slightly accumulated more or less N. Similarly, LP-V9 contained higher percentages of phosphorus and potassium compared to LP-V7 . Conversely, the amounts of secondary macro-nutrients, calcium and magnesium , in LP-V7 were within the normal range but greater than LP-V9, which showed Mg deficiency in the first year only. Regarding the micro-nutrients, their levels were mainly within or around the normal range at both vineyards and during both seasons, with some differences . For example, zinc was slightly higher in LP-V9, especially in the first year. On the contrary, manganese and chlorine were roughly 2 times higher in V7 . Similarly, soil analysis shoed a higher level of nitrogen, potassium and magnesium . However, no significant difference was observed in all other soil macro and micro-nutrients. During the two seasons of the study, we determined the total marketable yield and the number of clusters in both vineyards. Our data revealed a higher yield in V7 compared to V9 in 2016 and 2017, respectively. The lower yield in V9 can likely be attributed to the smaller number of clusters in V9 compared to V7 during 2016 and 2017. To monitor the changes in the biochemical composition of Scarlet Royal berries, V7 and V9 berries were periodically sampled at six time points from veraison until the end of the season . The obtained data showed that berry polyphenols exhibited discernible patterns in both vineyards, most notably during the ripening stage .

Of special interest were the tannin compounds, blueberry box which widely affect organoleptic properties such as astringency and bitterness . Our data showed that berries from both V7 and V9 vineyards maintained lower levels of tannin from veraison up to the middle of August . Subsequently, a significant gradual increment of tannin took place. However, only V9-berries showed consistent accumulation of tannin over the two studied seasons compared to V7-berries, where the significant induction occurred only during the first season. It is worth noting that the levels of tannin were lower in both vineyards during the second year compared to the first season. Nevertheless, they were more pronounced in V9-berries compared to V7-berries, with roughly 2- to 4.5-fold increases by the end of the harvesting time during the two seasons, respectively . The patterns of catechin and quercetin glycosides were inconsistent during both seasons, particularly within V7-berries . During the first year, for instance, the levels of catechin were similar in both vineyards, showing a dramatic increase only by the end of the season . In contrast, during the second year, such induction of catechin was exclusively restricted to V9-berries, starting from time S3 . For quercetin glycosides, V7-berries exhibited significantly higher amounts at early stages during both seasons relative to V9-berries . However, subsequent amounts were comparable in both vineyards during the first season only , but not in the second one, where V7-berries showed a significant drop at the last sample S6 . Interestingly, the levels of quercetin glycosides were roughly equal at the last V9-berries sample between both seasons despite such inconsistency. For total anthocyanins , the levels in early samples were comparable in both vineyards and seasons . Afterwards, their pattern started to vary between V7 and V9 within the same season, as well as from the first season to the second, as the nutrient amounts fluctuated as well . Nevertheless, TAC accumulation was positively correlated with the progress of ripening in V7-berries, but not V9-berries. To further confirm our data, we measured these phenolic compounds for the third time in mid-September of the next year .

Overall, the results showed that the patterns of tannins and TAC were reciprocally inverted between V7-berries and V9-berries as ripening advanced. In addition, both catechin and quercetin glycosides most likely followed the pattern of tannins despite their seasonal fluctuations. To further distinguish V7-berries and V9-berries and assess their astringency development, a panel test was performed using samples at three commercial harvest times . A group of 12 nontechnical panelists scored berry astringency on a scale from 1 to 7, where 1 is extremely low and 7 is extremely high. The panelists were trained using samples from contrasting standard varieties, including Flame Seedless and Crimson as non-astringent and Vintage Red known for its astringent taste . The results showed that V7-berries exhibited lower intensity of astringency compared to V9-berries . As ripening proceeded, astringency levels increased in V9-berries, but decreased in V7-berries. Moreover, we collected samples from clusters with various astringent taste and measured its tannins content. We were able to determine that the threshold level of tannins that causes the Scarlet Royal astringency taste is around 400 mg/L . Taking into account the levels of polyphenol compounds and the taste panel data together , it is evident that astringency development is positively associated with tannins’ accumulation throughout the ripening process of V9-berries. Nevertheless, organoleptic analysis revealed a significant difference in the berries of the two vineyards, particularly in terms of total soluble solids and titratable acidity . Notably, V9 berries exhibited higher titratable acidity and lower total soluble solids, especially in the later stages .

It’s worth noting that the weight of V9 berries is also higher than that of V7 .To better understand the molecular events associated with the induction of tannins and astringency upon ripening, the berry transcriptome profile was analyzed in both V7-berries and V9- berries at the late commercial harvest date . Following the quality and quantity check, extracted RNA from quadruplicate samples was deeply sequenced . Of the 19.7 to 24.4 million high-quality clean reads per replicate, 61.9% to 66.1% were mapped against the V. vinifera transcriptome . Hierarchical clustering of the RNAseq data showed explicit changes in the berry transcriptome profile between V7- berries and V9-berries . Samples were mainly separated along the first component , which was responsible for 97% of the variance, and was definitely associated with the site of cultivation; V7 and V9. In contrast, the second component was trivial, accounting for only 1% of the variance and was probably attributed to experimental error. Such results were expected, as berry samples came from the same cultivar, Scarlet Royal , and the only difference between them was the vineyard locations. To identify the differentially expressed genes in V7- berries and V9-berries at this specific time within the ripening window, the RNAseq data were analyzed using two different Bioconductor packages, DESeq2, and EdgeR . Subsequently, the DEGs with FDR < 0.05 and log2fold change > 1.5 or < –1.5 generated by both pipelines were considered . The pairwise comparison between berry transcriptomes resulted in 2134 DEGs, with 1514 up-regulated and 620 down-regulated . The data manifested the impact of the cultivation site on the transcriptional reprogramming of a large number of genes that ultimately affect berry quality. Most apparently, at the V9 vineyard, where roughly 2.5-fold higher number of berry transcripts were upregulated compared to V7 . Subsequently, the enrichment of Gene Ontology terms and Kyoto Encyclopedia of Genes and Genomes pathways were analyzed among the up- and down-regulated DEGs using the Vitis vinifera Ensembl GeneID . Among the significantly enriched GO terms, the up-regulated transcripts in V9-berries exhibited high enrichment in the molecular function GO terms for quercetin 3-O-glucosyltransferase activity and quercetin 7-Oglucosyltransferase activity . Additionally, the V9-berries induced DEGs were highly enriched in the biological process GO terms for the jasmonic acid signaling pathway and cellular response , Lphenylalanine metabolic process , L-phenylalanine biosynthetic process , and nitrogen compound metabolic process . Similarly, these DEGs were highly enriched in the KEGG pathways for the biosynthesis of secondary metabolites and phenylpropanoid biosynthesis . On the other hand, the down-regulated transcripts in V9-berries showed substantial augmentation in the MF GO terms for hormone binding , abscisic acid binding , and potassium ion transmembrane transporter activity . Correspondingly, the BP GO terms for hormone-mediated signaling pathway and response , auxin-activated signaling, cellular response, and homeostasis , abscisic acid-activated signaling, response, and cellular response , response to strigolactone , potassium ion transmembrane transport , and potassium ion transport , as well as the KEGG pathways for plant hormone signal transduction , brassinosteroid biosynthesis , blueberry package and carotenoid biosynthesis were highly enriched in the down-regulated genes of V9-berries . Overall, the transcriptome analysis pointed out the substantial changes in transcript abundance that coordinate and reflect the observed induction of tannins/astringency during the maturation and ripening of V9-berries compared to the V7-berries .To elucidate which fundamental processes were altered during tannins/astringency induction within berries, the Weighted Gene CoExpression Network Analysis was applied to construct coexpression networks. Forty-two modules were identified based on pairwise correlations among the 17553 non-lowly expressed genes . Subsequently, the biochemical data from both V7-berries and V9-berries were correlated to the WGCNA modules, and only 2 modules, M21 and M30, displayed substantial correlations with berry polyphenols, containing 5349 and 4559 genes, respectively .

The M21 module was positively linked with TAC , but negatively associated with tannins, catechin, and quercetin glycosides . On the contrary, the M30 module exhibited a positive correlation with tannins, catechin, and quercetin glycosides , but was negatively linked with TAC . The DEGs obtained from the two pipelines were assigned to both M21 and M30, yielding 604 and 1362 genes, respectively . Interestingly, the number of DEGs in each module, M21 and M30, was roughly equal to the down- and upregulated genes, respectively . To identify flavonoids/tannins-related genes that might result in such astringency diversity between V7-berries and V9-berries, hub genes were searched in the DEGs list of both modules . Only 8 hub genes were identified based on their transcript abundances in V9- berries and predicted functions. However, based on our previous work , we found another 11 genes that are significantlyexpressed but with a log2FoldChange less than 1.5, and they were included in our further analysis . The enrichment analysis of GO showed considerable enrichment in the BP GO terms for secondary metabolite biosynthetic process , flavonoid biosynthetic/metabolic process , L-phenylalanine metabolic/ catabolic process , phenylpropanoid metabolic process , phenylpropanoid biosynthetic process , chorismate biosynthetic/metabolic process , cinnamic acid biosynthetic/ metabolic process , anthocyanincontaining compound biosynthetic/metabolic process . The KEGG pathway analysis confirmed the BP GO terms, exhibiting enrichment for the biosynthesis of secondary metabolites , phenylpropanoid biosynthesis , flavonoid biosynthesis , and glutathione metabolism .To precisely elucidate their significance in the tannins/astringency diversity between V7-berries and V9-berries, we studied the expression levels of the 19 hub genes associated with the shikimic and flavonoids pathway. Except for the PAL1_1 gene , the analysis of their relative expression by real-time quantitative PCR showed a significant correlation with the Transcripts Per Million values for genes of interest, validating the transcriptomic data from both V7- and V9-berries . In general, all genes showed higher expression levels in V9-berries compared to V7-berries, but with different degrees of induction. Forinstance, the two genes involved in the shikimic acid pathway, chorismate synthase , and chorismate mutase , showed visibly higher accumulation abundance in V9-berries at the third harvesting time with approximately 6-fold and 3-fold increases, respectively, compared to V7-berries. Similarly, the upstream structural genes in the phenylpropanoids pathway, including phenylalanine ammonia lyase , trans-4- coumarate biosynthesis , and 4-coumaroyl:CoAligase 2 , were significantly induced by approximately 2- to 9-fold in V9-berries. Regarding flavonoids/PAs biosynthesis, chalcone synthase is considered a key enzyme in this pathway, converting p-coumaroyl-CoA to naringenin chalcone, which is later turned into naringenin by chalcone isomerase . Both genes were highly expressed in V9-berries . Naringenin is subsequently converted by flavonoid 3’-monooxygenase to dihydromyricetin and dihydroquercetin, which are further transformed by dihydroflavonol 4-reductase into leucodelphinidin and leucocyanidin, respectively . The expression levels of F3H and DFR also showed a commensurate induction with the upstream genes in V9-berries relative to V7-berries. Subsequently, leucoanthocyanidin dioxygenase and leucoanthocyanidin reductase catalyse the conversion of leucodelphinidin to delphinidin and -gallocatechin, respectively, as well as leucocyanidin to cyanidin and catechin, respectively. These three genes also exhibited a significant increase in V9-berries. Finally, the expression of genes encoding glutathione Stransferases , one of the most essential anthocyanin transporters, was significantly higher in V9-berries compared to V7-berries, with approximately 3- to 9.2-fold changes .