Josefina Bota

Effects of drought on photosynthesis in grapevines under field conditions: an evaluation of stomatal and mesophyll limitations

The effect of diffusional and photochemical limitations to photosynthesis was assessed in field-grown water-stressed grapevines (Vitis vinifera L.) by combined measurements of gas exchange and chlorophyll fluorescence. Drought was slowly induced, and the progressive decline of photosynthesis was examined in different grapevine cultivars along a continuous gradient of maximum mid-morning values of stomatal conductance (g), which were used as an integrative indicator of the water-stress conditions endured by the leaves. Initial decreases of g were accompanied by decreases of substomatal CO2 concentration (Ci), the estimated chloroplastic CO2 concentration (Cc) and net photosynthesis (AN), while electron transport rate (ETR) remained unaffected. With increasing drought, g, AN, Ci and Cc further decreased, accompanied by slight decreases of ETR and of the estimated mesophyll conductance (gmes). Severe drought led to strong reductions of both g and gmes, as well as of ETR. The apparent carboxylation efficiency and the compensation point for CO2 remained unchanged under severe drought when analysed on a Cc, rather than a Ci, basis, suggesting that previously reported metabolic impairment was probably due to decreased gmes.

Photosynthesis limitations during water stress acclimation and recovery in the drought-adapted Vitis hybrid Richter-110 (V. berlandieri×V. rupestris)

The hybrid Richter-110 (Vitis berlandierixVitis rupestris) has the reputation of being a genotype strongly adapted to drought. A study was performed with plants of R-110 subjected to sustained water-withholding to induce acclimation to two different levels of water stress, followed by rewatering to induce recovery. The goal was to analyse how photosynthesis is regulated during acclimation to water stress and recovery. In particular, the regulation of stomatal conductance (g(s)), mesophyll conductance to CO(2) (g(m)), leaf photochemistry (chlorophyll fluorescence and thermoluminescence), and biochemistry (V(c,max)) were assessed. During water stress, g(s) declined to 0.1 and less than 0.05 mol CO(2) m(-2) s(-1) in moderately and severely water-stressed plants, respectively, and was kept quite constant during an acclimation period of 1-week. Leaf photochemistry proved to be very resistant to the applied water-stress conditions. By contrast, g(m) and V(c,max) were affected by water stress, but they were not kept constant during the acclimation period. g(m) was initially unaffected by water stress, and V(c,max) even increased above control values. However, after several days of acclimation to water stress, both parameters declined below (g(m)) or at (V(c,max)) control values. For the latter two parameters there seemed to be an interaction between water stress and cumulative irradiance, since both recovered to control values after several cloudy days despite water stress. A photosynthesis limitation analysis revealed that diffusional limitations and not biochemical limitations accounted for the observed decline in photosynthesis during water stress and slow recovery after rewatering, both in moderately and severely stressed plants. However, the relative contribution of stomatal (SL) and mesophyll conductance (MCL) limitations changes during acclimation to water stress, from predominant SL early during water stress to similar SL and MCL after acclimation. Finally, photosynthesis recovery after rewatering was mostly limited by SL, since stomatal closure recovered much more slowly than g(m).

A ten-year study on the physiology of two Spanish grapevine cultivars under field conditions: effects of water availability from leaf photosynthesis to grape yield and quality

The effects of moderate irrigation, compared with non-irrigation, on leaf photosynthesis and transpiration, grape yield, and quality parameters, were studied over ten years in two Spanish cultivars (Tempranillo and Manto Negro) of field-grown grapevines (Vitis vinifera L.). The aim was to increase our knowledge of the relationships between water availability, canopy water losses, photosynthesis, and fruit yield and quality. A second aim was to analyse some of the mechanisms of photosynthetic down-regulation under drought, such as the capacity for RuBP regeneration and Rubisco activity.Moderate irrigation improved plant water status, leaf photosynthesis and transpiration. Considering the results over ten years, soil water availability (estimated as pre-dawn leaf water potential, ΨPD) largely determined leaf photosynthesis and leaf transpiration. Decreased photosynthesis was due to both stomatal and non-stomatal factors. The latter were related to decayed electron transport rate and reduced RuBP regeneration capacity, but not to decreased Rubisco activity.Moderate irrigation also improved grape yield, although this effect was much larger in Tempranillo than in Manto Negro. Moreover, the correlation between photosynthesis and grape yield was significant in Tempranillo, but not in Manto Negro. In contrast, the correlation between ΨPD and several parameters reflecting fruit quality (such as soluble solids and total polyphenol content) was significant only in Manto Negro. These results suggest that there is a close link between water availability and grape yield, mostly through water stress effects on photosynthesis. Drought effects on grape quality are linked to water availability but not to photosynthesis or yield.

Adjustments of water use efficiency by stomatal regulation during drought and recovery in the drought-adapted Vitis hybrid Richter-110 (V. berlandieri x V. rupestris).

The hybrid Richter-110 (Vitis berlandieri x Vitis rupestris) (R-110) has the reputation of being a genotype strongly adapted to drought. A study was performed with plants of R-110 subjected to water withholding followed by re-watering. The goal was to analyze how stomatal conductance (g(s)) is regulated with respect to different physiological variables under water stress and recovery, as well as how water stress affects adjustments of water use efficiency (WUE) at the leaf level. Water stress induced a substantial stomatal closure and an increase in WUE, which persisted many days after re-watering. The g(s) during water stress was mainly related to the content of ABA in the xylem and partly related to plant hydraulic conductivity but not to leaf water potential. By contrast, low g(s) during re-watering did not correlate with ABA contents and was only related to a sustained decreased hydraulic conductivity. In addition to a complex physiological regulation of stomatal closure, g(s) and rate of transpiration (E) were strongly affected by leaf-to-air vapor pressure deficit (VPD) in a way dependent of the treatment. Interestingly, E increased with increasing VPD in control plants, but decreased with increasing VPD in severely stressed plants. All together, the fine stomatal regulation in R-110 resulted in very high WUE at the leaf level. This genotype is revealed to be very interesting for further studies on the physiological mechanisms leading to regulation of stomatal responsiveness and WUE in response to drought.

Interactive effects of soil water deficit and air vapour pressure deficit on mesophyll conductance to CO2 in Vitis vinifera and Olea europaea

The present work aims to study the interactive effect of drought stress and high vapour pressure deficit (VPD) on leaf gas exchange, and especially on mesophyll conductance to CO(2) (g(m)), in two woody species of great agronomical importance in the Mediterranean basin: Vitis vinifera L. cv. Tempranillo and Olea europaea L. cv. Manzanilla. Plants were grown in specially designed outdoor chambers with ambient and below ambient VPD, under both well-irrigated and drought conditions. g(m) was estimated by the variable J method from simultaneous measurements of gas exchange and fluorescence. In both species, the response to soil water deficit was larger in g(s) than in g(m), and more important than the response to VPD. Olea europaea was apparently more sensitive to VPD, so that plants growing in more humid chambers showed higher g(s) and g(m). In V. vinifera, in contrast, soil water deficit dominated the response of g(s) and g(m). Consequently, changes in g(m)/g(s) were more related to VPD in O. europaea and to soil water deficit in V. vinifera. Most of the limitations of photosynthesis were diffusional and especially due to stomatal closure. No biochemical limitation was detected. The results showed that structural parameters played an important role in determining g(m) during the acclimation process. Although the relationship between leaf mass per unit area (M(A)) with g(m) was scattered, it imposed a limitation to the maximum g(m) achievable, with higher values of M(A) in O. europaea at lower g(m) values. M(A) decreased under water stress in O. europaea but it increased in V. vinifera. This resulted in a negative relationship between M(A) and the CO(2) draw-down between substomatal cavities and chloroplasts in O. europaea, while being positive in V. vinifera.

Effects of drought on light-energy dissipation mechanisms in high-light-acclimated, field-grown grapevines

The response of several light-energy dissipation mechanisms to water shortage was analysed in a 10-year study in field-grown, high-light-acclimated grapevines, and compared with those of greenhouse-grown, low-light-acclimated grapevines. Dissipation mechanisms, except leaf photochemistry, differ among cultivars and acclimate to the prevailing light conditions during growth. However, no additional acclimation to drought was observed. The dependence of the dissipation responses on stomatal conductance suggests that low CO2 availability in the chloroplasts during drought triggers variations in the energy dissipation pattern. In irrigated grapevines under high light, more than 50% of total absorbed energy is thermally dissipated. There is evidence that implicates the xanthophyll cycle as the main thermal dissipation processes. CO2 assimilation is the most important photochemical pathway of dissipation in irrigated plants, but is replaced by photorespiration when CO2 assimilation declines under mild drought. Under moderate to severe drought, both photosynthesis and photorespiration decline, and thermal dissipation increases to account for up to 90% of total dissipation. Involvement of other processes in light dissipation is minimal in grapevines. Even in severely-stressed leaves, the incidence of photoinhibition is very low, indicating that safe dissipation of absorbed energy is very effective in grapevines.

Effect of water stress on partitioning of 14C-labelled photosynthates in Vitis vinifera

The influence of fruits on export and distribution of photosynthates was studied in Vitis vinifera L. cv. Tempranillo using 14C labelling. Also, the influence of water stress was analysed in fruiting and fruitless plants of Tempranillo and fruiting plants of cv. Alfonso Lavallée. In fruitless plants of Tempranillo, reserve organs (trunk, roots and lower shoot) represented 80% of total plant dry matter (DM), and imported up to 90% of the total 14C exported from the fed leaf. Therefore, the distribution pattern of photosynthates in these plants reflected mainly the sink size. However, the presence of fruits in Tempranillo strongly stimulated 14C export and changed the distribution pattern of assimilates. Fruits imported up to 70-80% of the total 14C exported, while representing only 25% of the total plant DM. Therefore, the strength of fruits as carbon sinks was independent of sink size, and it is discussed on the basis of a water potential gradient theory.

Alterations in primary and secondary metabolism in Vitis vinifera ‘Malvasía de Banyalbufar’ upon infection with Grapevine leafroll‐associated virus 3

Plant defense mechanisms against pathogens result in differential regulation of various processes of primary and secondary metabolism. Imaging techniques, such as fluorescence imaging and thermography, are very valuable tools providing spatial and temporal information about these processes. In this study, effects of Grapevine leafroll-associated virus 3 (GLRaV-3) on grapevine physiology were analyzed in pot-grown asymptomatic plants of the white cultivar Malvasía de Banyalbufar. The virus triggered changes in the activity of photosynthesis and secondary metabolism. There was a decrease in the photorespiratory intermediates glycine and serine in infected plants, possibly as a defense response against the infection. The content of malate, which plays an important role in plant metabolism, also decreased. These results correlate with the increased non-photochemical quenching found in infected plants. On the other hand, the concentration of flavonols (represented by myricetin, kaempferol and quercetin derivatives) and hydroxycinnamic acids (which include derivatives of caffeic acid) increased following infection by the virus. These compounds could be responsible for the increase in multicolor fluorescence F440 (blue fluorescence) and F520 (green fluorescence) on the leaves, and changes in the fluorescence parameters F440/F680, F440/F740, F520/F680, F520/F740 and F680/F740. The combined analysis of chlorophyll fluorescence kinetics and blue-green fluorescence emitted by phenolics could constitute disease signatures allowing the discrimination between GLRaV-3 infected and non-infected plants at very early stage of infection, prior to the development of symptoms.

Exploring the genetic variability in water use efficiency: Evaluation of inter and intra cultivar genetic diversity in grapevines

Genetic improvement of crop Water Use Efficiency (WUE) is a general goal because the increasing water scarcity and the trend to a more sustainable agriculture. For grapevines, this subject is relevant and need an urgent response because their wide distribution in semi-arid areas. New cultivars are difficult to introduce in viticulture due to the narrow dependency of consumer appreciation often linked to a certain particular wine taste. Clones of reputed cultivars would presumably be more accepted but little is known on the intra-cultivar genetic variability of the WUE. The present work compares, on the basis of two field assays, the variability of intrinsic water use efficiency (WUEi) in a large collection of cultivars in contrast with a collection of clones of Tempranillo cultivar. The results show that clonal variability of WUEi was around 80% of the inter-cultivar, thus providing a first assessment on the opportunity for clonal selection by WUE. Plotting the WUEi data against stem water potential or stomatal conductance it was possible to identify cultivars and clones out of the confidence intervals of this linear regression thus with significantly higher and lower WUEi values. The present results contribute to open the expectative for a genetic improvement of grapevine WUE.

Combined effect of virus infection and water stress on water flow and water economy in grapevines

Water limitation is one of the major threats affecting grapevine production. Thus, improving water-use efficiency (WUE) is crucial for a sustainable viticulture industry in Mediterranean regions. Under field conditions, water stress (WS) is often combined with viral infections as those are present in major grape-growing areas worldwide. Grapevine leafroll-associated virus 3 (GLRaV-3) is one of the most important viruses affecting grapevines. Indeed, the optimization of water use in a real context of virus infection is an important topic that needs to be understood. In this work, we have focused our attention on determining the interaction of biotic and abiotic stresses on WUE and hydraulic conductance (Kh ) parameters in two white grapevine cultivars (Malvasia de Banyalbufar and Giró Ros). Under well-watered (WW) conditions, virus infection provokes a strong reduction (P < 0.001) in Kpetiole in both cultivars; however, Kleaf was only reduced in Malvasia de Banyalbufar. Moreover, the presence of virus also reduced whole-plant hydraulic conductance (Khplant ) in 2013 and 2014 for Malvasia de Banyalbufar and in 2014 for Giró Ros. Thus, the effect of virus infection on water flow might explain the imposed stomatal limitation. Under WS conditions, the virus effect on Kplant was negligible, because of the bigger effect of WS than virus infection. Whole-plant WUE (WUEWP ) was not affected by the presence of virus neither under WW nor under WS conditions, indicating that plants may adjust their physiology to counteract the virus infection by maintaining a tight stomatal control and by sustaining a balanced carbon change.