The physiology and proteomics of drought tolerance in maize: early stomatal closure as a cause of lower tolerance to short-term dehydration?

Understanding the response of a crop to drought is the first step in the breeding of tolerant genotypes. In our study, two maize (Zea mays L.) genotypes with contrasting sensitivity to dehydration were subjected to moderate drought conditions. The subsequent analysis of their physiological parameters revealed a decreased stomatal conductance accompanied by a slighter decrease in the relative water content in the sensitive genotype. In contrast, the tolerant genotype maintained open stomata and active photosynthesis, even under dehydration conditions. Drought-induced changes in the leaf proteome were analyzed by two independent approaches, 2D gel electrophoresis and iTRAQ analysis, which provided compatible but only partially overlapping results. Drought caused the up-regulation of protective and stress-related proteins (mainly chaperones and dehydrins) in both genotypes. The differences in the levels of various detoxification proteins corresponded well with the observed changes in the activities of antioxidant enzymes. The number and levels of up-regulated protective proteins were generally lower in the sensitive genotype, implying a reduced level of proteosynthesis, which was also indicated by specific changes in the components of the translation machinery. Based on these results, we propose that the hypersensitive early stomatal closure in the sensitive genotype leads to the inhibition of photosynthesis and, subsequently, to a less efficient synthesis of the protective/detoxification proteins that are associated with drought tolerance.     

Integrated drought responses of black poplar: how important is phenotypic plasticity?

Climate change is expected to increase drought frequency and intensity which will threaten plant growth and survival. In such fluctuating environments, perennial plants respond with hydraulic and biomass adjustments, resulting in either tolerant or avoidant strategies. Plants' response to stress relies on their phenotypic plasticity. The goal of this study was to explore physiology of young Populus nigra in the context of a time‐limited and progressive water deficit in regard to their growth and stress response strategies. Fourteen French 1‐year‐old black poplar genotypes, geographically contrasted, were subjected to withholding water during 8 days until severe water stress. Water fluxes (i.e. leaf water potentials and stomatal conductance) were analyzed together with growth (i.e. radial and longitudinal branch growth, leaf senescence and leaf production). Phenotypic plasticity was calculated for each trait and response strategies to drought were deciphered for each genotype. Black poplar genotypes permanently were dealing with a continuum of adjusted water fluxes and growth between two extreme strategies, tolerance and avoidance. Branch growth, leaf number and leaf hydraulic potential traits had contrasted plasticities, allowing genotype characterization. The most tolerant genotype to water deficit, which maintained growth, had the lowest global phenotypic plasticity. Conversely, the most sensitive and avoidant genotype ceased growth until the season's end, had the highest plasticity level. All the remaining black poplar genotypes were close to avoidance with average levels of traits plasticity. These results underpinned the role of plasticity in black poplar response to drought and calls for its wider use into research on plants' responses to stress.     

Water status of drought-resistant and drought-sensitive sorghum treated with ethephon

The effects of ethephon on stomatal resistance, water potential, osmotic potential, turgor potential, and ethylene production were determined on leaves of a drought-resistant (KS 65) and a drought-sensitive (IA 25) genotype of sorghum [Sorghum bicolor (L.) Moench] grown under wellwatered or drought-stressed conditions. With both sufficient and limited water supply, ethephon had no effect on the adaxial, abaxial, or total stomatal resistance of either genotype. For both water treatments, the adaxial stomatal resistance of the drought-sensitive genotype was higher than that of the drought-resistant genotype. Ethephon increased the amount of ethylene produced by the plants under both levels of water. For plants with sufficient water, water potentials of both genotypes were lowered by ethephon. Ethephon had no effect on the water potentials under drought or on the osmotic potentials under either water regime. With drought, the turgor potential of the drought-sensitive genotype, but not that of the drought-resistant, was increased by ethephon.     

Water status of drought-resistant and drought-sensitive sorghum treated with ethephon

The effects of ethephon on stomatal resistance, water potential, osmotic potential, turgor potential, and ethylene production were determined on leaves of a drought-resistant (KS 65) and a drought-sensitive (IA 25) genotype of sorghum [Sorghum bicolor (L.) Moench] grown under wellwatered or drought-stressed conditions. With both sufficient and limited water supply, ethephon had no effect on the adaxial, abaxial, or total stomatal resistance of either genotype. For both water treatments, the adaxial stomatal resistance of the drought-sensitive genotype was higher than that of the drought-resistant genotype. Ethephon increased the amount of ethylene produced by the plants under both levels of water. For plants with sufficient water, water potentials of both genotypes were lowered by ethephon. Ethephon had no effect on the water potentials under drought or on the osmotic potentials under either water regime. With drought, the turgor potential of the drought-sensitive genotype, but not that of the drought-resistant, was increased by ethephon.Contribution no. 90-147-J of the Journal Series of the Kansas Agricultural Experiment Station, Manhattan, Kansas, USA.     

Genetic variation in physiological responses of mungbeans (<Emphasis Type="Italic">Vigna radiata</Emphasis> (L.) Wilczek) to drought

Mungbean is a relatively drought tolerant leguminous crop with a short life cycle. Using leaf water loss (LWL) as a screen for drought tolerance, two mungbean genotypes exhibiting more than two–fold variation in leaf water loss were explored for the genetic variation in their physiological and molecular responses to drought. Efficient stomatal regulation together with better photosynthetic capacity constituted an important trait combination for drought adaptation in water saving low LWL genotype. The stomatal closure under drought was accompanied with a concomitant down-regulation of farnesyl transferase gene. However, cooler canopy temperature, a well branched root system coupled with a relatively higher proline accumulation in water spending high LWL genotype constituted another set of adaptive traits operating when exposed to deficit soil moisture conditions. We report drought induced down-regulation of proline dehydrogenase and the presence of 118 base pair intron in this gene. The high seed yield of low LWL genotype despite a hotter canopy might be attributed to higher net assimilation and quantum yield recorded under drought in this genotype. Thus, these interlinked features contribute to adaptive mechanisms of mungbeans which is widely grown in harsh environments exposed to drought and high temperatures.     

Screening for candidate bacterial biocontrol agents against soilborne fungal plant pathogens

Previous studies relating root systems and drought tolerance in oil palm focused mainly on biomass. Yet, total root length (TRL), total root surface area (TRS), and root distribution in the soil better determine water uptake. These morphological traits were studied on 3 oil palm genotypes displaying a contrasting drought tolerance. A new concept of potential root water extraction ratio (PRER) was developed using measured half-distances between roots and some assumptions about the distance of water migration from soil to root. PRER was determined in conjunction with soil moisture extraction efficiency (SMEE). The presumed tolerant genotype (T) had higher TRL, TRS and PRER than the susceptible genotype (S), whilst the performance of the control genotype (I) was intermediate. Surprisingly, during a period of moderate water deficit, T had a lower SMEE than S, which was interpreted successfully with PRER, as the result of a better access to a large volume of soil and of a slower drying out of the soil around the roots. PRER appears as a helpful indicator for comparing or ranking genotypes, and for addressing better the complexity of the genetic variability of drought tolerance.     

Drought Tolerance in Mung Bean is Associated with the Genotypic Divergence,Regulation of Proline,Photosynthetic Pigment and Water Relation

Drought is one of the critical conditions for the growth and productivity of many crops including mung bean (Vigna radiata L. Wilczek). Screening of genotypes for variations is one of the suitable strategies for evaluating crop adaptability and global food security. In this context, the study investigated the physiological and biochemical responses of four drought tolerant (BARI Mung-8, BMX-08010-2, BMX-010015, BMX-08009-7), and four drought sensitive (BARI Mung-1, BARI Mung-3, BU Mung-4, BMX-05001) mung bean genotypes under wellwatered (WW) and water deficit (WD) conditions. The WW treatment maintained sufficient soil moisture (22% ± 0.5%, i.e., 30% deficit of available water) by regularly supplying water. Whereas, the WD treatment was maintained throughout the growing period, and water was applied when the wilting symptom appeared. The drought tolerant (DT) genotypes BARI Mung-8, BMX-08010-2, BMX-010015, BMX-08009-7 showed a high level of proline accumulation (2.52–5.99 mg g⁻¹ FW), photosynthetic pigment (total chlorophyll 2.96–3.27 mg g⁻¹ FW at flowering stage, and 1.62–2.38 mg g⁻¹ FW at pod developing stage), plant water relation attributes including relative water content (RWC) (82%–84%), water retention capacity (WRC) (12–14) as well as lower water saturation deficit (WSD) (19%–23%), and water uptake capacity (WUC) (2.58–2.89) under WD condition, which provided consequently higher relative seed yield. These indicate that the tolerant genotypes gained better physiobiochemical attributes and adaptability in response to drought conditions. Furthermore, the genotype BMX- 08010-2 showed superiority in terms of those physio-biochemical traits, susceptibility index (SSI) and stress tolerance index (STI) to other genotypes. Based on the physiological and biochemical responses, the BMX-08010-2 was found to be a suitable genotype for sustaining yield under drought stress, and subsequently, it could be recommended for crop improvement through hybridization programs. In addition, the identified traits can be used as markers to identify tolerant genotypes for drought-prone areas.     

Photosynthesis of six barley genotypes as affected by water stress

The effect of water stress on plant water status and net photosynthetic gas exchange (PN) in six barley genotypes (Hordeum vulgare L.) differing in productivity and drought tolerance was studied in a controlled growth chamber. Osmotic adjustment (OA), PN, stomatal conductance (gs), and the ratio intercellular/ambient. CO2 concentration (Ci/Ca) were evaluated at four different levels of soil water availability, corresponding to 75, 35, 25 and 15 % of total available water. Variability in OA capacity was observed between genotypes: the drought tolerant genotypes Albacete and Alpha showed higher OA than drought susceptible genotypes Express and Mogador. The genotype Albacete exhibited also higher PN than the others at low water potential (Ψ). The ratios of PN/gs and Ci/Ca showed that differences in photosynthetic inhibition between genotypes at low Ψ were probably due to nonstomatal effects. In Tichedrett, a landrace genotype with a very extensive root development, OA was not observed, however, it exhibited a capacity to maintain its photosynthetic activity under water stress.     

Salicylic acid induced changes on antioxidant capacity,pigments and grain yield of soybean genotypes in water deficit condition

Salicylic acid (SA) plays an important role in the regulation of plant growth and development in response to water deficit. The effect of SA (0, 0.4 and 0.8?mM) on some physiological parameters of three soybean genotypes was investigated in three irrigation schedules included (85%, 65% and 45% of field capacity) during 2014–2015. Results showed that water deficit decreased stomatal conductance, leaf area index, relative water content, membrane stability index, yield components and grain yield particularly in L17 genotype. Activities of superoxide dismutase, ascorbate peroxidase and concentration of hydrogen peroxide, proline and total protein were increased in response to water deficit as well as SA applications. SA inhibited catalase activity resulting in increased hydrogen peroxide accumulation in soybean genotypes. Application of 0.4?mM SA decreased the adverse effects of water deficit in soybean genotypes by elevation of antioxidant enzymes activity and reducing malondialdehyde formation especially in Williams genotype.     

Effects of Drought on Photosynthetic Performance and Water Relations of Four Vigna Genotypes

The effect of drought on plant water relations and photosynthesis of Vigna glabrescens (Vg) and Vigna unguiculata (cvs. 1183, EPACE-1 and Lagoa), which differ in their drought resistance, was compared. With the increase of drought severity, Vg showed a more gradual stomatal closure and maintained significantly higher levels of stomatal conductance (g_s) and photosynthetic activity (P_N) than the other genotypes even when minimum relative water content (RWC) values were observed. Furthermore, Vg was the only genotype able to accumulate significant amounts of proline already under moderate water deficit, what could explain the lower osmotic potential (ψ_s) values observed in these plants. The three V. unguiculata cultivars presented a similar stomatal control under increasing water deficit. A mesophyllic impairment of photosynthetic capacity (P_max) was detected for cv. 1183 from the beginning of drought onset (85-75 % RWC) while in the Vg plants the values remained unaffected along the whole drought period, indicating that P_N decrease observed in this genotype is mainly a consequence of stomatal closure. Such P_max maintenance suggests the existence of a high mesophyllic ability to cope with increasing tissue dehydration in Vg. This revised version was published online in September 2006 with corrections to the Cover Date.     

Alterations in photosynthetic pigments, protein and osmotic components in cotton genotypes subjected to short-term drought stress followed by recovery

In order to assess drought tolerance mechanism in cotton, short-term drought-induced biochemical responses were monitored in two cotton (Gossypium hirsutum L.) genotypes contrasting their tolerance to water deficit. The seeds of two genotypes, namely GM 090304 (moderately drought tolerant) and Ca/H 631 (drought sensitive), were sown in pots containing soil, sand and peat in the ratio of 1:1:1, and irrigated every alternate day up to 45 days after sowing when each genotype was subjected to a cycle of water stress by withholding irrigation for 7 days. The stress cycle was terminated by re-watering the stressed plants for 7 days. The leaf of the drought tolerant genotype (GM 090304) maintained higher relative water content under water stress than that of the drought sensitive genotype (Ca/H 631). The levels of biochemical components, such as chlorophylls, carotenoids, total protein, free proline, total free amino acids, sugars, starch and polyphenols, were measured during the stress as well as the recovery periods. The chlorophylls, carotenoids, protein and starch contents decreased in drought stressed plants as compared to control and tended to increase when the plants were recovered from stress. The degree of decrease in chlorophylls, carotenoids and protein contents under drought was higher in the sensitive genotype (Ca/H 631) as compared to the moderately tolerant genotype (GM 090304). However, proline, total free amino acids, total sugars, reducing sugars and polyphenol contents were increased in drought stressed plants and tended to decrease during the period of recovery. Drought-induced increases in total free amino acids, proline, sugars and polyphenols were significantly higher in the moderately tolerant genotype (GM 090304) than in the sensitive genotype (Ca/H 631). These results suggest that proline, sugars and polyphenols act as main compatible solutes in cotton in order to maintain osmotic balance, to protect cellular macromolecules, to detoxify the cells, and to scavenge free radicals under water stress condition.     

Photosynthesis of six barley genotypes as affected by water stress

The effect of water stress on plant water status and net photosynthetic gas exchange (PN) in six barley genotypes (Hordeum vulgare L.) differing in productivity and drought tolerance was studied in a controlled growth chamber. Osmotic adjustment (OA), PN, stomatal conductance (gs), and the ratio intercellular/ambient. CO2 concentration (Ci/Ca) were evaluated at four different levels of soil water availability, corresponding to 75, 35, 25 and 15 % of total available water. Variability in OA capacity was observed between genotypes: the drought tolerant genotypes Albacete and Alpha showed higher OA than drought susceptible genotypes Express and Mogador. The genotype Albacete exhibited also higher PN than the others at low water potential (Ψ). The ratios of PN/gs and Ci/Ca showed that differences in photosynthetic inhibition between genotypes at low Ψ were probably due to nonstomatal effects. In Tichedrett, a landrace genotype with a very extensive root development, OA was not observed, however, it exhibited a capacity to maintain its photosynthetic activity under water stress. This revised version was published online in June 2006 with corrections to the Cover Date.     

Independent variation in photosynthetic capacity and stomatal conductance leads to differences in intrinsic water use efficiency in 11 soybean genotypes before and during mild drought

Intrinsic water use efficiency (WUE(intr)), the ratio of photosynthesis to stomatal conductance to water, is often used as an index for crop water use in breeding projects. However, WUE(intr) conflates variation in these two processes, and thus may be less useful as a selection trait than knowledge of both components. The goal of the present study was to determine whether the contribution of photosynthetic capacity and stomatal conductance to WUE(intr) varied independently between soybean genotypes and whether this pattern was interactive with mild drought. Photosynthetic capacity was defined as the variation in WUE(intr) that would occur if genotypes of interest had the same stomatal conductance as a reference genotype and only differed in photosynthesis; similarly, the contribution of stomatal conductance to WUE(intr) was calculated assuming a constant photosynthetic capacity across genotypes. Genotypic differences in stomatal conductance had the greatest effect on WUE(intr) (26% variation when well watered), and was uncorrelated with the effect of photosynthetic capacity on WUE(intr). Thus, photosynthetic advantages of 8.3% were maintained under drought. The maximal rate of Rubisco carboxylation, generally the limiting photosynthetic process for soybeans, was correlated with photosynthetic capacity. As this trait was not interactive with leaf temperature, and photosynthetic capacity differences were maintained under mild drought, the observed patterns of photosynthetic advantage for particular genotypes are likely to be consistent across a range of environmental conditions. This suggests that it is possible to employ a selection strategy of breeding water-saving soybeans with high photosynthetic capacities to compensate for otherwise reduced photosynthesis in genotypes with lower stomatal conductance.     

Differential responses of the enzymes involved in proline biosynthesis and degradation in drought tolerant and sensitive cotton genotypes during drought stress and recovery

The relative water content (RWC), free proline levels and the activities of enzymes involved in proline metabolism were studied in drought tolerant (Ca/H 680) and drought sensitive (Ca/H 148) genotypes of cotton (Gossypium hirsutum L.) during induction of water stress and posterior recovery. Water stress caused a significant increase in proline levels and P5CS activity in leaves of both tolerant and sensitive genotypes, whereas the activity of P5CR increased minimally and the activity of OAT remains unchanged. The activity of PDH decreased under drought stress in both the genotypes. The leaf of tolerant genotype maintained higher RWC, photosynthetic activity and proline levels, as well as higher P5CS and P5CR activities under water stress than that of drought sensitive genotype. The drought induced proline levels and activities of P5CS and P5CR declined and tend to be equal to their respective controls, during recovery, whereas the PDH activity tends to increase. These results indicate that induction of proline levels by up regulation of P5CS and down regulation of PDH may be involved in the development of drought tolerance in cotton.     

Protein composition and native state of pigments of thylakoid membrane of wheat genotypes differently tolerant to water stress

Protein composition and native state of chlorophylls were analyzed in two wheat (Triticum durum L.) genotypes with different tolerance to drought, Barakatli-95 (drought-tolerant) and Garagylchyg-2 (drought-sensitive), during water deficit. It is shown that the plants subjected to water deficit appear to have a slight increase in α-and β-subunits of CF₁ATP-synthase complex (57.5 and 55 kD, respectively) in Barakatli-95 and their lower content in Garagylchyg-2. Steady-state levels of the core antenna of PS II (CP47 and CP43) and light-harvesting Chl a/b-apoproteins (LHC) II in the 29.5–24 kD region remained more or less unchanged in both wheat genotypes. The synthesis of 36 kD protein and content of low-molecular-weight polypeptides (21.5, 16.5, and 14 kD) were noticeably increased in the tolerant genotype Barakatli-95. Drought caused significant changes in the carotenoid region of the spectrum (400–500 nm) in drought-sensitive genotype Garagylchyg-2 (especially in the content of pigments of the violaxanthin cycle). A shift of the main band from 740–742 to 738 nm is observed in the fluorescence spectra (77 K) of chloroplasts from both genotypes under water deficiency, and there is a stimulation of the ratio of fluorescence band intensity F687/F740. Published in Russian in Biokhimiya, 2006, Vol. 71, No. 2, pp. 223–228.     

Changes in antioxidant enzyme activities and gene expression in two muskmelon genotypes under progressive water stress

Responses of two muskmelon (Cucumis melo L.) genotypes (drought tolerant SC-15 and drought susceptible EC-564755) were analyzed at 0, 7, 14, and 21 d of progressive water stress. Although water deficit caused a significant decline in relative water content, the magnitude of reduction was lower in SC-15. Electrolyte leakage, hydrogen peroxide, and malonydialdehyde generation were higher in EC-564755, whereas accumulation of proline was higher in SC-15. Higher activities of antioxidant enzymes, such as catalase, superoxide dismutase, ascorbate peroxidase, guaiacol peroxidase, and glutathione reductase, and higher expression of the respective genes were recorded in SC-15 than in EC-564755. Expressions of DREB2C and DREB3 in SC-15 revealed a fluctuating pattern with down-regulation on days 7 and 21 of water stress, whereas up-regulation was observed on day 14. Concurrently, both genes in EC-564755 showed continuous down-regulation on days 7, 14, and 21 of water stress. Expressions of RD22 and dehydrin recorded on days 7, 14, and 21 were lower in SC-15. The cluster analysis showed that, these two genotypes had a clear distinction in physiological and biochemical properties and gene expressions under water stress and the genotype SC-15 had more efficient osmoprotectant mechanism than genotype EC-564755 under water deficit conditions.     

Proteomics analysis of sensitive and tolerant barley genotypes under drought stress

Drought is a severe environmental constraint to plant productivity and an important factor limiting barley yield. To investigate the initial response of barley to drought stress, changes in protein profile were analyzed using a proteomics technique. Three-day-old barley seedlings of sensitive genotype 004186 and tolerant genotype 004223 were given two treatments, one with 20 % polyethylene glycol and the second with drought induced by withholding water. After 3 days of treatments, proteins were extracted from shoots and separated by 2-dimensional polyacrylamide gel electrophoresis. Metabolism related proteins were decreased in the sensitive genotype under drought; however, they were increased in the tolerant genotype. Photosynthetic related proteins were decreased and increased among the three sensitive and three tolerant genotypes, respectively. In addition, amino acid synthesis and degradation related proteins were increased and decreased among the three tolerant genotypes. These results suggest that chloroplastic metabolism and energy related proteins might play a significant role in the adaptation process of barley seedlings under drought stress.     

Screening and Assessment of Selected Chilli (<i>Capsicum annuum</i> L.) Genotypes for Drought Tolerance at Seedling Stage

This study was undertaken to investigate oxidative stress tolerant mechanisms in chilli (Capsicum annuum L.) under drought genotypes through evaluating morphological, physiological, biochemical and stomatal parameters. Twenty genotypes were evaluated for their genetic potential to drought stress tolerant at seedling stage. Thirty days old seedlings were exposed to drought stress induced by stop watering for the following 10 days and rewatering for the following one week as recovery. Based on their survival performance, two tolerant genotypes viz. BD-10906 and BD-109012 and two susceptible genotypes viz. BD-10902 and RT-20 were selected for studying the oxidative stress tolerance mechanism. Drought reduced root and shoot length, dry weight, ratio, petiole weight and leaf area in both tolerant and susceptible genotypes, and a higher reduction was observed in susceptible genotypes. Lower reduction of leaf area and photosynthetic pigments were also found in tolerant genotypes. Moreover, tolerant genotypes showed higher recovery than susceptible genotypes after the removal of stress. A higher reduction of relative water content (RWC) may cause an imbalance between absorbed and transpirated water in susceptible genotypes. Higher accumulation of proline in tolerant genotypes might be helpful to for better osmotic maintenance than that in susceptible genotypes. Tolerant genotypes showed higher antioxidant activity as they showed DPPH radical scavenging percentage than the susceptible genotypes. Moreover, closer stomata in tolerant genotypes than susceptible ones helped to avoid dehydration in tolerant genotypes. Thus, the above morphological, physiological, biochemical and stomatal parameters helped to show better tolerance in chilli under drought stress.