Analysis of Natural Variation in Bermudagrass (Cynodon dactylon) Reveals Physiological Responses Underlying Drought Tolerance

Bermudagrass (Cynodon dactylon) is a widely used warm-season turfgrass and one of the most drought tolerant species. Dissecting the natural variation in drought tolerance and physiological responses will bring us powerful basis and novel insight for plant breeding. In the present study, we evaluated the natural variation of drought tolerance among nine bermudagrass varieties by measuring physiological responses after drought stress treatment through withholding water. Three groups differing in drought tolerance were identified, including two tolerant, five moderately tolerant and two susceptible varieties. Under drought stress condition, drought sensitive variety (Yukon) showed relative higher water loss, more severe cell membrane damage (EL), and more accumulation of hydrogen peroxide (H₂O₂) and malondialdehyde (MDA), while drought tolerant variety (Tifgreen) exhibited significantly higher antioxidant enzymes activities. Further results indicated that drought induced cell injury in different varieties (Yukon, SR9554 and Tifgreen) exhibited liner correlation with leaf water content (LWC), H₂O₂ content, MDA content and antioxidant enzyme activities. Additionally, Tifgreen plants had significantly higher levels of osmolytes (proline level and soluble sugars) when compared with Yukon and SR9554 under drought stress condition. Taken together, our results indicated that natural variation of drought stress tolerance in bermudagrass varieties might be largely related to the induced changes of water status, osmolyte accumulation and antioxidant defense system.     

Evaluation of physiological screening tests for breeding drought resistant triticale (x <Emphasis Type="Italic">Triticosecale</Emphasis> wittmack)

Effects of soil drought on crop yield of 4 strains and 7 cultivars of spring triticale was investigated under field condition. The Drought Susceptibility Index (DSI) was evaluated in a two year experiment by the determination of grain loss in conditions of two soil moisture levels (drought-D and irrigated-IR). In the experiment response to drought was evaluated by different screening tests (leaf gaseous exchange, leaf water potential, chlorophyll content and fluorescence, leaf injury by drought and by simulated drought and heat temperature and water loss by excited leaf. The DSI values and the results of screening tests showed the genetic variation in the degree of drought tolerance. The values of DSI enabled the ranking of the tested triticale genotypes with respect to their drought tolerance and allow to divide them into three groups of drought susceptibility. Large differences among studied forms were observed also in changes of leaf water potential, fluorescence and leaf injury. For plants in vegetative stage of growth the tested breeding forms were easily separated into groups of different drought tolerance. Changes of ψ, Fv/Fm and LI as a screening tests were the most suitable techniques for estimation of degree of drought tolerance for triticale. Laboratory screening tests (leaf injury by simulated drought (LI_DS) and high temperature (LI_HT) and water loss (WL) of excited leaf conducted for nonstressed plants in most cases were significantly correlated with DSI. The statistically significant correlation between leaf water potential (ψ) was observed only with leaf fluorescence (Fv/Fm). Changes of Fv/Fm were significantly correlated with ψ, LI and LI_HT for 50 °C. Index of leaf injury (LI) by soil drought were significantly correlated with Fv/Fm, LI_DS (−1.0, −1.5 MPa), LI_HT (45 and 50°C) and water loss (WL). The correlation coefficient between the tests LI_DS and LI_HT were most of the considered cases statistically significant which indicate that the mechanism of membranes injury resulted from simulated drought or high temperature were similar in triticale. Water loss (WL) of excited leaves was the most suitable test for screening drought tolerance in triticale population. Changes of gaseous exchange parameters were not useful as screening test in this research.     

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.     

Quantitative Trait Loci Mapping for ChlorophyⅡ Fluorescence and Associated Traits in Wheat （ Triticum aestivum）

Parameters of chlorophyll fluorescence kinetics （PCFKs） under drought stress condition are generally used to characterize instincts for dehydration tolerance in wheat （Triticum aestivum L.）. Therefore, it is important to map quantitative trait loci （QTLs） for PCFKs in wheat genetic improvement for drought tolerance. A doubled haploid （DH） population with 150 lines, derived from a cross between two common wheat varieties, Hanxuan 10 and Lumai 14, was used to analyze the correlation between PCFKs and chlorophyll content （CHIC） and to map QTLs at the grainfilling stage under conditions of both rainfed （drought stress, DS） and well-watered （WW）, respectively. QTLs for these traits were detected by QTLMapper version 1.0 based on the composite Interval mapping method of the mixed-linear model. The results showed a very significant positive correlation between Fv, Fm, Fv/Fm and Fv/Fo. The correlation coefficients were generally higher under WW than under DS. Also, there was a significant or a highly significant positive correlation between Fv, Fm, Fv/Fm, Fv/Fo and CHIC. The correlation coefficients were higher in the DS group than the WW group. A total of 14 additive QTLs （nine QTLs detected under DS and five QTLs under WW） and 25 pairs of eplstatlc QTLs （15 pairs detected under DS and 10 pairs under WW） for PCFKs were mapped on chromosomes 6A, 7A, 1B, 3B, 4D and 7D. The contributions of additive QTLs for PCFKs to phenotype variation were from 8.40% to 72.72%. Four additive QTLs （two QTLs detected under DS and WW apiece） controlling Chic were mapped on chromosomes 1A, 5A and 7A. The contributions of these QTLs for ChIC to phenotype variation were from 7.27% to 11.68%. Several QTL clusters were detected on chromosomes 1B, 7A and 7D, but no shared chromosomal regions for them were identified under different water regimes, indicating that these QTLs performed different expression patterns under rainfed and well-watered conditions.     

Evaluation of Wheat Chromosome Translocation Lines for High Temperature Stress Tolerance at Grain Filling Stage

High temperature (HT, heat) stress is detrimental to wheat (Triticum aestivum L.) production. Wild relatives of bread wheat may offer sources of HT stress tolerance genes because they grow in stressed habitats. Wheat chromosome translocation lines, produced by introgressing small segments of chromosome from wild relatives to bread wheat, were evaluated for tolerance to HT stress during the grain filling stage. Sixteen translocation lines and four wheat cultivars were grown at optimum temperature (OT) of 22/14°C (day/night). Ten days after anthesis, half of the plants were exposed to HT stress of 34/26°C for 16 d, and other half remained at OT. Results showed that HT stress decreased grain yield by 43% compared with OT. Decrease in individual grain weight (by 44%) was the main reason for yield decline at HT. High temperature stress had adverse effects on leaf chlorophyll content and Fv/Fm; and a significant decrease in Fv/Fm was associated with a decline in individual grain weight. Based on the heat response (heat susceptibility indices, HSIs) of physiological and yield traits to each other and to yield HSI, TA5594, TA5617, and TA5088 were highly tolerant and TA5637 and TA5640 were highly susceptible to HT stress. Our results suggest that change in Fv/Fm is a highly useful trait in screening genotypes for HT stress tolerance. This study showed that there is genetic variability among wheat chromosome translocation lines for HT stress tolerance at the grain filling stage and we suggest further screening of a larger set of translocation lines.     

Drought Responses of Foliar Metabolites in Three Maize Hybrids Differing in Water Stress Tolerance

Maize (Zea mays L.) hybrids varying in drought tolerance were treated with water stress in controlled environments. Experiments were performed during vegetative growth and water was withheld for 19 days beginning 17 days after sowing. Genotypic comparisons used measured changes of leaf water potential or results were expressed by time of treatment. Total dry matter of the drought tolerant hybrid on the final harvest was 53% less than that of the intermediate and susceptible maize hybrids when plants were water sufficient. This showed that maize hybrids selected for extreme drought tolerance possessed a dwarf phenotype that affected soil water contents and leaf water potentials. Changes of shoot and root growth, leaf water potential, net photosynthesis and stomatal conductance in response to the time of water stress treatment were diminished when comparing the drought tolerant to the intermediate or susceptible maize hybrids. Genotypic differences were observed in 26 of 40 total foliar metabolites during water stress treatments. Hierarchical clustering revealed that the tolerant maize hybrid initiated the accumulation of stress related metabolites at higher leaf water potentials than either the susceptible or intermediate hybrids. Opposite results occurred when changes of metabolites in maize leaves were expressed temporally. The above results demonstrated that genotypic differences were readily observed by comparing maize hybrids differing in drought tolerance based on either time of treatment or measured leaf water potential. Current findings provided new and potentially important insights into the mechanisms of drought tolerance in maize.     

Comparative physiological and metabolomic responses of four <Emphasis Type="Italic">Brachypodium distachyon</Emphasis> varieties contrasting in drought stress resistance

Brachypodium distachyon (Brachypodium) is not only a monocot grass species, but also a promising model organism of crop research. In this study, the drought resistance of four Brachypodium varieties was identified including drought stress-tolerant Bd1-1 and Bd21, drought stress-susceptible Bd3-1 and Bd18-1. Physiological assay showed that drought-tolerant varieties (Bd1-1 and Bd21) were more effective in maintenance of leaf water content, activation of catalase and peroxidase activities and accumulation of reduced glutathione, resulting in alleviated cell damage and lower reactive oxygen species level than drought-susceptible varieties (Bd3-1 and Bd18-1) in response to drought stress. In addition, 54 primary metabolites were differentially regulated among Brachypodium varieties and after drought stress treatment, indicating the complexity of Brachypodium response to drought stress. We also identified several commonly regulated metabolites especially some compatible solutes including proline and soluble sugars, which exhibited higher concentrations in the drought-tolerant varieties. Taken together, this study suggested that natural variation of Brachypodium varieties in response to drought stress might be connected with higher leaf water, enhanced accumulation of osmolyte and more effective antioxidant system, as well as the modulation of metabolic profiles under drought stress conditions.     

A comparative study on the protective role of trehalose and LEA proteins against abiotic stresses in transgenic Chinese cabbage (<Emphasis Type="Italic">Brassica campestris</Emphasis>) overexpressing<Emphasis Type="Italic">CaLEA</Emphasis> or<Emphasis Type="Italic">otsA</Emphasis>

Trehalose and LEA proteins, representative low MW chemicals that are synthesized under dehydration, are known to protect plants from drought stress. To compare their effectiveness on enhancing tolerance against various abiotic stresses, we generated transgenic Chinese cabbage plants overexpressingE. ctdi trehalose-6-phosphate synthase gene (otsA) or hot pepper (Capsicum annuum) LEA protein gene(CaLEA). Both transgenic plants exhibited altered phenotype including stunted growth and aberrant root development When subjected to drought, salt or heat stress, these plants showed remarkably improved tolerance against those stresses compared with nontransformants. After dehydration treatment, leaf turgidity and fresh weight was better maintained in both transgenic plants. GaUEA-plants performed somewhat better under dehydrated condition. When treated with 250 mM NaCI, both otsA-plants and CaLEA-plants remained equally healthier than nontransformants in maintaining leaf turgidity and delaying necrosis. Furthermore, leaf Chi content and Fv/Fm was maintained considerably higher in both transgenic plants than nontransformants. After heat-treatment at 45°C, both transgenic plants appeared much less damaged in external shape and PSII function, but LEA proteins were more protective. Our results indicate that although both trehalose and LEA proteins are effective in protecting plants against various abiotic stresses, LEA proteins seem to be more promising in generating stress-tolerant transgenic plants.     

Relationship between leaf water spin-lattice relaxation time and water relation parameters in three wheat cultivars

Pot culture studies were conducted using two drought-tolerant and one susceptible cultivar of wheat (Triticum aestivum L.) under three different moisture regimes. Proton spin-lattice relaxation time, T₁, leaf water content, LWC, leaf water potential, Ψ, solute potential, Ψ_s and turgor potential, Ψ_p were measured from 45 to 75 d after sowing at weekly intervals. The three cultivars did not differ significantly in their values of LWC, leaf water potential, and their components in the stressed and unstressed plants; but they did differ significantly in their T₁ values both under stressed and unstressed conditions on all days of measurement, with the drought tolerant cultivars having a higher T₁ compared to the susceptible cultivar. This suggests that leaf water T₁ is a better parameter for describing plant water status than the traditional water relation indices. The relation between Ψ and T₁ was logarithmic, indicating the similarity between T₁ and water activity of the cellular water.     

Which seed origin provides better tolerance to flooding and drought when restoring to face climate change?

Our goal was to establish the tolerance to flooding and drought of seedlings from a hydric gradient of different seed sources to provide recommendations for forest restoration in the face of climate change. We used Drimys winteri var. chilensis, a tree species that grows from extreme arid zones to wetlands along Chile, as the study subject. We expected that seedlings of xeric origin would perform better in drought conditions than populations from moist environments, and vice versa for flooding tolerance. We collected D. winteri seeds from xeric, mesic and wet environments. Seedlings at two development stages were submitted to an extreme flooding and drought treatment during 2 or 4 months in a common garden. After the flooding and drought assays finished, the number of surviving and damaged seedlings, lenticels and adventitious root presence, height, new leaves and specific leaf area, shoot/root ratio, water potential and/or chlorophyll fluorescence (Fv/Fm), were recorded. We found that flooding and drought affected almost all the parameters studied negatively. The xeric population seedlings, at both development stages studied, were the most tolerant to the drought and, unexpectedly, also to the flooding treatment. We recommend restoring with seedlings of xeric origin especially in arid areas where sudden flooding is frequent, as occurs in the Andes Mountains. In the face of climate change, we recommend carrying out common garden and field studies before advising which population origin should be used for restoration, since they do not always respond in accordance with expected patterns of local adaptation.     

Protective mechanism of desiccation tolerance in <Emphasis Type="Italic">Reaumuria soongorica</Emphasis>: Leaf abscission and sucrose accumulation in the stem

Reaumuria soongorica (Pall.) Maxim., a perennial semi-shrub, is widely found in semi-arid areas in northwestern China and can survive severe desiccation of its vegetative organs. In order to study the protective mechanism of desiccation tolerance in R. soongorica, diurnal patterns of net photosynthetic rate (Pn), water use efficiency (WUE) and chlorophyll fluorescence parameters of Photosystem II (PSII), and sugar content in the source leaf and stem were investigated in 6-year-old plants during progressive soil drought imposed by the cessation of watering. The results showed that R. soongorica was characterized by very low leaf water potential, high WUE, photosynthesis and high accumulation of sucrose in the stem and leaf abscission under desiccation. The maximum Pn increased at first and then declined during drought, but intrinsic WUE increased remarkably in the morning with increasing drought stress. The maximal photochemical efficiency of PSII (Fv/Fm) and the quantum efficiency of noncyclic electric transport of PSII(Φ_PSII) decreased significantly under water stress and exhibited an obvious phenomenon of photoinhibition at noon. Drought stressed plants maintained a higher capacity of dissipation of the excitation energy (measured as NPQ) with the increasing intensity of stress. Conditions of progressive drought promoted sucrose and starch accumulation in the stems but not in the leaves. However, when leaf water potential was less than −21.3 MPa, the plant leaves died and then abscised. But the stem photosynthesis remained and, afterward the plants entered the dormant state. Upon rewatering, the shoots reactivated and the plants developed new leaves. Therefore, R. soongorica has the ability to reduce water loss through leaf abscission and maintain the vigor of the stem cells to survive desiccation.     

Contribution of inorganic components to osmotic adjustment and leaf folding for drought tolerance in pearl millet

Pearl millet, Pennisetum glaucum , is capable of adapting to severely dry environmental conditions. In order to elucidate the mechanism of adaptation to highly dehydrated conditions, we selected both tolerant (IP8210) and susceptible (IP8949) accessions from a total of 15 pearl millet accessions and characterized their morphological and physiological responses to severe drought stress. When these selected accessions were stressed with a severe drought treatment, the leaves of IP8210 exhibited upright folding, a response that effectively reduces the evaporative surface area of the canopy. On the contrary, the leaves of IP8949 exhibited wilting and did not appear to adapt to the drought stress. In comparison with IP8949, the capacity of osmotic adjustment (OA) was greater in both younger leaves and stems of IP8210, while their decrease in relative water content was different. IP8210 accumulated higher concentrations of NO₃^– than IP8949 in response to drought stress. In addition to inorganic solutes, several organic components such as sucrose, glucose, quaternary ammonium compounds, and amino acids including proline were also accumulated. IP8210 tended to accumulate more amino acids, typically due to the accumulation of asparagine and proline, while IP8949 accumulated more soluble sugars. While it is possible that K⁺ and NO₃^– were the major components contributing to osmotic regulations, sugars and amino acids might also function as a cytoprotectant, in addition to their role as osmoprotectants. Collectively, these results demonstrate that the morphological adaptation of leaf folding, OA in both the younger leaves and the stem, and the accumulation of NO₃^– and amino acids during earlier stress period contribute to superior drought tolerance that was exhibited in IP8210 of pearl millet.     

Diffusion limitations and metabolic factors associated with inhibition and recovery of photosynthesis from drought stress in a C3 perennial grass species

Stomatal closure and metabolic impairment under drought stress limits photosynthesis. The objective of this study was to determine major stomatal and metabolic factors involved in photosynthetic responses to drought and recovery upon re‐watering in a C₃ perennial grass species, Kentucky bluegrass (Poa pratensis L.). Two genotypes differing in drought resistance, ‘Midnight’ (tolerant) and ‘Brilliant’ (sensitive), were subjected to drought stress for 15 days and then re‐watered for 10 days in growth chambers. Single‐leaf net photosynthetic rate (A), stomatal conductance (g_s) and transpiration rate (Tr) decreased during drought, with a less rapid decline in ‘Midnight’ than in ‘Brilliant’. Photochemical efficiency, Rubisco activity and activation state declined during drought, but were significantly higher in ‘Midnight’ than in ‘Brilliant’. The relationship between A and internal leaf CO₂ concentration (A/Ci curve) during drought and re‐watering was analyzed to estimate the relative influence of stomatal and non‐stomatal components on photosynthesis. Stomatal limitation (Ls %), non‐stomatal limitation (Lns %), CO₂ compensation point (CP) and dark respiration (Rd) increased with stress duration in both genotypes, but to a lesser extent in ‘Midnight’. Maximum CO₂ assimilation rate (A_max), carboxylation efficiency (CE) and mesophyll conductance (g_m) declined, but ‘Midnight’ had significantly higher levels of A_max, CE and g_m than ‘Brilliant’. Maximum carboxylation rate of Rubisco (V_cmax) and ribulose‐1,5‐bisphospate (RuBP) regeneration capacity mediated by maximum electron transport rate (J_max) decreased from moderate to severe drought stress in both genotypes, but to a greater extent in ‘Brilliant’ than in ‘Midnight’. After re‐watering, RWC restored to about 90% of the control levels in both genotypes, whereas A, g_s, Tr and Fv/Fm was only partially recovered, with a higher recovery level in ‘Midnight’ than in ‘Brilliant’. Rubisco activity and activation state restored to the control level after re‐watering, with more rapid increase in ‘Midnight’ than in ‘Brilliant’. The values of Ls, Lns, CP and Rd declined, and A_max, CE, V_cmax, J_max and g_m increased after re‐watering, with more rapid change in all parameters in ‘Midnight’ than in ‘Brilliant’. These results indicated that the maintenance of higher A and A_max under drought stress in drought‐tolerant Kentucky bluegrass could be attributed to higher Rubico activation state, higher CE and less stomatal limitation. The ability to resume metabolic activity (A_max, CE, Fv/Fm and Rubisco) was observed in the drought‐tolerant genotype and is the most likely cause for the increased recuperative ability of photosynthesis. Incomplete recovery of photosynthesis upon re‐watering could be attributable to lasting stomatal limitations caused by severe drought damage in both genotypes. Promoting rapid stomatal recovery from drought stress may be critical for plants to resume full photosynthetic capacity in C₃ perennial grass species.     

Physiological and biochemical traits of drought tolerance in Argania spinosa

The objective of this study was to understand and characterize the physiological and biochemical tolerance mechanisms of Argania spinosa under drought stress for selection tolerant ecotypes. Significant differences were observed among ecotypes in indices of leaf water status studied: stomatal conductance (g_s), predawn leaf water potential (Ψ_pd) and leaf relative water content. There was a significant decrease in these physiological traits with increasing degree of drought stress in all ecotypes. Drought stress significantly increased endogenous H₂O₂ and lipid peroxidation. Moderate and severe drought stress increased significantly the catalase, superoxide dismutase, peroxidase, polyphenoloxidase and lipoxygenase activities, depending on time. Their constitutive activities were higher in inland ecotypes than in coastal ecotypes. According to canonical discriminant analysis, the inland ecotypes were essentially distinguished from the coastal ecotypes by the following physiological and biochemical traits: Ψ_pd, g_s, polyphenol oxidase, superoxide dismutase and malonyldialdehyde. Inland ecotypes seem to be more tolerant to drought stress than coastal ecotypes.     

Dirunal leaf movement, chlorophyll fluorescence and carbon assimilation in soybean grown under different nitrogen and water availabilities

Diurnal heliotropic leaf movements, photosynthetic gas exchange, and the ratio of variable fluorescence to maximum fluorescence (Fv/Fm) of unrestrained and horizontally restrained leaves from soybean (Glycine max cv. Cumberland) plants grown in two different water and two different nitrogen treatments were measured. Leaves of plants grown in low water or low nitrogen availability treatments displayed more pronounced diaheliotropism (solar tracking) in the afternoon and a longer period of paraheliotropism (light avoiding) at midday relative to those of well-watered, high-nitrogen-grown plants. Photosaturated photosynthetic rates and the photon flux required to saturate photosynthesis were reduced by water stress and nitrogen deficiency. Compared to horizontal leaves, irradiance on orienting leaves was nearer to the breakpoint of the photosynthetic light response curve, where photosynthesis is co-limited by ribulose biphosphate regeneration and carboxylation. This would increase the carbon return on investments of nitrogen into photosynthesis. A positive linear relationship between Fv/Fm and quantum yield of photosynthesis was measured. Leaves of low-nitrogen-grown plants had earlier and more prolonged reductions in Fv/Fm at midday compared to leaves of high nitrogen grown plants of the same water treatment. Within the same water and nitrogen treatment, horizontally restrained leaves had lower midday Fv/Fm in relation to orienting leaves. Nitrogen deficiency and water stress enhanced this difference such that horizontally restrained leaves of low water and low nitrogen grown plants had earlier and longer midday depressions in Fv/Fm.