Deciphering drought‐induced metabolic responses and regulation in developing maize kernels

Drought stress conditions decrease maize growth and yield, and aggravate preharvest aflatoxin contamination. While several studies have been performed on mature kernels responding to drought stress, the metabolic profiles of developing kernels are not as well characterized, particularly in germplasm with contrasting resistance to both drought and mycotoxin contamination. Here, following screening for drought tolerance, a drought‐sensitive line, B73, and a drought‐tolerant line, Lo964, were selected and stressed beginning at 14 days after pollination. Developing kernels were sampled 7 and 14 days after drought induction (DAI) from both stressed and irrigated plants. Comparative biochemical and metabolomic analyses profiled 409 differentially accumulated metabolites. Multivariate statistics and pathway analyses showed that drought stress induced an accumulation of simple sugars and polyunsaturated fatty acids and a decrease in amines, polyamines and dipeptides in B73. Conversely, sphingolipid, sterol, phenylpropanoid and dipeptide metabolites accumulated in Lo964 under drought stress. Drought stress also resulted in the greater accumulation of reactive oxygen species (ROS) and aflatoxin in kernels of B73 in comparison with Lo964 implying a correlation in their production. Overall, field drought treatments disordered a cascade of normal metabolic programming during development of maize kernels and subsequently caused oxidative stress. The glutathione and urea cycles along with the metabolism of carbohydrates and lipids for osmoprotection, membrane maintenance and antioxidant protection were central among the drought stress responses observed in developing kernels. These results also provide novel targets to enhance host drought tolerance and disease resistance through the use of biotechnologies such as transgenics and genome editing.     

Improvement of heat and drought photosynthetic tolerance in wheat by overaccumulation of glycinebetaine

Within their natural habitat, crops are often subjected to drought and heat stress, which suppress crop growth and decrease crop production. Causing overaccumulation of glycinebetaine (GB) has been used to enhance the crop yield under stress. Here, we investigated the response of wheat (Triticum aestivum L.) photosynthesis to drought, heat stress and their combination with a transgenic wheat line (T6) overaccumulating GB and its wild-type (WT) Shi4185. Drought stress (DS) was imposed by controlling irrigation until the relative water content (RWC) of the flag leaves decreased to between 78 and 82%. Heat stress (HS) was applied by exposing wheat plants to 40°C for 4 h. A combination of drought and heat stress was applied by subjecting the drought-stressed plants to a heat stress as above. The results indicated that all stresses decreased photosynthesis, but the combination of drought and heat stress exacerbated the negative effects on photosynthesis more than exposure to drought or heat stress alone. Drought stress decreased the transpiration rate (Tr), stomatal conductance (Gs) and intercellular CO₂ concentration (Ci), while heat stress increased all of these; the deprivation of water was greater under drought stress than heat stress, but heat stress decreased the antioxidant enzyme activity to a greater extent. Overaccumulated GB could alleviate the decrease of photosynthesis caused by all stresses tested. These suggest that GB induces an increase of osmotic adjustments for drought tolerance, while its improvement of the antioxidative defense system including antioxidative enzymes and antioxidants may be more important for heat tolerance.     

Drought stimulation by hypocotyl exposure altered physiological responses to subsequent drought stress in peanut seedlings

Drought stress occurring at the seedling stage of peanut (Arachis hypogaea L.) plants is a limiting factor resulting in considerable reductions in production. Plants can improve their resistance to subsequent stresses after experiencing an initial stress. The aim of this study was to explore the possible role of drought priming by hypocotyl exposure in alleviating subsequent severe drought stress in peanut. Hypocotyl exposure in peanut seedlings as a drought stimulus induced xerophytophysiological regulation, shown by induced osmotic adjustment, activated antioxidant enzymes, anthocyanin accumulation, up-regulation of Gdi-15 and fewer amyloplasts. The seedlings primed by hypocotyl exposure showed improved leaf water retention and reduced proline content when exposed to subsequent drought stress. The alleviated oxidative damage and lower antioxidant enzyme activities indicated rapid acclimation following past hypocotyl exposure and further defenses against subsequent drought stress by retaining ‘memories’ to enable more rapid or stronger physiological responses. The improved leaf photosynthesis and low photosynthetic hysteresis as drought ended indicated a positive effect of drought priming in peanut seedlings. The peanut seedlings ‘remembered’ the xerophytophysiological responses caused by the prior drought stimulation from hypocotyl exposure and displayed quicker and more potent physiological responses to following drought stress. The results showed that hypocotyl exposure could help peanut seedlings survive the severe environments that occurred in the later growth stages.     

Seed biopriming with drought tolerant isolates of Trichoderma harzianum promote growth and drought tolerance in Triticum aestivum

Green house study was aimed to investigate the effect of seed biopriming with drought tolerant isolates of Trichoderma harzianum, viz. Th 56, 69, 75, 82 and 89 on growth of wheat under drought stress and to explore the mechanism underlying plant water stress resilience in response to Trichoderma inoculation. Measurements of relative water content, osmotic potential, osmotic adjustment, leaf gas exchange, chlorophyll fluorescence and membrane stability index were performed. In addition, analysis of the phenolics, proline, lipid peroxidation and measurements of phenylalanine ammonia‐lyase activity were carried out. Seed biopriming enhanced drought tolerance of wheat as drought induced changes like stomatal conductance, net photosynthesis and chlorophyll fluorescence were delayed. Drought stress from 4 to 13 days of withholding water induced an increase in the concentration of stress induced metabolites in leaves, while Trichoderma colonisation caused decrease in proline, malondialdehyde (MDA) and hydrogen peroxide (H₂O₂), and an increase in total phenolics. A common factor that negatively affects plants under drought stress conditions is accumulation of toxic reactive oxygen species (ROS), and we tested the hypothesis that seed biopriming reduced damages resulting from accumulation of ROS in stressed plants. The enhanced redox state of colonised plants could be explained by higher l ‐phenylalanine ammonia‐lyase (PAL) activity in leaves after 13 days of drought stress in Trichoderma treated plants. Similar activity was induced in untreated plants in response to drought stress but to a lower extent in comparison to treated plants. Our results support the hypothesis that seed biopriming in wheat with drought tolerant T. harzianum strains increased root vigour besides performing the process of osmoregulation. It ameliorates drought stress by inducing physiological protection in plants against oxidative damage, due to enhanced capacity to scavenge ROS and increased level of PAL, a mechanism that is expected to augment tolerance to abiotic stresses.     

Expression of the Nicotiana protein kinase (NPK1) enhanced drought tolerance in transgenic maize

Drought is one of the most important abiotic stresses affecting the productivity of maize. Previous studies have shown that expression of a mitogen-activated protein kinase kinase kinase (MAPKKK) gene activated an oxidative signal cascade and led to the tolerance of freezing, heat, and salinity stress in transgenic tobacco. To analyse the role of activation of oxidative stress signalling in improving drought tolerance in major crops, a tobacco MAPKKK (NPK1) was expressed constitutively in maize. Results show that NPK1 expression enhanced drought tolerance in transgenic maize. Under drought conditions, transgenic maize plants maintained significantly higher photosynthesis rates than did the non-transgenic control, suggesting that NPK1 induced a mechanism that protected photosynthesis machinery from dehydration damage. In addition, drought-stressed transgenic plants produced kernels with weights similar to those under well-watered conditions, while kernel weights of drought-stressed non-transgenic control plants were significantly reduced when compared with their non-stressed counterparts.     

Physio‐biochemical assessment and expression analysis of genes associated with drought tolerance in sugarcane (Saccharum spp. hybrids) exposed to GA3 at grand growth stage

• Drought is one of the most serious environmental factors limiting production of sugarcane worldwide. In order to assess the influence of gibberellins (GA₃) on drought and plant growth, along with associated physio‐biochemical attributes, expression of eight drought‐responsive genes were quantified and analysed.
• At grand growth stage (120 DAP) two sugarcane varieties (CoLk94184, CoPK05191) were exposed to drought by withholding irrigation. GA₃ (35 ppm) was applied using battery‐operated uniform controlled dispensing sprayer twice at 1‐week intervals on 2‐week drought‐stressed plants. Physio‐biochemical attributes including antioxidant enzyme activities were estimated following standard protocols. RT‐PCR was performed to visualise the drought‐associated gene expression patterns.
• Drought triggered a reduction in RWC and chlorophyll content but these recovered when droughted plants were exposed to GA₃. Proline content increased many fold in both varieties under stress, but decreased under the influence of GA₃. There was a mixed response of antioxidant enzyme activity, which distinctly declined after GA₃ exposure, together with a lesser reduction in dry matter content over that of control plants. With increasing stress, expression of pyrroline‐5‐carboxylase synthetase (P5CS) and betaine‐aldehyde dehydrogenase genes was observed, selectively up‐regulated in CoPK05191. Expression of proline oxidase/transporter was high in CoPK05191 but diminished along with proline content after exposure to GA_3. CoLk94184 showed no significant difference in P5CS gene expression under stress condition, whereas expression of betaine‐aldehyde dehydrogenase gene was unchanged in response to stress.
• Results demonstrated that exposure of droughted plants to GA₃ not only led to recovery of activity of drought‐associated physio‐biochemical attributes, but also minimised impact on cane dry weight and quality. Further, GA₃ application caused differential gene expression that possibly triggers increased responsiveness towards drought tolerance in sugarcane.

     

Exogenously applied polyamines increase drought tolerance of rice by improving leaf water status,photosynthesis and membrane properties

Drought stress hampers rice performance principally by disrupting the plant–water relations and structure of biological membranes. This study appraised the role of polyamines (PAs) in improving drought tolerance in fine grain aromatic rice (Oryza sativa L.). Three PAs [putrescine (Put), spermidine (Spd) and spermine (Spm)] were used each at 10 μM as seed priming (by soaking seeds in solution) and foliar spray. Primed and non-primed seeds were sown in plastic pots with normal irrigation in a phytotron. At four-leaf stage, plants were subjected to drought stress by bringing the soil moisture down to 50% of field capacity by halting water supply. For foliar application, 10 μM solutions each of Put, Spd and Spm were sprayed at five-leaf stage. Results revealed that drought stress severely reduced the rice fresh and dry weights, while PAs application improved net photosynthesis, water use efficiency, leaf water status, production of free proline, anthocyanins and soluble phenolics and improved membrane properties. PAs improved drought tolerance in terms of dry matter yield and net photosynthesis was associated with the maintenance of leaf water status and improved water use efficiency. Among the antioxidants, catalase activity was negatively related to H₂O₂ and membrane permeability, which indicated alleviation of oxidative damage on cellular membranes by PAs application. Foliar application was more effective than the seed priming, and among the PAs, Spm was the most effective in improving drought tolerance.     

Proteins and Metabolites Regulated by Trinexapac-ethyl in Relation to Drought Tolerance in Kentucky Bluegrass

Plants have developed various mechanisms in adaptation to water deficit stress, including growth retardant to reduce water loss. Previous studies reported that plants treated with a growth inhibitor, trinexapac-ethyl (TE), had improved drought tolerance. The objective of this study was to determine alterations in proteins and metabolite accumulation associated with drought tolerance improvement in a perennial grass species, Kentucky bluegrass (Poa pratensis), induced by TE application. Plants were treated with TE [1.95 ml l⁻¹ (v:v); a.i. TE = 0.113%] through foliar spray for 14 days, and then subjected to drought stress by withholding irrigation for 15 days in growth chambers. TE-treated plants exhibited significantly higher relative water content and photosynthetic capacity and lower membrane leakage than nontreated plants under drought stress, suggesting TE-enhanced drought tolerance in Kentucky bluegrass. Physiological improvement in drought tolerance through TE application was associated with the increased accumulation of various proteins and metabolites, including ferritin, catalase, glutathione-S-transferase, Rubisco, heat shock protein 70, and chaperonin 81, as well as fatty acids (palmitic acid, α-linolenic acid, linoleic acid, and octadecanoic acid). Our results suggest that TE may regulate metabolic processes for antioxidant defense, protective protein synthesis, photorespiration, and fatty acid synthesis, and thereby contribute to better drought tolerance in Kentucky bluegrass.     

Drought increases freezing tolerance of both leaves and xylem of Larrea tridentata

Drought and freezing are both known to limit desert plant distributions, but the interaction of these stressors is poorly understood. Drought may increase freezing tolerance in leaves while decreasing it in the xylem, potentially creating a mismatch between water supply and demand. To test this hypothesis, we subjected Larrea tridentata juveniles grown in a greenhouse under well‐watered or drought conditions to minimum temperatures ranging from ?8 to ?24 °C. We measured survival, leaf retention, gas exchange, cell death, freezing point depression and leaf‐specific xylem hydraulic conductance (k_l). Drought‐exposed plants exhibited smaller decreases in gas exchange after exposure to ?8 °C compared to well‐watered plants. Drought also conferred a significant positive effect on leaf, xylem and whole‐plant function following exposure to ?15 °C; drought‐exposed plants exhibited less cell death, greater leaf retention, higher k_l and higher rates of gas exchange than well‐watered plants. Both drought‐exposed and well‐watered plants experienced 100% mortality following exposure to ?24 °C. By documenting the combined effects of drought and freezing stress, our data provide insight into the mechanisms determining plant survival and performance following freezing and the potential for shifts in L. tridentata abundance and range in the face of changing temperature and precipitation regimes.     

An osmotin from the resurrection plant Tripogon loliiformis (TlOsm) confers tolerance to multiple abiotic stresses in transgenic rice

Osmotin is a key protein associated with abiotic and biotic stress response in plants. In this study, an osmotin from the resurrection plant Tripogon loliiformis (TlOsm) was characterized and functionally analyzed under abiotic stress conditions in T. loliiformis as well as in transgenic Nicotiana tabacum (tobacco) and Oryza sativa (rice) plants. Real‐time PCR analysis on mixed elicitor cDNA libraries from T. loliiformis showed that TlOsm was upregulated a 1000‐fold during the early stages of osmotic stresses (cold, drought, and salinity) in both shoots and roots but downregulated in shoots during heat stress. There was no change in TlOsm gene expression in roots of heat‐stressed plants and during plant development. The plasma membrane localization of TlOsm was showed in fluorescent‐tagged TlOsm tobacco plants using confocal laser scanning microscopic analysis. Transgenic rice plants expressing TlOsm were assessed for enhanced tolerance to salinity, drought and cold stresses. Constitutively expressed TlOsm in transgenic rice plants showed increased tolerance to cold, drought and salinity stress when compared with the wild‐type and vector control counterparts. This was evidenced by maintained growth, retained higher water content and membrane integrity, and improved survival rate of TlOsm‐expressing plants. The results thus indicate the involvement of TlOsm in plant response to multiple abiotic stresses, possibly through the signaling pathway, and highlight its potential applications for engineering crops with improved tolerance to cold, drought and salinity stress.     

Impact of drought stress on growth and quality of miscanthus for biofuel production

Miscanthus has a high potential as a biomass feedstock for biofuel production. Drought tolerance is an important breeding goal in miscanthus as water deficit is a common abiotic stress and crop irrigation is in most cases uneconomical. Drought may not only severely reduce biomass yields, but also affect biomass quality for biofuel production as cell wall remodeling is a common plant response to abiotic stresses. The quality and plant weight of 50 diverse miscanthus genotypes were evaluated under control and drought conditions (28 days no water) in a glasshouse experiment. Overall, drought treatment decreased plant weight by 45%. Drought tolerance – as defined by maintenance of plant weight – varied extensively among the tested miscanthus genotypes and ranged from 30% to 110%. Biomass composition was drastically altered due to drought stress, with large reductions in cell wall and cellulose content and a substantial increase in hemicellulosic polysaccharides. Stress had only a small effect on lignin content. Cell wall structural rigidity was also affected by drought conditions; substantially higher cellulose conversion rates were observed upon enzymatic saccharification of drought‐treated samples with respect to controls. Both cell wall composition and the extent of cell wall plasticity under drought varied extensively among all genotypes, but only weak correlations were found with the level of drought tolerance, suggesting their independent genetic control. High drought tolerance and biomass quality can thus potentially be advanced simultaneously. The extensive genotypic variation found for most traits in the evaluated miscanthus germplasm provides ample scope for breeding of drought‐tolerant varieties that are able to produce substantial yields of high‐quality biomass under water deficit conditions. The higher degradability of drought‐treated samples makes miscanthus an interesting crop for the production of second‐generation biofuels in marginal soils.     

Impact on physiology and malting quality of barley exposed to heat,drought and their combination during different growth stages under controlled environment

Drought and heat stress are two major abiotic stresses that tend to co‐occur in nature. Recent climate change models predict that the frequency and duration of periods of high temperatures and moisture‐deficits are on the rise and can be detrimental to crop production and hence a serious threat for global food security. In this study we examined the impact of short‐term heat, drought and combined heat and drought stress on four barley varieties. These stresses were applied during vegetative stage or during heading stages. The impact on root and shoot biomass as well as seed yields were analyzed. This study demonstrated that sensitivity to combined stress was generally greater than heat or drought individually, and greater when imposed at heading than at the vegetative stages. Micromalted seeds collected from plants stressed during heading showed differences in malt extract, beta‐glucan content and percent soluble protein. Screening barley germplasm during heading stage is recommended to identify novel sources of tolerance to combined stress. Apart from seed yield, assessing the seed quality traits of concern for the stakeholders and/or consumers should be an integral part of breeding programs for developing new barley varieties with improved heat and drought stress tolerance.     

缓慢干旱下春小麦叶片质膜脂肪酸组成、H~+-ATPase肥及5′-AMPase活力的变化

研究了田间缓慢干旱胁迫下,抗旱性不同的两个小麦(Triticum aestivum)品种的生长状况、质膜极性脂脂肪酸组成以及质膜关键酶活力的变化。在小麦生长发育的早期,干旱胁迫使其叶片质膜极性脂脂肪酸不饱和度下降、质膜微囊消耗O_2的速率升高、膜蛋白含量降低、H~+-ATPase(EC 3.6.1.35)活力下降、5′-AMPase(EC 3.1.3.5)活力大幅度升高;在小麦发育的后期,随着干旱的持续,小麦叶片质膜的极性脂脂肪酸不饱和度不变或升高、质膜微囊消耗O_2的速率降低、膜蛋白含量与H~+-ATPase活力升高、5′-AMPase活力下降。以上结果表明,小麦在发育的早期阶段对干旱较敏感,其细胞质膜流动性降低、细胞中能荷贮备降低;而在后期,则又表现出对干旱的适应。这些结果将有助于阐明自然干旱条件下植物的抗旱机制。     

Physiological responses of wheat to drought stress and its mitigation approaches

Drought is a polygenically controlled stress and a major agricultural risk that reduces crop productivity and limits the successful insight of land potential throughout the world. This review article has been divided into two parts, i.e., effect of drought stress on physiology of wheat and potential drought mitigation approaches. In the first part, physiological responses of wheat to stress were discussed. Cell membrane stability, relative water content, early maturity, decreased leaf area, small plant size, increased dry weight and root–shoot ratio, and the whole-plant transpiration rate response to enhanced atmospheric vapor pressure deficit are physiological traits associated with drought tolerance in wheat. Reduction of relative water content closes stomata and thereby reduces stomatal conductance. Osmotic adjustment improves drought tolerance by allowing cell enlargement, plant growth, and stomata to stay partially open and by maintaining CO₂ fixation under severe water deficit. The wheat plant accumulates several organic and inorganic solutes in its cytosol to lessen its osmotic potential for maintenance of cell turgor. Drought affects photosynthesis negatively by changing the inner structure of chloroplasts, mitochondria, and chlorophyll content and minerals. Destruction of the photosystem II (PSII) oxygen releasing complex and reaction center can disturb production and use of electrons, causing lipid peroxidation of cell membrane through the production of reactive oxygen species. In the second part, drought mitigation approaches were discussed. Seed, drought, bacterial, and hormonal priming are common approaches used to lessen the effects of water deficit. Physiological trait-based breeding, molecular breeding, marker-assisted backcrossing, aerial phenotyping, water budgeting, and resource allocation are modern approaches used to develop drought tolerant wheat cultivars. Wheat genotypes produced as a result of a combination of all these methodologies will increase food security regarding the currently changing climate.     

缓慢干旱下春小麦叶片质膜脂肪酸组成、H+-ATPase及5′-AMPase活力的变化

 研究了田间缓慢干旱胁迫下,抗旱性不同的两个小麦(Triticum aestivum)品种的生长状况、质膜极性脂脂肪酸组成以及质膜关键酶活力的变化。在小麦生长发育的早期,干旱胁迫使其叶片质膜极性脂脂肪酸不饱和度下降、质膜微囊消耗O2的速率升高、膜蛋白含量降低、H+-ATPase (EC 3.6.1.35)活力下降、5'-AMPase (EC 3.1.3.5)活力大幅度升高;在小麦发育的后期,随着干旱的持续,小麦叶片质膜的极性脂脂肪酸不饱和度不变或升高、质膜微囊消耗O2的速率降低、膜蛋白含量与H+-ATPase活力升高、5'-AMPase活力下降。以上结果表明,小麦在发育的早期阶段对干旱较敏感,其细胞质膜流动性降低、细胞中能荷贮备降低;而在后期,则又表现出对干旱的适应。这些结果将有助于阐明自然干旱条件下植物的抗旱机制。