Both stress avoidance and stress tolerance are responsible for stress resistance, understood as the ability to cope with adverse environmental conditions, by keeping a balance between growth, reproduction, and activation of suitable coping strategies
[7]. This kind of resistance can be called
biological resistance, which is the strategy of an individual plant to tolerate and survive stress conditions. An example of
biological resistance is also the stressor escape strategy which occurs in stress-sensitive plants. From the perspective of plant users, crop resistance to environmental stresses should be defined as the ability to cope with stress conditions thanks to defence responses (stress tolerance and/or avoidance) which enables the maintenance of stable and good quality yields. Therefore, it can be called
agricultural resistance.
2. Plant Responses to Drought
Drought is a meteorological term defined as a period of little or no rainfall, which reduces the amount of water in the soil, and is usually accompanied by high evaporative demand, leading to continuous loss of water by transpiration. It is considered the most frequent climate-related constraint in many regions of the world
[8][9]. This stress factor generates the state of stress (strain) in plant cells, which is the reduction in water content (dehydration, water deficit), adversely affecting plant physiological activity, growth, reproduction, and crop productivity
[10]. The level of dehydration depends on stress severity and duration, as well as on adaptive traits protecting against water loss (smaller leaves, leaves covered with cuticle or tomentose, as well as leaf folding) and supporting water uptake from deeper soil layers (extensive vertically orientated root system). Another example of an adaptive trait protecting from water loss is stomatal behaviour (stomata open at night and closed during the day) in crassulacean acid plants (CAM) having an alternative route of carbon assimilation which occurs during the night
[11][12].
Based on the ability to maintain stable leaf hydration under water deficit conditions the water management strategy of plants is classified as isohydric or anisohydric
[13]. Isohydric species (‘water savers’) maintain nearly constant RWC through precise control of stomatal behaviour. These plants respond to drought by a rapid decrease in stomatal conductance (g
s) and restriction of excessive water loss without a reduction in leaf area but at the same time show a decrease in photosynthetic activity. In contrast, anisohydric plants (‘water wasters’) show a decrease in leaf water content and strong leaf area reduction but keep stomata open and maintain a high photosynthetic rate
[14][15][16]. The extent of tissue dehydration is a signal triggering, directly or through ABA increase, the activation of appropriate metabolic and physiological changes responsible for plants’ adjustment to drought
[17]. Even a slight decrease in RWC triggers upstream signalling events, leading to ABA accumulation and stomatal closure
[18]. ABA is also involved in several downstream events responsible for the maintenance of tissue hydration (dehydration avoidance strategy), which include osmotic adjustment, comprising the accumulation of organic osmotic compounds (proline, glycine-betaine, soluble proteins, carbohydrates) in leaves and roots
[10][17]. The strategy of dehydration avoidance (isohydric behaviour) allows plants to sustain physiological functions under stress conditions and recover after stress termination. This strategy is effective in plants exposed to the mild or moderate drought that does not last very long but affects carbon starvation under prolonged drought
[19]. Moreover, when stomata are closed plants absorb more light than can be used in carbon fixation, which triggers the generation of reactive oxygen species (ROS), affecting secondary stress and damage of PSII, leading to further weakness of photosynthesis
[6][20]. What is more, during long-term drought the ability of plants to maintain stomatal closure may be weakened due to a decrease in ABA level and plant behaviour changes to anisohydric
[21]. The response to drought in anisohydric plants (barley, wheat, sunflower) is mainly regulated hydraulically. The maintenance of stomatal conductance in these plants is supported by the capacity for osmotic adjustment, controlled by the dehydration signal, which enables plants to extract water from the soil to maintain tissue hydration
[13][19]. In anisohydric wheat genotypes, the level of ABA in leaves did not change under water deficit conditions, while in roots it increased but only after 21 days of stress
[22]. Therefore, it is possible that, along with tissue dehydration, ABA may also play a role in the response of anisohydric species to prolonged drought. The stomatal conductance in anisohydric plants is also maintained by undisturbed water movement through cell membrane aquaporins responsible for roots’ ability to conduct water
[23].
In plants exposed to severe and long-term drought, dehydration cannot be avoided, and activation of dehydration tolerance mechanisms becomes important. Dehydration has a deleterious effect on cell membranes and disrupts many biochemical and physiological processes
[4][24]. A frequently used indicator of dehydration tolerance is the cell membrane injury index or membrane stability index, which shows the ability to maintain membrane integrity at a given level of dehydration
[25][26]. The dehydration tolerance mechanisms enable plants to maintain membrane integrity and cell homeostasis and to regain physiological activity after stress cessation
[6][20]. These mechanisms are controlled by ABA-dependent and -independent pathways and include the synthesis of protective proteins (LEA proteins, dehydrins, chaperons) and compatible compounds (proline, glycine-betaine, proline-betaine, trehalose, raffinose mannitol, pinitol) involved in enzyme and membrane protection
[4][17][20][27][28]. Dehydration-induced disturbance of the respiratory metabolic pathway exhibits the generation of ROS, leading to a state of oxidative stress
[4][24][29][30][31]. Moreover, in drought-stressed plants, the enhanced build-up of ROS is caused by photosynthesis disruption and increased photorespiration due to the limitation of CO
2 uptake
[31][32]. Overproduction of ROS (secondary stress), which includes superoxide radicals (O
2•−), hydrogen peroxide (H
2O
2), hydroxyl radical (OH
•), and singlet oxygen (
1O
2), is harmful to organelles through lipid peroxidation and damage to nucleic acids and proteins
[4][8][24]. To overcome oxidative damage, plants possess enzymatic and non-enzymatic ROS-scavenging systems. Enzymatic antioxidants include superoxide dismutase (SOD), catalase (CAT), and peroxidases (POX). The non-enzymatic components of the antioxidative system comprise ascorbic acid, α-tocopherol, flavonoid, glutathione, carotenoids, proline, and phenolic compounds which mitigate oxidative damage by direct reduction of ROS activity and by working together with antioxidant enzymes
[31][33]. Additionally, alternative oxidase (AOX) is involved in the avoidance of excess generation of ROS in mitochondrial electron transport chains
[32]. ABA plays a pivotal role in activating antioxidant enzymes and the synthesis of low molecular ROS scavengers
[27][28]. Upregulation of the antioxidant system may also be controlled by JA, SA, and BRs
[29][30][31][32][33][34][35][36][37]. Thanks to the efficient antioxidative system, plants can keep ROS at non-toxic levels, and these molecules are thought to act as signals for activation of stress defence responses
[32][38].
Plant responses to drought are governed by a sophisticated regulatory system working at the molecular level. The decrease in turgor pressure leads to tension changes in plasma membranes, which are perceived by membrane proteins including receptor-like kinases (RLKs), histidine kinases (HKs), and integrin-like proteins (ILPs) working as osmotic stress sensors. ATHK1 is an
Arabidopsis thaliana His kinase is postulated to play a role in water stress perception triggering the mitogen-activated protein kinase (MAPK) signalling cascade both in ABA-dependent and ABA-independent regulatory systems
[39]. A crucial role in the signal transduction route is played by transcription factors (TFs) that bind to TF binding sites (TFBS) in the promoter region and regulate gene expression. TF families involved in plants’ response to drought include bZip (AREB/ABF), AP2/ERF (DREB/CBF), MYB/MYC, WRKY, and NAC
[8][40]. In the ABA-dependent pathway, the perception of ABA by receptor proteins is the primary event that triggers downstream signalling cascades to induce final physiological responses. The receptors for this hormone are small soluble cytosol/nucleus-localized pyrabactin resistance (PYR)/PYR-like (PYL)/regulatory components of ABA receptor (RCAR) proteins. The interaction of ABA with PYR/PYL/RCARs affects the deactivation of protein phosphatase enzymes (PP2Cs), which are constitutive negative regulators of ABA-induced responses. The inhibition of PP2Cs leads to auto-phosphorylation of the protein kinases SnRK2s, which induces stomatal closure and stimulates nuclear targets that trigger the expression of various water stress associated genes due to activation of TFs
[40]. ABA-dependent gene expression systems involve activation of b-ZIP (AREBs/ABFs), MYC/MYB, as well as NAC transcription factors
[41]. In ABA-independent responses to drought, the dehydration signal from the cell surface to the nucleus is mediated by calcium, JA, and ROS
[40]. Water deficit leads to membrane destabilization and Ca
2+ influx into the cytoplasm. The calcium signal is detected and transduced through calmodulin (CaM), calcium dependent protein kinases (CDPK), and calcineurin B-like proteins (CBLs) and interacts with the MAPK cascade, leading to the activation of TFs (DREB, NAC) and expression of genes coding the synthesis of functional proteins (LEA proteins, chaperones, dehydrins, enzymes of osmolyte biosynthesis). JA, on the other hand, is engaged in the activation of the MYC2 transcription factor, which triggers the expression of stress-responsive genes
[40]. Furthermore, JA along with ROS acts as a stress-signalling unit activating the expression of genes involved in the activation of enzymatic and non-enzymatic scavenging events
[40][42]. The widespread plant response to drought is proline accumulation due to the stimulation of its synthesis from glutamate catalyzed by pyrroline-5-carboxylate synthetase (P5CS) and pyrolino-5-carboxylate reductase (P5CR)
[43][44]. Synthesis of this amino acid under drought is driven by both ABA-dependent and ABA-independent signalling pathways engaged in triggering the expression of
P5CS and
P5CR genes regulated by many TFs, which are also related to responses to drought controlled by other growth regulators
[45].
Important components of the stress-factor-induced regulatory system are epigenetic modifications which are independent of DNA sequence changes. These changes include chromatin remodellings such as DNA methylation and histone modifications altering the structure and accessibility of chromatin, leading to changes in gene expression at the transcriptional and post-transcriptional levels
[46]. Drought-stress-induced changes in DNA methylation have been observed in diverse plant species. These changes were related to the expression of genes encoding transcription factors and were involved in drought resistance mechanisms or were linked to drought sensitivity
[47][48][49]. It was found that changes in DNA methylation (demethylation) in water deficit stressed rice were responsible for proline accumulation via the upregulation of proline metabolism-related gene expression
[50]. In addition to DNA methylation, drought-induced histone modifications (methylation, acetylation) are involved in controlling gene expression in stressed plants
[51]. It was observed that drought stress triggered histone H3 lisyne4 tri-methylation (H3K4 me3) in the gene body region of nine cis-epoxycarotenoid dioxygenase 3 (NCED3), which is a key enzyme involved in ABA synthesis. Additionally, some studies reported the increase in H3K4me3 and H3 lisyne9 acetylation (H3K9Ac) in the promotor region of such genes as
RD29A,
RD29B,
RD22, and
RELATED TO AP2.4 (
RAP2.4) encoding synthesis of LEA proteins. The abundance of histone modification and the number of genes expressed depend on stress duration and degree
[48][51]. Most of the epigenetic modifications are removed when the stress is relieved, but some of them persist, enabling plants to remember past stress and to prepare for future recurrent stress events which occur during plant life. This is so-called “plant stress memory”, which can also be transferred to further generations during sexual and vegetative reproduction
[47][49][52]. Integral components of the stress response at the molecular level also involved in memory pathways are non-coding small RNAs (miRNAs, siRNAs), which can trigger DNA methylation and histone modifications. Plants exposed to drought can memorize stress events through DNA and histone modifications for specific gene expression thanks to the up- and downregulation of small RNAs responsible for the increased resistance to future stress events through the control of TFs, ROS, and hormone levels
[49][52].
3. Drought Coping Strategies and Resistance
The ability to avoid or tolerate dehydration is crucial in dealing with drought at cellular and tissue levels (
biological resistance), which allows plants to survive during water scarcity conditions and recovery. The tolerance and avoidance mechanisms were developed during evolution to adjust to environmental conditions but usually do not have beneficial effects on agricultural production. Plants can withstand drought without any visible signs of dehydration and/or dehydration damage, but their growth and yield may be lower than expected. This is an unwanted side effect of plant adjustment to stress, which has a negative impact on biomass accumulation and yield (
agricultural resistance). The activation of coping mechanisms is connected to increased energy and nutrient consumption, which results in the allocation of less energy and assimilates into growth processes, leading to yield reduction
[10][53]. Furthermore, many traits associated with drought resistance have a dual effect (positive or negative) on plant productivity which depends on stress intensity and timing as well as on climatic conditions such as light intensity and evaporative demand
[54]. The dehydration avoidance strategy, such as stomatal closure, reduces water loss from leaves. However, at the same time, it causes the restriction of CO
2 uptake, ROS generation, damage of PSII, and the inhibition of photosynthesis, resulting in the reduction of crop production
[6][20][55][56][57]. Moreover, changes in the hormonal balance, which is a part of the coping strategy consisting of an increase in the levels of ABA, JA, Et, and SA, and a decrease in CKs, AUXs, and GAs, may also bring about photosynthesis inhibition, growth restriction, leaf senescence acceleration, and leaf fall, negatively affecting yield
[34][58][59]. Therefore, there is a conflict between plant coping strategies (avoidance, tolerance) and resistance to drought essential for agricultural production. In an agricultural perspective, drought-resistant plants are those that maintain growth and stable yield during water-limited conditions. The priority in breeding research focusing on improving drought resistance is to obtain crop genotypes that can cope with drought stress without growth and yield reduction. Therefore, the research on plant stress physiology should concentrate on finding those features of coping strategies that ensure growth maintenance and stable yield.
The source of traits valuable in developing new drought-resilient crop varieties may be wild genotypes and landraces originating from rainfed areas
[8]. Another promising approach is the introduction of new crop species able to cope under water-limited conditions and maintain stable growth. An interesting species in this regard is quinoa (
Chenopodium quinoa Willd.), which originated in the Andean region. It has begun to be called ‘the 21st-century crop’, and recently it has been introduced into cultivation in many regions of the world. Quinoa has received special attention due to its high nutritional composition of seeds and strong natural ability to cope with drought
[60][61]. There is wide diversity among quinoa genotypes in the traits of drought coping strategy (
biological resistance) and resistance assessed based on the seed yields (
agricultural resistance). The drought response mechanisms in quinoa to endure water deficits include accelerated root growth, high water-use efficiency (WUE), osmotic adjustment, turgor maintenance, increased synthesis of osmoprotectants such as amino acid proline, and soluble sugars (glucose, trehalose), ABA biosynthesis, antioxidant defence, heat-shock, and LEA protein synthesis
[62].
A promising drought resistance strategy for crops is the ability to optimize water use, along with sustained high photosynthetic activity, which is an essential component of plant productivity
[63]. It may be achieved by triggering varied metabolic and physiological responses of the dehydration avoidance strategy, which includes the modification of root conductivity and architecture, regulation of stomatal behaviour allowing the maintenance of photosynthetic CO
2 fixation, as well as protection against non-stomatal photosynthesis limitation
[64][65][66].
One recently considered approach in attaining crop resistance to drought is focused on a better understanding of the role of plant growth regulators (PGRs) in the coping strategy along with the mitigation of the negative effect of drought on productivity and yield. PGRs play an important role in triggering, directly or through specific signal cascades, a wide range of metabolic and physiological responses of plants to drought. Many of these responses, which are components of the drought stress coping strategy, are the result of positive or negative interactions between diverse PGRs
[58][67][68]. Broadening knowledge about the impact of drought on the fluctuation of the level of PGRs and about the crosstalk between them in triggering appropriate responses seems to be essential in identifying components of drought coping strategies, which permit undisturbed growth and stable yield. The hormone that plays a key role in the plant response to drought is ABA, commonly called a “stress hormone”. An increased level of this PGR in drought-stressed plants acts as a signal that regulates multiple responses at physiological and biochemical levels
[3][28]. It was suggested that the interaction between plant hormones (ABA, AUXs, CKs, and ET) may play an important role in the diverse drought responses of sensitive and resistant wheat lines
[69]. The resistant wheat line was able to maintain growth and was characterized by lower yield reduction under drought. This line was temporarily anisohydric and closed the stomata only at a higher level of drought which correlated with the repression of ABA synthesis. At the same time, it could activate defence responses (ROS protection, LEA proteins, and cuticle synthesis) and trigger the expression of photosynthetic genes as well as genes involved in AUXs, CKs and Et metabolism, and signalling. However, the drought-sensitive wheat line was isohydric, had a higher ABA level, closed stomata at the start of stress and began photosynthesis inhibition. Certain recently obtained results of research focused on the crosstalk between ABA, CKs, and BRs at physiological and molecular levels seem to be promising in finding drought coping strategies that prevent yield reduction
[70][71].