Supratim Basu

Cross-talk between abscisic acid-dependent and abscisic acid-independent pathways during abiotic stress

Salinity, drought and low temperature are the common forms of abiotic stress encountered by land plants. To cope with these adverse environmental factors, plants execute several physiological and metabolic responses. Both osmotic stress (elicited by water deficit or high salt) and cold stress increase the endogenous level of the phytohormone abscisic acid (ABA). ABA-dependent stomatal closure to reduce water loss is associated with small signaling molecules like nitric oxide, reactive oxygen species and cytosolic free calcium, and mediated by rapidly altering ion fluxes in guard cells. ABA also triggers the expression of osmotic stress-responsive (OR) genes, which usually contain single/multiple copies of cis-acting sequence called abscisic acid-responsive element (ABRE) in their upstream regions, mostly recognized by the basic leucine zipper-transcription factors (TFs), namely, ABA-responsive element-binding protein/ABA-binding factor. Another conserved sequence called the dehydration-responsive element (DRE)/C-repeat, responding to cold or osmotic stress, but not to ABA, occurs in some OR promoters, to which the DRE-binding protein/C-repeat-binding factor binds. In contrast, there are genes or TFs containing both DRE/CRT and ABRE, which can integrate input stimuli from salinity, drought, cold and ABA signaling pathways, thereby enabling cross-tolerance to multiple stresses. A strong candidate that mediates such cross-talk is calcium, which serves as a common second messenger for abiotic stress conditions and ABA. The present review highlights the involvement of both ABA-dependent and ABA-independent signaling components and their interaction or convergence in activating the stress genes. We restrict our discussion to salinity, drought and cold stress.

Amelioration of salinity stress by exogenously applied spermidine or spermine in three varieties of indica rice differing in their level of salt tolerance

We present here the comparative protective potentiality of exogenously applied polyamines (PAs), namely spermidine (Spd) and spermine (Spm), in mitigating NaCl toxicity and inducing short-term salinity tolerance in three indica rice varieties, namely M-1-48 (salt-sensitive), Nonabokra (salt-tolerant) and Gobindobhog (highly sensitive). The retardation in root length or shoot length and toxic Na(+) accumulation or K(+) loss, the considerable increment in malondialdehyde/H(2)O(2) accumulation or lipoxygenase activity, all of which were particularly noteworthy in M-1-48 and Gobindobhog during salinity stress, was appreciably reduced by co-treatment with Spd or Spm. Both the PAs also inhibited the extent of salt-induced protein carbonylation in all the varieties and enhanced protease activity, especially in Gobindobhog. The prevention of chlorophyll degradation was better with Spd in Nonabokra and Gobindobhog. While the salt-induced increase in anthocyanin or reducing sugar level was further prompted by Spd or Spm in all the varieties, the proline content was elevated by Spd particularly in Gobindobhog. During salinity stress, both the PAs were effective in lowering the putrescine accumulation in M-1-48 and Gobindobhog, and strikingly increasing the Spm level in all the varieties, the highest being in Gobindobhog. In addition, they enhanced the activity of peroxidases and compensated for the decreased catalase activity in all the varieties. Thus the two PAs could recuperate all the three varieties from salt-induced damages to different degrees. The salt injuries, encountered in M-1-48 and Gobindobhog, both of which showed greater susceptibility to salinity stress, were more pronouncedly alleviated and counteracted by the PAs, than the salt-tolerant Nonabokra. The reversal of inhibitory effect of salinity stress was conferred by preventing growth inhibition or various forms of cellular damages, maintaining proper K(+)/Na(+) balance or triggering the level of osmolytes and activity of antioxidant enzymes. Our communication offers a referenced evidence for an understanding of the mechanism by which higher PAs relieve the damages particularly in salt-sensitive rice varieties.

Differential antioxidative responses of indica rice cultivars to drought stress.

The present study investigated the linkages between drought stress, oxidative damages and variations in antioxidants in the three rice varieties IR-29 (salt-sensitive), Pokkali (salt-tolerant) and aromatic Pusa Basmati (PB), to elucidate the antioxidative protective mechanism governing differential drought tolerance. Water deficit, induced by 20% (w/v) polyethylene glycol (PEG-6000), provoked severe damages in IR-29 and PB in the form of huge chlorophyll degradation and elevated H2O2, malondialdehyde and lipoxygenase (LOX, EC 1.13.11.12) levels as compared to Pokkali. The protein oxidation was more conspicuous in IR-29. Increment in antioxidants, particularly flavonoids and phenolics was several folds higher over control in Pokkali, while much lesser in IR-29 and PB. The activity of catalase (CAT, EC 1.11.1.6) and superoxide dismutase (SOD, EC 1.15.1.1) were decreased in IR-29 and PB, but unaltered in Pokkali. However, marked drought-induced increase in guaiacol peroxidase (GPX, EC 1.11.1.7) activity was noted in both IR-29 and PB. Induction in radical scavenging activity, being the maximum in IR-29, and increased reducing power ability in all the cultivars, accompanied with drought stress, were observed as a defense mechanism. The novelty of our work is that it showed the aromatic rice PB behaving more closely to IR-29 in greater susceptibility to dehydration stress, while the salt-tolerant Pokkali also showed effective drought tolerance properties.

Coordinated regulation of photosynthesis in rice increases yield and tolerance to environmental stress

Plants capture solar energy and atmospheric carbon dioxide (CO2) through photosynthesis, which is the primary component of crop yield, and needs to be increased considerably to meet the growing global demand for food. Environmental stresses, which are increasing with climate change, adversely affect photosynthetic carbon metabolism (PCM) and limit yield of cereals such as rice (Oryza sativa) that feeds half the world. To study the regulation of photosynthesis, we developed a rice gene regulatory network and identified a transcription factor HYR (HIGHER YIELD RICE) associated with PCM, which on expression in rice enhances photosynthesis under multiple environmental conditions, determining a morpho-physiological programme leading to higher grain yield under normal, drought and high-temperature stress conditions. We show HYR is a master regulator, directly activating photosynthesis genes, cascades of transcription factors and other downstream genes involved in PCM and yield stability under drought and high-temperature environmental stress conditions.

Plant adaptation to drought stress

Plants in their natural habitats adapt to drought stress in the environment through a variety of mechanisms, ranging from transient responses to low soil moisture to major survival mechanisms of escape by early flowering in absence of seasonal rainfall. However, crop plants selected by humans to yield products such as grain, vegetable, or fruit in favorable environments with high inputs of water and fertilizer are expected to yield an economic product in response to inputs. Crop plants selected for their economic yield need to survive drought stress through mechanisms that maintain crop yield. Studies on model plants for their survival under stress do not, therefore, always translate to yield of crop plants under stress, and different aspects of drought stress response need to be emphasized. The crop plant model rice ( Oryza sativa) is used here as an example to highlight mechanisms and genes for adaptation of crop plants to drought stress.

Comparative analysis of some biochemical responses of three indica rice varieties during polyethylene glycol-mediated water stress exhibits distinct varietal differences

Extensive investigation into plant response and adaptation to diverse osmotic stresses like high salt/dehydration/low temperature, involving a broad spectrum of cellular physiological and biochemical changes, is essential to unravel intrinsic mechanism to mitigate against such stresses. In our previous communications, we conducted biochemical analyses of indica rice varieties, subjected to exogenous salt/abscisic acid-mediated oxidative stress. The aim of this study was to compare differential biochemical responses of the salt-sensitive (IR-29), salt-tolerant (Pokkali) and aromatic (Pusa Basmati or PB) rice varieties during polyethylene glycol (PEG)-induced dehydration stress. The greater susceptibility of IR-29 and PB, to water scarcity, was reflected by the higher toxic Na+ and putrescine accumulation, considerable decrease in (reduced/oxidized) glutathione, maximal increment in protease activity and greater downregulation of nitrate reductase activity. On the other hand, Pokkali appeared to suffer lesser damages as evidenced from much lower endogenous Na+ but higher K+, Ca2+ and Mg2+ accumulation, registering the highest levels of osmolytes like glycinebetaine and higher polyamines (spermidine and spermine) accounting to improved relative water content, higher (reduced/oxidized) glutathione, maximal induction of the enzyme phenylalanine ammonia-lyase and practically unhindered nitrate reductase activity, following PEG treatment. The highest induction of sugars and proline in IR-29 and PB probably played the osmoprotective/antioxidative functions, enabling to a certain extent to heighten their lipoxygenase inhibition or H2O2 scavenging potential, more than Pokkali, to ward off oxidative damages and sustain survival under critical dehydrated situations. Thus, the salt-tolerant Pokkali also showed prominent dehydration-tolerance properties, whereas the aromatic rice PB, almost identical in their biochemical responses to IR-29, showed greater sensitivity to PEG-mediated water deficit.

Rice GROWTH UNDER DROUGHT KINASE Is Required for Drought Tolerance and Grain Yield under Normal and Drought Stress Conditions

A drought stress-inducible receptor-like kinase is important for plant growth and grain yield in rice. Rice (Oryza sativa) is the primary food source for more than one-half of the world’s population. Because rice cultivation is dependent on water availability, drought during flowering severely affects grain yield. Here, we show that the function of a drought-inducible receptor-like cytoplasmic kinase, named GROWTH UNDER DROUGHT KINASE (GUDK), is required for grain yield under drought and well-watered conditions. Loss-of-function gudk mutant lines exhibit sensitivity to salinity, osmotic stress, and abscisic acid treatment at the seedling stage, and a reduction in photosynthesis and plant biomass under controlled drought stress at the vegetative stage. The gudk mutants interestingly showed a significant reduction in grain yield, both under normal well-watered conditions and under drought stress at the reproductive stage. Phosphoproteome profiling of the mutant followed by in vitro assays identified the transcription factor APETALA2/ETHYLENE RESPONSE FACTOR OsAP37 as a phosphorylation target of GUDK. The involvement of OsAP37 in regulating grain yield under drought through activation of several stress genes was previously shown. Our transactivation assays confirmed that GUDK is required for activation of stress genes by OsAP37. We propose that GUDK mediates drought stress signaling through phosphorylation and activation of OsAP37, resulting in transcriptional activation of stress-regulated genes, which impart tolerance and improve yield under drought. Our study reveals insights around drought stress signaling mediated by receptor-like cytoplasmic kinases, and also identifies a primary regulator of grain yield in rice that offers the opportunity to improve and stabilize rice grain yield under normal and drought stress conditions.

Effects of exogenous abscisic acid on some physiological responses in a popular aromatic indica rice compared with those from two traditional non-aromatic indica rice cultivars

The poor productivity and local confinement of indigenous aromatic rice varieties are mostly due to their susceptibility to salinity/drought/abscisic acid (ABA)-mediated abiotic stresses. It is thus essential to study the effects of several stress factors on their physiological parameters so as to improve their tolerance mechanism and enhance their global demand. Previously, we studied the effect of salinity stress on the physiological and molecular responses of the common aromatic rice Gobindobhog. The objective of this study was to understand the influence of exogenous ABA on some biochemical parameters in Gobindobhog, and comparison with those from non-aromatic M-1-48 and Nonabokra rice. The highest endogenous hydrogen peroxide content and membrane lipid peroxidation (increased malondialdehyde and lipoxygenase activity) were found in ABA-treated Gobindobhog leaves. While the catalase activity was down regulated the most in ABA-treated Gobindobhog leaves, the guaiacol peroxidase activity was induced maximally, indicating the protective role of peroxidase rather than catalase, during ABA-induced oxidative damages. The antioxidant, anthocyanin, showed the highest level in ABA-treated Nonabokra. Enhanced cysteine, following ABA exposure and the highest levels of reducing sugars, total amino acids, proline, and polyamines (putrescine and spermidine) recorded in Gobindobhog, probably served to shield from ABA-induced stress injuries, whereas the spermine levels were comparable in ABA-treated Nonabokra and Gobindobhog. The aroma content, intensified after ABA treatment, was markedly noted in Gobindobhog. Thus, the systematic examination of ABA-mediated stress revealed the most prominent oxidative damages in Gobindobhog, even higher than M-1-48, with a concomitant enhancement in peroxidase system and particularly osmolyte or polyamine levels to ensure its sustenance.

Expression Profiling of Abiotic Stress-Inducible Genes in response to Multiple Stresses in Rice (Oryza sativa L.) Varieties with Contrasting Level of Stress Tolerance

The present study considered transcriptional profiles and protein expression analyses from shoot and/or root tissues under three abiotic stress conditions, namely, salinity, dehydration, and cold, as well as following exogenous abscisic acid treatment, at different time points of stress exposure in three indica rice varieties, IR-29 (salt sensitive), Pokkali, and Nonabokra (both salt tolerant). The candidate genes chosen for expression studies were HKT-1, SOS-3, NHX-1, SAPK5, SAPK7, NAC-1, Rab16A, OSBZ8, DREBP2, CRT/DREBP, WRKY24, and WRKY71, along with the candidate proteins OSBZ8, SAMDC, and GST. Gene expression profile revealed considerable differences between the salt-sensitive and salt-tolerant rice varieties, as the expression in the latter was higher even at the constitutive level, whereas it was inducible only by corresponding stress signals in IR-29. Whether in roots or shoots, the transcriptional responses to different stressors peaked following 24 h of stress/ABA exposure, and the transcript levels enhanced gradually with the period of exposure. The generality of stress responses at the transcriptional level was therefore time dependent. Heat map data also showed differential transcript abundance in the three varieties, correlating the observation with transcript profiling. In silico analysis of the upstream regions of all the genes represented the existence of conserved sequence motifs in single or multiple copies that are indispensable to abiotic stress response. Overall, the transcriptome and proteome analysis undertaken in the present study indicated that genes/proteins conferring tolerance, belonging to different functional classes, were overrepresented, thus providing novel insight into the functional basis of multiple stress tolerance in indica rice varieties. The present work will pave the way in future to select gene(s) for overexpression, so as to generate broad spectrum resistance to multiple stresses simultaneously.

Identification of trans-acting factors regulating SamDC expression in Oryza sativa.

Abiotic stress affects the growth and productivity of crop plants; to cope with the adverse environmental conditions, plants have developed efficient defense machinery comprising of antioxidants like phenolics and flavonoids, and osmolytes like polyamines. SamDC is a key enzyme in the polyamine biosynthesis pathway in plants. In our present communication we have done in silico analysis of the promoter region of SamDC to look for the presence of different cis-regulatory elements contributing to its expression. Based on the presence of different cis-regulatory elements we completed comparative analysis of SamDC gene expression in rice lamina of IR-29 and Nonabokra by qPCR in response to the abiotic stress treatments of salinity, drought, cold and the biotic stress treatments of ABA and light. Additionally, to explore the role of the cis-regulatory elements in regulating the expression of SamDC gene in plants we comparatively analyzed the binding of rice nuclear proteins prepared from IR-29 and Nonabokra undergoing various stress treatments. The intensity of the complex formed was low and inducible in IR-29 in contrast to Nonabokra. Southwestern blot analysis helped in predicting the size of the trans-acting factors binding to these cis-elements. To our knowledge this is the first report on the comprehensive analysis of SamDC gene expression in rice and identification of the trans-acting factors regulating its expression.