Shashibhushan Gahir

Bio: Shashibhushan Gahir is an academic researcher from University of Hyderabad. The author has contributed to research in topics: Guard cell & Biotic stress. The author has an hindex of 2, co-authored 4 publications receiving 63 citations.

Mechanism of Stomatal Closure in Plants Exposed to Drought and Cold Stress.

Abstract: Drought is one of the abiotic stresses which impairs the plant growth/development and restricts the yield of many crops throughout the world. Stomatal closure is a common adaptation response of plants to the onset of drought condition. Stomata are microscopic pores on the leaf epidermis, which regulate the transpiration/CO2 uptake by leaves. Stomatal guard cells can sense various abiotic and biotic stress stimuli from the internal and external environment and respond quickly to initiate closure under unfavorable conditions. Stomata also limit the entry of pathogens into leaves, restricting their invasion. Drought is accompanied by the production and/or mobilization of the phytohormone, abscisic acid (ABA), which is well-known for its ability to induce stomatal closure. Apart from the ABA, various other factors that accumulate during drought and affect the stomatal function are plant hormones (auxins, MJ, ethylene, brassinosteroids, and cytokinins), microbial elicitors (salicylic acid, harpin, Flg 22, and chitosan), and polyamines . The role of various signaling components/secondary messengers during stomatal opening or closure has been a matter of intense investigation. Reactive oxygen species (ROS), nitric oxide (NO), cytosolic pH, and calcium are some of the well-documented signaling components during stomatal closure. The interrelationship and interactions of these signaling components such as ROS, NO, cytosolic pH, and free Ca2+ are quite complex and need further detailed examination.

Abscisic Acid-Induced Stomatal Closure: An Important Component of Plant Defense Against Abiotic and Biotic Stress.

Abstract: Abscisic acid (ABA) is a stress hormone that accumulates under different abiotic and biotic stresses. A typical effect of ABA on leaves is to reduce transpirational water loss by closing stomata and parallelly defend against microbes by restricting their entry through stomatal pores. ABA can also promote the accumulation of polyamines, sphingolipids, and even proline. Stomatal closure by compounds other than ABA also helps plant defense against both abiotic and biotic stress factors. Further, ABA can interact with other hormones, such as methyl jasmonate (MJ) and salicylic acid (SA). Such cross-talk can be an additional factor in plant adaptations against environmental stresses and microbial pathogens. The present review highlights the recent progress in understanding ABA's multifaceted role under stress conditions, particularly stomatal closure. We point out the importance of reactive oxygen species (ROS), reactive carbonyl species (RCS), nitric oxide (NO), and Ca2+ in guard cells as key signaling components during the ABA-mediated short-term plant defense reactions. The rise in ROS, RCS, NO, and intracellular Ca2+ triggered by ABA can promote additional events involved in long-term adaptive measures, including gene expression, accumulation of compatible solutes to protect the cell, hypersensitive response (HR), and programmed cell death (PCD). Several pathogens can counteract and try to reopen stomata. Similarly, pathogens attempt to trigger PCD of host tissue to their benefit. Yet, ABA-induced effects independent of stomatal closure can delay the pathogen spread and infection within leaves. Stomatal closure and other ABA influences can be among the early steps of defense and a crucial component of plants' innate immunity response. Stomatal guard cells are quite sensitive to environmental stress and are considered good model systems for signal transduction studies. Further research on the ABA-induced stomatal closure mechanism can help us design strategies for plant/crop adaptations to stress.

Protein Phosphatases in Guard Cells: Key Role in Stomatal Closure and Opening

Abstract: Protein phosphatases (PPs) are indispensable for growth, development, and various signaling pathways in plants. Opening or closure of stomata, the tiny pores on leaf surface, is essential for optimizing plant performance, under abiotic stress. PPs are important links in the signal transduction events to connect signal perception with subsequent steps of ion efflux/influx leading to the decrease/increase in turgor of the guard cells. Of the diverse spectrum of PPs in plants, only a few are involved in stomatal movements, particularly protein phosphatase 2C (PP2C), protein phosphatase 2A (PP2A), protein phosphatase 1 (PP1), and protein phosphatase 7 (PP7). PP2C plays a pivotal role in abscisic acid (ABA)-induced stomatal closure as it forms a ternary complex with ABA and receptors, switching on the Open STomata 1 (OST1) kinase by phosphorylation, which in turn triggers events leading to ion efflux. On the other hand PP1 participates during stomatal opening by blue light perceived by phototropins (Phots) or cryptochromes (Crys). On sensing the blue light, stomatal phototropin can modulate PP1 by a blue light-activated kinase. As a consequence, H+-ATPase is activated, followed by membrane hyperpolarization leading to K+ influx, increased turgor, and opening. The tripartite interaction of ABA, ABA receptors, and PP2C provides the missing link to complete the chain of perception of ABA and its regulatory effects. The presence of ABA is perceived by receptors PYLs and stabilized by complex with PP2C. This tripartite interaction activates OST1 kinase and in turn phosphorylates NADPH oxidase to produce ROS and initiate events of closure. Arabidopsis mutants were instrumental in demonstrating the roles of PPs in signal transduction of stomatal guard cells. This chapter concludes with remarks on existing gaps in our knowledge and suggestions for further studies.

Abscisic Acid-Induced Stomatal Closure: An Important Component of Plant Defense Against Abiotic and Biotic Stress.

Abstract: Abscisic acid (ABA) is a stress hormone that accumulates under different abiotic and biotic stresses. A typical effect of ABA on leaves is to reduce transpirational water loss by closing stomata and parallelly defend against microbes by restricting their entry through stomatal pores. ABA can also promote the accumulation of polyamines, sphingolipids, and even proline. Stomatal closure by compounds other than ABA also helps plant defense against both abiotic and biotic stress factors. Further, ABA can interact with other hormones, such as methyl jasmonate (MJ) and salicylic acid (SA). Such cross-talk can be an additional factor in plant adaptations against environmental stresses and microbial pathogens. The present review highlights the recent progress in understanding ABA's multifaceted role under stress conditions, particularly stomatal closure. We point out the importance of reactive oxygen species (ROS), reactive carbonyl species (RCS), nitric oxide (NO), and Ca2+ in guard cells as key signaling components during the ABA-mediated short-term plant defense reactions. The rise in ROS, RCS, NO, and intracellular Ca2+ triggered by ABA can promote additional events involved in long-term adaptive measures, including gene expression, accumulation of compatible solutes to protect the cell, hypersensitive response (HR), and programmed cell death (PCD). Several pathogens can counteract and try to reopen stomata. Similarly, pathogens attempt to trigger PCD of host tissue to their benefit. Yet, ABA-induced effects independent of stomatal closure can delay the pathogen spread and infection within leaves. Stomatal closure and other ABA influences can be among the early steps of defense and a crucial component of plants' innate immunity response. Stomatal guard cells are quite sensitive to environmental stress and are considered good model systems for signal transduction studies. Further research on the ABA-induced stomatal closure mechanism can help us design strategies for plant/crop adaptations to stress.

Physiological mechanisms of exogenous calcium on alleviating salinity-induced stress in rice (Oryza sativa L.)

Abstract: Being more sensitive to salt stress among the cereals, growth of rice (Oryza sativa L.) has been habitually affected by salinity. Although, several practices have evolved to sustain the growth of rice under salinity, the enormous role of calcium (Ca2+) as a signalling molecule in salt stress mitigation is still arcane. Considering this fact, an experiment was performed aiming to explicate the mechanism of salt-induced growth inhibition in rice and its alleviation by exogenous Ca2+. At germination stage, 10 mM and 15 mM CaCl2 primed rice (cv. Binadhan-10 & Binadhan-7) seeds were grown in petri dishes for 9 days under 100 mM NaCl stress. At seedling stage, 9-day-old rice seedlings grown on sand were exposed to 100 mM NaCl alone and combined with 10 mM and 15 mM CaCl2 for 15 days. This research revealed that salinity radically slowed down growth of rice seedlings and Ca2+ treatment noticeably improved growth performances. At germination stage, 10 mM CaCl2 treatment significantly increased the final germination percentage, germination rate index (in Binadhan-7), shoot, root length (89.20, 67.58% in Bindhan-10 & 84.72, 31.15% in Bindhan-7) and biomass production under salinity. Similarly, at seedling stage, 10 mM CaCl2 supplementation in salt-stressed plants enhanced shoot length (42.17, 28.76%) and shoot dry weight (339.52, 396.20%) significantly in Binadhan-10 & Binadhan-7, respectively, but enhanced root dry weight (36.76%) only in Binadhan-10. In addition, 10 mM CaCl2 supplementation on salt-stressed seedlings increased the chlorophyll and proline content, and oppressed the accretion of reactive oxygen species thus protecting from oxidative damage more pronouncedly in Binadhan-10 than Binadhan-7 as reflected by the elevated levels of catalase and ascorbate peroxidase activity. The 15 mM CaCl2 somehow also enhanced some growth parameters but overall was less effective than 10 mM CaCl2 to alleviate salt stress, and sometimes showed negative effect. Therefore, supplementary application of calcium-rich fertilizers in saline prone soils can be an effective approach to acclimatize salt stress and cultivate rice successfully.

Plant growth under suboptimal water conditions: early responses and methods to study them

Abstract: Drought stress forms a major environmental constraint during the life cycle of plants, often decreasing plant yield and in extreme cases threatening survival. The molecular and physiological responses induced by drought have been the topic of extensive research during the past decades. Because soil-based approaches to studying drought responses are often challenging due to low throughput and insufficient control of the conditions, osmotic stress assays in plates were developed to mimic drought. Addition of compounds such as polyethylene glycol, mannitol, sorbitol, or NaCl to controlled growth media has become increasingly popular since it offers the advantage of accurate control of stress level and onset. These osmotic stress assays enabled the discovery of very early stress responses, occurring within seconds or minutes following osmotic stress exposure. In this review, we construct a detailed timeline of early responses to osmotic stress, with a focus on how they initiate plant growth arrest. We further discuss the specific responses triggered by different types and severities of osmotic stress. Finally, we compare short-term plant responses under osmotic stress versus in-soil drought and discuss the advantages, disadvantages, and future of these plate-based proxies for drought.

Inactivating transcription factor OsWRKY5 enhances drought tolerance through abscisic acid signaling pathways.

Abstract: During crop cultivation, water-deficit conditions retard growth, thus reducing crop productivity. Therefore, uncovering the mechanisms behind drought tolerance is a critical task for crop improvement. Here, we show that the rice (Oryza sativa) WRKY transcription factor OsWRKY5 negatively regulates drought tolerance. We determined that OsWRKY5 was mainly expressed in developing leaves at the seedling and heading stages, and that its expression was reduced by drought stress and by treatment with NaCl, mannitol, and abscisic acid (ABA). Notably, the genome-edited loss-of-function alleles oswrky5-2 and oswrky5-3 conferred enhanced drought tolerance, measured as plant growth under water-deficit conditions. Conversely, the overexpression of OsWRKY5 in the activation-tagged line oswrky5-D resulted in higher susceptibility under the same conditions. Loss of OsWRKY5 activity increased sensitivity to ABA, thus promoting ABA-dependent stomatal closure. Transcriptome deep sequencing and RT-qPCR analyses demonstrated that the expression of abiotic stress-related genes including rice MYB2 (OsMYB2) was upregulated in oswrky5 knockout mutants and downregulated in oswrky5-D mutants. Moreover, dual-luciferase, yeast one-hybrid, and chromatin immunoprecipitation assays showed that OsWRKY5 directly binds to the W-box sequences in the promoter region of OsMYB2 and represses OsMYB2 expression, thus downregulating genes downstream of OsMYB2 in the ABA signaling pathways. Our results demonstrate that OsWRKY5 functions as a negative regulator of ABA-induced drought stress tolerance, strongly suggesting that inactivation of OsWRKY5 or manipulation of key OsWRKY5 targets could be useful to improve drought tolerance in rice cultivars.