A novel micronutrients and methyl jasmonate cocktail of elicitors via seed priming improves drought tolerance by mitigating oxidative stress in rice (Oryza sativa L.).

Drought is a major limiting factor for rice (Oryza sativa L.) production globally, and a cost-effective seed priming technique using bio-elicitors has been found to have stress mitigating effects. Till date, mostly phytohormones have been preferred as bio-elicitors, but the present study is a novel attempt to demonstrate the favorable role of micronutrients-phytohormone cocktail, i.e., iron (Fe), zinc (Zn), and methyl jasmonate (MJ) via seed priming method in mitigating the deleterious impacts of drought stress through physio-biochemical and molecular manifestations. The effect of cocktail/priming was studied on the relative water content, chlorophyll a/b and carotenoid contents, proline content, abscisic acid (ABA) content, and on the activities of ascorbate peroxidase (APX), superoxide dismutase (SOD), NADPH oxidase (Nox), and catalase (CAT). The expressions of drought-responsive genes OsZn-SOD, OsFe-SOD, and Nox1 were found to be modulated under drought stress in contrasting rice genotypes -N-22 (Nagina-22, drought-tolerant) and PS-5 (Pusa Sugandh-5, drought-sensitive). A progressive rise in carotenoids (10-19%), ABA (18-50%), proline (60-80%), activities of SOD (27-62%), APX (46-61%), CAT (50-80%), Nox (16-30%), and upregulated (0.9-1.6-fold) expressions of OsZn-SOD, OsFe-SOD, and Nox1 genes were found in the primed plants under drought condition. This cocktail would serve as a potential supplement in modern agricultural practices utilizing seed priming technique to mitigate drought stress-induced oxidative burst in food crops.

Expression pattern of candidate genes and their correlation with various metabolites of abscisic acid biosynthetic pathway under drought stress in rice.

Drought hampers global rice production. Abscisic acid (ABA) plays versatile roles under different environmental stresses. While the link between drought and ABA is known, its effect on ABA biosynthesis genes and metabolites is unclear. This study explored the impact of drought on various metabolites, namely beta-carotene, zeaxanthin, antheraxanthin, violaxanthin, neoxanthin, and candidate genes viz. zeaxanthin epoxidase (ZEP) and 9-cis epoxycarotenoid dioxygenase (NCED) of ABA biosynthesis pathway in rice cultivars (N22 and IR64) at anthesis {65 DAT (Days after transplanting)} with different stress levels. In stressed plants, zeaxanthin significantly increased (92%), while the concentration of beta-carotene, antheraxanthin, violaxanthin and neoxanthin decreased as drought stress progressed. The concentration of metabolites in roots was notably lower than in leaves in both genotypes. The ZEP expression was upregulated in roots (8.24-fold) under drought stress. Among five NCED isoforms, NCED3 showed significant upregulation (7.29-fold) in leaf and root tissue. NCED1 was significantly downregulated as stress progressed and was negatively correlated with ABA accumulation. NCED2, NCED4 and NCED5 showed no significant change in their expression. Drying and rolling of rice leaves was observed after imparting drought stress. The findings revealed that drought stress significantly influenced the expression of candidate genes and the concentration of metabolites of the ABA biosynthesis pathway. There was a significantly higher accumulation of ABA in N22 leaves (47%) and roots (30%) compared to IR64. The N22, a drought-tolerant genotype, exhibited significantly higher concentrations of intermediates and demonstrated increased expression of ZEP and NCED3, potentially contributing to its resilience against drought.

Recent advances in enhancing the production of long chain omega-3 fatty acids in microalgae.

Omega-3 fatty acids have gained attention due to numerous health benefits. Eicosapentaenoic (EPA) and docosahexaenoic acid (DHA) are long chain omega-3 fatty acids produced from precursor ALA (α-linolenic acid) in humans but their rate of biosynthesis is low, therefore, these must be present in diet or should be taken as supplements. The commercial sources of omega-3 fatty acids are limited to vegetable oils and marine sources. The rising concern about vegan source, fish aquaculture conservation and heavy metal contamination in fish has led to the search for their alternative source. Microalgae have gained importance due to the production of high-value EPA and DHA and can thus serve as a sustainable and promising source of long chain omega-3 fatty acids. Although the bottleneck lies in the optimization for enhanced production that involves strategies viz. strain selection, optimization of cultivation conditions, media, metabolic and genetic engineering approaches; while co-cultivation, use of nanoparticles and strategic blending have emerged as innovative approaches that have made microalgae as potential candidates for EPA and DHA production. This review highlights the possible strategies for the enhancement of EPA and DHA production in microalgae. This will pave the way for their large-scale production for human health benefits.

Comparative proteome analysis of phosphorus-responsive genotypes reveals the proteins differentially expressed under phosphorous starvation stress in rice.

Phosphorus (P)-deficiency is one of the major nutrient constraints for global rice production. P-deficiency tolerance in rice involves complex regulatory mechanisms. To gain insights into the proteins involved in phosphorus acquisition and use efficiency in rice, proteome analysis of a high-yielding rice cultivar Pusa-44 and its near-isogenic line (NIL)-23 harboring a major phosphorous uptake (Pup1) QTL, grown under control and P-starvation stress, was performed. Comparative proteome profiling of shoot and root tissues from the plants grown hydroponically with P (16 ppm, +P) or without P (0 ppm, -P) resulted in the identification of 681 and 567 differentially expressed proteins (DEPs) in shoot of Pusa-44 and NIL-23, respectively. Similarly, 66 and 93 DEPs were identified in root of Pusa-44 and NIL-23, respectively. These P-starvation responsive DEPs were annotated to be involved in metabolic processes like photosynthesis, starch-, sucrose-, energy-metabolism, transcription factors (mainly ARF, ZFP, HD-ZIP, MYB), and phytohormone signaling. Comparative analysis of the expression patterns observed by proteome analysis with that reported at the transcriptome level indicated the Pup1 QTL-mediated post-transcriptional regulation plays an important role under -P stress. Thus, the present study describes the molecular aspect of the regulatory functions of Pup1 QTL under P-starvation stress in rice, which might help develop an efficient rice cultivar with enhanced P acquisition and assimilation for better performance in P-deficient soil.

Insight of PBZ mediated drought amelioration in crop plants.

Water scarcity is a significant environmental limitation to plant productivity as drought-induced crop output losses are likely to outnumber losses from all other factors. In this context, triazole compounds have recently been discovered to act as plant growth regulators and multi-stress protectants such as heat, chilling, drought, waterlogging, heavy metals, etc. Paclobutrazol (PBZ) [(2RS, 3RS)-1-(4-chlorophenyl)- 4, 4-dimethyl-2-(1H-1, 2, 4-trizol-1-yl)-pentan-3-ol)] disrupts the isoprenoid pathway by blocking ent-kaurene synthesis, affecting gibberellic acid (GA) and abscisic acid (ABA) hormone levels. PBZ affects the level of ethylene and cytokinin by interfering with their biosynthesis pathways. Through a variety of physiological responses, PBZ improves plant survival under drought. Some of the documented responses include a decrease in transpiration rate (due to reduced leaf area), higher diffusive resistance, relieving reduction in water potential, greater relative water content, less water use, and increased antioxidant activity. We examined and discussed current findings as well as the prospective application of PBZ in regulating crop growth and ameliorating abiotic stresses in this review. Furthermore, the influence of PBZ on numerous biochemical, physiological, and molecular processes is thoroughly investigated, resulting in increased crop yield.

Isolation and characterization of drought and ABA responsive promoter of a transcription factor encoding gene from rice.

Water deficit is a significant impediment to enhancing rice yield. Genetic engineering tools have enabled agriculture researchers to develop drought-tolerant cultivars of rice. A common strategy to achieve this involves expressing drought-tolerant genes driven by constitutive promoters such as CaMV35S. However, the use of constitutive promoters is often limited by the adverse effects it has on the growth and development of the plant. Additionally, it has been observed that monocot-derived promoters are more successful in driving gene expression in monocots than in dicots. Substitution of constitutive promoters with stress-inducible promoters is the currently used strategy to overcome this limitation. In the present study, a 1514 bp AP2/ERF promoter that drives the expression of a transcription factor was cloned and characterized from drought-tolerant Indian rice genotype N22. The AP2/ERF promoter was fused to the GUS gene (uidA) and transformed in Arabidopsis and rice plants. Histochemical GUS staining of transgenic Arabidopsis plants showed AP2/ERF promoter activity in roots, stems, and leaves. Water deficit stress and ABA upregulate promoter activity in transformed Arabidopsis and rice. Quantitative PCR for uidA expression confirmed induced GUS activity in Arabidopsis and rice. This study showed that water deficit inducible Os-AP2/ERF-N22 promoter can be used to overcome the limitations of constitutive promoters. Transformants overexpressing Os-AP2/ERF-N22 showed higher relative water content, membrane stability index, total chlorophyll content, chlorophyll stability index, wax content, osmotic potential, stomatal conductance, transpiration rate, photosynthetic rate and radical scavenging activity. Drought tolerant (N22) showed higher expression of Os-AP2/ERF-N22 than the susceptible (MTU1010) cultivar. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01246-9.

Phosphorus homeostasis: acquisition, sensing, and long-distance signaling in plants.

Phosphorus (P), an essential nutrient required by plants often becomes the limiting factor for plant growth and development. Plants employ various mechanisms to sense the continuously changing P content in the soil. Transcription factors, such as SHORT ROOT (SHR), AUXIN RESPONSE FACTOR19 (ARF19), and ETHYLENE-INSENSITIVE3 (EIN3) regulate the growth of primary roots, root hairs, and lateral roots under low P. Crop improvement strategies under low P depend either on improving P acquisition efficiency or increasing P utilization. The various phosphate transporters (PTs) are involved in the uptake and transport of P from the soil to various plant cellular organelles. A plethora of regulatory elements including transcription factors, microRNAs and several proteins play a critical role in the regulation of coordinated cellular P homeostasis. Among these, the well-established P starvation signaling pathway comprising of central transcriptional factor phosphate starvation response (PHR), microRNA399 (miR399) as a long-distance signal molecule, and PHOSPHATE 2 (PHO2), an E2 ubiquitin conjugase is crucial in the regulation of phosphorus starvation responsive genes. Under PHR control, several classes of PHTs, microRNAs, and proteins modulate root architecture, and metabolic processes to enable plants to adapt to low P. Even though sucrose and inositol phosphates are known to influence the phosphorus starvation response genes, the exact mechanism of regulation is still unclear. In this review, a basic understanding of P homeostasis under low P in plants and all the above aspects are discussed.

Effects of excessive use of mobile phone technology in India on human health during COVID-19 lockdown.

OBJECTIVE: The global health crisis in the form of COVID-19 has forced people to shift their routine activities into a remote environment with the help of technology. The outbreak of the COVID-19 has caused several organizations to be shut down and forced them to initiate work from home employing technology. Now more than ever, it's important for people and institutions to understand the impact of excessive use of mobile phone technology and electronic gadgets on human health, cognition, and behavior. It is important to understand their perspective and how individuals are coping with this challenge in the wake of the COVID-19 pandemic. The investigation is an effort to answer the research question: whether dependency on technology during lockdown has more effects on human health in comparison to normal times. METHODS: The study included participants from India (n = 122). A questionnaire was framed and the mode of conducting the survey chosen was online to maintain social distancing during the time of the Pandemic. The gathered data was statistically analysed employing RStudio and multiple regression techniques. RESULTS: The statistical analysis confirms that lockdown scenarios have led to an increase in the usage of mobile phone technology which has been confirmed by around 90% of participants. Moreover, 95% of the participants perceive an increased risk of developing certain health problems due to excessive usage of mobile phones and technology. It has been evaluated that participants under the age group 15-30 years are highly affected (45.9%) during lockdown due to excessive dependence on technology. And, amongst different professions, participants involved in online teaching-learning are the most affected (42.6%). CONCLUSION: The findings indicate that dependency on technology during lockdown has more health effects as compared to normal times. So, it is suggested that as more waves of pandemics are being predicted, strategies should be planned to decrease the psychological and physiological effects of the overuse of technology during lockdown due to pandemics. As the lockdown situation unfolds, people and organization functioning styles should be rolled back to the limited dependency on technology.

Targeted knockdown of ribulose-1, 5-bisphosphate carboxylase-oxygenase in rice mesophyll cells.

We generated antisense constructs targeting two of the five Rubisco small subunit genes (OsRBCS2 and 4) which account for between 30-40 % of the RBCS transcript abundance in leaf blades. The constructs were driven by a maize phosphoenolpyruvate carboxylase (PEPC) promoter known to have enriched expression in mesophyll cells (MCs). In the resulting lines leaf, Rubisco protein content was reduced by between 30-50 % and CO2 assimilation rate was limited under photorespiratory and non-photorespiratory conditions. A relationship between Rubisco protein content and CO2 assimilation rate was found. This was associated with a significant reduction in dry biomass accumulation and grain yield of between 37-70%. In addition to serving as a resource for reducing Rubisco accumulation in a cell-preferential manner, these lines allow us to characterize gene function and isoform specific suppression on photosynthesis and growth. Our results suggest that the knockdown of multiple genes is required to completely reduce Rubisco accumulation in MCs.

Correlation between expression and activity of ADP glucose pyrophosphorylase and starch synthase and their role in starch accumulation during grain filling under drought stress in rice.

ADP glucose pyrophosphorylase (AGPase, EC 2.7.7.27) and starch synthase (SS, EC 2.4.1.21) are key regulatory enzymes involved in the starch biosynthesis. Comprehensive analysis of transcription levels of ADP-glucose pyrophosphorylase and starch synthase genes was performed in leaves, roots, and developing grains of drought susceptible (IR64) and drought-tolerant (N22) cultivars under applied water deficit stress (WDS). AGPase and SS genes are differentially regulated in leaves, roots, and grains under the drought stress. The expression pattern of SS and AGPase genes was correlated with the activity of both AGPase, SS, and starch content of developing grains under the drought. Drought stress reduced transitory starch in leaves and enhanced storage starch in developing grains. An increase in the activity of AGPase in developing grains was due to induced expression of ADP glucose pyrophosphorylase large subunit 3 (AGPL3) in N22 and both ADP glucose pyrophosphorylase small subunit 2 (AGPS2) & ADP glucose pyrophosphorylase large subunit 3 (AGPL3) in IR64 and a positive correlation was established with starch content. Similarly, an increase in the SS activity in developing grains was due to induced expression of soluble starch synthase (SSIIB, SSIVA, and SSIVB) in N22 and SSIVB in IR64.

Starch accumulation in rice grains subjected to drought during grain filling stage.

Drought stress during the grain filling stage severely affects the quality and quantity of starch in rice grains. The enzymes such as ADP-glucose pyrophosphorylase (AGPase, EC 2.7.7.27) and starch synthase (SS, EC 2.4.1.21) are the key regulatory enzymes involved in the starch biosynthesis. In this study, the activity of the AGPase and starch synthase (SS) was correlated with the qualitative and quantitative parameters such as sucrose, starch, amylose, amylopectin, and resistant starch in leaves, roots, and grains of drought tolerant (N22) and drought susceptible (IR64) cultivars under applied water deficit stress (WDS). Drought stress enhanced the remobilization of stored starch from leaves to developing rice grains which was positively correlated with a decrease in the starch and starch synthase activity in leaves. Starch accumulation in developing grains was positively correlated with an increase in the AGPase and SS activity under drought. It was found that starch, amylopectin, and sucrose content in developing grains increased under water deficit stress (WDS), while amylose content decreased in both the varieties. However, in leaves, the SS activity decreased while AGPase activity was found to be increased under WDS in both varieties. Decreased starch content in matured grains was due to shortening of grain filling stage as drought stress caused early plant senescence. Yield reduction under drought was more in susceptible variety IR64 as compared to tolerant genotype N22.

Regulation of phytosterol biosynthetic pathway during drought stress in rice.

Plants respond to drought stress in the form of various physio-biochemical and molecular changes at both cellular and molecular levels. Drought stress causes the destruction of cell membranes by disintegration of membrane lipids. One of the major groups of membrane lipids that plays important role in preserving the integrity of cell membranes is phytosterols. HMG-CoA reductase (HMGR) is the principal enzyme in the biosynthesis of plant sterols, synthesized via mevalonic acid pathway. Phospholipid: sterol acyltransferase (PSAT) is another important enzyme that plays an important role in turnover of phytosterols into steryl esters and helps maintain homeostasis of membrane lipids. In this study, the expression of both HMGR and PSAT genes in drought sensitive (IR64) and drought tolerant (N22) rice cultivars under applied drought conditions were found to be elevated. The increase in expression of these genes was proportional to the level of severity of applied drought stress. This is substantiated by the negative correlation of HMGR and PSAT expression to relative water content (RWC) and membrane stability index (MSI). Expression of PSAT was also found to be positively correlated to ABA content and HMGR expression.

Elicitor-Induced Biochemical and Molecular Manifestations to Improve Drought Tolerance in Rice (Oryza sativa L.) through Seed-Priming.

Rice (Oryza sativa L.) is one of the major grain cereals of the Indian subcontinent which face water-deficit stress for their cultivation. Seed-priming has been reported to be a useful approach to complement stress responses in plants. In the present study, seed-priming with hormonal or chemical elicitor [viz. methyl jasmonate (MJ), salicylic acid (SA), paclobutrazol (PB)] showed significant increase in total phenolic content, antioxidant activity and expression of Rice Drought-responsive (RD1 and RD2) genes (of AP2/ERF family) in contrasting rice genotypes (Nagina-22, drought-tolerant and Pusa Sugandh-5, drought-sensitive) under drought stress. However, decrease in lipid peroxidation and protein oxidation was observed not only under the stress but also under control condition in the plants raised from primed seeds. Expression analyses of RD1 and RD2 genes showed upregulated expression in the plants raised from primed seeds under drought stress. Moreover, the RD2 gene and the drought-sensitive genotype showed better response than that of the RD1 gene and the drought-tolerant genotype in combating the effects of drought stress. Among the elicitors, MJ was found to be the most effective for seed-priming, followed by PB and SA. Growth and development of the plants raised from primed seeds were found to be better under control and drought stress conditions compared to that of the plants raised from unprimed seeds under the stress. The present study suggests that seed-priming could be one of the useful approaches to be explored toward the development of simple, cost-effective and farmer-friendly technology to enhance rice yield in rainfed areas.

Expression analysis of bZIP transcription factor encoding genes in response to water deficit stress in rice.

In plants, basic region/leucine zipper motif (bZIP) transcription factors regulate several developmental processes and activate genes in response to biotic and abiotic stresses. Role of stress responsive bZIP transcription factors was studied in paddy in relation to different stages of development and water deficit stress (WDS) in a drought tolerant cultivar N22 and susceptible IR 64. Further, relative water content (RWC), membrane stability index (MSI) and abscisic acid (ABA) content were measured as indices of WDS at different stages of development and levels of stress. Expression of stress responsive bZIP transcription factors was directly correlated to developmental stage and WDS and indirectly to RWC, MSI and ABA content.

Cloning and characterization of a water deficit stress responsive transcription factor gene from Oryza sativa L.

Understanding the biochemical and molecular basis of drought mechanism in rice is important as drought is one of the major causes affecting rice crop adversely. A 1017 bp gene sequence encoding AP2/ERF family TF was isolated from Oryza sativa sp. Indica cv N22 encoding a protein of 338 amino acid residues, with a molecular weight of 36.58 kDa, and no intron in the ORF. The gene was named as AP2/ERF-N22(2) different from the drought responsive gene AP2/ERF-N22 that we reported earlier. AP2/ERF-N22(2) has entirely different characteristics from that of AP2/ERF-N22. It has a single AP2 domain of 55 amino acid residues and a cluster of acidic amino acid residues at the C-terminal region, which could function as a trans-activation domain. Presence of NLS indicates that it is a nuclear localized transcription factor encoding gene. It falls in group VI L, sharing characteristic similarities. Arabidopsis members of group VI L have been shown to be involved in response to cytokinin under drought stress.

Role of phytosterols in drought stress tolerance in rice.

Phytosterols are integral components of the membrane lipid bilayer in plants. They regulate membrane fluidity to influence its properties, functions and structure. An increase in accumulation of phytosterols namely campesterol, stigmasterol and ß-sitosterol was observed in rice as seedlings matured. The levels of the major phytosterol, ß-sitosterol in N22 (drought tolerant) rice seedlings was found to increase proportionately with severity of drought stress. Its levels were 145, 216, 345 and 364 µg/g FW after subjecting to water stress for 3, 6, 9 and 12 days respectively, while for IR64 (drought susceptible), levels were 137, 198, 227 and 287 µg/g FW at the same stages. Phytosterols were also found to increase with maturity as observed at 30, 50 and 75 days after planting. The activity of HMG-CoA reductase (EC 1.1.1.34) which is considered to be a key limiting enzyme in the biosynthesis of phytosterols was 0.55, 0.56, 0.78 and 0.85 µmol/min/L at 3, 6, 9 and 12 days of water stress in N22 and 0.31, 0.50, 0.54 and 0.65 µmol/min/L in case of IR64 respectively. The elevation in the levels of phytosterols as well as the activity of HMG-CoA reductase during drought stress indicates the role of phytosterols in providing tolerance to stress.

Role of gamma-oryzanol in drought-tolerant and susceptible cultivars of rice (Oryza sativa L.).

Drought-tolerant cultivars and their phytochemical composition, which has a role in providing drought tolerance are gaining importance. In this study, rice bran oil and semi-purified oryzanol (SPO) obtained from five rice (Oryza sativa L.) cultivars, namely P1401 and PB1 (drought-susceptible) and N22, PNR381 and APO (drought-tolerant) were analyzed for the gamma-oryzanol content, an antioxidant present in considerable amount in the rice bran. The higher level of gamma-oryzanol and its antioxidant activity was observed in drought-tolerant cultivars (N22, PNR381 and APO) as compared to drought-susceptible (PB1 and P1401), suggesting the role of gamma-oryzanol in drought tolerance, as antioxidants are known to play an important role by scavenging free radicals. The total antioxidant activity of gamma-oryzanol might be attributed to 24-methylene cycloartanyl ferulate, a major component of gamma-oryzanol. By enhancing the level of active oryzanol components identified in this study by genetic and molecular means could impart increased drought tolerance.

Cloning and characterization of a gene encoding novel zinc finger protein transcription factor induced under water deficit stress from rice (Oryza sativa) cv. N-22.

A gene OsZnI encoding Cys3/His1-type zinc finger protein was isolated from the water stress-induced cDNA library of rice (Oryza sativa) cv. N-22, an early maturing, deep-rooted, drought-tolerant genotype adapted to upland conditions. The in-silico analysis revealed an insert of 800 bp with an ORF of 663 nucleotides, encoding 221 amino acids. OsZnI had three distinct features--nuclear localization signal (NLS) present in Arg152-Arg168, Zn finger domain between 185-193 amino acids and 12 amino acids conserved domain in 71-82 amino acids homologous to LEA motif, and belonged to C-type family of Zn finger protein. OsZnI showed induced expression under water deficit stress.

Cloning and characterization of drought responsive partial gene sequence(s) from Oryza sativa L. subsp. Indica.

Differential display gels were run for the drought tolerant (N-22) and drought susceptible (Panidhan) genotypes of rice (Oryza sativa) to identify the genes showing differential expression with respect to moisture stress. Differential cDNA products were cloned in PCR-Trap vector and analyzed for differential expression by Northern hybridization. Two clones namely R4A and R7G were found to be associated with water deficit stress (WDS). Sequencing revealed an insert of 244 bp in the clone R4A. BLASTN and FASTA results showed that R4A had maximum homology with a full-length cDNA clone: 002-110-H10 and OSJNBa006109.12 protein. GO classification suggested that it had beta-glucosidase motif which had been implicated in ABA mobilization and thereby ABA dependent gene expression.

Isolation of stress responsive Psb A gene from rice (Oryza sativa l.) using differential display.

Differential display (DD) experiments were performed on drought-tolerant rice (Oryza sativa L.) genotype N22 to identify both upregulated and downregulated partial cDNAs with respect to moisture stress. DNA polymorphism was detected between drought-stressed and control leaf tissues on the DD gels. A partial cDNA showing differential expression, with respect to moisture stress was isolated from the gel. Northern blotting analysis was performed using this cDNA as a probe and it was observed that mRNA corresponding to this transcript was accumulated to high level in rice leaves under water deficit stress. At the DNA sequence level, the partial cDNA showed homology with psb A gene encoding for Dl protein.