Free Radicals Mediated Redox Signaling in Plant Stress Tolerance.

Abiotic and biotic stresses negatively affect plant cellular and biological processes, limiting their growth and productivity. Plants respond to these environmental cues and biotrophic attackers by activating intricate metabolic-molecular signaling networks precisely and coordinately. One of the initial signaling networks activated is involved in the generation of reactive oxygen species (ROS), reactive nitrogen species (RNS), and reactive sulfur species (RSS). Recent research has exemplified that ROS below the threshold level can stimulate plant survival by modulating redox homeostasis and regulating various genes of the stress defense pathway. In contrast, RNS regulates the stress tolerance potential of crop plants by modulating post-translation modification processes, such as S-nitrosation and tyrosine nitration, improving the stability of protein and DNA and activating the expression of downstream stress-responsive genes. RSS has recently emerged as a new warrior in combating plant stress-induced oxidative damage by modulating various physiological and stress-related processes. Several recent findings have corroborated the existence of intertwined signaling of ROS/RNS/RSS, playing a substantial role in crop stress management. However, the molecular mechanisms underlying their remarkable effect are still unknown. This review comprehensively describes recent ROS/RNS/RSS biology advancements and how they can modulate cell signaling and gene regulation for abiotic stress management in crop plants. Further, the review summarizes the latest information on how these ROS/RNS/RSS signaling interacts with other plant growth regulators and modulates essential plant functions, particularly photosynthesis, cell growth, and apoptosis.

Investigating NAC Transcription Factor Role in Redox Homeostasis in Solanum lycopersicum L.: Bioinformatics, Physiological and Expression Analysis under Drought Stress.

NAC transcription factors regulate stress-defence pathways and developmental processes in crop plants. However, their detailed functional characterization in tomatoes needs to be investigated comprehensively. In the present study, tomato hybrids subjected to 60 and 80 days of drought stress conditions showed a significant increase in membrane damage and reduced relative water, chlorophyll and proline content. However, hybrids viz., VRTH-16-3 and VRTH-17-68 showed superior growth under drought stress, as they were marked with low electrolytic leakage, enhanced relative water content, proline content and an enhanced activity of enzymatic antioxidants, along with the upregulation of NAC and other stress-defence pathway genes. Candidate gene(s) exhibiting maximum expression in all the hybrids under drought stress were subjected to detailed in silico characterization to provide significant insight into its structural and functional classification. The homology modelling and superimposition analysis of predicted tomato NAC protein showed that similar amino acid residues were involved in forming the conserved WKAT domain. DNA docking discovered that the SlNAC1 protein becomes activated and exerts a stress-defence response after the possible interaction of conserved DNA elements using Pro72, Asn73, Trp81, Lys82, Ala83, Thr84, Gly85, Thr86 and Asp87 residues. A protein-protein interaction analysis identified ten functional partners involved in the induction of stress-defence tolerance.

Integrating speed breeding with artificial intelligence for developing climate-smart crops.

INTRODUCTION: In climate change, breeding crop plants with improved productivity, sustainability, and adaptability has become a daunting challenge to ensure global food security for the ever-growing global population. Correspondingly, climate-smart crops are also the need to regulate biomass production, which is imperative for the maintenance of ecosystem services worldwide. Since conventional breeding technologies for crop improvement are limited, time-consuming, and involve laborious selection processes to foster new and improved crop varieties. An urgent need is to accelerate the plant breeding cycle using artificial intelligence (AI) to depict plant responses to environmental perturbations in real-time. MATERIALS AND METHODS: The review is a collection of authorized information from various sources such as journals, books, book chapters, technical bulletins, conference papers, and verified online contents. CONCLUSIONS: Speed breeding has emerged as an essential strategy for accelerating the breeding cycles of crop plants by growing them under artificial light and temperature conditions. Furthermore, speed breeding can also integrate marker-assisted selection and cutting-edged gene-editing tools for early selection and manipulation of essential crops with superior agronomic traits. Scientists have recently applied next-generation AI to delve deeper into the complex biological and molecular mechanisms that govern plant functions under environmental cues. In addition, AIs can integrate, assimilate, and analyze complex OMICS data sets, an essential prerequisite for successful speed breeding protocol implementation to breed crop plants with superior yield and adaptability.

Functional characterization of two WD40 family proteins, Alr0671 and All2352, from Anabaena PCC 7120 and deciphering their role in abiotic stress management.

WD40 domain-containing proteins are one of the eukaryotes' most ancient and ubiquitous protein families. Little is known about the presence and function of these proteins in cyanobacteria in general and Anabaena in particular. In silico analysis confirmed the presence of WD40 repeats. Gene expression analysis indicated that the transcript levels of both the target proteins were up-regulated up to 4 fold in Cd and drought and 2-3 fold in heat, salt, and UV-B stress. Using a fluorescent oxidative stress indicator, we showed that the recombinant proteins were scavenging reactive oxygen species (ROS) (4-5 fold) more efficiently than empty vectors. Chromatin immunoprecipitation analysis (ChIP) and electrophoretic mobility shift assay (EMSA) revealed that the target proteins function as transcription factors after binding to the promoter sequences. The presence of kinase activity (2-4 fold) in the selected proteins indicated that these proteins could modulate the functions of other cellular proteins under stress conditions by inducing phosphorylation of specific amino acids. The chosen proteins also demonstrated interaction with Zn, Cd, and Cu (1.4-2.5 fold), which might stabilize the proteins' structure and biophysical functions under multiple abiotic stresses. The functionally characterized Alr0671 and All2352 proteins act as transcription factors and offer tolerance to agriculturally relevant abiotic stresses.

Alr0671 and All2352 are novel WD40 proteins of Anabaena capable of regulating biochemical functions and abiotic stress tolerance by acting as a transcription factor and mediating DNA-protein interaction.

Regulation of antioxidant defense and glyoxalase systems in cyanobacteria.

Oxidative stress is common consequence of abiotic stress in plants as well as cyanobacteria caused by generation of reactive oxygen species (ROS), an inevitable product of respiration and photosynthetic electron transport. ROS act as signalling molecule at low concentration however, when its production exceeds the endurance capacity of antioxidative defence system, the organisms suffer oxidative stress. A highly toxic metabolite, methylglyoxal (MG) is also produced in cyanobacteria in response to various abiotic stresses which consequently augment the ensuing oxidative damage. Taking recourse to the common lineage of eukaryotic plants and cyanobacteria, it would be worthwhile to explore the regulatory role of glyoxalase system and antioxidative defense mechanism in combating abiotic stress in cyanobacteria. This review provides comprehensive information on the complete glyoxalase system (GlyI, GlyII and GlyIII) in cyanobacteria. Furthermore, it elucidates the recent understanding regarding the production of ROS and MG, noteworthy link between intracellular MG and ROS and its detoxification via synchronization of antioxidants (enzymatic and non-enzymatic) and glyoxalase systems using glutathione (GSH) as common co-factor.

Heterologous expression of cyanobacterial PCS confers augmented arsenic and cadmium stress tolerance and higher artemisinin in Artemisia annua hairy roots.

The present study provides the first report of heterologous expression of phytochelatin synthase from Anabaena PCC 7120 (anaPCS) into the hairy roots of Artemisia annua. Transformed hairy roots of A. annua expressing anaPCS gene showed better tolerance to heavy metals, viz., arsenic (As) and cadmium (Cd) owing to 143 and 191% more As- and Cd-accumulation, respectively, as compared to normal roots with a bioconcentration factor (BCF) of 9.7 and 21.1 for As and Cd, respectively. Under As and Cd stresses, transformed hairy roots possessed significantly higher amounts of phytochelatins and thiols probably due to the presence of both AaPCS (Artemisia annua PCS) and anaPCS. In addition, artemisinin synthesis was also induced in transformed hairy roots under heavy metals stresses. In-silico analysis revealed the presence of conserved motifs in both AaPCS and anaPCS sequences as well as structural modelling of PCS functional domain was conducted. Interaction of AaPCS and anaPCS proteins with CdCl2 and sodium arsenate gene ontology analysis gave insights to anaPCS functioning in transformed hairy roots of A. annua. The study provides transformed hairy roots of A. annua as an efficient tool for effective phytoremediation with added advantages of artemisinin extraction from hairy roots used for phytoremediation. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s11816-021-00682-5.

Biotechnological strategies for enhancing heavy metal tolerance in neglected and underutilized legume crops: A comprehensive review.

Contamination of agricultural land and water by heavy metals due to rapid industrialization and urbanization including various natural processes have become one of the major constraints to crop growth and productivity. Several studies have reported that to counteract heavy metal stress, plants should be able to maneuver various physiological, biochemical and molecular processes to improve their growth and development under heavy metal stress. With the advent of modern biotechnological tools and techniques it is now possible to tailor legume and other plants overexpressing stress-induced genes, transcription factors, proteins, and metabolites that are directly involved in heavy metal stress tolerance. This review provides an in-depth overview of various biotechnological approaches and/or strategies that can be used for enhancing detoxification of the heavy metals by stimulating phytoremediation processes. Synthetic biology tools involved in the engineering of legume and other crop plants against heavy metal stress tolerance are also discussed herewith some pioneering examples where synthetic biology tools that have been used to modify plants for specific traits. Also, CRISPR based genetic engineering of plants, including their role in modulating the expression of several genes/ transcription factors in the improvement of abiotic stress tolerance and phytoremediation ability using knockdown and knockout strategies has also been critically discussed.

Impact of green synthesized WcAgNPs on in-vitro plant regeneration and withanolides production by inducing key biosynthetic genes in Withania coagulans.

KEY MESSAGE: Withania coagulans (L.) Dunal bio-synthesized silver nanoparticles (WcAgNPs) worked as an abiotic elicitor or auto-catalyst that enhanced root regeneration and withanolides production in in-vitro regenerated W. coagulans. Rapid development in the production / consumption of silver nanoparticles (AgNPs) raised serious concern over its effects on the growth of natural plant community. The knowledge related to impact of AgNPs on plant growth and biocompatibility is increasing day by day, but comprehensive mechanism and gaps regarding their impacts on plant health have yet to be addressed. In the present study, we investigated the impact of Withania coagulans biosynthesized AgNPs (WcAgNPs) on in-vitro plant growth and withanolides production. Obtained results showed that the low concentrations of WcAgNPs significantly induced the plant growth by regulating oxidative stress via anti-oxidative defense system. Physiological, morphology and anatomical features also reflected healthy plant growth under low WcAgNPs exposure. While higher concentrations of WcAgNPs have a negative impact on W. coagulans plant growth due to induced lipid peroxidation, ROS accumulation, and root cell death. At lower concentrations, WcAgNPs have shown a positive effect on in-planta withanolides biosynthesis stimulating withanolide A and withaferin A up to 11.15-22.8-fold, respectively. Furthermore, the expression of withanolides biosynthetic genes were also quantified upon WcAgNPs exposure and terpenes biosynthetic genes showed over-expression. Thus, the present study concludes that the lower concentrations of WcAgNPs positively induced plant growth via improved root organogenesis and also have potential to act as an elicitor for withanolides production.

Interactive role of salicylic acid and nitric oxide on transcriptional reprogramming for high temperature tolerance in lablab purpureus L.: Structural and functional insights using computational approaches.

Salicylic acid (SA) and nitric oxide (NO) are considered as putative plant growth regulators that are involved in the regulation of an array of plant's growth and developmental functions under environmental fluctuations when applied at lower concentrations. The possible involvement of NO in SA induced attenuation of high temperature (HT) induced oxidative stress in plants is however, still vague and need to be explored. Therefore, the present study aimed to investigates the biochemical and physiological changes induced by foliar spray of SA and NO combinations to ameliorate HT induced oxidative stress in Lablab purpureus L. Foliar application of combined SA and NO significantly improved relative water content (27.8 %), photosynthetic pigment content (67.2 %), membrane stability (45 %), proline content (1.0 %), expression of enzymatic antioxidants (7.1-18 %) along with pod yield (1.0 %). Heat Shock Factors (HSFs) play crucial roles in plants abiotic stress tolerance, however there structural and functional classifications in L. purpureus L. is still unknown. So, In-silico approach was also used for functional characterization and homology modelling of HSFs in L. purpureus. The experimental findings depicted that combine effect of SA and NO enhances tolerance in HT stressed L. purpureus L. plants by regulating physiological functions, antioxidants, expression and regulation of stress-responsive genes via transcriptional regulation of heat shock factor.

Epigenetic control of UV-B-induced flavonoid accumulation in Artemisia annua L.

MAIN CONCLUSION: UV-B-induced flavonoid biosynthesis is epigenetically regulated by site-specific demethylation of AaMYB1, AaMYC, and AaWRKY TF-binding sites inAaPAL1promoter-causing overexpression ofAaPALgene inArtemisia annua. The present study was undertaken to understand the epigenetic regulation of flavonoid biosynthesis under the influence of ultraviolet-B radiation using Artemisia annua L. as an experimental model. In-vitro propagated and acclimatized plantlets were treated with UV-B radiation (2.8 W m-2; 3 h), which resulted in enhanced accumulation of total flavonoid and phenolics content as well as eleven individual flavonoids measured through HPLC-DAC. Expression of eight genes (phenylanaline ammonia lyase, cinnamate-4-hydroxylase, 4-coumarate: CoA ligase; chalcone synthase, chalcone isomerase, cinnamoyl reductase, flavonoid-3'-hydroxylase, and flavones synthase) from upstream and downstream flavonoid biosynthetic pathways was measured through RT-PCR and RT-Q-PCR and all were variably induced under UV-B irradiation. Among them, AaPAL1 transcript and its protein were most significantly upregulated. Global DNA methylation analysis revealed hypomethylation of genomic DNA in A. annua. Further epigenetic characterization of promoter region of AaPAL1 revealed cytosine demethylation at five sites, which in turn caused epigenetic activation of six transcription factor-binding sites including QELEMENT, EBOXBNNAPA/MYCCONSENSUSAT, MYBCORE, MYBCOREATCYCB1, and GCCCORE. MYB transcription factors are positive regulators of flavonoid biosynthesis. Epigenetic activation of transcription-enhancing cis-regulatory elements in AaPAL1 promoter and subsequent overexpression of AaMYB1 and AaMYC and AaWRKY transcription factors under UV-B irradiation may probably be the reason for higher AaPAL1 expression and hence greater biosynthesis of flavonoids in A. annua L. The present study is the first report that provides mechanistic evidence of epigenetic regulation of flavonoid biosynthesis under UV-B radiation in A. annua L.

Response of Lablab purpureus L. to high temperature stress and role of exogenous protectants in mitigating high temperature induced oxidative damages.

Present study was conducted to explore the role of exogenous salicylic acid (SA), sodium nitroprusside (SNP), abscisic acid (ABA) and proline (PRO) in mitigating high-temperature (HT) induced oxidative stress in different Lablab purpureus L. cultivars. The attempt was made to examine whether these phytohormones, when applied exogenously, were able to regulate plant morpho-physiological behavior by modulating genes and proteins involved in antioxidant defense system. The HT stress induced membrane damage, degraded chlorophyll, generated redox metabolites and significantly reduced growth and biomass in all the cultivars. Among all the four treatments, foliar application of SA and SNP were most effective in the regulation of growth and physiological processes of the cultivars compared to ABA and PRO applications. Thus, signifying the protective role of SA and SNP in mitigation of HT induced oxidative stress and conferring HT stress tolerance in the cultivars. Gene expression and leaf proteome analysis revealed that these phytohormones were also involved in regulation of defense related gene expression, stress inducible proteins and de novo synthesis of specific proteins under HT stress. The experimental findings depict that foliar applications of SA and SNP enhances HT stress tolerance in lablab cultivars by modulating antioxidant defense system and by regulating bio-physical growth more effectively as compared to ABA and PRO application.

Salicylic acid and nitric oxide alleviate high temperature induced oxidative damage in Lablab purpureus L plants by regulating bio-physical processes and DNA methylation.

Salicylic acid (SA) and sodium nitroprusside (SNP, NO donor) modulates plant growth and development processes and recent findings have also revealed their involvement in the regulation of epigenetic factors under stress condition. In the present study, some of these factors were comparatively studied in hyacinth bean plants subjected to high temperature (HT) environment (40-42 °C) with and without exogenous application of SA and SNP under field condition. Exogenous application of SA and SNP substantially modulated the growth and biophysical process of hyacinth bean plants under HT environment. Exogenous application of SA and SNP also remarkably regulated the activities of antioxidant enzymes, modulated mRNA level of certain enzymes, improves plant water relation, enhance photosynthesis and thereby increasing plant defence under HT. Coupled restriction enzyme digestion-random amplification (CRED-RA) technique revealed that many methylation changes were "dose dependent" and HT significantly increased DNA damages as evidenced by both increase and decrease in bands profiles, methylation and de-methylation pattern. Thus, the result of the present study clearly shows that exogenous SA and SNP regulates DNA methylation pattern, modulates stress-responsive genes and can impart transient HT tolerance by synchronizing growth and physiological acclimatization of plants, thus narrowing the gaps between physio-biochemical and molecular events in addressing HT tolerance.

Molecular Approach Coupled with Biochemical Attributes to Elucidate the Presence of DYMV in Leaf Samples of Lablab purpureus. L Genotypes.

A laboratory study was delineated to ascertain the impact and the extent of Dolichos yellow mosaic virus (DYMV) on biochemical constituents and various enzyme levels in the leaves of hyacinth bean. DYMV-infected leaves of all the genotypes used in the study revealed significant and consistent changes in activities of CAT, APX, PPO, DHAR, and MDHAR paralleled with a compelling hike in proline levels. Unlike that in non-infected leaves of the genotypes VRSEM-301 and VRSEM-749, VRSEM-894 and VRSEM-855, the enzyme level did not alter much which extended equally with increased phenolics, suggesting a well-coordinated generation of free radicals thereby suppressing oxidative stress in the latter. The genotypes were also evaluated at molecular level for elucidating the presence of the virus by using five sets of primer pairs. Two primers viz., DAC1 and DAC2 witnessed the presence of the virus in both non-infected and infected leaves. The difference in the appearance and/or disappearance of bands according to non-infected to infect reverberates the variation between genotypes in defense against infection.

HIV service delivery models towards 'Zero AIDS-related Deaths': a collaborative case study of 6 Asia and Pacific countries.

BACKGROUND: In the Asia-Pacific region, limited systematic assessment has been conducted on HIV service delivery models. Applying an analytical framework of the continuum of prevention and care, this study aimed to assess HIV service deliveries in six Asia and Pacific countries from the perspective of service availability, linking approaches and performance monitoring for maximizing HIV case detection and retention. METHODS: Each country formed a review team that provided published and unpublished information from the national HIV program. Four types of continuum were examined: (i) service linkages between key population outreach and HIV diagnosis (vertical-community continuum); (ii) chronic care provision across HIV diagnosis and treatment (chronological continuum); (iii) linkages between HIV and other health services (horizontal continuum); and (iv) comprehensive care sites coordinating care provision (hub and heart of continuum). RESULTS: Regarding the vertical-community continuum, all districts had voluntary counselling and testing (VCT) in all countries except for Myanmar and Vietnam. In these two countries, limited VCT availability was a constraint for referring key populations reached. All countries monitored HIV testing coverage among key populations. Concerning the chronological continuum, the proportion of districts/townships having antiretroviral treatment (ART) was less than 70% except in Thailand, posing a barrier for accessing pre-ART/ART care. Mechanisms for providing chronic care and monitoring retention were less developed for VCT/pre-ART process compared to ART process in all countries. On the horizontal continuum, the availability of HIV testing for tuberculosis patients and pregnant women was limited and there were sub-optimal linkages between tuberculosis, antenatal care and HIV services except for Cambodia and Thailand. These two countries indicated higher HIV testing coverage than other countries. Regarding hub and heart of continuum, all countries had comprehensive care sites with different degrees of community involvement. CONCLUSIONS: The analytical framework was useful to identify similarities and considerable variations in service availability and linking approaches across the countries. The study findings would help each country critically adapt and adopt global recommendations on HIV service decentralization, linkages and integration. Especially, the findings would inform cross-fertilization among the countries and national HIV program reviews to determine county-specific measures for maximizing HIV case detection and retention.

Risk of HIV infection among men having sex with men in Kathmandu Valley, Nepal.

Despite extensive distribution of free condoms and lubricants to prevent HIV transmission among men having sex with men (MSM) in Nepal, the prevalence of HIV and risky sexual behaviors remain high. The influence of individual-level, social-capital, and social-structural factors on HIV risk has been insufficiently explored in MSM. The authors assessed association of these factors with HIV risk among 150 MSM enrolled using snowball sampling in the Kathmandu Valley. HIV risk was calculated on the basis of number and type of sexual partners and condom use during anal sex. Multivariate analysis showed a high risk of HIV infection was significantly associated with being involved in sex work, having no knowledge of male sexually transmitted infection (STI) symptoms, and having a history of STI symptoms. HIV prevention could be made more effective by targeting MSM who are involved in sex work and by improving their knowledge of male STI symptoms and early diagnosis and treatment.

An unusual cause of status epilepticus.

A 24-year-old female presented with status epilepticus following ingestion of a pyrethroid insecticide Deltamethrin. The pathophysiology, clinical features, and management of pyrethroid poisoning are discussed in this article.