Comparison of biochemical, anatomical, morphological, and physiological responses to salinity stress in wheat and barley genotypes deferring in salinity tolerance

A greenhouse hydroponic experiment was performed using salt-tolerant (cv. Suntop) and -sensitive (Sunmate) wheat cultivars and a salt-tolerant barley cv. CM72 to evaluate how cultivar and species differ in response to salinity stress. Results showed that wheat cv. Suntop performed high tolerance to salinity, being similar tolerance to salinity with CM72, compared with cv. Sunmate. Similar to CM72, Suntop recorded less salinity induced increase in malondialdehyde (MDA) accumulation and less reduction in plant height, net photosynthetic rate (Pn), chlorophyll content, and biomass than in sensitive wheat cv. Sunmate. Significant time-course and cultivar-dependent changes were observed in the activities of antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR) in roots and leaves after salinity treatment. Higher activities were found in CM72 and Suntop compared to Sunmate. Furthermore, a clear modification was observed in leaf and root ultrastructure after NaCl treatment with more obvious changes in the sensitive wheat cv. Sunmate, rather than in CM72 and Suntop. Although differences were observed between CM72 and Suntop in the growth and biochemical traits assessed and modified by salt stress, the differences were negligible in comparison with the general response to the salt stress of sensitive wheat cv. Sunmate. In addition, salinity stress induced an increase in the Na+ and Na+/K+ ratio but a reduction in K+ concentrations, most prominently in Sunmate and followed by Suntop and CM72

Mechanistic insights into potassium‐conferred drought stress tolerance in cultivated and Tibetan wild barley : differential osmoregulation, nutrient retention, secondary metabolism and antioxidative defense capacity

Keeping the significance of potassium (K) nutrition in focus, this study explores the genotypic responses of two wild Tibetan barley genotypes (drought tolerant XZ5 and drought sensitive XZ54) and one drought tolerant barley cv. Tadmor, under the exposure of polyethylene glycol‐induced drought stress. The results revealed that drought and K deprivation attenuated overall plant growth in all the tested genotypes; however, XZ5 was least affected due to its ability to retain K in its tissues which could be attributed to the smallest reductions of photosynthetic parameters, relative chlorophyll contents and the lowest Na+/K+ ratios in all treatments. Our results also indicate that higher H+/K+‐ATPase activity (enhancement of 1.6 and 1.3‐fold for shoot; 1.4 and 2.5‐fold for root), higher shoot K+ (2 and 2.3‐fold) and Ca2+ content (1.5 and 1.7‐fold), better maintenance of turgor pressure by osmolyte accumulation and enhanced antioxidative performance to scavenge ROS, ultimately suppress lipid peroxidation (in shoots: 4% and 35%; in roots 4% and 20% less) and bestow higher tolerance to XZ5 against drought stress in comparison with Tadmor and XZ54, respectively. Conclusively, this study adds further evidence to support the concept that Tibetan wild barley genotypes that utilize K efficiently could serve as a valuable genetic resource for the provision of genes for improved K metabolism in addition to those for combating drought stress, thereby enabling the development of elite barley lines better tolerant of abiotic stresses

Stress signaling convergence and nutrient crosstalk determine zinc-mediated amelioration against cadmium toxicity in rice

Consumption of rice (Oryza sativa L.) is one of the major pathways for heavy metal bioaccumulation in humans over time. Understanding the molecular responses of rice to heavy metal contamination in agriculture is useful for eco-toxicological assessment of cadmium (Cd) and its interaction with zinc (Zn). In certain crops, the impacts of Cd stress or Zn nutrition on the biophysical chemistry and gene expression have been widely investigated, but their molecular interactions at transcriptomic level, particularly in rice roots, are still elusive. Here, hydroponic investigations were carried out with two rice genotypes (Yinni-801 and Heizhan-43), varying in Cd contents in plant tissues to determine their transcriptomic responses upon Cd15 (15 µM) and Cd15+Zn50 (50 µM) treatments. High throughput RNA-sequencing analysis confirmed that 496 and 2407 DEGs were significantly affected by Cd15 and Cd15+Zn50, respectively, among which 1016 DEGs were commonly induced in both genotypes. Multitude of DEGs fell under the category of protein kinases, such as calmodulin (CaM) and calcineurin B-like protein-interacting protein kinases (CBL), indicating a dynamic shift in hormonal signal transduction and Ca2+ involvement with the onset of treatments. Both genotypes expressed a mutual regulation of transcription factors (TFs) such as WRKY, MYB, NAM, AP2, bHLH and ZFP families under both treatments, whereas genes econding ABC transporters (ABCs), high affinity K+ transporters (HAKs) and Glutathione-S-transferases (GSTs), were highly up-regulated under Cd15+Zn50 in both genotypes. Zinc addition triggered more signaling cascades and detoxification related genes in regulation of immunity along with the suppression of Cd-induced DEGs and restriction of Cd uptake. Conclusively, the effective integration of breeding techniques with candidate genes identified in this study as well as economically and technologically viable methods, such as Zn nutrient management, could pave the way for selecting cultivars with promising agronomic qualities and reduced Cd for sustainable rice production

Spatio-Temporal Variation in the Phyllospheric Microbial Biodiversity of Alternaria Alternata-Infected Tobacco Foliage

Phyllospheric microbial composition of tobacco (Nicotiana tabacum L.) is contingent upon certain factors, such as the growth stage of the plant, leaf position, and cultivar and its geographical location, which influence, either directly or indirectly, the growth, overall health, and production of the tobacco plant. To better understand the spatiotemporal variation of the community and the divergence of phyllospheric microflora, procured from healthy and diseased tobacco leaves infected by Alternaria alternata, the current study employed microbe culturing, high-throughput technique, and BIOLOG ECO. Microbe culturing resulted in the isolation of 153 culturable fungal isolates belonging to 33 genera and 99 bacterial isolates belonging to 15 genera. High-throughput sequencing revealed that the phyllosphere of tobacco was dominantly colonized by Ascomycota and Proteobacteria, whereas, the most abundant fungal and bacterial genera were Alternaria and Pseudomonas. The relative abundance of Alternaria increased in the upper and middle healthy groups from the first collection time to the third, whereas, the relative abundance of Pseudomonas, Sphingomonas, and Methylobacterium from the same positions increased during gradual leaf aging. Non-metric multi-dimensional scaling (NMDs) showed clustering of fungal communities in healthy samples, while bacterial communities of all diseased and healthy groups were found scattered. FUNGuild analysis, from the first collection stage to the third one in both groups, indicated an increase in the relative abundance of Pathotroph-Saprotroph, Pathotroph-Saprotroph-Symbiotroph, and Pathotroph-Symbiotroph. Inclusive of all samples, as per the PICRUSt analysis, the predominant pathway was metabolism function accounting for 50.03%. The average values of omnilog units (OUs) showed relatively higher utilization rates of carbon sources by the microbial flora of healthy leaves. According to the analysis of genus abundances, leaf growth and leaf position were the important drivers of change in structuring the microbial communities. The current findings revealed the complex ecological dynamics that occur in the phyllospheric microbial communities over the course of a spatiotemporal varying environment with the development of tobacco brown spots, highlighting the importance of community succession

Resemblance and Difference of Seedling Metabolic and Transporter Gene Expression in High Tolerance Wheat and Barley Cultivars in Response to Salinity Stress

To elucidate inter-specific similarity and difference of tolerance mechanism against salinity stress between wheat and barley, high tolerant wheat cv. Suntop and sensitive cv. Sunmate and tolerant barley cv. CM72 were hydroponically grown in a greenhouse with 100 mM NaCl. Glutathione, secondary metabolites, and genes associated with Na+ transport, defense, and detoxification were examined to discriminate the species/cultivar difference in response to salinity stress. Suntop and CM72 displayed damage to a lesser extent than in Sunmate. Compared to Sunmate, both Suntop and CM72 recorded lower electrolyte leakage and reactive oxygen species (ROS) production, higher leaf relative water content, and higher activity of PAL (phenylalanine ammonia-lyase), CAD (cinnamyl alcohol dehydrogenase), PPO (polyphenol oxidase), SKDH (shikimate dehydrogenase), and more abundance of their mRNA under salinity stress. The expression of HKT1, HKT2, salt overly sensitive (SOS)1, AKT1, and NHX1 was upregulated in CM72 and Suntop, while downregulated in Sunmate. The transcription factor WRKY 10 was significantly induced in Suntop but suppressed in CM72 and Sunmate. Higher oxidized glutathione (GSSG) content was accumulated in cv. CM72 and Sunmate, but increased glutathione (GSH) content and the ratio of GSH/GSSG were observed in leaves and roots of Suntop under salinity stress. In conclusion, glutathione homeostasis and upregulation of the TaWRKY10 transcription factor played a more important role in wheat salt-tolerant cv. Suntop, which was different from barley cv. CM72 tolerance to salinity stress. This new finding could help in developing salinity tolerance in wheat and barley cultivars

Pan-transcriptomic Profiling Demarcates Serendipita Indica-Phosphorus Mediated Tolerance Mechanisms in Rice Exposed to Arsenic Toxicity

Abstract Inadvertent accumulation of arsenic (As) in rice (Oryza sativa L.) is a concern for people depending on it for their subsistence, as it verily causes epigenetic alterations across the genome as well as in specific cells. To ensure food safety, certain attempts have been made to nullify this highest health hazard encompassing physiological, chemical and biological methods. Albeit, the use of mycorrhizal association along with nutrient reinforcement strategy has not been explored yet. Mechanisms of response and resistance of two rice genotypes to As with or without phosphorus (P) nutrition and Serendipita indica (S. indica; S.i) colonization were explored by root transcriptome profiling in the present study. Results revealed that the resistant genotype had higher auxin content and root plasticity, which helped in keeping the As accumulation and P starvation response to a minimum under alone As stress. However, sufficient P supply and symbiotic relationship switched the energy resources towards plant’s developmental aspects rather than excessive root proliferation. Higher As accumulating genotype (GD-6) displayed upregulation of ethylene signaling/biosynthesis, root stunting and senescence related genes under As toxicity. Antioxidant defense system and cytokinin biosynthesis/signaling of both genotypes were strengthened under As + S.i + P, while the upregulation of potassium (K) and zinc (Zn) transporters depicted underlying cross-talk with iron (Fe) and P. Differential expression of phosphate transporters, peroxidases and GSTs, metal detoxification/transport proteins, as well as phytohormonal metabolism were responsible for As detoxification. Taken together, S. indica symbiosis fortified with adequate P-fertilizer can prove to be effective in minimizing As acquisition and accumulation in rice plants

Physio-ultrastructural footprints and iTRAQ-based proteomic approach unravel the role of Piriformospora indica-colonization in counteracting cadmium toxicity in rice

Due to its immense capability to concentrate in rice grain and ultimately in food chain, cadmium (Cd) has become the cause of an elevated concern among agriculturists, scientists and the environmental activists. Symbiotic association of Piriformospora indica (P. indica) has been characterized as a potential aid in combating heavy metal stress in plants for sustainable crop production but our scant knowledge regarding ameliorative tendency of P. indica against Cd, specifically in rice, necessitates an in-depth investigation. This study aimed at elaborating the underlying mechanisms involved in P. indica-mediated tolerance against Cd stress in two rice genotypes, IR8 and ZX1H, varying in Cd accumulation pattern. Either colonized or un-inoculated with P. indica, seedlings of both genotypes were subjected to Cd stress. The results showed that P. indica colonization significantly supported plant biomass, photosynthetic attributes and chlorophyll contents in Cd stressed plants. P. indica colonization sustained chloroplast integrity and reduced Cd translocation (46% and 64%), significantly lowering malondialdehyde (MDA) content (11.3% and 50.4%) compared to uninoculated roots under Cd stress in IR8 and ZX1H, respectively. A genotypic difference was evident when a 2-fold enhancement in root peroxidase (POD) activity was recorded in P. indica colonized IR8 plants as compared to ZX1H. The root proteomic analysis was performed using isobaric tags for relative and absolute quantification (iTRAQ) and the results showed that P. indica alleviates Cd stress in rice via down-regulation of key glycolysis cycle enzymes in a bid to reduce energy consumption by the plants and possibly re-directing it to Cd defense response pathways; and up-regulation of glutamine synthetase, a key enzyme in the L-Arg-dependent pathway for nitric oxide (NO) production, which acts as a stress signaling molecule, thus conferring tolerance by reduction of NO-mediated modification of essential proteins in response to Cd stress. Conclusively, both the tested genotypes benefited from P. indica symbiosis at varying levels by an enhanced detoxification capacity and signaling efficiency in response to stress. Hence, a step forward towards the employment of an environmentally sound and self-renewing approach holding the hope for a healthy future

Cadmium-zinc cross-talk delineates toxicity tolerance in rice via differential genes expression and physiological / ultrastructural adjustments

Understanding the physiological and molecular response of crop genotypes could be useful in eco-toxicological evaluation with cadmium (Cd) and could be a strategy to solve heavy metal contamination in agriculture. This study corroborates unique patterns of Cd accumulation and molecular mechanisms adopted by plants to acquire Cd tolerance and counteractive effects of zinc (Zn) against Cd toxicity. Two rice (Oryza sativa) genotypes (Heizhan 43 and Yinni 801) differing in cadmium tolerance and its accumulation in plant tissues were investigated hydroponically using two Cd levels [Cd10 (10 μM L−1) and Cd15 (15 μM L−1)] and two Zn levels [Zn25 (25 μM L−2) and Zn50 (50 μM L−1)] and their combinations. Cadmium toxicity rendered substantial reduction in plant height, biomass, chlorophyll contents and photosynthesis as compared to the control plants after 15 days of treatment. Supplementation of Zn evidently ameliorated Cd toxicity by minimizing the reduction in plant growth, chlorophyll contents and photosynthetic attributes (Pn, gs, Ci, and Tr). Comparatively, lower accumulation of Cd in Yinni 801 under combined treatments revealed a preferential uptake of Zn in this genotype. A cross-talk among Cd, Zn, Fe, Ca and K correlated with fluctuating gs, Ci and Tr. Both genotypes also differed in morphological alterations of cell membrane, chloroplasts and appearance of enlarged plastoglobuli along with distorted mitochondria. An increased ascorbate peroxidase activity in roots of Yinni 801 presented a defensive strategy. Relative expression of Cd and Zn ion transporter genes also confirmed the genotypic background of phenotypic divergence. The OsLCT1 and OsHMA2 expression was significant in Heizhan 43, indicating possible translocation of Cd from shoot to grains contrary to Yinni 801, which accumulated Cd in shoot and showed stunted growth. Zn supplementation promises tolerance to Cd in Yinni 801 by differential expression of putative genes for Cd translocation with minimum ultrastructural modifications by maintaining physiological functions in contrast to Heizhan 43

Zinc alleviates cadmium toxicity by modulating photosynthesis, ROS homeostasis, and cation flux kinetics in rice

Understanding of cadmium (Cd) uptake mechanism and development of lower Cd crop genotypes are crucial for combating its phytotoxicity and meeting 70% increase in food demand by 2050. Bio-accumulation of Cd continuously challenges quality of life specifically in regions without adequate environmental planning. Here, we investigated the mechanisms operating in Cd tolerance of two rice genotypes (Heizhan-43 and Yinni-801). Damage to chlorophyll contents and PSII, histochemical staining and quantification of reactive oxygen species (ROS), cell viability and osmolyte accumulation were studied to decipher the interactions between Cd and zinc (Zn) by applying two Cd and two Zn levels (alone as well as combined). Cd2+ and Ca2+ fluxes were also measured by employing sole Cd100 (100 μmol L−1) and Zn50 (50 μmol L−1), and their combination with microelectrode ion flux estimation (MIFE) technique. Cd toxicity substantially reduced chlorophyll contents and maximal photochemical efficiency (Fv/Fm) compared to control plants. Zn supplementation reverted the Cd-induced toxicity by augmenting osmoprotectants and interfering with ROS homeostasis under combined treatments, particularly in Yinni-801 genotype. Fluorescence microscopy indicated a unique pattern of live and dead root cells, depicting more damage with Cd10, Cd15 and Cd15+Zn50. Our results confer that Cd2+ impairs the uptake of Ca2+ whereas, Zn not only competes with Cd2+ but also Ca2+, thereby modifying ion homeostasis in rice plants. This study suggests that exogenous application of Zn is beneficial for rice plants in ameliorating Cd toxicity in a genotype and dose dependent manner by minimizing ROS generation and suppressing collective oxidative damage. The observations confer that Yinni-801 performed better than Heizhan-43 genotype mainly under combined Zn treatments with low-Cd, presenting Zn fortification as a solution to increase rice production

Phosphorus and Serendipita indica synergism augments arsenic stress tolerance in rice by regulating secondary metabolism related enzymatic activity and root metabolic patterns

The multifarious problems created by arsenic (As), for collective environment and human health, serve a cogent case for searching integrative agricultural approaches to attain food security. Rice (Oryza sativa L.) acts as a sponge for heavy metal(loid)s accretion, specifically As, due to anaerobic flooded growth conditions facilitating its uptake. Acclaimed for their positive impact on plant growth, development and phosphorus (P) nutrition, ‘mycorrhizas’ are able to promote stress tolerance. Albeit, the metabolic alterations underlying Serendipita indica (S. indica; S.i) symbiosis-mediated amelioration of As stress along with nutritional management of P are still understudied. By using biochemical, RT-qPCR and LC-MS/MS based untargeted metabolomics approach, rice roots of ZZY-1 and GD-6 colonized by S. indica, which were later treated with As (10 µM) and P (50 µM), were compared with non-colonized roots under the same treatments with a set of control plants. The responses of secondary metabolism related enzymes, especially polyphenol oxidase (PPO) activities in the foliage of ZZY-1 and GD-6 were enhanced 8.5 and 12-fold, respectively, compared to their respective control counterparts. The current study identified 360 cationic and 287 anionic metabolites in rice roots, and the commonly enriched pathway annotated by Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis was biosynthesis of phenylalanine, tyrosine and tryptophan, which validated the results of biochemical and gene expression analyses associated with secondary metabolic enzymes. Particularly under As+S.i+P comparison, both genotypes exhibited an upregulation of key detoxification and defense related metabolites, including fumaric acid, L-malic acid, choline, 3,4-dihydroxybenzoic acid, to name a few. The results of this study provided the novel insights into the promising role of exogenous P and S. indica in alleviating As stress