Evaluating the imazethapyr herbicide mediated regulation of phenol and glutathione metabolism and antioxidant activity in lentil seedlings.

The imidazolinone group of herbicides generally work for controlling weeds by limiting the synthesis of the aceto-hydroxy-acid enzyme, which is linked to the biosynthesis of branched-chain amino acids in plant cells. The herbicide imazethapyr is from the class and the active ingredient of this herbicide is the same as other herbicides Contour, Hammer, Overtop, Passport, Pivot, Pursuit, Pursuit Plus, and Resolve. It is commonly used for controlling weeds in soybeans, alfalfa hay, corn, rice, peanuts, etc. Generally, the herbicide imazethapyr is safe and non-toxic for target crops and environmentally friendly when it is used at low concentration levels. Even though crops are extremely susceptible to herbicide treatment at the seedling stage, there have been no observations of its higher dose on lentils (Lens culinaris Medik.) at that stage. The current study reports the consequence of imazethapyr treatment on phenolic acid and flavonoid contents along with the antioxidant activity of the phenolic extract. Imazethapyr treatment significantly increased the activities of several antioxidant enzymes, including phenylalanine ammonia lyase (PAL), phenol oxidase (POD), glutathione reductase (GR), and glutathione-s-transferase (GST), in lentil seedlings at doses of 0 RFD, 0.5 RFD, 1 RFD, 1.25 RFD, 1.5 RFD, and 2 RFD. Application of imazethapyr resulted in the 3.2 to 26.31 and 4.57-27.85% increase in mean phenolic acid and flavonoid content, respectively, over control. However, the consequent fold increase in mean antioxidant activity under 2, 2- diphenylpicrylhdrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assay system was in the range of 1.17-1.85 and 1.47-2.03%. Mean PAL and POD activities increased by 1.63 to 3.66 and 1.71 to 3.35-fold, respectively, in agreement with the rise in phenolic compounds, indicating that these enzyme's activities were modulated in response to herbicide treatment. Following herbicide treatments, the mean thiol content also increased significantly in corroboration with the enhancement in GR activity in a dose-dependent approach. A similar increase in GST activity was also observed with increasing herbicide dose.

MAPK signaling pathway orchestrates and fine-tunes the pathogenicity of Colletotrichum falcatum.

Colletotrichum falcatum is the causal organism of red rot, the most devastating disease of sugarcane. Mitogen-activated protein kinase (MAPK) signaling pathway plays pivotal role in coordinating the process of pathogenesis. We identified eighteen proteins implicated in MAPK signaling pathway in C. falcatum, through nanoLCMS/MS based proteomics approach. Twelve of these proteins were the part of core MAPK signaling pathway, whereas remaining proteins were indirectly implicated in MAPK signaling. Majority of these proteins had enhanced abundance in C. falcatum samples cultured with host sugarcane stalks. To validate the findings, core MAPK pathway genes (MAPKKK-NSY1, MAPK 17-MAPK17, MAPKKK 5-MAPKKK5, MAPK-HOG1B, MAPKKK-MCK1/STE11, MAPK-MST50/STE50, MAPKK-SEK1, MAPKK-MEK1/MST7/STE7, MAPKK-MKK2/STE7, MAPKKK-MST11/STE11, MAPK 5-MPK5, and MAPK-MPK-C) were analyzed by qPCR to confirm the real-time expression in C. falcatum samples cultured with host sugarcane stalks. The results of qPCR-based expression of genes were largely in agreement with the findings of proteomics. String association networks of MAPKK- MEK1/MST7/STE7, and MAPK- MPK-C revealed strong association with plenty of assorted proteins implicated in the process of pathogenesis/virulence. This is the novel and first large scale study of MAPK proteins in C. falcatum, responsible for red rot epidemics of sugarcane various countries. KEY MESSAGE: Our findings demonstrate the pivotal role of MAPK proteins in orchestrating the pathogenicity of Colletotrichum falcatum, responsible devastating red rot disease of sugarcane. SIGNIFICANCE: Our findings are novel and the first large scale study demonstrating the pivotal role of MAPK proteins in C. falcatum, responsible devastating red rot disease of sugarcane. The study will be useful for future researchers in terms of manipulating the fungal pathogenicity through genome editing.

Harnessing Rhizospheric Microbes for Eco-friendly and Sustainable Crop Production in Saline Environments.

Soil salinization is a global issue that negatively impacts crop yield and has become a prime concern for researchers worldwide. Many important crop plants are susceptible to salinity-induced stresses, including ionic and osmotic stress. Approximately, 20% of the world's cultivated and 33% of irrigated land is affected by salt. While various agricultural practices have been successful in alleviating salinity stress, they can be costly and not environment-friendly. Therefore, there is a need for cost-effective and eco-friendly practices to improve soil health. One promising approach involves utilizing microbes found in the vicinity of plant roots to mitigate the effects of salinity stress and enhance plant growth as well as crop yield. By exploiting the salinity tolerance of plants and their associated rhizospheric microorganisms, which have plant growth-promoting properties, it is possible to reduce the adverse effects of salt stress on crop plants. The soil salinization is a common problem in the world, due to which we are unable to use the saline land. To make proper use of this land for different crops, microorganisms can play an important role. Looking at the increasing population of the world, this will be an appreciated effort to make the best use of the wasted land for food security. The updated information on this issue is needed. In this context, this article provides a concise review of the latest research on the use of salt-tolerant rhizospheric microorganisms to mitigate salinity stress in crop plants.

Drought and salinity stresses induced physio-biochemical changes in sugarcane: an overview of tolerance mechanism and mitigating approaches.

Sugarcane productivity is being hampered globally under changing environmental scenarios like drought and salinity. The highly complex nature of the plant responses against these stresses is determined by a variety of factors such as genotype, developmental phase of the plant, progression rate and stress, intensity, and duration. These factors influence plant responses and can determine whether mitigation approaches associated with acclimation are implemented. In this review, we attempt to summarize the effects of drought and salinity on sugarcane growth, specifically on the plant's responses at various levels, viz., physiological, biochemical, and metabolic responses, to these stresses. Furthermore, mitigation strategies for dealing with these stresses have been discussed. Despite sugarcane's complex genomes, conventional breeding approaches can be utilized in conjunction with molecular breeding and omics technologies to develop drought- and salinity-tolerant cultivars. The significant role of plant growth-promoting bacteria in sustaining sugarcane productivity under drought and salinity cannot be overlooked.

Stable and reproducible expression of bacterial ipt gene under the control of SAM-specific promoter (pKNOX1) with interference of developmental patterns in transgenic Peperomia pellucida plants.

Reproducible and stable transgene expression is an important goal in plant basic research and applications. Hence, we report the first stable expression of bacterial transgenes in medicinal plant, Peperomia pellucida (L.) Kunth. Two key elements relevant to the dynamic expression of the bacterial cytokinin biosynthesis gene, ipt (isopentenyltransferase) were examined. First, by designing a specific expression cassette driven by a tissue-specific promoter for the required levels of gene expression in the particular function of development, and second by using P. pellucida as a model plant due to its short developmental cycle that supported expedient tracking of transgene expression in the progeny. Transgenic frequencies of ipt gene obtained from different expression cassettes of pKNOX1 for tissue-specific promoter in shoot apical meristem were compared with the cauliflower mosaic virus (CaMV35S) promoter (p35S), a constitutive promoter investigated for T3 generation. It was clearly shown that transgenic plants with pKNOX1 showed percentage of survivals in T3 at about 2.2 folds more than those of p35S-transgenic. Transgenic P. pellucida under controllable expression of pKNOX1 showed increased leaf and seed size with a high percentage of fertile seed, whereas transgenic plants with p35S showed phenotypic features of bushy and small leaves, sterile pollen and lower reproductive fitness. Quantitative examination of ipt-positive gene expression in T3 generation of transformants with pKNOX1 were 100% (line k-14) and 50% (line k-20), while 33.3% was observed in transgenic line c-11 with p35S. Interestingly, the endogenous cytokinin biosynthesis gene (ipt3) was significantly upregulated (2-3 folds higher) in pKNOX1-transformants. The overall relative mRNA expression of bacterial ipt gene and overproducing of cytokinin contents (t-ZR and 2-iP) detected in p35S-transformants caused abnormality and low percentages of transgene reproducible Interestingly, pKNOX1-transgenic plants tended to maintain chlorophyll contents 4-5 folds and extending the developmental cycle to 12.4 weeks (wk), which was 2 folds more than wildtype (5.8 wk) and p35S-transformants (7.4 wk). The promotor effect on stable and reproducible transgene-expressions demonstrated prominent features of P. pellucida and also empowered further omics studies.

A synthetic cytokinin influences the accumulation of leaf soluble sugars and sugar transporters, and enhances the drought adaptability in rice.

Oryza sativa cv. PTT1 (Pathumthani1) was treated with phenyl-urea-based synthetic cytokinin under drought stress. Soluble sugar contents were examined in rice flag leaves at tillering and grain-filling stages. The same leaf samples were used to analyze the differential abundance intensities of proteins related to metabolism and transport of soluble sugars, and the process of senescence. The results showed drought-induced accumulation of hexose sugars (glucose and fructose) in rice flag leaves, which could be corroborated with enhanced accumulation of MST8 under drought stress. On the other hand, cytokinin-treated plants maintained the normal contents of hexose sugar in their flag leaves under drought stress, alike well-watered plants. In the case of sucrose, cytokinin treatment reduced its accumulation at tillering stage, but the results were reversed at the grain-filling stage, where the cytokinin-treated plants maintained significantly higher contents of sucrose under drought stress. Growth stage dependent variations in sucrose contents corroborated with the accumulation of SPS (SPS1, SPS2, and SPS5) proteins, implicated in sucrose biosynthesis. In our study, among the proteins involved in sucrose transport, SUT1 transporter was induced by drought stress at both the growth stages, whereas SUT2 transporter accumulated equally in all the treatments. However, cytokinin treatment reversed the effect of drought on the accumulation of SUT1. Similarly, SWEET5, and SWEET13 proteins, which were induced by drought stress treatment, were inhibited by cytokinin treatment. However, the accumulation SWEET6, SWEET7, and SWEET15 was not influenced by the treatment of cytokinin in the flag leaves of rice. In addition, cytokinin treatment reduced the leaf wilting, enhanced the fresh weight and grain yield, and curtailed the accumulation of proteins involved in drought-induced senescence. In conclusion, the cytokinin treatment had a positive agro-economic impact on the rice plants and provided better drought adaptability.

A Synthetic Cytokinin Improves Photosynthesis in Rice under Drought Stress by Modulating the Abundance of Proteins Related to Stomatal Conductance, Chlorophyll Contents, and Rubisco Activity.

Drought susceptible rice cultivar PTT1 (Pathumthani1) was treated with drought (-72 kPa) and CPPU (N-2-(chloro-4-pyridyl)-N-phenyl urea) @ 5 mg/L at tillering and grain-filling stages. Plants were tested for the effect of synthetic cytokinin on the parameters influencing the process of photosynthesis. Exogenous spray of CPPU improved the stomatal conductance of rice leaves, which was severely reduced by drought. The abundance intensities of proteins, associated with the stomatal conductance (ZEP, NCED4, PYL9, PYL10, ABI5, SnRK4, Phot1, and Phot2), were also in agreement with the positive impact of CPPU on the stomatal conductance under drought stress. Among the photosynthetic pigments, Chl b contents were significantly reduced by drought stress, whereas CPPU treated plants retained the normal contents of Chl b under drought stress. Subsequently, we examined the abundance intensities of chlorophyll synthase and HCR proteins, implicated in the biosynthesis of chlorophyll pigments and the conversion of Chl b to Chl a, respectively. The results indicated a drought-mediated suppression of chlorophyll synthase. However, CPPU treated plants retained normal levels of chlorophyll synthase under drought stress. In addition, drought stress induced HCR proteins, which might be the cause for reduced Chl b contents in drought stressed plants. Further, CPPU treatment helped the plants sustain photosynthesis at a normal rate under drought stress, which was comparable with well-watered plants. The results were further confirmed by examining the abundance intensities of two key proteins, RAF1 and Rubisco activase, implicated in the assembly and activation of Rubisco, respectively. CPPU treatment reversed the drought mediated suppression of these proteins at both of the growth stages of rice under drought stress. Based on the results, it can be suggested that synthetic cytokinins help the plants sustain photosynthesis at a normal rate under drought stress by positively influencing the determinants of photosynthesis at a molecular level.

The Mode of Cytokinin Functions Assisting Plant Adaptations to Osmotic Stresses.

Plants respond to abiotic stresses by activating a specific genetic program that supports survival by developing robust adaptive mechanisms. This leads to accelerated senescence and reduced growth, resulting in negative agro-economic impacts on crop productivity. Cytokinins (CKs) customarily regulate various biological processes in plants, including growth and development. In recent years, cytokinins have been implicated in adaptations to osmotic stresses with improved plant growth and yield. Endogenous CK content under osmotic stresses can be enhanced either by transforming plants with a bacterial isopentenyl transferase (IPT) gene under the control of a stress inducible promoter or by exogenous application of synthetic CKs. CKs counteract osmotic stress-induced premature senescence by redistributing soluble sugars and inhibiting the expression of senescence-associated genes. Elevated CK contents under osmotic stress antagonize abscisic acid (ABA) signaling and ABA mediated responses, delay leaf senescence, reduce reactive oxygen species (ROS) damage and lipid peroxidation, improve plant growth, and ameliorate osmotic stress adaptability in plants.

Protein modeling and molecular dynamics simulation of SlWRKY4 protein cloned from drought tolerant tomato (Solanum habrochaites) line EC520061.

WRKY genes are members of one of the largest families of plant transcription factors and play an important role in response to biotic and abiotic stresses, and overall growth and development. Understanding the interaction of WRKY proteins with other proteins/ligands in plant cells is of utmost importance to develop plants having tolerance to biotic and abiotic stresses. The SlWRKY4 gene was cloned from a drought tolerant wild species of tomato (Solanum habrochaites) and the secondary structure and 3D modeling of this protein were predicted using Schrödinger Suite-Prime. Predicted structures were also subjected to plot against Ramachandran's conformation, and the modeled structure was minimized using Macromodel. Finally, the minimized structure was simulated in the water environment to check the protein stability. The behavior of the modeled structure was well-simulated and analyzed through RMSD and RMSF of the protein. The present work provides the modeled 3D structure of SlWRKY4 that will help in understanding the mechanism of gene regulation by further in silico interaction studies.