Publications of NIPGR Scientists

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Author: search.filters.author.Senthil-Kumar, MuthappaHas files: No

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    Novel method for rapid screening of chickpea for combined dry root rot disease and osmotic stress
    (Springer Nature Publishing AG, 2026) Ranjan, Shubhashish; Chavan, Chaitali Narendra; Senthil-Kumar, Muthappa
    Chickpea (Cicer arietinum L.), confronts substantial challenges from the emerging pathogenic fungus Macrophomina phaseolina (Tassi) Goid, causing dry root rot (DRR) disease. Chickpea plants severely affected by combined DRR and drought stress. Currently sick plot and sick pot method are utilized for germplasm screening to identify tolerant genotypes. These methods are time-consuming; therefore, we propose a novel methodology for the rapid screening of chickpea under combined DRR and osmotic stress conditions. This chapter introduces an adept high-throughput phenotyping methodology, conducted within controlled laboratory conditions, aiming to investigate the interaction between osmotic stress and DRR disease in chickpea crops. The methodology employs an innovative pouch technique for screening combined stress, providing a streamlined temporal investigation process and precise control over stress parameters. The incorporation of polyethylene glycol (PEG) enables the simultaneous imposition of osmotic stress alongside pathogen infection, making the methodology versatile for studying combined stress scenarios. This approach fills a gap in concurrent stress imposition techniques, enhancing germplasm screening by identifying genotypes with varying susceptibility and resistance levels. Thus, we suggest use of high-throughput phenotyping in combination genome-wide association study (GWAS) can take combined stress resistance breeding in chickpea at next level to combat food security and climate change.
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    A blotting paper technique for the screening of chickpea genotypes against dry root rot disease
    (Springer Nature Publishing AG, 2026) Durgadevi, Athimoolam; Pandey, Prachi; Senthil-Kumar, Muthappa
    Dry root rot (DRR) disease is a major threat to chickpea production across the world. This disease is caused by a soil-borne necrotrophic fungal pathogen, Macrophomina phaseolina. The use of disease-resistant varieties paves the way to conquer the disease spread. Though chickpea germplasm with rich genetic diversity is available around the world, its response to DRR is still unexplored. In turn, this demands screening and identification of resistant genotypes for crop protection against the disease. Here we describe an improved blotting paper technique for the large-scale screening of chickpea genotypes for DRR resistance. The method is quick, cost-effective, less labour-intensive, and thus optimized for high-throughput screening and can be efficiently used to screen a large number of chickpea genotypes for resistance against DRR.
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    TAIL-PCR for the recovery of Tnt1 flanking sequences in chickpea: a tool for functional genomics studies
    (Springer Nature Publishing AG, 2026) Chauhan, Chetan; Ranjan, Shubhashish; Jangid, Vinod Kumar; Sinharoy, Senjuti; Senthil-Kumar, Muthappa
    Thermal asymmetric interlaced polymerase chain reaction (TAIL-PCR) is a powerful technique for amplifying genomic regions flanking Tnt1-retortransposon insertions in plants. Here, we present a TAIL-PCR protocol for amplifying Tnt1-flanking genomic sequences in chickpea using Tnt1-transformed hairy roots as the starting material. The amplified products can be cloned and sequenced for the precise mapping of Tnt1-integration sites in the chickpea genome. This method enables the functional characterization of chickpea genes governing root-specific traits and can be easily adapted for flanking sequence tag recovery in chickpea Tnt1-mutant populations.
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    A method for isolation and transfection of arabidopsis protoplast for sucrose feeding assay
    (Springer Nature Publishing AG, 2026) Anjali, Anjali; Senthil-Kumar, Muthappa
    Protoplasts serve as a powerful system to study various plant physiological processes and to understand crucial signaling pathways within the plant system. The isolation of Arabidopsis protoplasts is a well-established technique and being utilized for wide range of assays. The method described herein includes the precise cutting of Arabidopsis leaves using extraction buffer containing cellulase and macerozyme. Further we present a polyethylene glycol [PEG]-mediated protoplast transformation method. Here, we have elucidated the methodology for isolation of protoplast for the AtSWEET-mediated sucrose uptake assay in control, and Pseudomonas syringe pv tomato DC3000 (Pst DC3000) treated leaves by utilizing GC/MS analysis. Our approach includes certain modifications to the previously published protoplast isolation technique, streamlining the process and providing a more accessible alternative to the highly specialized Xenopus oocyte uptake assay.
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    Plant biotic stress: tools and techniques for crop protection
    (Springer Nature Publishing AG, 2026) Acharya, Krishna Ramakrishnan; Chilakala, Aswin Reddy; Senthil-Kumar, Muthappa
    As climate change continues to impact crop yields, developing strategies to enhance plant tolerance to biotic stress has become increasingly important. This requires a thorough evaluation of the tools and methodologies used to manipulate and study biotic stress tolerance. It is crucial to comprehensively understand both conventional and modern techniques, as well as their effectiveness in addressing the specific needs of the crop under study. Detecting diseases at the early stages of plant development can prevent significant losses in large-scale cultivations. Two broad approaches commonly used to mitigate biotic stresses are eliminating causative agents such as fungi, bacteria, nematodes, viruses, or pests, and imparting resistance to the plant. Although there are similarities in the tools and techniques used to address different biotic stresses, each scenario requires dedicated case studies. It is also essential to stay up to date with the latest developments in plant biotechnology to incorporate a cross-disciplinary approach in conducting and validating experiments. This chapter provides an overview of methods covered in this book ranging from molecular breeding to nondestructive techniques that help achieve the goal of safeguarding plant health.
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    Extracellular acidification assay to evaluate the effectiveness of antifungal agents on the pathogenicity of Macrophomina phaseolina
    (Springer Nature Publishing AG, 2026) Acharya, Krishna Ramakrishnan; Chilakala, Aswin Reddy; Senthil-Kumar, Muthappa
    Macrophomina phaseolina is a fungus that causes dry root rot disease and considerable yield loss worldwide. Fungi exhibit various ways of absorbing nutrients through their plasma membrane, such as free or facilitated diffusion, diffusion channels, or active transport. Glucose, as a preferred carbon source, activates the plasma membrane H+-ATPase, resulting in the release of protons. Consequently, the protons, along with the organic acid metabolites released into the extracellular environment, acidify the cell surroundings. This decrease in pH cues the fungus to shift from saprotrophic to necrotrophic growth, facilitating host invasion. Sustainable dry root rot disease management often relies on the employment of antifungal agents from various biological sources. Despite the discovery of numerous antifungal agents, only a limited number have been evaluated for their efficacy against this phytopathogenic fungus. This scarcity of testing is primarily due to the limitations of existing methods, which often lack standardisation and reproducibility. This chapter introduces a rapid and sensitive method to assess the antifungal activity of various agents against M. phaseolina. By measuring extracellular pH changes after treatment in the presence of a nutrient source, we can determine the inhibitory concentrations of these agents and evaluate their potential for controlling fungal pathogenicity in plants.
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    Recent advances in plant gene silencing methods
    (Springer Nature Publishing AG, 2022) Pandey, Prachi; Mysore, Kirankumar S.; Senthil-Kumar, Muthappa
    With the increasing understanding of fundamentals of gene silencing pathways in plants, various tools and techniques for downregulating the expression of a target gene have been developed across multiple plant species. This chapter provides an insight into the molecular mechanisms of gene silencing and highlights the advancements in various gene silencing approaches. The prominent aspects of different gene silencing methods, their advantages and disadvantages have been discussed. A succinct discussion on the newly emerged microRNA-based technologies like microRNA-induced gene silencing (MIGS) and microRNA-mediated virus-induced gene silencing (MIR-VIGS) are also presented. We have also discussed the gene-editing system like CRISPR-Cas. The prominent bottlenecks in gene silencing methods are the off-target effects and lack of universal applicability. However, the tremendous growth in understanding of this field reflects the potentials for improvements in the currently available approaches and the development of new widely applicable methods for easy, fast, and efficient functional characterization of plant genes.
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    High-throughput analysis of gene function under multiple abiotic stresses using leaf disks from silenced plants
    (Springer Nature Publishing AG, 2022) Yamunarani, Ramegowda; Ramegowda, Venkategowda; Senthil-Kumar, Muthappa; Mysore, Kirankumar S.
    The high throughputness and affordability of “omics” technologies is leading to the identification of a large number of abiotic stress genes, with many of them responsive to multiple stresses. In vivo functional characterization of these genes under multiple stresses is challenging but essential to develop resilient crops for the changing climate. Here we describe a high-throughput Virus-Induced Gene Silencing-based methodology for functional analysis of genes under multiple abiotic stresses using leaf disks. Leaves with maximal silencing, which is localized to only a few leaves and to a short period, can be effectively used for multiple stress imposition and stress affect quantification.