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    Isolation of cytoskeleton and cytoskeleton-bound polysome, and identification of cytoskeletal proteins from a grain legume
    (Springer Nature Publishing AG, 2026) Kumar, Sunil; Chakraborty, Sohela; Chakraborty, Subhra; Chakraborty, Niranjan
    The plant cytoskeleton is an essential component of cellular architecture, enabling various critical metabolic processes, including cell division, differentiation, expansion and intracellular transport. It consists primarily of three distinct filamentous structures: microtubules, microfilaments and intermediate filaments. These structures are not static; they undergo continuous remodeling in response to environmental signals and developmental cues, which allow plants to adapt to changing conditions. The microtubules and actin filaments have previously been successfully isolated from various plant tissues, contributing to our understanding of their functions. Among the diverse plant families, legumes (Fabaceae) stand out as the third largest, encompassing approximately 20,000 species. They hold significant agricultural importance, ranking second to cereals in global crop production. To fully grasp the developmental and adaptive processes in legumes, it is essential to identify and understand their regulatory components. This chapter focuses on the isolation of cytoskeletal proteins from chickpea, a prominent grain legume, facilitating biochemical and proteomic analyses that may uncover new insights into the functioning of the cytoskeleton in legumes.
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    Identification of tRNA-derived fragments in legumes
    (Springer Nature Publishing AG, 2026) Arora, Simran; Aftab, Sahrish; Shree, Tanu; Kumar, Shailesh
    The tRNA-derived noncoding RNAs (tncRNAs) belong to the novel class of noncoding RNAs, acting as important components of genome regulatory circuits. In planta, the mechanism of generation and function of tncRNAs is not fully elucidated. Production of important leguminous plants like chickpea, Medicago and soybean is majorly hampered due to different biotic and abiotic stresses. Identification and characterization of tncRNAs in legumes may open a new paradigm for molecular biologists to make novel tools for the improved varieties of legumes for sustainable agriculture. The first step in the study of tncRNAs is to identify and annotate them in small RNA sequencing datasets. Here, we have demonstrated the tncRNA Toolkit for the identification and annotation of tncRNAs in a small RNA sequencing dataset of the important legume crop chickpea.
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    Genome editing in legumes: Current status
    (Springer Nature, 2025) Sharma, Aman; Chattopadhya, Debasis
    Legume crops are indispensable for global food security due to their high protein content and phytonutraceutical properties. Despite their importance, legume crops suffer from poor crop yields due to limited areas of cultivation, various biotic and abiotic stress factors, as well as poor agriculture infrastructure. The increasing global population as well as aggravating climate change has further worsened the situation. The recent revolution in genome sequencing and annotation has opened the gateway for legume crop improvement through advanced molecular breeding tools such as genomic selection, genetic engineering, and genome editing. Gene editing by CRISPR-Cas9 and its derivative systems is a powerful tool to make specific genetic changes at precise positions in the genome facilitating rapid crop improvement through precision breeding. In this chapter, an overview of the basic machinery of CRISPR-Cas9 and allied gene editing systems is provided followed by a comprehensive outline of the execution of a basic CRISPR-Cas9 gene editing experiment including various tools and techniques for construct designing and result analysis. The application of CRISPR-Cas9-mediated gene editing in several legume species is also discussed. We have also discussed the major constraints and bottlenecks faced during the practical implementation of genome editing techniques in legume crops.