Browsing by Author "Mandlik, Rushil"

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A CRISPR-Cas9 library to target putative redundant gene sets facilitates their functional exploration in grain development in rice
(Springer Nature Publishing AG, 2025) Yadav, Banita; Sardar, Shaswati; Yadav, Anil; Kumari, Annapurna; Gautam, Mohini; Mandlik, Rushil; Arora, Simran; Kumar, Shailesh; Jewaria, Pawan Kumar; Sonah, Humira; Deshmukh, Rupesh; Chinnusamy, Viswanathan; Ram, Hasthi
Advent of CRISPR-Cas9 library approach has revolutionized the field of high throughput targeted mutagenesis in plants. By identifying an sgRNA spacer that can target multiple paralogous genes in a genome, higher-order knockout plants can be developed. Using this concept, we developed ten CRISPR-Cas9 pool libraries and generated higher-order knockout plants in rice. Towards this, firstly we identified genome-wide sets of genes which are co-expressed and have high sequence similarity and can be targeted by a single sgRNA. Based on the expression pattern, these genes were divided into ten groups, and subsequently ten CRISPR-Cas9 plasmid libraries were developed. One such library designed against seed-expressed genes was transformed into rice and higher-order knockout plants were developed. Genotyping revealed that around 90% T0 plants had editing, and among the edited plants majority of them were higher-order knockouts. Phenotypic analysis in the next generation discovered functions of several seed specific genes in grain length, width, number and 100-grain weight. By analyzing single and double mutants for two Agenet domain-containing proteins, we have discovered an epistatic interaction between them for grain development. Further application of our approach will help to uncover hidden functions of the targeted genes and accelerate functional genomics research in rice. The CRISPR-Cas9 library is a useful approach to generate higher-order knockout mutants and identify functions of the targeted genes in rice.
Regulation of metalloid uptake in plants by transporters and their solute specificity
(Elsevier B.V., 2023) Sharma, Yogesh; Soni, Praveen; Raturi, Gaurav; Mandlik, Rushil; Rachappanavar, Vinay Kumar; Kumar, Manish; Salvi, Prafull; Tripathi, Durgesh Kumar; Ram, Hasthi; Deshmukh, Rupesh
Metalloids are a class of elements having properties like metals and non-metals which act as beneficial as well as hazardous for plant growth. Here, the precise role and molecular mechanism involved in the uptake and transport of different metalloids to different plant tissues is discussed. We have also described the efforts made to engineer the metalloid transport i.e. influx/efflux of metalloids that improve the uptake of beneficial metalloids and reduce hazardous metalloids. Metalloids like boron (B) and silicon (Si) play a beneficial role in plant growth and development. Optimum levels of these metalloids improve plant growth and immunity by various direct and indirect effects. On the other hand, metalloids like arsenic (As) and germanium (Ge) are detrimental for plants even at lower concentrations. The presence of these toxic entities inside cells disrupts cellular homeostasis by affecting the molecular, biochemical and physiological processes. The prospect to increase the uptake of beneficial metalloids and limit the hazardous metalloids and the challenges associated with the structural analogy and common transport mechanism is also discussed. The molecular insights into the biochemical and physiological aspects of metalloid transport and detoxification mechanisms will be helpful to exploit metalloid-derived benefits for crop improvement and accomplish food safety.