Browsing by Author "Jewaria, Pawan Kumar"

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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.
Deletion of the PtrDJ1C gene leads to increased branching in poplar
(Elsevier B.V., 2025) Du, Jingxia; Shao, Chunxue; Wang, Dong; Feng, Zizhuo; Cui, Chuwen; Li, Ruili; Jewaria, Pawan Kumar; Wang, Xuanyi; Xiao, Jianwei; Wang, Xinwei
The PtrDJ1C gene is essential for poplar growth and early chloroplast development. Disruption of PtrDJ1C expression results in an albino leaf phenotype and increased branching. However, the underlying mechanism for the increased branching remains unknown. In this study, we employed integrated approaches to investigate the function of PtrDJ1C in the branch-increasing phenotype. Our results revealed that levels of indole-3-acetic acid (IAA), gibberellin (GA), and abscisic acid (ABA) were significantly reduced in ptrdj1c mutants, while cytokinin (CK) levels were slightly increased. Transcriptomic and proteomic analyses identified several key genes and proteins involved in hormone regulation and branching development that were differentially expressed. Specifically, the expression levels of TAA, ZEP, and GA20ox—genes involved in IAA, GA, and ABA biosynthesis—were significantly reduced in ptrdj1c, while IPT and LOG, which regulate CK synthesis, were upregulated. Moreover, immunoblot analysis further validated reduced levels of key biosynthetic enzymes for IAA, GA, and ABA, alongside increased levels of IPT and LOG enzymes. Interestingly, our findings suggest that hormone signaling pathways act in concert with the transcription factor WUSCHEL (WUS) to synergistically promote branching development. These results provide novel insight into the regulatory role of PtrDJ1C in hormone balance and its downstream effects on poplar branching.
Photosynthetic adaptation in poplar under abiotic and biotic stress: integrating molecular, physiological, and biotechnological perspectives
(MDPI AG, 2025) Wang, Dong; Jewaria, Pawan Kumar; Xiao, Jianwei
In the context of global climate change, the carbon storage and sequestration capacity of terrestrial ecosystems is of increasing concern. Poplars are widely planted because of their fast growth and environmental adaptability. We reviewed the effects of abiotic and biotic stresses on photosynthesis in poplar, focusing on the damage caused by adversity conditions to photosynthetic apparatus, which leads to decreased carbon dioxide (CO2) assimilation and an increase in reactive oxygen species (ROS)-induced oxidative damage. The mechanisms of photosynthesis response to stress in poplar are reviewed, especially the role of genes regulation in regulating photosynthetic efficiency. These findings are particularly important for improving the resilience of poplar under changing environmental conditions. In addition, we discussed a range of strategies to enhance photosynthesis in poplar under stress, such as genetic engineering and synthetic biology. These approaches provide theoretical guidance for improving the resilience of poplar and insights for improving other crops facing similar challenges.
Reduced RG- II pectin dimerization disrupts differentialgrowth by attenuating hormonal regulation
(American Association for the Advancement of Science, 2025) Jewaria, Pawan Kumar; Aryal, Bibek; Begum, Rifat Ara; Wang, Yaowei; Sancho-Andrés, Gloria; Baba, Abu Imran; Yu, Meng; Li, Xiaojuan; Lin, Jinxing; Fry, Stephen C.; Verger, Stephane; Russinova, Eugenia; Jonsson, Kristoffer; Bhalerao, Rishikesh P.
Defects in cell wall integrity (CWI) profoundly affect plant growth, although, underlying mechanisms are not well understood. We show that in Arabidopsis mur1 mutant, CWI defects from compromising dimerization of RG-II pectin, a key component of cell wall, attenuate the expression of auxin response factors ARF7-ARF19. As a result, polar auxin transport components are misexpressed, disrupting auxin response asymmetry, leading to defective apical hook development. Accordingly, mur1 hook defects are suppressed by enhancing ARF7 expression. In addition, expression of brassinosteroid biosynthesis genes is down-regulated in mur1 mutant, and supplementing brassinosteroid or enhancing brassinosteroid signaling suppresses mur1 hook defects. Intriguingly, brassinosteroid enhances RG-II dimerization, showing hormonal feedback to the cell wall. Our results thus reveal a previously unrecognized link between cell wall defects from reduced RG-II dimerization and growth regulation mediated via modulation of auxin-brassinosteroid pathways in early seedling development.