Browsing by Author "Rani, Manjusha"

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    An improved Agrobacterium-mediated transformation method for genome editing using CRISPR/Cas9 in elite indica rice (Oryza sativa L.)
    (Springer Nature, 2026-04-07) Behera, Laxmipreeya; Samal, Kailash Ch.; Parameswaran C,; Agrawal, Pawan Kumar; Achary, V. Mohan Murali; Dash, Manasi; Mishra, Ashok; Rani, Manjusha; Masika, Fred Bwayo; Goud, Gurunatham Sai Deekshith; Kesawat, Mahipal Singh; Samantaray, Sanghamitra
    Rice feeds nearly half of the world’s population and underpins global food security. Climate change now poses a major threat to rice productivity worldwide. Genome editing has reshaped crop improvement strategies. Among these tools, the Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated protein (CRISPR/Cas) system stands out for its precision, efficiency, and scalability. However, Agrobacterium-mediated transformation efficiency is often low, particularly in indica rice varieties. Here, we optimized an Agrobacterium-mediated transformation protocol for indica rice cultivars. The method was established in Lalat and MTU-1010. Seed-derived embryogenic calli were used to introduce the thermosensitive genic male sterile (OsTMS5) gene. A CRISPR/Cas9 vector carrying a gRNA and the selectable marker hptII was used for transformation. Callus induction reached 96.87% in MTU-1010 and 93.30% in Lalat MS medium supplemented with 3 mg/L 2,4-D and 0.5 mg/L BAP. In contrast, regeneration efficiency was higher in Lalat (90.28%) than in MTU-1010 (87.51%) on MS medium supplemented with 0.25 mg/L NAA, 0.5 mg/L kinetin, and 2 mg/L BAP. In addition, PCR analysis further verifies the integration of the transgene. Subsequently, the transformation efficiency was 37.20% in Lalat and 29.62% in MTU-1010. Therefore, this protocol provides a robust platform for gene function analysis and trait editing in rice. Its application may accelerate yield improvement and enhance stress tolerance under changing climatic conditions.
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    Genome-wide survey and expression analysis of peptides containing tyrosine sulfation (PSY) gene family in Cicer arietinum L.
    (Springer Nature, 2026-02-21) Kesawat, Mahipal Singh; Kumar, Vinay; Manohar, Swati; Sohail, Aqib; Rani, Manjusha; Chung, Sang-Min; Kumar, Deepak; Lenka, Sangram K.; Masika, Fred Bwayo
    Background Plant growth and developmental processes are tightly regulated by small secreted peptides, however, the functions and mechanisms of Tyrosine Sulfation-containing Peptides (PSY) remain unclear. In chickpea, knowledge of PSY genes family is limited. Results This study employed comprehensive bioinformatics approaches to identify and characterize seven CaPSY genes in the chickpea genome. The analyses encompass chromosomal localization, evolutionary relationships, gene structure, conserved motif identification, promoter architecture, prediction of PSY-targeting miRNAs, and expression profiling. Chromosomal mapping revealed that CaPSY genes are confined to four specific chromosomes rather than being evenly distributed across the genome. Phylogenetic analysis resolved nine distinct groups, each further subdivided into subgroups. Additionally, CaPSY genes were found to contain one to two introns. Amino acid sequence comparisons demonstrated that each CaPSY gene consistently harbors a PSY domain in its C-terminal end. Promoter analysis of CaPSY genes revealed the presence of multiple hormone-responsive elements, including ABRE, SARE, AuxRE, and MeJARE, as well as stress-related elements such as the drought-responsive MBS, suggesting potential regulatory roles in development and stress adaptation. Further, the expression patterns of CaPSY were evaluated in multiple tissues as well as in response to abiotic stresses. The results indicated differential expression of CaPSY genes among tissues and under multiple abiotic stress conditions. We further detected several miRNAs likely to target CaPSY genes and assessed how they are expressed in different tissues. Conclusion Thus, these findings serve as a crucial resource for basic and applied research, enabling advancements in chickpea productivity and stress tolerance via precise genome editing and innovative breeding methods.