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Debnath, A. J.

Publications and source records attributed to Debnath, A. J..

3 recordsLinked to original sources

Both tobacco TA29 and sesame GN13 promoters provide anther-specific GUS expression in transgenic sesame (Sesamum indicum L.)

Yield improvement is one of the most concerning areas of the "Queen of Oilseed" sesame (Sesamum indicum L.) for its successful commercialisation. Heterosis breeding is an alternate approach for improving sesame compared to the time- and labour-consuming conventional breeding. However, tedious hand emasculation and pollination processes restrict the implementation of commercial heterosis on sesame. The unavailability of male sterile, restorer, and maintainer lines further complicates the problem. Biotechnological gene manipulation can silence anther-specific vital gene(s) leading to male sterility. Anther-specific gene study is therefore crucial to reach such a goal. In this study, we have cloned two established anther-specific promoters: sesame SiBGproplus (hereafter GN13, NCBI accession no. KT246471) and tobacco TA29 (NCBI accession no. X52283) in the plant expression vector pCAMBIA2301 producing respective GN13::GUS and TA29::GUS chimeric vectors. We recovered putatively transgenic T0 sesame lines using conventional Agrobacterium-mediated transformation with 1.76% and 1.71% frequencies, respectively. The T0 lines were segregated at the 3:1 Mendelian segregation ratio for kanamycin resistance and generated T1 transgenic lines by self-fertilisation. Most of the T1 transgenics had a single copy of transgene integration. The histological assay revealed the tapetum-specific GUS expression in the T1 transgenic lines; GUS expression was undetected in other tested plant parts. The results depict successful anther/tapetum-specific expression of two promoters in transgenic sesame lines. In further research, these promoters could be used for anther-specific cytotoxic gene expression, RNAi, or CRISPR-mediated mutational approaches to destroy androecium selectively and exhibit male sterility in sesame, resulting in heterosis-mediated improvement.

plant biology↗

Optimisation of explant-specific isolation, culture, and micro-callus induction of sesame (Sesamum indicum L.) protoplasts

The low yield of sesame (Sesamum indicum L.) compared to other oilseed crops hinders its successful commercialisation process. Sesame yield improvement using molecular-biotechnological tools involving gene manipulation is a better alternative than traditional breeding because it requires less time, effort, and labour. Protoplasts are plant cells devoid of the cell wall. Protoplast systems are deployed as versatile cell-based tools for tissue culture, genomics, transcriptomics, proteomics, metabolomics, and epigenetics studies leading to crop improvement. Inadequate reports of sesame protoplasts restrict its potential use in this crop. In the present study, we are reporting the successful isolation and purification of sesame protoplasts from four different sesame explants: hypocotyl, internode, leaf, and hypocotyl-derived callus using the one-step enzymatic digestion method. We have carefully optimised enzyme combinations, digestion durations, temperatures, and shaking speeds for every tested explant to obtain the highest protoplast yield and viability. Maximum yield (9.9 x 106 protoplasts/gm fresh weight) and viability (92.1%) were achieved from callus explants. We purified isolated protoplasts by floating them over a 20% (w/v) sucrose solution. Among tested media, callus-derived protoplasts divided rapidly in Murashige and Skoog broth medium supplemented with 17.77 M 6-benzylaminopurine (BAP) and 0.54 M -naphthalene acetic acid (NAA). In the same medium, protoplasts divided into the 2-cell stage after 2-3 days of culture and progressed to the micro-callus stage at 0.24 {+/-} 0.05% frequency within 16-20 days of culture. Sesame protoplasts could be an excellent system for investigating potential avenues for biotechnological improvements of sesame including transient gene expression and CRISPR-based studies.

plant biology↗

A comparative analysis between two flax varieties indicates lignan-mediated salt stress adaptiveness

WITHDRAWAL STATEMENTThe authors have withdrawn their manuscript because of the necessary revisions to the manuscript that may reorient the study findings. Its publication will be possible only after the peer review process in a scientific journal. Therefore, the authors do not wish this work to be cited as reference for the project. If you have any questions, please contact the corresponding author

plant biology↗