Development of reproducible regeneration and transformation system for Sesamum indicum
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ORIGINAL ARTICLE
Development of reproducible regeneration and transformation system for Sesamum indicum T. Gayatri1 · Asitava Basu1 Received: 30 July 2019 / Accepted: 11 September 2020 © Springer Nature B.V. 2020
Abstract Sesamum indicum is an important oilseed crop with beneficiary nutrients such as essential fatty acids, proteins, and lignan. S. indicum crop improvement using conventional breeding methods was disastrous. Moreover, no reproducible regeneration and transformation protocol for large-scale generation of independent primary transformed plant lines of S. indicum virtually exists. Therefore, in the present study, a reproducible regeneration and transformation method was developed for S. indicum. Direct shoot regeneration (94.44%) was achieved using plumule tips as an explant source and when maintained on regeneration medium (BM11) an amalgamation of half the strength of MS macronutrients and B5 macronutrients together with MS micronutrients and vitamins. Maximum number of shoots (~ 16–19 shoots/explant) was obtained on BM11 medium supplemented with TDZ (1 mg/L) and BAP (0.1 mg/L). High root induction efficiency (~ 97%) was observed on RM4 medium consisted of half MS macronutrients, SH micronutrients and vitamins. The developed protocol was found effective for other commercially valuable genotypes such as Co 1, Phule til and Tilottama as well. Successful regeneration was subsequently extended to transformation of reporter gene (gfp: green fluorescent protein) as revealed by molecular analyses includes Southern hybridization, northern hybridization, real time PCR and histological study of the transformants developed in the present study. Based on the Southern analysis the calculated transformation frequency was found to be 1.33%. The presently developed regeneration and transformation system might facilitate any desired improvement of sesame crop. Key message The present study demonstrated an improved regeneration and transformation method for S. indicum, which is genotype independent and can undergo desired modifications. Keywords Direct regeneration · Indirect regeneration · Agrobacterium-mediated transformation method · Green fluorescence protein
Introduction
Communicated by K X Tang. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11240-020-01931-1) contains supplementary material, which is available to authorized users. * Asitava Basu [email protected]; [email protected] 1
Advanced Laboratory for Plant Genetic Engineering, Advanced Technology Development Centre, Indian Institute of Technology-Kharagpur, Kharagpur, West Bengal 721302, India
Sesame (Sesamum indicum L.), an oldest oilseed crop, is widely cultivated in tropical and subtropical regions belong to the Sesamum genus of the Pedaliaceae family (Ashri 2010), is rich in oil (48.5%), proteins (20%), carbohydrate (7.78%) and fibre (9.4%) (Nzikou et al. 2009). Sesame is considered as a valuable source of an essential fatty acid including linoleic acid. Recently, it has gain
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