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Genetic transformation of cultivated jute (Corchorus capsularis L.) by particle bombardment using apical meristem tissue and development of stable transgenic plant

Author:
Bhattacharyya, Jagannath, Chakraborty, Anirban, Roy, Souri, Pradhan, Subrata, Mitra, Joy, Chakraborty, Monami, Manna, Anulina, Sikdar, Narattam, Chakraborty, Saikat, Sen, Soumitra Kumar
Source:
Plant cell, tissue, and organ culture 2015 v.121 no.2 pp. 311-324
ISSN:
0167-6857
Subject:
Corchorus capsularis, DNA, Western blotting, apical meristems, culture media, explants, gene expression, genetic engineering, genetic transformation, indole butyric acid, inheritance (genetics), jute, quantitative polymerase chain reaction, seedlings, shoots, transgenes, transgenic plants
Abstract:
An in vitro plant regeneration and genetic transformation protocol was established in jute (Corchorus capsularis L. var JRC321). One-day-old apical, meristematic tissues of germinating seedlings were used as explants. Multiple shoots were regenerated from each explant using Murashige and Skoog basal medium containing 1.78 µM benzylamino purine and 4.92 µM indole-3-butyric acid. Transformation was carried out in three independent sets (each set comprising of three independent experiments each comprising three replications with 35 explants per replication) using the bialaphos resistance gene (bar), synthetically designed for high level plant expression. The positive transformants containing the bar gene were selected in growth medium containing 2.5 mg/l bialaphos. Polymerase chain reaction (PCR), Southern and northern blots, real-time quantitative PCR, western blot and enzymatic assay of five putative transformants from three independent sets provided evidence for full-length gene integration into the genomic DNA of transformed jute, as well as high level expression of the transgene. Analysis of the T₁plants revealed a stable inheritance of the transgene through the progenies. The data presented in this report showed considerable advancement in jute transformation and should improve future genetic engineering strategies to be employed for improvement of this very important fibre crop.
Agid:
1240333