TY - JOUR
T1 - Genetic transformation of cultivated jute (Corchorus capsularis L.) by particle bombardment using apical meristem tissue and development of stable transgenic plant
AU - Bhattacharyya, Jagannath
AU - Chakraborty, Anirban
AU - Roy, Souri
AU - Pradhan, Subrata
AU - Mitra, Joy
AU - Chakraborty, Monami
AU - Manna, Anulina
AU - Sikdar, Narattam
AU - Chakraborty, Saikat
AU - Sen, Soumitra Kumar
N1 - Publisher Copyright:
© 2015, Springer Science+Business Media Dordrecht.
PY - 2015/5/1
Y1 - 2015/5/1
N2 - 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 T1 plants 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.
AB - 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 T1 plants 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.
KW - Bar gene
KW - Bialaphos
KW - Herbicide resistance
KW - Jute transformation
KW - Particle bombardment
KW - Transgenic plants
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U2 - 10.1007/s11240-014-0702-2
DO - 10.1007/s11240-014-0702-2
M3 - Article
AN - SCOPUS:84940006217
SN - 0167-6857
VL - 121
SP - 311
EP - 324
JO - Plant Cell, Tissue and Organ Culture
JF - Plant Cell, Tissue and Organ Culture
IS - 2
ER -