Articles liés à Genetic modification of Brassica juncea: Agrobacterium...

Genetic modification of Brassica juncea: Agrobacterium mediated transformation of Brassica juncea with antimicrobial Wasabi defensin gene - Couverture souple

Khan Bangash, Sajid Ali; Khattak, Sahir Hameed; Swati, Zahoor Ahmad

 
9783846539170: Genetic modification of Brassica juncea: Agrobacterium mediated transformation of Brassica juncea with antimicrobial Wasabi defensin gene
Couverture souple
ISBN 10 : 3846539171 ISBN 13 : 9783846539170
Editeur : LAP LAMBERT Academic Publishing, 2011

Synopsis

Brassica juncea is one of the most important oilseed crop. We have developed an efficient and reproducible protocol for Agrobacterium-mediated transformation of B. juncea variety NIFA RAYA with Wasabi defensin gene to produce transgenic plants. This gene was isolated from the leaves of Wasabia japonica. We find out that co-cultivation of explants with the transformed agrobacterium inoculum having an OD ranged from 0.6-1 for 15-20 minutes give high transformation efficiency. These explants were then regenerated on MS medium containing 1-6 mgL-1 BAP, 0.1-0.6 mgL-1 NAA and 300 mgL-1 cefotoxim sodium with subsequent application of 10-100 mgL-1 kanamycin. Our results indicated that medium containing 3 mgL-1 BAP and 0.3 mgL-1 NAA was found to be the best hormone combination for callus formation whereas BAP 3 mgL-1 and NAA 0.5 mgL-1 was found to be the best combination for shoot regeneration. In all cases supplementing the medium with 20 μM AgNO3 had positive effect on callus formation and shoot regeneration. The transgenic nature of the transformed callus and regenerated shoots was confirmed via PCR analysis.

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Présentation de l'éditeur

Brassica juncea is one of the most important oilseed crop. We have developed an efficient and reproducible protocol for Agrobacterium-mediated transformation of B. juncea variety NIFA RAYA with Wasabi defensin gene to produce transgenic plants. This gene was isolated from the leaves of Wasabia japonica. We find out that co-cultivation of explants with the transformed agrobacterium inoculum having an OD ranged from 0.6-1 for 15-20 minutes give high transformation efficiency. These explants were then regenerated on MS medium containing 1-6 mgL-1 BAP, 0.1-0.6 mgL-1 NAA and 300 mgL-1 cefotoxim sodium with subsequent application of 10-100 mgL-1 kanamycin. Our results indicated that medium containing 3 mgL-1 BAP and 0.3 mgL-1 NAA was found to be the best hormone combination for callus formation whereas BAP 3 mgL-1 and NAA 0.5 mgL-1 was found to be the best combination for shoot regeneration. In all cases supplementing the medium with 20 μM AgNO3 had positive effect on callus formation and shoot regeneration. The transgenic nature of the transformed callus and regenerated shoots was confirmed via PCR analysis.

Biographie de l'auteur

Sajid Ali Khan Bangash, BSc (Hons)Plant breeding and genetics(silver medalist, M.Phil Biotechnology, Institute of Biotechnology and Genetic Engineering, Khyber Pakhtunkhwa Agricultural University Peshawar, Pakistan.

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Éditeur
LAP LAMBERT Academic Publishing
Date d'édition
2011
Langue
anglais
ISBN 10 
3846539171
ISBN 13 
9783846539170
Reliure
Broché
Nombre de pages
60
Coordonnées du fabricant
non disponible
Personne responsable
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