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Genotype independent regeneration and agrobacterium-mediated genetic transformation of sweet potato (Ipomoea batatas L.)

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Title Genotype independent regeneration and agrobacterium-mediated genetic transformation of sweet potato (Ipomoea batatas L.)
 
Creator Shekhar, Shubhendu
Agrawal, Lalit
Buragohain, Alak Kumar
Datta, Asis
Chakraborty, Subhra
Chakraborty, Niranjan
 
Subject Sweet potato
Somatic embryogenesis
Selectable marker
GUS expression
 
Description Development of an efficient genotype independent regeneration and genetic transformation system in sweet potato continues to be of great interest. Agrobacterium‐mediated genetic transformation protocol was established in two different cultivars of sweet potato using Agrobacterium strain EHA105 harbouring binary plasmid pBI121 containing GUS and nptII genes. The internodal stem segments from 30‐day‐old micropropogated plants were used as explant with different combinations of media and hormones. MS and LS media with various concentrations of growth regulators proved to be non‐responsive and the infecundity was severe with the addition of cytokinins. Nonetheless, MS with 2,4‐D and TDZ gave a good percentage of callusing but with low differentiation. In different concentrations of NAA, significant amount of callusing was observed but percentage of rooting remained low in both the genotypes. Gamborg’s B5 supplemented with NAA proved to be the most suitable media and hormone combination, which yielded shoot formation after 8 ‐ 10 weeks with a regenera‐ tion efficiency of 40 ‐ 70%. Stable integration of transgene was confirmed by PCR analysis. Furthermore, qRT‐PCR analysis was performed to assess the transcript accumulation in addition to the GUS enzymatic assay in the transgenic lines.
This work was supported by grants (BT/PR/3123/16/250/02) from the Department
of Biotechnology (DBT), Govt. of India. The authors thank Divya Rathi for critical
reading of the manuscript, and Jasbeer Singh for technical assistance.
 
Date 2015-11-19T08:30:32Z
2015-11-19T08:30:32Z
2013
 
Type Article
 
Identifier Plant Tissue Cult. Biotech., 23(1): 87-100
1818-8745
http://172.16.0.77:8080/jspui/handle/123456789/371
http://www.banglajol.info/index.php/PTCB/article/view/15565
http://dx.doi.org/10.3329/ptcb.v23i1.15565
 
Language en_US
 
Publisher Bangladesh Association for Plant Tissue Culture and Biotechnology