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Regeneration of sweet potato transgenic plants with oryzacystatin-I (OCI) gene

Published online by Cambridge University Press:  12 February 2007

Jiang Sheng-Jun ,
Liu Qing-Chang ,
Zhai Hong ,
Wu Li-Sha  and
Wang Yu-Ping
Jiang Sheng-Jun
Affiliation:
Key Laboratory of Crop Genomics and Genetic Improvement, Ministry of Agriculture, China Agricultural University, Beijing 100094, China
Liu Qing-Chang*
Affiliation:
Key Laboratory of Crop Genomics and Genetic Improvement, Ministry of Agriculture, China Agricultural University, Beijing 100094, China
Zhai Hong
Affiliation:
Key Laboratory of Crop Genomics and Genetic Improvement, Ministry of Agriculture, China Agricultural University, Beijing 100094, China
Wu Li-Sha
Affiliation:
Key Laboratory of Crop Genomics and Genetic Improvement, Ministry of Agriculture, China Agricultural University, Beijing 100094, China
Wang Yu-Ping
Affiliation:
Key Laboratory of Crop Genomics and Genetic Improvement, Ministry of Agriculture, China Agricultural University, Beijing 100094, China
*
*Corresponding author: Email: [email protected]
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Abstract

Regeneration of sweet potato (Ipomoea batatas cv. Lizixiang) transgenic plants with oryzacystatin-I (OCI) gene was achieved using an Agrobacterium tumefaciens-mediated method. A. tumefaciens strain LBA4404, harbouring a binary vector pBinh with neomycin phosphotransferase II and OCI genes, was used. After 3 days of subculture, Lizixiang embryogenic suspension cultures were co-cultivated with LBA4404 (OD600nm=0.5) for 4 days. Next, the infected suspension cultures were first cultured for 5 days in MS medium with 2 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) and 300 mg/l carbencillin (Carb) but without kanamycin (Kan) and then transferred to MS medium supplemented with 2 mg/l 2,4-D, 50 mg/l Kan and 300 mg/l Carb for the selection culture. Four weeks after selection, 200 Kan-resistant cell aggregates (∼1 mm in size) from the embryogenic suspension cultures were transferred to MS solid medium supplemented with 2 mg/l 2,4-D, 50 mg/l Kan and 300 mg/l Carb, and eight embryogenic calluses were obtained. After transferring to MS medium supplemented with 1 mg/l ABA, 50 mg/l Kan and 100 mg/l Carb, these embryogenic calluses formed 13 plantlets via somatic embryogenesis. PCR and PCR–Southern blot analysis indicated that seven of the 13 plantlets were transgenic.

Keywords

Ipomoea batatastransgenic plantsoryzacystatin-I gene
Type
Research Article
Information
Chinese Journal of Agricultural Biotechnology , Volume 1 , Issue 2 , August 2004 , pp. 99 - 102
Copyright
Copyright © China Agricultural University and Cambridge University Press 2004

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