文献类型：期刊文献

英文题名：Improved Genetic Transformation of Jatropha curcas Using Phosphomannose Isomerase as Biosafe Selectable Marker

作者：Chen, Jienan[1,2,4] Liu, Bobin[1,2,3] Lu, Mengzhu[1,2,3] Zhao, Shutang[3] Zhang, Lin[1,2]

第一作者：Chen, Jienan

通信作者：Chen, JN[1]

机构：[1]Cent South Univ Forestry & Technol, Minist Forestry Bioethanol Res Ctr, Changsha 410004, Hunan, Peoples R China;[2]Cent South Univ Forestry & Technol, Inst Biol & Environm Sci & Technol, Changsha 410004, Hunan, Peoples R China;[3]Chinese Acad Forestry, Res Inst Forestry, State Key Lab Tree Genet & Breeding, Beijing 100091, Peoples R China;[4]Transpoints Inc, Gainesville, FL 32614 USA

年份：2015

卷号：9

期号：1

起止页码：9-15

外文期刊名：JOURNAL OF BIOBASED MATERIALS AND BIOENERGY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000356206600003)】；

基金：This work was supported by grants from the National Special Research Program for Public Causes (Nos. 201404604, 200904038) administered through the Ministry of Forestry of China and the National High Technology Research and Development Program of China ("863" Program) (No. 2007AA10Z182).

语种：英文

外文关键词：Jatropha curcas; pmi Gene; Selectable Marker; Genetic Transformation.

摘要：An Agrobacteriuim-mediated Jatropha curcas transformation procedure has been optimized around high frequency leaf disc regeneration using phosphomannose isomerase (pmi) gene as biosafe selectable marker. A mannose to sucrose ratio of (20 g/10 g) . L-1 was found to be optimal as selection pressure. Cephalothin as a bacteriostat was optimized at 500 mg L-1 for inhibition of Agrobacteriuim overgrowth without destructive effect on callus induction. The effect of acetosyringone concentration, inoculation duration, and co-cultivation time was evaluated and found to be most desirable at 0.02 mM, 10 min and 2 days, respectively. Sonication treatment had a suppressing effect on resistant shoots induction. The transformation efficiency with mannose selection was twice of that with kanamycin selection. PCR amplification identified 6 positive plants from 12 independent mannose-resistant plants. This efficient and biosafe transformation system is of significance to facilitating the genetic improvement of J. curcas as a promising biodiesel plant and also to alleviating public concerns on the wide use of transgenic energy plants.