文献类型：期刊文献

英文题名：Optimization of the conditions for Casuarina cunninghamiana Miq. genetic transformation mediated by Agrobacterium tumefaciens

作者：Jiang, Qingbin[1] Ma, Yingzi[2] Zhong, Chonglu[1] Zeng, Bingshan[1] Zhang, Yong[1] Pinyopusarerk, Khongsak[3] Bogusz, Didier[4] Franche, Claudine[4]

第一作者：Jiang, Qingbin

通信作者：Zhong, CL[1]

机构：[1]Chinese Acad Forestry, Res Inst Trop Forestry, Guangzhou 510520, Guangdong, Peoples R China;[2]Cent South Univ Forestry & Technol, Coll Life Sci & Technol, Changsha 410004, Hunan, Peoples R China;[3]CSIRO Natl Res Collect Australia, Black Mt Labs, Canberra, ACT 2600, Australia;[4]Inst Rech Dev, UMR DIADE, Equipe Rhizogenese, F-34394 Montpellier 5, France

年份：2015

卷号：121

期号：1

起止页码：195-204

外文期刊名：PLANT CELL TISSUE AND ORGAN CULTURE

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000351524900017)】；

基金：This study was financially supported by the 12th National Five-Year Plan for Forestry Project (2012BAD01B0603), the Ministry of Finance of China through a Specific Program for National Non-profit Scientific Institutions (RITFYWZX201203) and Guangdong Province Forestry Science & Technology Innovation Project (2014KJCX017).

语种：英文

外文关键词：Actinorhizal plant; Casuarina cunninghamiana; Agrobacterium tumefaciens; Genetic transformation; Frankia

摘要：Using epicotyl fragments of the actinorhizal tree Casuarina cunninghamiana and the disarmed strain of Agrobacterium tumefaciens C58C1 (pGV2260) containing the pBIN19-35S-GUSINT binary vector, a method for the genetic transformation of C. cunninghamiana was established. Transformed cells were initially selected for 2 weeks on nutrient medium supplemented with kanamycin 20 mg L-1 during callus induction, and then subcultured with 50 mg L-1 until adventitious bud and shoot differentiation. Different factors involved in the early stages of the T-DNA transfer process were studied. Agrobacterium-mediated DNA delivery was most successful when epicotyl fragments excised from 45-day-old seedlings were co-cultivated with an exponentially growing culture of A. tumefaciens at an OD600nm of 0.3, for 5 days in the presence of 50 mu M of acetosyringone. Kanamycin resistant calli were observed on 88.89 % of the explants and transgenic rooted C. cunninghamiana plants were obtained in 6 months. Evidence of genetic transformation was demonstrated by -glucuronidase histochemical assays and polymerase chain reaction analyses. The possibility to obtain transgenic nitrogen-fixing nodules after inoculation by the soil actinomycete Frankia was established.