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, Chonglu

机构：[1] Research Institute of Tropical Forestry, Chinese Academy of Forestry, Guangzhou, 510520, China; [2] College of Life Science and Technology, Central South University of Forestry and Technology, Changsha, 410004, China; [3] Black Mountain Laborartories, CSIRO National Research Collections Australia, Canberra, ACT, 2600, Australia; [4] Equipe Rhizogenèse, UMR DIADE, Institut de Recherche pour le Développement, 911 avenue Agropolis, BP64501, Montpellier Cedex 5, 34394, 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：EI(收录号：20150100392761);Scopus(收录号：2-s2.0-84925488507)

语种：英文

外文关键词：Cultivation - Seed - Bacteria - Nitrogen fixation - Plants (botany) - Binary trees

摘要：Using epicotyl fragments of the actinorhizal tree Casuarinacunninghamiana 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?μM 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. ? 2014, Springer Science+Business Media Dordrecht.