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根癌农杆菌介导茶树转基因体系的建立
项威,史成颖,贺志荣,徐燕
0
(安徽农业大学教育部和农业部茶叶生物化学与生物技术重点实验室,合肥230036)
摘要:
利用茶树[Camellia sinensis(L.)O. Kuntze]品种“农抗早”的叶片进行培养,诱导形成愈伤组织。选择生长旺盛、致密的愈伤组织块与根癌农杆菌EHA105进行共培养,通过潮霉素(Hygromycin)抗性筛选、GUS染色及PCR的验证,获得部分转基因愈伤组织。在添加100 μmol·L-1的乙酰丁香酮(AS)处理后,转化率约为10%。同时试验结果证明,AS可以提高农杆菌的转化效率,增加渗透压却不能提高转化率。
关键词:  茶树  转基因  组织培养  愈伤组织  根癌农杆菌
DOI:
投稿时间:2012-05-02
基金项目:国家自然科学基金项目(31070614)和安徽农业大学人才引进基金共同资助。
Establishment of an agrobacterium-mediated transformation system for tea plant (Camellia sinensis)
XIANG Wei,SHI Cheng-ying,HE Zhi-rong,XU Yan
(Key Laboratory of Tea Biochemistry and Biotechnology,Ministry of Education and Agriculture, Anhui Agricultural University, Hefei 230036)
Abstract:
Calli were induced from leaves of tea cultivar ‘Nongkangzao’, and those growing vigorously with compact tissue were chosen for agrobacterium-mediated transformation by EHA105. The transgenic callus was screened using hygromycin. The resistant callus was confirmed by RT-PCR and GUS dyeing. The results showed that transgenic callus was obtained using this method, and the conversion rate was about 10% after adding 100 μmol·L-1 acetosyringone (AS). The results also showed that the efficiency of agrobacterium-mediated transformation could be effectively improved by adding AS, but the increase in osmotic pressure had no effect .
Key words:  Camellia sinensis   transgenosis  cell tissue culture  calli  Agrobacterium

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