为了明确不同农杆菌菌种对茶多酚的耐受性能变化幅度,以及驯化培养对提高农杆菌茶多酚耐受性、从而提高茶树遗传转化中遗传转化率的作用与可行性,本文比较了茶多酚对四种农杆菌生长的抑制率,进行了耐酚菌株的驯化培养,最后比较了驯化菌株与原始菌株的转化能力。结果表明：茶多酚对几种农杆菌均有明显的生长抑制作用,但不同菌种的耐受性能差异较大;通过驯化培养得到的耐酚菌株EHA105AP,其耐酚性能提高了90.89%,同时,其在茶树叶片和愈伤中的GUS瞬时表达率也分别提高了43.85%和57.55%。
In order to make clear the variation of polyphenol-resisting characteristics of different kinds of Agrobacterium as well as the potential in improving the polyphenol-resisting ability and the Agrobacterium-mediated genetic transformation rate by domestication culture, the inhibition effect of tea polyphenols on four kinds of Agrobacterium strains, the polyphenol-resisting demestication of Agrobacterium strains and the improvement of genetic transformation ability in demesticated polyphenol-resisting strain were studied. The results showed that tea polyphenols had obvious inhibition on the growth of different kinds of Agrobacterium, but it varied greatly among these studied Agrobacterium. A polyphenol-resisting strain EHA105AP was obtained by domestication culture procedure, the polyphenol-resisting ability of which enhanced 90.89% compared with that of the original. Meanwhile, the rate of transient expression of GUS gene induced by the domesticated strain increased 43.85% and 57.55% respectively in callus and leaf explants case.
[1] 张广辉, 梁月荣, 陆建良. 发根农杆菌介导的茶树发根高频诱导与遗传转化[J]. 茶叶科学, 2006, 26(1): 1~10.
[2] 奚彪, 刘祖生, 梁月荣, 等. 发根农杆菌介导的茶树的遗传转化[J]. 茶叶科学, 1997, 17(增): 155~156.
[3] Zebra M, Banerjee S, Mathur A K, et al. Induction of hairy roots in tea(Camellia sinensis L.)[J].Current Science,1996,70(1):84~86.
[4] Mondal, Bhattacharya, Ahuja, et al. Transgenic tea [Camellia sinensis(L.)O. Kuntze cv. Kangra Jat] plants obtained by Agrobacterium-Mediated transformation of somatic embryos[J]. Plant Cell Reports, 2001, 20(8): 712~720.
[5] 赵东, 刘祖生, 陆建良, 等. 根癌农杆菌介导茶树转化研究[J]. 茶叶科学, 2001, 21(2): 108~111.
[6] 骆颖颖, 梁月荣. Bt基因表达载体的构建及对茶树遗传转化的研究[J]. 茶叶科学, 2000, 20(2): 42~47.
[7] 吴珊, 梁月荣, 陆建良, 等. 茶树农杆菌转化系统和基因枪转化系统的优化[J]. 茶叶科学, 2003, 23(1): 6~10.
[8] 王丹, 朱常香, 郑成超, 等. 根癌农杆菌介导的马铃薯遗传转化条件的优化[J]. 山东农业大学学报(自然科学版), 2002, 33(1): 23~27.
[9] 毛清黎, 施兆鹏, 李玲, 等. 茶叶儿茶素对发根农杆菌的抑制作用及抗酚菌株筛选研究[J]. 茶叶科学, 2007(3): 243~247.
[10] 王关林, 方宏筠. 植物基因工程原理与技术[M]. 北京: 科学出版社, 1998, 776~777.
[11] RuebS, Hensgens L A M. Improved histochemical staining for β-D-glucuronidase activity in monocotyledonous plants[J]. Rice Genetics Newsletter, 1989(6): 168~169.
[12] Tang W, Luo H, Newton R J.Effects of antibiotics on the elimation of Agrobacterium tumefaciens from loblolly pine(Pinus taeda)zygotic embtyo explants and on transgenic plant regenetation[J]. Plant Cell, Tissue and Organ Culture, 2004, 79(1): 71~81.
[13] 陶均, 李玲. 农杆菌转化的分子生物学[J]. 植物生理学通讯, 2002, 8(6): 639~644.
浙公网安备 33019902000101号 
