Cloning of a chitinase gene and the acquisition of transgenic potato plants

Abstract

Abstract: The antagonistic ability of potato against fungal disease was enhanced by acquiring transgenic potato plants via gene engineering. A chitinase gene was cloned via PCR products from cDNA of Trichoderma atroviride. This gene was recombined into the plant expression vector p33ECR and transferred into micro tubers from flasks of two varieties (Favorita and Shepody) of potato via Agrobacterium tumefaciens-mediated transformation. After infection and co-culture, 18 transformed plants were acquired through Kanamycin and Carbenicillin screening tests. After PCR tests for transgenic plants, 67% of the plants (12 plantlets) were positive. This will give support for obtaining high biologically effective potato varieties in future research.

CLC Number:

ZHANG Ru, LI Jin-hua, CHAI Zhao-xiang, ZHANG Jun-lian, ZHANG Jin-wen , WANG Di. Cloning of a chitinase gene and the acquisition of transgenic potato plants[J]. Acta Prataculturae Sinica, 2009, 18(6): 51-58.

References

[1] 于品华. 饲用型马铃薯新品系“杂3单选-5”选育研究[J]. 草业学报, 2004, 13(2): 115-117.
[2] 夏启中, 张明菊. 植物抗真菌病害基因及其应用研究[J]. 武汉职业技术学院学报, 2005, (3): 62-66.
[3] 孙慧生. 马铃薯育种学[M]. 北京: 中国农业出版社, 2003.
[4] 李金花, 柴兆祥, 王蒂, 等. 甘肃马铃薯贮藏期真菌性病害病原菌的分离鉴定[J]. 兰州大学学报(自然科学版), 2007, 43(2): 39-42.
[5] 张俊莲, 王丽, 王蒂, 等. 转根癌农杆菌介导的AtNHXI基因马铃薯的获得[J]. 作物学, 2007, 33(7): 1067-1072.
[6] 郭金芳, 潘俊松, 王琛, 等.病程相关蛋白与植物抗病性关系的研究及其在草坪草抗病育种中的应用[J].草业学报, 2008, 17(6): 156-163.
[7] 惠有为, 孙勇, 潘亚妮, 等. 木霉菌在植物真菌病害防治上的作用[J]. 西北农业学报, 2003, 12(3): 96-99.
[8] 杨合同, 唐文华, Ryder M. 木霉菌与植物病害的生物防治[J]. 山东科学, 1999, 12(4): 7-16.
[9] Si H J, Xie C H, Liu J. An efficient protocol for Agrobacterium-mediated transformation with microtuber and the introduction of an antisense class I patatin gene into potato[J]. Acta Agronomica Sinca, 2003, 29(6): 801-805.
[10] 张茹, 李金花, 柴兆祥, 等. 甘肃河西马铃薯根际生防木霉菌对接骨木镰刀菌的拮抗筛选及鉴定[J]. 草业学报, 2009, 18(2): 138-145.
[11] 萨姆布鲁克.分子克隆实验指南[M].北京: 科学出版社, 2002.
[12] 李梅云, 王革, 李天飞, 等. 烟草主要真菌病类生防木霉的筛选[J]. 西南农业大学学报, 2001, 23(1): 10-12.
[13] 杨依军, 王勇, 杨秀荣, 等. 拮抗木霉菌在生物防治中的作用[J]. 天津农业科学, 2000, 6(3): 29-33.
[14] 徐同, 柳良好. 木霉几丁质酶及其对植物病原真菌的拮抗作用[J]. 植物病理学报, 2002, 32(2): 97-102.
[15] 王伟, 赵谦, 杨微. 木霉菌对土传病原尖孢镰刀菌拮抗作用[J]. 中国生物防治, 1997, 13(1): 46-47.
[16] Bell D K, Well H D, Markham C R. In vitro antagonism of Trichoderma species against fungal pathogens[J]. Phytopath, 1982, 2(4): 279-382.
[17] 东丽, 李杰, 朱延明. 抗真菌、抗渗透胁迫基因多价植物表达载体构建及对黄瓜遗传转化的研究[J]. 农业科技通讯, 2008, 3: 37-41.
[18] 贾晓霞, 张金文, 王汉宁, 等. 抗真菌和抗除草剂基因多价植物表达载体构建及对烟草遗传转化的研究[J]. 草业学报, 2009, 18(1): 86-93.
[19] 王丽, 杨宏羽, 张俊莲, 等. 根癌农杆菌介导马铃薯试管薯转化体系的优化及AtNHX1基因的导入[J]. 西北植物学报, 2008, 28(6): 1088-1094.