国槐遗传转化体系的优化*

doi:10.11707/j.1001-7488.20090503

Scientia Silvae Sinicae ›› 2009, Vol. 12 ›› Issue (5): 20-26.doi: 10.11707/j.1001-7488.20090503

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An effective Transformation System of Sophora japonica

Zhang Xiaoying^1,2,Gan Jing³,Yin Weilun¹,Zhu Zhen⁴,Wang Huafang¹

（1．College of Biological Science and Biotechnology，Beijing Forestry University Beijing 100083；2．Beijing Langshan Nursery Beijing 100039; 3. Beijing Gardening and Greening Bureau Beijing 100029; 4. Institute of Genetics and Developmental Biology, Chinese Academy of Sciences Beijing 100101）

Received:2008-12-23 Revised:1900-01-01 Online:2009-05-25 Published:2009-05-25

Abstract

Abstract:

This paper investigated several factors affecting transformation frequency of Sophora japonica in establishing the modified sck gene transfer system process with Agrobacterium tumefaciens-mediated leaf disc transformation method. The results showed that S. japonica leaves were required to pre-culture on MS medium for 5 d in the dark. Agrobacterium strains, GV3101, were selected, and the Agrobacterium was needed to co-cultivation for 3 d. Agrobacterium bacterium concentration was about 0.7 in OD600, and the infection time was 10 min. Addition of 200μmol·L^-1acetosyringone (AS) to the co-culture medium before infection significantly promoted S. japonica transformation. Antibiotic, 500 mg·L^-1 Cef, was best for inhibiting Agrobacterium growth. PCR and Southern blotting confirmed that sck gene had been integrated into the S. japonica genome.

Key words: Sophora japonica, Agrobacterium tumefaciens, acetosyringone, genetic transformation, Semiothisa cinerearia, insect-resistant genes

Zhang Xiaoying;Gan Jing;Yin Weilun;Zhu Zhen;Wang Huafang. An effective Transformation System of Sophora japonica[J]. Scientia Silvae Sinicae, 2009, 12(5): 20-26.

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An effective Transformation System of Sophora japonica

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