国槐离体再生及抗虫基因sck的转导

doi:10.11707/j.1001-7488.20060907

林业科学 ›› 2006, Vol. 42 ›› Issue (9): 34-38.doi: 10.11707/j.1001-7488.20060907

国槐离体再生及抗虫基因sck的转导

张晓英¹ 王华芳¹ 朱祯² 王天祥¹ 尹伟伦¹

1.北京林业大学生物科学与技术学院,北京100083；2.中国科学院遗传与发育生物学研究所,北京100101

收稿日期:2005-03-10 修回日期:1900-01-01 出版日期:2006-09-25 发布日期:2006-09-25

In Vitro Regeneration and Agrobacterium Transformation of Sophora japonica with sck Gene

Zhang Xiaoying¹,Wang Huafang¹,Zhu Zhen²,Wang Tianxiang¹,Yin Weilun¹

1.College of Biological Sciences and Biotechnology, Beijing Forestry University Beijing 100083; 2. Institute of Genetics and Developmental Biology, Chinese Academy of Sciences Beijing 100101

Received:2005-03-10 Revised:1900-01-01 Online:2006-09-25 Published:2006-09-25

摘要/Abstract

摘要：

以国槐叶片作为外植体,筛选出诱导不定芽分化的最佳培养基;利用根癌农杆菌介导法将经过修饰的广谱抗虫基因豇豆胰蛋白酶抑制剂基因sck(Signal-CpTI-KDEL)导入国槐。国槐叶片与农杆菌共培养结束后,转移至筛选培养基MS+BA 3 mg·L^-1+IAA 0.05 mg·L^-1+G418 8 mg·L^-1+Cef 500 mg·L^-1上。不定芽在培养基1/2MS+IBA 1.0 mg·L^-1+G418 10 mg·L^-1+Cef 100 mg·L^-1上进一步培养获得完整再生植株。通过PCR和Southern blotting检测证明,修饰的豇豆胰蛋白酶抑制剂基因sck已成功导入国槐细胞。

关键词: 国槐, 根癌土壤农杆菌, 豇豆胰蛋白酶抑制剂基因sck, 基因转导

Abstract:

Sophora japonica is a valuable gardening tree that has been planted in the North of China. However, a problem hampering the application of its full potential is susceptibility to insect. In this study, taking leaves of Sophora japonica as the explants, the optimum medium for the induction of adventitious bud differentiation was selected. Modified cowpea trypsin inhinitor(CpTI)gene sck(Signal-CpTI-KDEL), an insecticidal gene, was introduced into S. japonica by Agrobacterium-mediated genetic transformation. After cocultivation of leaves of S. japonica with Agrobacterium tumefaciens harbored sck gene, kanamycin resistance shoots were transferred to selective MS medium containing 3 mg·L^-1 BA, 0.05 mg·L^-1IAA, 8 mg·L^-1 G418 and 500 mg·L^-1 cefotaxime. Transgenic plants were obtained after rooting on half-strength MS medium supplemented with 1.0 mg·L^-1 indole-3-butyrique acide (IBA), 10 mg·L^-1 G418 and 100 mg·L^-1 cefotaxime. PCR and Southern blotting confirmed that Modified CpTI gene(sck)was successfully transferred into S. japonica mediated by A. tumefaciens.

Key words: Sophora japonica, Agrobacterium tumefaciens, cowpea trypsin inhinitor(CpTI) gene sck, gene transformation

张晓英王华芳朱祯王天祥尹伟伦. 国槐离体再生及抗虫基因sck的转导[J]. 林业科学, 2006, 42(9): 34-38.

Zhang Xiaoying;Wang Huafang;Zhu Zhen;Wang Tianxiang;Yin Weilun. In Vitro Regeneration and Agrobacterium Transformation of Sophora japonica with sck Gene[J]. Scientia Silvae Sinicae, 2006, 42(9): 34-38.

[1]	王开芳, 吴德军, 臧真荣, 翟国峰, 王守国, 胡丁猛, 任飞. 国槐良种'鲁槐1号’[J]. 林业科学, 2014, 50(9): 189-189.
[2]	卜庆梅;柏新富;朱建军. 4种园林树木叶绿素荧光参数对脱水的响应及耐旱特性分析[J]. 林业科学, 2011, 47(10): 37-43.
[3]	张晓英甘敬尹伟伦朱祯王华芳. 国槐转雪花莲凝集素基因及抗蚜性[J]. 林业科学, 2010, 46(2): 51-55.
[4]	张晓英.甘敬尹伟伦朱祯王华芳. 国槐遗传转化体系的优化*[J]. 林业科学, 2009, 12(5): 20-26.
[5]	邱延昌张秀省黄勇杨重军于守超苗中芹. 国槐新品种‘聊红’槐[J]. 林业科学, 2008, 44(5): 173-173.
[6]	张川红沈应柏尹伟伦潘青华赵毓桂. 盐胁迫对几种苗木生长及光合作用的影响[J]. 林业科学, 2002, 38(2): 27-31.
[7]	冷平生马世超李树蓉郭平. 增施CO₂气肥对国槐幼苗生长与生理特性的影响[J]. 林业科学, 2002, 38(1): 44-49.
[8]	薛皎亮谢映平李艳芳. 城市污染对国槐树体VB₂含量及瘤坚大球蚧种群影响的研究[J]. 林业科学, 2001, 37(2): 69-73.
[9]	唐燕平刘桂华. 锈色粒肩天牛卵期预测预报的研究[J]. 林业科学, 2000, 36(6): 86-89.
[10]	谢映平薛皎亮刘红霞刘贤谦李景平. 城市空气污染物SO_2和Pb在大球蚧及其寄主国槐体内积累的研究[J]. , 1998, 34(1): 45-49.

国槐离体再生及抗虫基因sck的转导

In Vitro Regeneration and Agrobacterium Transformation of Sophora japonica with sck Gene

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 10

编辑推荐

Metrics

本文评价

作者中心

期刊获奖

引证指标

联系方式