• 主题
高级搜索  >
历史搜索 清空历史
首页> 中文期刊> 《草业学报》 >农杆菌介导Chi-linker-Glu融合基因和bar基因转化玉米茎尖的研究

农杆菌介导Chi-linker-Glu融合基因和bar基因转化玉米茎尖的研究

         
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Most maize disease are caused by fungi pathogens, such as northern leaf blight, southern leaf blight, ear and stalk rots, and so on. In normal years, this disease makes maize yield lost by 10% and this percentage may be increased by 30%-40% in its popular years. However, it is very difficult for conventional breeding to develop an elite maize material with the resistance to all the fungi disease. Therefore, we can use transgenic method to transfer some genes related to the resistance of these disease into maize in order to obtain resistance materials. Chitinase and β-1,3-glucanase are two important enzymes which play a key role in the hydrolysis reaction of fungal cell wall, so it is useful in the control of fungi disease, meanwhile, the herbicide resistant gene bar are usually used as a selectable marker in maize transformation. The purpose of this research is to introduce the fused gene of chitinase and β-1,3-glucanase and bar as well into elite maize inbred Zheng58 for improving its resistance to fungi disease. The wounded shoot apical meristem (SAM) of germinated seedlings were used as the material for Agrobacterium mediated transformation. We report here: 1) An optimized transformation system for Agrobacterium mediated transformation of shoot apical meristem, using the optimal concentration of bacterial culture (the value of OD600 was 0. 6) for infection, addition of 150 μmol/L acetosyringone (AS) in the bacterial suspension, the whole infection process was carried out in a vacuum desiccators with a negative pressure of 50 kPa for 12 minutes. 2) Identified by herbicide screening procedure and PCR detection, 13 transformed plants were obtained among the 32 herbicide-resistant plants, and the overall transformation was 2. 6%. The preliminary evidences showed that the foreign genes had been introduced into the maize genome. This method circumvented the long period of tissue culture step and limitation of different seasons, in addition, many elite inbred lines which are recalcitrant in callus induction can be efficiently transformed by this method.%真菌性病害和田间杂草严重影响着玉米产量及饲用品质,几丁质酶和β-1,3葡聚糖酶催化病原真菌细胞壁的水解反应,二者协同作用能够有效地抑制病原真菌生长.为了获得兼具真菌病及除草剂抗性的玉米种质新材料,本研究通过对农杆菌介导的玉米茎尖遗传转化体系的优化,将几丁质酶、β -1,3葡聚糖酶的融合基因以及bar基因导入优良玉米自交系郑58,用200 mg/L的Basta除草剂进行筛选,并对抗性植株进行了PCR检测.结果表明,LBA4404菌液浓度((OD600值)为0.6、乙酰丁香酮浓度为150 μmol/L、50 kPa负压侵染12 min为最佳的遗传转化条件；对除草剂筛选后的32株抗性植株进行PCR鉴定,有13株为阳性植株,初步鉴定结果表明目的基因已经整合进玉米基因组中.

著录项

  • 来源
    《草业学报》 |2012年第5期|69-76|共8页
  • 作者

    方永丰; 李永生; 彭云玲; 王芳; 王威; 穆延召; 王汉宁;

  • 作者单位

    甘肃省作物遗传改良与种质创新重点实验室,甘肃兰州730070;

    甘肃省干旱生境作物学重点实验室,甘肃兰州730070;

    甘肃农业大学农学院,甘肃兰州730070;

    甘肃省干旱生境作物学重点实验室,甘肃兰州730070;

    甘肃农业大学农学院,甘肃兰州730070;

    甘肃农业大学农学院,甘肃兰州730070;

    甘肃农业大学农学院,甘肃兰州730070;

    甘肃省干旱生境作物学重点实验室,甘肃兰州730070;

    甘肃省作物遗传改良与种质创新重点实验室,甘肃兰州730070;

    甘肃省干旱生境作物学重点实验室,甘肃兰州730070;

    甘肃农业大学农学院,甘肃兰州730070;

  • 原文格式 PDF
  • 正文语种 chi
  • 中图分类 饲料;植物基因工程;
  • 关键词

    玉米; 真菌性病害; 融合基因; 农杆菌介导;

相似文献

  • 中文文献
  • 外文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号