• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Plant Biotechnology >A high-throughput Agrobacterium tumefaciens-mediated transformation system for molecular breeding and functional genomics of rice (Oryza sativa L.)
【24h】

A high-throughput Agrobacterium tumefaciens-mediated transformation system for molecular breeding and functional genomics of rice (Oryza sativa L.)

机译：高通量根癌农杆菌介导的水稻分子育种和功能基因组学转化系统

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

摘要

Agrobacterium tumefaciens -mediated transformation is the preferred method for genetic transformation of rice. Here we provide a comprehensive and high-through-put protocol for Agrobacterium -mediated transformation of rice for generating the large numbers of transgenic plants that are required for functional analysis and for evaluating traits of agronomic importance. In a project designed to produce nitrogen use efficient rice plants, we refined/improved several factors including optimization of conditions for inducing vir genes prior to transformation, infection and co-cultivation medium for better interaction of target tissues with Agrobacterium . These optimizations supported not only the survival of co-cultured calli in high frequency, but also the production of multiple resistant cell lines per co-cultured embryogenic and nodular callus. In addition, improvements in plant tissue culture, selection and regeneration media has enabled us to produce large numbers of transgenic rice plants containing genes of agronomical importance. Partial desiccation and ABA treatment to hygromycin-resistant callus tissue significantly ( P <0.05) enhanced both regeneration frequency and regeneration of transgenic plantlets per callus tissue. Regeneration frequency was further improved by optimizing the concentration of copper sulphate in the regeneration medium. The majority of the transgenic plants obtained using this improved protocol displayed a normal phenotype and the gene of interest was inherited by the progeny in a Mendelian fashion. Homozygous plants of selected lines showed stable phenotypes both in soil and hydroponic conditions even after five generations. Availability of such a high-throughput Agrobacterium -mediated transformation system will improve future opportunities for rice genetics and functional genomics study.
机译：根癌农杆菌介导的转化是水稻遗传转化的优选方法。在这里，我们提供了一种用于农杆菌介导的水稻转化的全面且高通量的方案，用于产生功能分析和评估农艺学重要特征所需的大量转基因植物。在一个旨在利用氮素高效利用水稻植物的项目中，我们完善/改进了几个因素，包括优化转化前诱导vir基因的条件，感染和共同培养培养基，以使目标组织与农杆菌更好地相互作用。这些优化不仅支持共培养的愈伤组织在高频率下的存活，而且还支持每个共培养的胚性和结节性愈伤组织产生多个抗性细胞系。此外，植物组织培养，选择和再生培养基的改进使我们能够生产出大量含有农艺学重要基因的转基因水稻。对潮霉素抗性愈伤组织进行部分干燥和ABA处理可显着提高（P <0.05），提高了每个愈伤组织的再生频率和转基因植株的再生。通过优化再生介质中硫酸铜的浓度，进一步提高了再生频率。使用该改进方案获得的大多数转基因植物表现出正常的表型，并且目的基因以孟德尔方式被子代遗传。所选品系的纯合植物即使在五代后在土壤和水培条件下也表现出稳定的表型。这种高通量农杆菌介导的转化系统的可用性将为水稻遗传学和功能基因组学研究提供更多机会。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 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