• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>British Biotechnology Journal >Establishment of Agrobacterium-MediatedTransformation System in Sweet Potato(Ipomoea batatas) by Culture of Leaf Segmentsfor Functional Analysis of ASG-1, an Apomixis-Specific Gene
【24h】

Establishment of Agrobacterium-MediatedTransformation System in Sweet Potato(Ipomoea batatas) by Culture of Leaf Segmentsfor Functional Analysis of ASG-1, an Apomixis-Specific Gene

机译：通过叶节的培养建立农杆菌介导的甘薯（Ipomoea batatas）转化系统，以分析无融合生殖特异基因ASG-1

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

摘要

Aims: In order to do the functional analysis of apomixis-specific gene (ASG-1), which was isolated from apomictic guineagrass, the sweet potato was used to establish an Agrobacterium-mediated transformation system.Study Design: At first, plant regeneration was achieved from the culture of leaf segments of sweet potato. Based on it, a binary vector pSMA35H2-NG for transformation of ASG-1 was used for establishment of a suitable procedure for plant regeneration of transformants. Place and Duration of Study: Faculty of Environmental and Horticultural Science, Minami Kyushu University, between June 2009 and December 2012.Methodology: The leaf segments were used for somatic embryogenesis and plantlets regeneration. For the preliminary transformation, a GUS gene set in pSMA35H2-NG was introduced into the Agrobacterium strain GV3101/PMP9, and the Agrobacterium was used to infect the callus derived from leaf segments of sweet potato “Miyazakibeni” and the callus derived from seeds of rice “Nipponbare”. For the plasmid construction, the GUS was replaced by ASG-1, named as pSMA35H2/ASG1. The resultant plasmid was mobilized into Agrobacterium strain GV3101/PMP9 for transformation. For detection of ASG-1, DNAs of the transgenic plantlets were used for PCR, using the primers designed according to ASG-1 and hygromycin, respectively. Results: 1) When the leaf segments were sterilized with sodium hypochlorite solution of 0.3% and 0.4% for 15 min, 100% of surviving rates was achieved. And the segments cultured on Murashige and Skoog (1962) gave 100% of callus formation rates. 2) When the calli were placed onto Komamine and Nomura (1998) medium for differentiation, somatic embryogenesis was obtained with white color and grain-like tissue, and plantlets with multiple shoot-like tissues were obtained from the somatic embryo. 3) For the preliminary transformation, the calli showed GUS blue spots gradually on the surface. 4) When the pSMA35H2/ASG1 was used to the transformation of the embryogenic calli, the plantlets were developed through multiple shoots. 5) The specific bands of ASG-1 and hygromycin were observed from the PCR products of the plantlets’ DNAs, respectively. Conclusion: Overall the above results, the procedure using the binary vector pSMA35H2/ASG1 containing ASG-1 revealed, as the first case, that Agrobacterium-mediated transformation system in sweet potato was established using the culture of leaf segments in this study.
机译：目的：为了对从无融合生殖的豚鼠中分离的无融合生殖特异基因（ASG-1）进行功能分析，将甘薯用于建立农杆菌介导的转化系统。研究设计：首先，进行植物再生从甘薯叶段的培养中获得。在此基础上，将用于ASG-1转化的二元载体pSMA35H2-NG用于建立合适的转化体植物再生程序。研究地点和持续时间：南九州大学环境与园艺科学学院，2009年6月至2012年12月。方法：将叶段用于体细胞胚发生和小植株再生。为了进行初步转化，将pSMA35H2-NG中设置的GUS基因导入农杆菌菌株GV3101 / PMP9中，并用农杆菌感染源自番薯“ Miyazakibeni”叶节的愈伤组织和来源于水稻种子的愈伤组织。 “日本ippo”。对于质粒构建，GUS被ASG-1取代，命名为pSMA35H2 / ASG1。将所得质粒动员到农杆菌菌株GV3101 / PMP9中进行转化。为了检测ASG-1，分别使用根据ASG-1和潮霉素设计的引物，将转基因植株的DNA用于PCR。结果：1）将叶段分别用0.3％和0.4％的次氯酸钠溶液灭菌15分钟，存活率达到100％。在Murashige和Skoog（1962）上培养的片段给出了100％的愈伤组织形成率。 2）将愈伤组织置于Komamine和Nomura（1998）培养基上进行分化时，获得了具有白色和颗粒状组织的体细胞胚发生，并且从体细胞胚中获得了具有多个芽状组织的小植株。 3）对于初步转化，愈伤组织在表面上逐渐显示出GUS蓝点。 4）当将pSMA35H2 / ASG1用于胚发生愈伤组织的转化时，通过多次芽使小植株发育。 5）从小苗DNA的PCR产物中分别观察到ASG-1和潮霉素的特异性条带。结论：总的来说，以上结果表明，使用含ASG-1的二元载体pSMA35H2 / ASG1进行的程序揭示，作为第一种情况，本研究利用叶段的培养建立了农杆菌介导的甘薯转化体系。

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