Rapid Detection of Bt Transgenic Sugarcane Using Root Tips by PCR

来源 :Agricultural Biotechnology | 被引量 : 0次 | 上传用户:cassyqc
下载到本地 , 更方便阅读
声明 : 本文档内容版权归属内容提供方 , 如果您对本文有版权争议 , 可与客服联系进行内容授权或下架
论文部分内容阅读
[Objective]This study aimed to establish an effective detection technology for rapidly identifying Bt transgenic sugarcane,promptly removing non-Bt transgenic plants and increasing breeding efficiency of Bt transgenic plants,thereby improving breeding efficiency of insect-resistant sugarcane cultivars in China.[Method]Approximately 1 mg of root tips and other materials were crushed,placed in the bottom of PCR tubes,added successively with solution I and mineral oil,headed at 95 ℃ for 15 min,added with solution II,and finally added with solution III containing GBt primer for PCR. After separated by using electrophoresis on agarose gel,amplification products were observed and photographed under a gel imaging system. [Result]Various materials treated with different concentrations of solution led to different amplification results. Specifically,the appropriate concentration of NaOH in solution I was 0. 1- 0. 2 mol /L; the appropriate pH value of solution II was 2. No bands were amplified from mature leaves,tender leaves and old roots of Bt transgenic sugarcane,while root tips successfully amplified 545bp target bands. The established method was used to identify and analyze 63 hybrid seedlings of YT 91- 976 × SBR216,results showed that a total of 17 samples exhibited 545 bp bands,accounting for approximately 27% of the total number of hybrid seedlings,indicating that these 17 seedlings were Bt transgenic sugarcane plants. [Conclusion]The rapid detection method established in this study is conducive to identifying Bt transgenic sugarcane and improving breeding efficiency of insect-resistant sugarcane lines,which provides basis for screening of other transgenic materials. [Objective] This study aimed to establish effective detection technology for rapidly identifying Bt transgenic sugarcane, promptly removing non-Bt transgenic plants and increasing breeding efficiency of Bt transgenic plants, so improving breeding efficiency of insect-resistant sugarcane cultivars in China. [Method ] Approximately 1 mg of root tips and other materials were crushed, placed in the bottom of PCR tubes, added successively with solution I and mineral oil, headed at 95 ° C for 15 min, added with solution II, and finally added with solution III containing GBt primer for PCR. After separated by using electrophoresis on agarose gel, amplification products were observed and photographed under a gel imaging system. [Result] Various materials treated with different concentrations of solution led to different amplification results. in solution I was 0. 1- 0. 2 mol / L; the appropriate pH value of solution II was 2. No bands were amplified from mature leaves, tender leaves and old roots of Bt transgenic sugarcane, while root tips successfully amplified 545 bp target bands. The established method was used to identify and analyze 63 hybrid seedlings of YT 91-976 × SBR216, results showed that a total of 17 samples exhibited 545 bp bands, accounting for approximately 27% of the total number of hybrid seedlings, indicating that these 17 seedlings were Bt transgenic sugarcane plants. [Conclusion] The rapid detection method established in this study is conducive to identifying Bt transgenic sugarcane and improving breeding efficiency of insect-resistant sugarcane lines, which provides basis for screening of other transgenic materials.
其他文献
本文通过对荣华二采区10
托妮?莫里森是第一位荣获“诺贝尔文学奖”的黑人女性作家。《家》是莫里森的第十部小说,于2012年出版。这部小说展现了20世纪50年代非裔美国黑人的命运。这部小说以种族主义
关联理论是一种语言交际理论,它给翻译提供了一个理论框架:翻译是一个对原语进行阐释的明示-推理过程,译者根据译文读者的期待,然后再把自己对原文作者交际意图的了解、译文语言
全球化语境的大背景下,社会各行业对翻译人才特别是口译人才的需求日益增加,单一的英语专业和基础技能型人才已不能完全满足国际社会发展的需求,传统的翻译教学理念和教学模式也
我国是农业大国,但是种植方法设备简单,管理技术品种相对落后;传统的棚内的湿度、灌溉等均是通过人工控制的,不可避免有测控精度低、劳动强度大及由于测控不及时等弊端,容易
该文就生猪养殖场粪污排放情况及对环境污染作了简要介绍,对生猪养殖环境污染的治理进行了分析,为生态养殖生猪提供一些借鉴。
名称是特殊的语言符号,也是典籍的重要组成部分。在典籍英译中,名称的译法选择关系到文化的传播效果。本文运用传播学和符号学理论描述了名称英译的编码解码过程,论证了名称的符
针对目前世界各国正竞相研究的第三次网络新浪潮——网格技术 ,阐述了网格的基本概念、技术特征以及网格应用前景和今后主要研究方向 ,讨论了生物研究和军事领域对网格技术的
传统童话起源于民间传说,反映了人类集体的记忆和人类生存的经验。但是和其它传统文学一样,许多传统童话被打上父权文化的烙印。童话故事中的男女主人公形象正符合了父权社会对
以1989年、1994年、2001年和2010年莆田东圳水库周边地区遥感数据为基础,利用ERDAS软件生成研究区的景观类型图,计算分析东圳水库周边地区的土地利用动态变化。结果表明:研究