焦朝霞;邵建柱;孙建设;

• 1:河北农业大学园艺学院

摘要(Abstract)：

针对田间苹果成熟组织富含多酚、多糖和大量次生代谢产物的特点,建立了适合田间苹果不同组织的高效RNA提取方法,以不同季节田间生长的苹果叶片和皮层为材料,用此方法均能在2 h左右完成超过10个样品的总RNA提取,质量高、完整性好的总RNA,可用于田间苹果病毒RT-PCR检测,获得良好效果。经反复大量试验证实,该方法适合田间苹果不同组织的RNA快速提取,操作简便、快捷、高效,且使其不受季节限制,有效地提高了田间果树病毒检测以及其他分子生物学研究的效率。

关键词(KeyWords)： 田间苹果;成熟组织;RNA;高效提取

Abstract:

Keywords:

基金项目(Foundation): 农业部公益性行业科研专项(nyhyzx07-024);农业部国家现代苹果产业技术体系功能岗位专家项目(08-01-02-03)

作者(Author): 焦朝霞;邵建柱;孙建设;

Email:

DOI:

参考文献(References)：

• [1]RUAN Xiao-feng,WILLIELM J,YANG Yong.Improved RNA virusextraction from fruit trees by silica capture[J].Journal of Fruit Sci-ence,2004,21(4):388-390.阮小凤,WILLIELM J,杨勇.果树病毒RNA的二氧化硅提取法[J].果树学报,2004,21(4):388-390.
• [2]ZHAO Ying,NIU Jian-xin.Research on detection effect of fruit treeviruses using highly effective RNA isolation method designed[J].Biotechnology,2008,18(3):31-33.赵英,牛建新.自行设计高效RNA提取方法对果树病毒检测效果的研究[J].生物技术,2008,18(3):31-33.
• [3]WANG Zhuang-wei,QU Shen-chun,ZHANG Zhen,ZHANG Jun-yi.A fast method for total RNA extraction from the tissue culture mate-rial of Malus sp[J].Journal of Fruit Science,2004,21(4):385-387.王壮伟,渠慎春,章镇,张君毅.苹果属RNA高效快速提取新方法[J].果树学报,2004,21(4):385-387.
• [4]CHENG Jian-hong,ZHANG Yu-gang,LI Tian-zhong.Quick SDSmethod for RNA isolation from apple and other plant tissues withroom temperature Centrifugation[J].Acta Horticulturae Sinica,2006,33(3):470.成建红,张玉刚,李天忠.室温离心SDS法快速提取苹果等植物组织RNA[J].园艺学报,2006,33(3):470.
• [5]HOU Yi-long,ZHANG Kai-chun,WU Lu-ping,HU Wen-yu,LINke,FENG Xiao-yuan,XING Fu-you.New methods for total RNAextraction in fruit trees[J].Journal of Shenyang Agricultural Univer-sit,2002,33(2):122-125.侯义龙,张开春,吴禄平,胡文玉,林珂,冯晓元,邢福有.果树组织中总RNA提取的新方法[J].沈阳农业大学学报,2002,33(2):122-125.
• [6]HE Mei,LI Hai-sheng,ZHAO Ying,NIU Jian-xin,MA Bing-gang.A method of rapidly extracting total RNA from kurle aroma pear[J].Journal of Shihezi University,2004,22(6):474-476.何梅,李海生,赵英,牛建新,马兵刚.库尔勒香梨总RNA的快速提取方法[J].石河子大学学报,2004,22(6):474-476.
• [7]BAI Yun-feng,GUO Zhi-hua,BAI Dong-mei,WANG Xiao-qi,ZHANG Wei-feng.An improved method for extraction and identifi-cation of potato RNAs[J].Acta Horticulturae Sinica,2007,34(4):1059-1062.白云凤,郭志华,白冬梅,王小琦,张维锋.马铃薯总RNA提取和鉴定方法的改进[J].园艺学报,2007,34(4):1059-1062.
• [8]JAMES D.A simple and reliable protocol for the detection of applestem grooving virus by RT-PCR and in a multiplex PCR assay[J].Journal of Virological Methods,1999,83:1-9.
• [9]KUNDU J K.The occurrence of apple stem pitting virus and applestem grooving virus within field-grown apple cultivars evaluated byRT-PCR[J].Plant Protect Science,2003,39(3):88-92.
• [10]LI Hong,WANG Xin-li.The difficulties in the isolation of RNAfrom plant tissues and their resolving strategies[J].Biotechnologyinformation,1999(1):36-39.李宏,王新力.植物组织RNA提取的难点及对策[J].生物技术通报,1999(1):36-39.
• [11]YIN Hui,CHEN Li,LI Xiao-yan,CHEN Qiu-ming,YI Ming-fang.Analysis and improvement of high-quality RNA extraction in leavesof lily[J].Journal of China Agricultural University,2008,13(4):41-45.尹慧,陈莉,李晓艳,陈秋明,义鸣放.百合叶片总RNA提取方法比较及优化[J].中国农业大学学报,2008,13(4):41-45.