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园艺学报 ›› 2008, Vol. 35 ›› Issue (8): 1209-1214.

• 研究简报 • 上一篇    下一篇

萝卜中一种新dsRNA的全长cDNA克隆及序列分析

李露露1;李力强1;乔爱民2;陈亮1;陈集双1*   

  1. ( 1浙江理工大学生物工程研究所 杭州 310018;2仲恺农业技术学院农业与园林学院 广州 510225)
  • 收稿日期:2008-03-04 修回日期:2008-05-27 出版日期:2008-08-25 发布日期:2008-08-25
  • 通讯作者: 陈集双

Full-length cDNA Clone and Sequence Analysis of A New Double-stranded RNA from Radish

LI Lu-lu1, LI Li-qiang1, QIAO Ai-min2,CHEN Liang1, and CHEN Ji-shuang1*   

  1. (1Institute of Bioengineering, Zhejiang Sci-Tech University, Hangzhou 310018 China; 2College of Agriculture and Landscape Architecture, Zhongkai University of Agriculture and Technology, Guangzhou, 510225 China)
  • Received:2008-03-04 Revised:2008-05-27 Online:2008-08-25 Published:2008-08-25
  • Contact: CHEN Ji-shuang

摘要: 从一点红萝卜品种(Raphanus sativus-root cv. Yidianhong)叶片中提取dsRNA,应用SPAT方法对各dsRNA条带进行cDNA克隆并测序,除获得先前报道的5条序列(RasR1-RasR5)以外,还得到一条新的全长序列,将其定名为RasR 6(EU285027)。序列分析结果表明:RasR 6全长为1 778 bp,其正义链编码1个由502个氨基酸组成、分子量约为55.1 kDa的蛋白质。该序列与前人报道的双分病毒科5个病毒序列具有相似性,且它们均编码双分病毒科病毒的外壳蛋白(CP,Capsid protein)。核苷酸序列比对结果显示:RasR 6与同时来源于萝卜的RasR 1和RasR 2的5'UTR序列高度同源,且其3'末端具有poly A结构,而与RasR 3、RasR 4和RasR 5的UTR则没有明显的相似性。因此,我们推测RasR 6与RasR 1、RasR 2同属于双分病毒RasV 1(Raphanus sativus virus 1),它可能与RasR 2共同编码该病毒的CP或作为RasV 1的卫星RNA存在。

关键词: 萝卜, 双链RNA, 双分病毒科, Raphanus sativus virus 1

Abstract:

Double stranded RNA (dsRNA) were extracted from leaf tissues of radish (Raphanus sativus-root cv. Yidianhong) grown in Hangzhou China. Complementary DNAs corresponding to the dsRNA segments were obtained by using a modified single primer amplification technique (SPAT), and followed by cloning and sequencing. The results revealed that a new dsRNA sequence, different from all the previously reported sequences RasR1-RasR5, was obtained. And the newly detected RasR 6 (EU285027) was found to be 1778 bp in length. It predicted that the positive strand of RasR 6 potentially encoded a protein of about 55.1 kDa, containing 502 amino acids. Five members of Partitiviridae, all of which encoded putative viral CPs, were found to have some identity with RasR 6. Multiple alignments showed that RasR 6 were highly conserved at the 5(UTR regions with RasR 1 and RasR 2 previously reported from the same radish varities, but had no similarity with RasR 3-RasR 5. Thus, it could be presumed that both RasR 6, RasR 1 and RasR 2 should belong to Raphanus sativus virus 1(RasV 1), and RasR 6 encods CP together with RasR 2, or is a satellite dsRNA of this virus.

Key words: Raphanus, double-stranded RNA, Partitiviridae, Raphanus sativus virus 1

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