中国农业科学 ›› 2007, Vol. 40 ›› Issue (5): 1079-1084 .

• 研究简报 • 上一篇    

SCAR分子标记方法检测家蚕品种和纯度的研究

唐 斌,徐世清,戴璇颖,陈息林,司马杨虎,王玉军,吴阳春   

  1. 苏州大学生命科学学院
  • 收稿日期:2005-04-29 修回日期:1900-01-01 出版日期:2007-05-10 发布日期:2007-05-10
  • 通讯作者: 徐世清

Study on the Method of SCAR to Identify the Variety and Purity of Silkworm, Bombyx mori L.

  

  1. 苏州大学生命科学学院
  • Received:2005-04-29 Revised:1900-01-01 Online:2007-05-10 Published:2007-05-10

摘要: 【目的】家蚕(Bombyx mori L.)品种和纯度的鉴别,普遍依靠幼虫斑纹或茧形等形态鉴别方法,该方法易受环境影响,结果时效性和准确性差。【方法】筛选RAPD随机引物,稳定扩增出亲本品种的DNA特异性片段,转换成SCAR标记。 【结果】利用微量DNA抽提法,从家蚕主要品种苏5、苏6及其F1孵化前日卵快速提取单粒卵基因组DNA,筛选出RAPD随机引物S42,稳定扩增出苏5和苏6两个亲本品种的DNA特异性片段R5和R6,克隆和测序确认其大小分别为255bp和343bp。Blastn分析显示,R5的nt7~192与家蚕的一个非长末端逆转录转座子(AB002270.1)的nt183~368片段碱基序列有高达91%的相似性,无缺口序列。R6的nt5~340与家蚕的一个WGS(BAAB01113163.1)的nt675~337有91%的一致性,其中存在13个碱基的缺口序列。将2个亲本的RAPD标记转换成SCAR标记,分别利用合成的S42-255-2(P5-2)和S42-343-2(P6-2)SCAR标记引物,能够准确、快速地鉴别所标记的苏5和苏6及其F1蚕品种。【结论】SCAR分子标记方法能够检测家蚕品种和纯度。

关键词: 家蚕, 分子标记, SCAR, 品种, 纯度

Abstract: Sericulture production is used to the first filial generation. The rule method to identify silkworm variety and purity with morphological characters of larval pattern or cocoon shape is easily affected by environment. Thus,the prescription and accuracy is low. The genome DNA of silkworm was extracted from single blue-stage-egg of mainly silkworm varieties in JiangSu province of SU5, SU6 and their F1with rapid method. The specific bands R5 and R6 of RAPD were amplify traced each from of SU5 and SU6 variety, with selected random primer of S42. The sequences were 255bp and 343bp respectively after cloned and sequenced. Then analyze the sequences by courtesy of database on-line. The results showed the nt7-192 of R5 and the nt183-368 of the non-LTR retrotransposon of silkworm has 91% consistency. Accordingly, 91% of nt675-337 of one of whole genome shotgun sequence(WGS)of silkworm shows the same sequence as the nt5-340 of R5, which include 13 gaps. Further, transform the RAPD mark of two parental varieties to SCAR mark. Thus, SU5, SU6 and their F1 were identified accurately and fast, with the synthetic specific primers of S42-255-2(P5-2)and S42-343-2(P6-2).

Key words: Silkworm (Bombyx mori L.), Molecular mark, SCAR (sequence characterized amplified regions), Variety, Purity