现代农业科技
現代農業科技
현대농업과기
XIANDAIHUA NONGYE
2013年
23期
21-24
,共4页
赵莉%孔小卫%汪强%林勇翔%田东丰
趙莉%孔小衛%汪彊%林勇翔%田東豐
조리%공소위%왕강%림용상%전동봉
芝麻%保持系%不育株%RAPD反应体系%建立
芝痳%保持繫%不育株%RAPD反應體繫%建立
지마%보지계%불육주%RAPD반응체계%건립
sesame%maintainer line%infertile plant%RAPD reaction system%optimization
以临时保持系WB51220、两用系0176A不育株、可育株0176B和皖芝1号等4份材料的叶片为研究材料,对其基因组DNA的提取及对RAPD反应体系进行优化。结果表明,改良的CTAB法获得的芝麻基因组DNA片段大小经电泳检测满足RAPD等遗传多样性分析要求;筛选出RAPD最佳反应体系:20μL反应体系含有1.5 U Taq DNA聚合酶,0.25 mmoL/L dNTP,2.0 mmoL/L Mg2+,0.75μmoL/L引物;4个不同品种的叶片基因组DNA的电泳条带之间有着明显差异,他们共有的条带为1900、1800、1000、900 bp;不同的是,临时保持系WB51220比两用系0176A不育株少了1个2000 bp的条带,可育株0176B比两用系0176A不育株少了1个450 bp条带,皖芝1号比可育株0176B少了1个750 bp和1个1200 bp的条带。
以臨時保持繫WB51220、兩用繫0176A不育株、可育株0176B和皖芝1號等4份材料的葉片為研究材料,對其基因組DNA的提取及對RAPD反應體繫進行優化。結果錶明,改良的CTAB法穫得的芝痳基因組DNA片段大小經電泳檢測滿足RAPD等遺傳多樣性分析要求;篩選齣RAPD最佳反應體繫:20μL反應體繫含有1.5 U Taq DNA聚閤酶,0.25 mmoL/L dNTP,2.0 mmoL/L Mg2+,0.75μmoL/L引物;4箇不同品種的葉片基因組DNA的電泳條帶之間有著明顯差異,他們共有的條帶為1900、1800、1000、900 bp;不同的是,臨時保持繫WB51220比兩用繫0176A不育株少瞭1箇2000 bp的條帶,可育株0176B比兩用繫0176A不育株少瞭1箇450 bp條帶,皖芝1號比可育株0176B少瞭1箇750 bp和1箇1200 bp的條帶。
이림시보지계WB51220、량용계0176A불육주、가육주0176B화환지1호등4빈재료적협편위연구재료,대기기인조DNA적제취급대RAPD반응체계진행우화。결과표명,개량적CTAB법획득적지마기인조DNA편단대소경전영검측만족RAPD등유전다양성분석요구;사선출RAPD최가반응체계:20μL반응체계함유1.5 U Taq DNA취합매,0.25 mmoL/L dNTP,2.0 mmoL/L Mg2+,0.75μmoL/L인물;4개불동품충적협편기인조DNA적전영조대지간유착명현차이,타문공유적조대위1900、1800、1000、900 bp;불동적시,림시보지계WB51220비량용계0176A불육주소료1개2000 bp적조대,가육주0176B비량용계0176A불육주소료1개450 bp조대,환지1호비가육주0176B소료1개750 bp화1개1200 bp적조대。
The genomic DNA extraction and the RAPD-PCR reaction conditions were optimized when using genomes from sesame leaves of temporary maintainer line WB51220,AB Line infertile plant 0176A,fertile plant 0176B,and Wanzhi No.1 as templates. The results indicated that the genomic DNAs extracted from sesame leaves by improved CTAB DNA extraction method satisfied the requirement of genetic diversity analysis. The best reaction condition for RAPD analysis was as follows:Taq DNA polymerase 1.5 U,dNTP 0.25 mmoL/L,Mg2+2.0 mmoL/L,and random primer 0.75μmoL/L in a total volume of 20μL. Electrophoresis results indicated that bands amplified from the genomic DNAs of four varieties had four common bands in length of 1900,1800,1000,and 900 bp,respectively. The differences were that new bands in length of 2000 bp and 450 bp were amplified from AB line infertile plant 0176A when compared with temporary maintainer line WB51220 and fertile plant 0176B,respectively. And two new bands in length of 750 bp and 1200 bp were amplified from fertile plant 0176B when compared with Wanzhi NO.1.