摘要
蓖麻种子大小因品种不同差异很大,目前尚未见与蓖麻种子大小性状发育相关的分子生物学研究。以32个不同大小的蓖麻种子作为研究材料,用U25(55)均匀试验设计法对种子大小性状的RAPD-PCR反应体系和条件进行优化。最佳反应体系为:25μL体系中10×Buffer 2.7μL、d NTP 2.4μL、Mg2+2.4μL、DNA 1.7μL、引物1.9μL、Taq polymerase 2.0μL、dd H2O 11.9μL。最佳反应条件为:94℃150 s;94℃45 s,39℃65 s,72℃90 s,35个循环;72℃10 min;4℃保存。用SB-002引物(碱基序列(5'-3'):TGCCGAGCTG),采用最佳的反应体系和条件在最大粒蓖麻种子中筛选到一个差异条带,测序结果在NCBI中进行比对,发现与高产粳稻基因组DNA、mRNA或c DNA同源性达到100%,因此推断所获差异条带与蓖麻种子产量有关,为蓖麻种子大小性状相关分子机理研究奠定了基础。
Castor seed size varies greatly due to breed,has yet to see molecular biology research on castor bean size traits associated with the development. In this experiment,32 castor seeds of different sizes as research material,with U25（ 55） uniform design for seed size traits RAPD-PCR reaction system and conditions were optimized. The optimal reaction system： 25 μL system,10 × Buffer 2. 7 μL,d NTP 2. 4 μL,Mg2 ＋2. 4 μL,DNA 1. 7 μL,primers 1. 9μL,Taq polymerase 2. 0 μL,dd H2 O 11. 9 μL. The best reaction conditions were： 94 ℃ 150 s; 94 ℃ 45 s,39 ℃65 s,72 ℃ 90 s,35 cycles; 72 ℃ 10 min; 4 ℃ preservation. With SB-002 primer（ nucleotide sequence（ 5＇-3＇） ：TGCCGAGCTG）,using the best reaction system and conditions,screening a different band in the largest grain castor seeds,sequencing results were compared in the NCBI found with high yield rice genome DNA,mRNA or cDNA homology of 100%. Therefore,It can be concluded that the different band relevant to castor seed yield,laid the foundation for the relevant molecular mechanisms of castor seed size traits studied.
出处
《华北农学报》
CSCD
北大核心
2015年第5期108-114,共7页
Acta Agriculturae Boreali-Sinica
基金
国家自然科学基金项目(30760123
31160290)
国家农业部公益性行业(农业)科研专项(201003057)
内蒙古自治区高等学校"青年科技英才支持计划"(NJYT-14-A10)
市校合作项目(SXZD2012018)
内蒙古自治区高校蓖麻产业工程技术研究中心开放基金项目(BMYJ2011009)
