摘要
为探讨蓖麻果刺性状相关分子机理,以9个果实有刺和5个果实无刺的蓖麻为研究对象,对蓖麻果刺性状进行RAPD分析,结果表明:用单因素法优化后的蓖麻果刺相关性状的RAPD-PCR反应体系为Taq polymerase0.9μL、10×Buffer 2.5μL、dNTP 2.5μL、Mg2+1.3μL、SBS126号Primer 1.4μL、DNA 1.4μL、ddH2O 15.0μL,Total25μL;优化后的反应条件为94℃4 min 30 s;94℃45 s,38℃45 s,72℃45 s,35个循环;72℃5 min;4℃保存。利用优化后的反应体系与反应条件,在果实有刺材料中得到了约1 800 bp的片段,测序结果表明,9个材料的条带上游同源性非常差,但是距离下游约28 bp之前有长度为112 bp的序列完全相同,推断蓖麻果实有刺性状的发育可能与包含组氨酸的磷酸转移蛋白2有关。
We selected nine fruit thorns and five fruit non-thorns of castor as the research object using the RAPD marker for the thorn traits analysis. The results were as follows：the RAPD-PCR reaction system with opti-mized by single factor was 0. 9 μL of Taq polymerase,2. 5 μL of Taq polymerase 10 ＆#215; Buffer,2. 5 μL of each dNTP,1. 3 μL of Mg2＋,1. 4 μL of 10 pmol/L primer SBS126,1. 4 μL of DNA,ddH2 O 15. 0 μL,Total 25 μL;And the related amplification condition was 1 cycle of 4 min 30 s at 94 ℃;35 cycles of 45 s at 94 ℃,45 s at 38 ℃and 45 s at 72 ℃,followed by a final cycle of 5 min at 72 ℃,then save in 4 ℃. Using the optimized reaction system and condition,several 1 800 bp fragments amplified by SBS126 was found in all the fruit thorn material,and sequen-cing results showed that the upstream homology of fragment was very low,whereas the downstream has the identical sequence with about 112 bp length. Basing on these results,we deduced that the development of fruit thorn may be associated with histidine-containing phosphotransfer protein 2,and this laid a foundation for research on the molecu-lar mechanism of fruit thorn traits for castor.
出处
《华北农学报》
CSCD
北大核心
2014年第1期83-88,共6页
Acta Agriculturae Boreali-Sinica
基金
国家自然科学基金项目(30760123
31160290
31060194)
国家民委项目(10NM02)
内蒙古自然科学基金项目(2010BS0511)
内蒙古人才基金项目
内蒙古民族大学市校合作项目(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)
