摘要
建立适于荔枝的SCoT反应体系,并利用SCoT体系对‘紫娘喜’、‘无核荔’及其杂交后代进行真假杂种鉴定和遗传多样性分析。采用正交设计方法对影响荔枝SCoT-PCR反应的rTaq酶、Mg2＋浓度、模板DNA用量、dNTPs浓度和引物等因素进行体系优化。优化的荔枝SCoT反应体系为：20μL反应体系中含有模板DNA 30 ng,rTaq酶1 U,引物0.75μmol/L,Mg2＋3 mmol/L,dNTPs 0.3 mmol/L。最适退火温度为50.6℃。利用该体系对‘紫娘喜’、‘无核荔’及其杂交后代进行遗传分析,结果显示,32条引物共扩增出280个位点,多态性位点有131个,占总位点数46.78%。4条引物就可以将60株杂交后代鉴定出来。UPGMA聚类分析显示,SCoT标记能将30株杂交后代区分开,遗传相似系数在0.406-0.867,在相似系数0.68时,30株杂交后代可聚为3类。优化建立的荔枝SCoT体系结果稳定,重复性好,为荔枝遗传育种提供了新的技术支持。
The present study was aimed to establish and optimize SCoT-PCR amplification system of litchi,and conduct the identification and genetic diversity analysis of‘Ziniangxi＇,‘Wuheli＇, and their hybrids.Orthogonal design was applied to optimize litchi SCoT-PCR amplification system in five factors, includingrTaq DNA polymerase, Mg2 ＋, DNA template, dNTPs and primers. The optimum reaction system（20 μL）contained 30 ng template DNA, 1 U rTaq polymerase, 0.75 μmol/L primers, 3 mmol/LMg2＋and 0.3 mmmol/LdNTPs. The most suitable annealing temperature of primers was 50.6℃. Genetic analysis of‘Ziniangxi＇, ‘Wuheli＇, and their hybrids were carried out using this SCoT-PCR system. A total of 280 loci were obtainedusing 32 SCoT primers and 131 loci were polymorphic. The real hybrids could be identified by SCoT. 60hybrids can be identified by four SCoT primers. UPGMA cluster analysis and genetic similarity coefficientshowed that the similarity coefficient of 30 hybrids ranged from 0.406 to 0.867, and 30 progenies could begrouped into three distinct families based on similarity of 0.68, indicating that it possessed rich geneticdiversity. The optimum SCoT-PCR system of litchi was stable and very useful for genetic diversity analysis oflitchi, which would possess considerable potential for litchi breeding.
出处
《中国农学通报》
CSCD
2014年第13期147-156,共10页
Chinese Agricultural Science Bulletin
基金
国家现代农业产业技术体系建设专项资金资助项目"国家荔枝龙眼产业技术体系"(CARS-33-04)
