摘要
选用国家柿种质资源圃中24个牛心柿和22个野柿为实验材料，采用L16（4^5）正交试验设计，建立了最优SCoT-PCR反应体系（20μL反应体系中，含Mg^2＋ 2．0mmol／L，dNTPs0．2mmol／L，Taq聚合酶l.0U，引物0．5μmol／L，DNA模板80ng），并对不同引物的退火温度进行了优化，以探讨SCoT标记在柿亲缘关系鉴定和品种分类方面的应用。结果显示：（1）10条SCoT引物对46份材料共扩增出176个位点，其中多态性位点170个，多态率达96．6％；46份材料的遗传相似系数在0．619～O．852之间。（2）UPGMA聚类分析显示，SCoT标记能将46份材料完全区分开，其中：红花雄野毛柿、红花雌野毛柿单独被聚为一组，独立于牛心柿和野柿之外，与形态特征观察结果一致；璧山牛心柿与野柿聚为一组，在形态特征上璧山牛心柿也更趋于野柿，且璧山牛心柿为四倍体，与其他牛心柿（六倍体）不同，璧山牛心柿应属于野生柿。研究表明，SCoT标记是研究柿种质资源遗传多样性的有效手段；46份材料存在着较高的遗传多样性，在一定程度上反映了品种的地理分布情况，也说明地理因素对柿的亲缘关系有很大影响。
In this experiment, a L16 （4s）orthogonal design was used for the optimal SCoT-PCR reaction sys- tem with 24 Niuxinshi and 22 Wild persimmon accessions from National Field Genebank for Persimmon. The optimal PCR reaction mixture of SCoT-PCR system was 2.0 mmol/L Mg^2＋ ,0.2 mmol/L dNTPs, 1.0 U Taq polymerase,0. 5 μmol/L primer and 80 ng template DNA in 20 μL mixtures. Furthermore,the an- nealing temperature of each primer was optimized, to explore relative identification and variety classification of persimmon with SCoT markers. The results showed that： （1）A total of 176 loci were detected with 10 SCoT primers in 46 accessions, 170 of which were polymorphic with a polymorphism of 96.6%. The genetic similarity ranged from 0. 619 to 0. 852. （2）The cluster analysis results showed that the 46 materials could be completely separated by using SCoT markers, Yemaoshi with staminate red flower and pistillate red flower were gathered for a group,they have far relationships with Niuxinshi group and Wild persimmon group,which is consistent with the morphologic observation. Bishan Niushishi was clustered on Wild per- simmon group, which was based on its morphological characteristics showed the same as wild types. Bishan Niushishi is tetraploid, different from others which are hexaploid. Bishan Niuxinshi should belong to theWild persimmon. The research indicated that SCoT markers can be effectively used to evaluate the genetic diversity of persimmon germplasm. 46 persimmon accessions have high genetic diversity, reflecting the geo- graphical distribution of species to a certain extent, and also show geographical factors have a significant impact on the genetic relationship for persimmon.
出处
《西北植物学报》
CAS
CSCD
北大核心
2014年第3期473-480,共8页
Acta Botanica Boreali-Occidentalia Sinica
基金
国家公益性行业(农业)科研专项(201203047)
农业部农作物种质资源保护项目(NB2012-2130135-21)
关键词
SCoT标记
柿
体系优化
遗传多样性
SCoT marker
Diospyros kaki Thunb.
system optimization
genetic diversity
