刘厚宇,吴敏芳,宋莎,吴亚维,乔光.樱桃属(Cerasus)REMAP标记的建立及贵州种质的亲缘关系分析[J].中国南方果树,2020,49(2): |
樱桃属(Cerasus)REMAP标记的建立及贵州种质的亲缘关系分析 |
Establishment of REMAP marker on Cerasus and analysis of genetic relatedness for Guizhou cherry germplasms |
投稿时间:2019-10-18 修订日期:2019-11-09 |
DOI:10.13938/j.issn.1007-1431.20190588 |
中文关键词: 樱桃属 REMAP 技术体系 聚类分析 |
英文关键词:Cerasus REMAP maker Technical system Cluster analysis |
基金项目:国家自然科学基金项目(面上项目,重点项目,重大项目) |
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中文摘要: |
为丰富樱桃属分子标记,我们利用樱桃LTR引物和ISSR引物,筛选REMAP引物组合,建立了适合樱桃的REMAP标记的技术体系,并利用该标记对40个贵州地方樱桃种质亲缘关系进行分析。结果显示,适宜樱桃REMAP-PCR体系为: 总体系25 μL,MgCl2 2.0 mmol·L-1、 Taq酶0.75 U 、引物0.4 mmol·L-1 、模板DNA10 ng、dNPT 0.4 mmol·L-1,8%非变性PAGE对PCR扩增产物检测效果最好。利用上述体系筛选出10对REMAP引物组合,扩增出234个位点,平均多态性比率是92.29%。聚类分析显示,以相似性系数0.69为阈值,可以将供试材料分为3类:33种贵州地方栽培品种聚为一类,玛瑙红樱桃与4种野樱桃聚在一类,“红皮”樱桃与“青皮”樱桃聚在一类。 |
英文摘要: |
To develop a new molecular marker in cherry, LTR primers in combination of ISSR primers were used to screem REMAP primer, subsequently REMAP-PCR molecular marker system was established herein. Further, the genetic relatedness among the 40 germplasms of Chinese cherry (Cerasus pseudocerasus Lind1) originated from in Guizhou Province were illuminated. The result showed that the optimum REMAP-PCR system was totally in 25 μL, composed with 2.0 mmol L-1 MgCl2, 0.75 U Taq DNA polymerase, 0.4 mmol L-1 primer, 10 ng template DNA and 0.4 mmol L-1 dNTP, respectively. PAGE with concentration of 8% non-denaturing polyacrylamide gel elctrophoresis was the optimal PCR detection method. In optimization of REMAP-PCR reaction system, 10 REMAP primers was screened out, produced a total of 234 bands, average of polymorphism rate was 92.29%. UPGMA clustering result indicated REMAP primers could devided 40 germplasms into three groups at similarity coefficient of 0.69. Four wild species clustered together with ‘Manaohong’ . ‘Hongpi’ and ‘Qingpi’ clustered together closely and had far genetic distance with other germplasms. 33 cultivated species clustered together with same group. |
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