宽皮柑橘单核苷酸多态性的高分辨率熔解曲线分型  

Genotyping Single Nucleotide Polymorphisms in Mandarin Cultivars Using High Resolution Melting Analysis

作　　者：吴波[1,2,3] 杨润婷[1] 朱世平[2] 钟云[3] 姜波[3] 曾继吾[3] 钟广炎[3] 

机构地区：[1]西南大学园艺园林学院,重庆400715 [2]中国农业科学院柑桔研究所,重庆400712 [3]广东省农业科学院果树研究所,农业部热带南亚热带果树生物学与遗传资源利用重点实验室,广州510640

出　　处：《园艺学报》2012年第4期777-782,共6页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(30971992);国家现代产业(柑橘)技术体系项目(CARS-27);长江学者和创新团队发展计划项目(IRT0976)

摘　　要：高分辨率熔解曲线分析(High resolution melting analysis,HRM)可以检测单碱基改变引起的DNA双链熔解温度(Tm)值变化,从而可以对样本在单核苷酸多态性分子标记(Single nucleotide polymor-phism,SNP)上进行基因分型。通过分析NCBI数据库中宽皮柑橘的表达序列标签(Expressed sequence tag,EST)数据鉴别SNP位点,并用小片段扩增法高分辨率熔解曲线分型技术(High resolution melting analysis of small amplicons)分析11个宽皮柑橘(Citrus reticulata)品种以及柳橙(Citrus sinensis Osbeck var.‘Liucheng’)的5个SNP位点的基因型。结果显示,小片段扩增法高分辨率熔解曲线分型可以快速、清楚地分辨纯合与杂合基因型,在校正温度差异后也可以很好地分辨同一个SNP位点不同的纯合型。统计分析表明样本在所有SNP位点上均存在多态性,5个SNP位点的平均多态性信息含量(PIC)为0.3190,显示样本在这组SNP位点上具有较高的杂合率。High resolution melting analysis（HRM）is capable of detecting changes in melting temperature（Tm）of double strand DNA sequences caused by single nucleotide changes,and is suitable to genotype genetic samples using known single nucleotide polymorphic site（sSNPs）. Citrus reticulata ESTs from NCBI EST database were searched for SNPs,and 5 SNPs were selected to genotype 11 mandarin and 1 sweet orange cultivars using HRM genotyping of small amplicons. The results showed that,at a given SNP site,a heterozygous genotype could be easily and clearly distinguished from a homozygous genotype by HRM,and 2 different homozygous genotypes could also be easily identified when temperature was calibrated. All 5 SNPs were found polymorphic in analyzed citrus samples,with an average polymorphic information content（PIC）value of 0.3190,indicating that these samples are highly heterozygous at these SNP sites.

关 键 词：宽皮柑橘 高分辨率熔解曲线 单核苷酸多态性 基因分型 

分 类 号：S666.2[农业科学—果树学]