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

ACTA HORTICULTURAE SINICA ›› 2012, Vol. 39 ›› Issue (4): 777-782.

• Technologies and Methods • Previous Articles Next Articles

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

WU Bo1,2,3，YANG Run-ting1，ZHU Shi-ping2，ZHONG Yun3，JIANG Bo3，ZENG Ji-wu3，and ZHONG Guang-yan3,*

（1College of Horticulture and Landscape，Southwest University，Chongqing 400715，China；2Citrus Research Institute，Chinese Academy of Agricultural Sciences，Chongqing 400712，China；3Key Laboratory of Tropical and Subtropical Fruit Biology and Genetic Resource Utilization，Ministry of Agriculture，Fruit Tree Institute，Guangdong Academy of Agricultural Sciences，Guangzhou 510640，China）

Online:2012-04-25 Published:2012-04-25

Abstract

Abstract: High resolution melting analysis（HRM）is capable of detecting changes in melting temperature（T_m）of double strand DNA sequences caused by single nucleotide changes，and is suitable to genotype genetic samples using known single nucleotide polymorphic sites（SNPs）. 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 givenSNP 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.

Key words: mandarin, high resolution melting, single nucleotide polymorphism, genotyping

CLC Number:

S 666.2

WU Bo, YANG Run-Ting, ZHU Shi-Ping, ZHONG Yun, JIANG Bo, ZENG Ji-Wu, ZHONG Guang-Yan. Genotyping Single Nucleotide Polymorphisms in Mandarin Cultivars Using High Resolution Melting Analysis[J]. ACTA HORTICULTURAE SINICA, 2012, 39(4): 777-782.

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

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