利用PCR产物混池重测序检测梨褐皮关联SLAF标签多态性  

Detection of SLAF Tag Polymorphism Associated with Pear Russet Peel by PCR Product Pool Resequencing

作　　者：蒋爽[1] 王晓庆[1] 施春晖[1] 李水根[1] 骆军[1] JIANG Shuang;WANG Xiaoqing;SHI Chunhui;LI Shuigen;LUO Jun(Shanghai Key Laboratory of Protected Horticultural Technology,Forestry and Fruit Tree Research Institute,Shanghai Academy of Agricultural Sciences,Shanghai 201403,China)

机构地区：[1]上海市农业科学院林木果树研究所,上海市园艺重点实验室,上海201403

出　　处：《园艺学报》2024年第4期707-714,共8页Acta Horticulturae Sinica

基　　金：上海市农委科技兴农项目(沪农科推字2019第1-3号)。

摘　　要：在已知梨褐皮关联的SLAF标签(Marker286117)序列上存在4个SNP位点,共有4个基因型(a~d)的基础上,使用PCR加混池重测序技术鉴定该SLAF标签在64个梨品种中的多态性。共获得7.1 Gb原始数据,通过编译的SNP-rseq和Calculate程序对数据进行处理,64个梨样品获得的平均read数量为117642条,满足分析需求。在该SLAF标签上新发现2个基因型(e和f)。6个基因型中c为原始类型,在栽培种砂梨和白梨中鉴定到b、d和e基因型;a和f基因型则出现在拥有西方梨血统的样品中。对基因型和表型统计后发现,c基因型存在时,梨果皮为褐色,不含c基因型时为绿色。该规律不仅适用于东方梨,在西方梨的一些样品中也适用。混池重测序技术降低了测序成本,相关技术手段能够应用于简化重测序后续的验证和标记辅助育种。Previous studies have shown that there are four SNP loci on the sequence of the russet peel associated SLAF tag(Marker286117),with a total of four genotypes(a,b,c,d)in Pyrus.In this study,PCR plus mixing pool re-sequencing was used to identify the polymorphisms of the SLAF tag in 64 pear cultivars.The results showed that a total of 7.1 Gb of raw data was obtained by re-sequencing,and the data were processed by two programs SNP-rseq and Calculate.The average number of reads obtained from 64 pear accessions was 117642,which met the analysis needs.Two new genotypes(e and f)were found on this SLAF locus.Among six genotypes,genotype c was the original type,and genotypes b,d and e were produced in cultivated pears of P.pyrifolia and P.bretschneideri.Genotypes a and f appeared in samples of Western pear.When genotype and phenotype statistics showed that only genotype c existed,the peel of the pear fruit was russet,and those without genotype c were green.This rule applies not only to Eastern pears,but also to some Western pears.In this study,mixed-pool was used to reduce the cost of sequencing.This method could be applied to simplify the subsequent validation of re-sequencing and marking-assisted breeding.

关 键 词：梨 SLAF标签 SNP 多态性 重测序 

分 类 号：S661.2[农业科学—果树学]