Detection of SLAF Tag Polymorphism Associated with Pear Russet Peel by PCR Product Pool Resequencing
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 117 642,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.
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