甜樱桃S基因型PCR鉴定技术研究

Identification of S-allele in sweet cherry(Prunus avium) by allele-specific PCR

绝大部分甜樱桃品种自交不亲和,因此自交不亲和基因型的鉴定对于生产具有重要的意义。以甜樱桃主栽品种为试材,建立基于PCR技术的甜樱桃品种S基因型鉴定技术。试验根据已发表的樱桃S基因序列设计了2对引物组合BFP73/74,BFP93/94,结合S1、S5基因的特异扩增引物,利用扩增片段长度的不同,就可以对樱桃品种的S基因型进行鉴定。通过对已知S基因型品种基因组的扩增,最终建立了基于PCR技术的甜樱桃品种S基因型鉴定技术。

Sweet cherry exhibits the monofactorial gametophytic self-incompatibility system which is similar to other self-incompatible fruit tree species of Rosaceae. Determination of correct pollen incompatibility groups and assignment of cultivars to the groups are essential for good crop production. The incompatibility groups in sweet cherry have been identified by controlled pollination tests and/or pollen tube growth tests, but these tests is time-consuming and prone to be affected by environmental factors. The allele-specific PCR technique was established to identify the S-genotype of sweet cherry cuhivars in this study The PCR based identification technology of 7 known self-incompatibility alleles of sweet cherry was established. Two pairs of consensus primers BFP73/74 and BFP93/94 were designed from published cDNA sequences of S1 to S6 S- RNase to reveal length variation of the first and the second introns.With the consensus and allele-Specific primers of S1 and S5, most sweet cherry cuhivars＇ S-genotype could be visualized directly on an agarose gel. The use of the consensus and allele-specific primers was demonstrated by detecting genotypes that have been published recently. This method is a rapid and convenient technique that can be used on vegetative material.