基于序列分析的13个中国梨品种S基因型鉴定(英文)

    为鉴定中国梨S-RNase基因及其S基因型,根据日本梨S-RNase基因保守区设计引物,对13个中国梨品种进行基因组PCR特异扩增,并对PCR产物克隆、测序、序列分析.结果鉴定出13个S等位基因,其中11个S基因与GenBank中已知的S1, S7, S12, S15, S16, S18, S19, S22, S27, S29, S34-RNases相同,另外2个则为新的S-RNase基因,命名为S37-RNase和S38-RNase,GenBank接受号分别为DQ839238、DQ839239.13个S等位基因中,在推导氨基酸水平上,S18与S27相似性最低,为58%,S7与S27,S12与S19,S15与S37及S15与S38间相似性最高,为94%.经分析,S19为中国梨中出现频率最高的等位基因,对其DNA序列进行限制性酶切位点分析,建立了一种快速、经济鉴定S19的方法,该方法无需测序而仅使用特异的限制性内切酶AflⅡ对PCR产物进行酶切消化.根据分子鉴定结果,13个中国梨品种S基因型被确定为:'冰糖'(S16S19), '六棱' (S16S19), '锦香'(S34S37), '鹅酥' (S15S38), '蜜梨'(S19S29), '甜橙子'(S7S12), '大青皮'(S19S34), '秋白' (S19S34), '紫酥' (S19S34), '花长把'(S19S22), '灌阳雪梨'(S18S27), '早蜜'(S19S29),'青面'(S1S18).

Determination of S-genotypes of 13 Chinese pear cultivars based on sequence analysis

To identify S-RNase genes and establish the correlation between the S-RNase genes and S-genotypes of Chinese pears, 13 cultivars were investigated. Genomic PCR with primers derived from the conserved nucleotide sequence of Japanese S-RNases, was carried out. As a result, 13 S-alleles were amplified. Comparison of the nucleotide and amino acid sequences between the 13 S-alleles and S-alleles deposited under Genbank databases revealed that 11 S-alleles corresponded to S1, S7, S12, S15, S16, S18, S19, S22, S27, S29 and S34-RNases, and the other two were new S-alleles (designated as S37 and S38). A pairwise comparison among the 13 S-alleles revealed an identity of 58% (between S18 and S27) to 94% (between S7 and S27, S12 and S19, S15 and S37, S15 and S38) at the deduced amino acid level. In addition, S19 was found to be the most frequent S-allele in Chinese pears. Based on restriction sites analysis, a rapid and economical method was developed for the identification of S19 without going to the effort of sequencing but using the digestion with S19-specific restriction endonuclease AflⅡ. The 13 cultivars were genotyped as follows:'Bingtang'(S16S19), 'Liuleng' (S16S19), 'Jinxiang'(S34S37), 'Esu' (S15S38), 'Mili'(S19S29), 'Tianchengzi'(S7S12), 'Daqingpi'(S19S34), 'Qiubai' (S19S34), 'Zisu' (S19S34), 'Huachangba'(S19S22), 'Guanyangxueli'(S18S27), 'Zaomi'(S19S29) and 'Qingmian' (S1S18). Elucidation of S-genotypes of Chinese pears would contribute to higher and stable yields in the orchards and a more efficient breeding program.