蛇龙珠等酿酒葡萄品种羧酸酯酶基因片段的SNP分析

目的]利用单核苷酸多态笥(SNP)分析蛇龙珠等酿酒葡萄的单核苷酸多态性及遗传亲缘关系,为今后研究酿酒葡萄的抗性机制以及培育抗病性葡萄提供参考。方法]以酿酒葡蛇龙珠8个新株系(E01~E08)、品丽珠、赤霞珠为材料,以欧亚种黑比诺为对照,对羧酸酯酶基因(CXEs)片段进行克隆和序列分析,研究蛇龙珠等酿酒葡萄之间的遗传差异。结果]序列对比显示,该基因片段在编码区和非编码区均存在不同程度的碱基替换。11份葡萄材料的基因片段共发现87个多态性位点,平均74 bp左右的序列长度上能够检测到一个SNP位点,核苷酸多样度(Pi)0.050 76±0.011 49,单倍型多样度(Hd)1.000±0.039,表现出丰富的遗传多样性。3种中性检验方法比较序列变异模式,结果符合中性生化模型。通过聚类分析可将蛇龙珠E02、E03聚为一类,E07、E08聚为一类,其遗传距离和地理距离一致。该序列片段存在一些突变,这些突变能将蛇龙珠8个新株系与其他酿酒葡萄品种明显区分。结论]CXEs基因片段的突变位点可作为遗传标记,利用SNP方法可分析葡萄的遗传多样性和亲缘关系。

The SNP Analysis of Carboxylesterases in Cabernet Gernischt (Vitis vinifera L.) and Other Wine Grape Cultivars

Objective] The aim was to analyze mononucleotide polymorphism and genetic relationships of Cabernet Gernischt (Vitis vinifera L.) and other wine grape cultivars,provide reference for study on resistance mechanism of wine grape and cultivation of disease resistant grape. Method] The difference among Carboxylesterase gene fragments was analyzed to investigate the genetic diversity of Cabernet Gernischt and other wine grape cultivars.Eight new strains of Cabernet Gernischt (E01-E08),Cabernet Sauvignon,Cabernet Franc were used as experimental mate-rials and Pinot Noir ( V.vinifera) were taken as control materials,Carboxylesterase gene fragments were cloned and analyzed. Result] The analy-sis of Carboxylesterase gene fragments sequence alignment showed that there were different degrees of base substitutions between the samples in the coding and non-coding regions.A total of 87 SNPs were detected in the sequenced fragments,a SNP locus was found on every 4 bp sequence on average.Nucleotide diversity (Pi) =0.050 76 ( ±0.011 49),Haplotype diversity (Hd)=1.000 ( ±0.039) which give a high leve1 of poly-morphism.Three neutrality tests were used to compare the patterns of sequence variations,the result indicates conform to neutral mutation.The clustering dendrogram was constructed by Neighbour Joining (NJ),Cabernet Gernischt E02 (north of qixia) and E03 (south of qixia) classified into one group,E07(development zone) and E08 (development zone) classified into one group,genetic distance in accord with geographic dis-tance.The results of the experiment showed that there were several mutations in the Carboxylesterase gene fragments might be selected to be the molecular marker to distinguish Cabernet Gernischt eight new strains and other wine grape varieties.Conclusion]The mutation site of CXEs gene fragment can be used as a kind of genetic marker,SNP method can identify genetic relationships of grape.