山葡萄及其杂种Myb相关基因的基因型及VvmybA1片段的分析

Genotyping Myb-related genes in Vitis amurensis and its hybrids and characterizing the VvmybA1 fragment sequences

【目的】确定不同类型山葡萄及其杂种的Myb相关基因的基因型,分析其VvmybA1基因片段的差异,探讨山葡萄之间的遗传差异。【方法】以10个单性花山葡萄株系(品种)、3个两性花山葡萄株系(品种)、7个山欧杂种品种为材料,以欧美杂种‘巨峰’及欧亚种品种‘黑比诺’作参照,对调控花色苷合成的多个Myb转录因子的基因型进行鉴定,通过克隆VvmybA1基因片段并进行序列比对,对不同材料的VvmybA1基因片段进行进化树的构建。【结果】山葡萄中只有VvmybA1基因;欧亚种‘黑比诺’、欧美杂种‘巨峰’和山欧杂种‘北玫’和‘北方’中有VvmybA1a基因;山欧杂种‘北红’中有VvmybA1和VvmybA3基因;山欧杂种‘北醇’、‘北玫’、‘北方’和‘公酿一号’中有VvmybA1、VvmybA2和VvmybA3基因。山欧杂种‘左红一’和‘左优红’中存在VvmybA1和VlMybA2基因。山葡萄及山欧杂种VvmybA1基因片段的序列对比显示,在该基因的第3个外显子区域不存在碱基的插入和缺失但存在较多的碱基替换。【结论】不同类型山葡萄在Myb相关基因的基因型上具有一致性,都只有VvmybA1基因,并且VvmybA1基因片段序列在第3个外显子区域存在较大差异,表现出丰富的遗传多样性。两性花山葡萄株系‘双庆’应该是山葡萄内部变异产生的。不同的山欧杂种在Myb相关基因的基因型上具有较大差异。

【Objective】 The aim of the study was to identify the genotype of Myb-related genes in Vitis amurensis and its hybrids, and analyse the difference among VvmybA1 gene fragments to investigate the genetic diversity in V. amurensis. 【Method】 Ten unisexual samples and 3 hermaphroditic samples in V. amurensis, 5 hybrids of V. amurensis × V. vinifera were used as experimental materials and ‘Kyoho'grape(V. labrusca ×V. vinifera) and ‘Pinot Noir'(V. vinifera) were taken as control materials. The genotypes of Myb-related transcription factors regulating the composition of anthocyanins and clone VvmybA1gene fragments were identified to analyse sequence alignment. And the phylogenetic tree was constructed with the sequences of VvmybA1 gene fragments. 【Result】 V. amurensis only has VvmybA1 gene. There is VvmybA1a gene in ‘Pinot Noir'(V. vinifera), ‘Kyoho'(V. labrusca ×V. vinifera) and two hybrids of V. amurensis × V. vinifera, ‘Beimei' and ‘Noir de Nord'. There are VvmybA1 and VvmybA3 genes in‘Beihong', while there are VvmybA1, VvmybA2 and VvmybA3 genes in other four hybrids of V. amurensis ×V. vinifera, ‘Beichun', ‘Beimei', ‘Noir de Nord' and ‘Gongniang 1'. There are VvmybA1 and VlmybA2 genes in two hybrids of V. amurensis × V. vinifera, ‘Zuohong 1' and ‘Zuoyouhong'. The analysis of VvmybA1 gene fragments sequence alignment shows that there is no insertion or deletion, but a lot of base substitutions between the samples in the third exon. 【Conclusion】 The genotype of Myb-related genes in different V. amurensis is coincident and there is only VvmybA1 gene. The obvious difference in the third exon shows abundant genetic diversity. The herrmaphrodite sample of V. amurensis,‘Shuangqing' may be the production of the internal variation in V. amurensis. There are significant differences among the genotypes of Myb-related genes in the hybrids of V. amurensis × V. vinifera.