葡萄果实香气物质的QTL定位及关联分析研究
发布时间：2018-11-22 08:05
【摘要】：葡萄果实中香气物质是由多基因控制的复杂性状,其不仅受到栽培技术、环境条件等因素影响,更与品种基因型和遗传因素密切相关,利用传统的遗传研究方法较难从本质上揭示果实香气的遗传机制。而基于系谱的连锁作图和基于自然群体的关联分析是现今开展植物数量性状研究的主要方法。本研究以母本'87-1' 和父本'9-22'杂交获得的149株杂交后代为作图群体,采用SSR和SRAP两种分子标记技术相结合,构建欧亚种葡萄的分子遗传图谱,并对葡萄果实中主要香气成分进行了 QTL定位研究。另一方面,以92个品种资源群体为试材,基于与萜类物质合成相关的VvDXS和α-TPS基因,对基因在不同品种资源群体中的多态性与葡萄果实香气特征进行关联分析,发掘功能性SNP位点。试验主要结果如下:1.通过对杂交群体'87-1' × '9-22'组合和品种群体进行香气物质含量的测定,在杂交群体中发现芳樟醇、香茅醇、橙花醇、香叶醇、香叶酸几种物质存在广泛的超亲现象,而αα-萜品醇较少出现超亲现象;在品种群体香气物质的检测中,发现不同香气物质的分布存在较大差异,其中芳樟醇含量集中在0-5μg/kg和高于25μg/kg的两个极端区域;香茅醇、橙花醇在各含量区间分布较为均匀;αα-萜品醇、香叶醇、香叶酸香气物质含量主要集中在0-5μg/kg的区间内。2.以杂交群体'87-1' × '9-22'为作图群体,构建了遗传图谱,共获得母本特有位点156个,父本特有位点142个,双亲共有位点215个,双亲遗传图谱均形成19个连锁群。其中,母本'87-1'遗传图谱包含308个标记,覆盖总图距2017.9cM,位点间平均距离为6.55 cM;父本'9-22'遗传图谱包含295个标记,覆盖总图距1803.3 cM,位点间平均距离为6.11cM。3.应用MapQTL 5.0定位软件进行六种香气物质QTL定位。在所构建的遗传图谱中共计发现了与香气物质相关的QTL位点48个。其中,LG3和LG5两个连锁群获得了较多的QTL位点。位于连锁群LG3标记VMC1A5与VMC2E7之间定位Lin14-1、Lin15-1、α-Ter15-1、Cit14-1、Ner15-2 和 Ger15-1 六个与香气物质相关的 QTL 位点。通过标记物理位置比对,发现在该区间内存在与萜类物质代谢路径相关的HDR基因。另一方面,位于连锁群LG5标记VLG5-C-1与UDV106之间定位Lin14-3、α-Ter15-2、Ner15-3、Ger15-2和Geracid14-1五个与香气物质相关的QTL位点。并且发现在该区间内存在与萜类物质代谢路径相关的DXS基因。4.通过对VvDXS基因中22个SNP位点与6种香气物质进行关联分析,最终得到了位点P852和P1678变异与香气物质含量存在显著关联。其中,位点P852等位基因为G/T杂合时,其对应品种的芳樟醇、αα-萜品醇、橙花醇、香叶酸四种香气物质含量较高。位点P1678的等位基因为A/A时,该品种表现为浓香型,且芳樟醇、α-萜品醇、香茅醇、橙花醇、香叶醇、香叶酸含量较高。该位点为A/G基因型对应的品种极可能不属于无香型品种。5.通过对α-TPS基因与αα-萜品醇之间的关联分析,本研究发现了 S1556位点的突变会会显著影响其所对应基因型的葡萄果实中α-萜品醇含量。位点S1556的等位基因为C/T时对应品种有较高α-萜品醇含量的。
[Abstract]:The aroma substance in the grape fruit is a complex character controlled by the multi-gene, which is not only influenced by the factors such as the cultivation technology and the environmental conditions, but also is closely related to the genotype and the genetic factor of the variety, and the genetic mechanism of the fruit aroma is difficult to be revealed from the essence by using the traditional genetic research method. The linkage mapping based on the genealogy and the correlation analysis based on the natural population are the main methods to carry out the research on the quantitative characters of the plants nowadays. The study uses female parent '87-1.' and male parent '9-22 The molecular genetic map of the Eurasian grape was constructed by the combination of SSR and SRAP, and the main aroma components in the fruit of the grape were studied by using the combination of SSR and SRAP. On the other hand, on the other hand, 92 varieties of resource groups were used as the test materials, and based on the VvDXS and the P-TPS genes related to the synthesis of the wild type substances, the association analysis of the polymorphism of the gene in different resource groups and the aroma characteristics of the grape fruits was carried out, and the functional SNP sites were excavated. The main results of the test are as follows: 1. by comparing the hybridization population' 87-1 'to the hybrid population' 87-1 ', to make a difference' 9-22 'The composition and the variety group carry out the determination of the content of the aroma substances, and in the hybridization group, the aromatic camphor alcohol, the citronellol, the orange flower alcohol, the geraniol and the aromatic folic acid are found to have a wide super-affinity phenomenon, It is found that there is a great difference in the distribution of the different aroma substances, in which the content of the linalool is in the range of 0-5 & mu; g/ kg and the two extreme regions higher than 25 & mu; g/ kg; the citronellol and the orange flower alcohol are distributed more uniformly in the content intervals; and the alcitronol, the geraniol, the content of the aroma of the fragrant and folic acid is mainly in the interval of 0-5 & mu; g/ kg. in that hybridization group' 87-1, '%' 9-22 The genetic map was constructed for the mapping group, 156 male parent-specific sites, 142 male parent-specific sites, and 215 parent-specific sites, and 19 linkage groups were formed in both the parent and the two-parent genetic maps. wherein the female parent '87-1' The genetic map contains 308 markers, covering the general plot distance of 2017. 9cM, the average distance between the sites is 6.55 cM, and the male parent '9-22' The genetic map contains 295 markers, covering the general plot distance of 1803. 3cM, and the average distance between the sites is 6.11cM.3. Six kinds of aroma substance QTLs were located by using the MapQTL 5.0 location software. A total of 48 QTL sites associated with the aroma compounds were found in the constructed genetic map. Among them, two linkage groups of LG3 and LG5 have more QTL sites. Lo14-1, Lin15-1, L-Ter15-1, Cit14-1, Nere15-2 and Ger15-1 QTLs associated with the aroma substances are located between the linkage group LG3 marker VMC1A5 and the VMC2E7. By marking the physical location ratio pair, it was found that there was an HDR gene associated with the metabolic pathway of the opioid substance within the interval. On the other hand, five QTLs related to the aroma substance are located between the linkage group LG5-labeled VLG5-C-1 and the UDV 106, and five QTLs related to the aroma substance are located between the LG5-C-1 and the UDV 106. and it was found that there was a DXS gene in this interval associated with the metabolic pathway of the opioid. The relationship between the mutation of P852 and P1678 and the content of the aroma substance was obtained by the correlation analysis of 22 SNP sites in the VvDXS gene and 6 kinds of aroma substances. Among them, when the allele of the locus P852 is G/ T, the content of the aromatic compounds of the corresponding variety is higher than the content of the four kinds of aromatic substances. When the allele of the site P1678 is A/ A, the variety shows a strong flavor, and the content of the linalool, the alcitol, the citronellol, the citronellol, the geraniol and the aromatic folic acid is high. The corresponding variety of the A/ G genotype may not belong to the non-flavor type. The study found that the mutation of the S1556 site could significantly affect the content of the p-hydroxybenzene in the grape fruit of the corresponding genotype. The allele of the locus S1556 is C/ T, and the corresponding variety has a higher content of the high-yield-equivalent.
【学位授予单位】：沈阳农业大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：S663.1
[Abstract]:The aroma substance in the grape fruit is a complex character controlled by the multi-gene, which is not only influenced by the factors such as the cultivation technology and the environmental conditions, but also is closely related to the genotype and the genetic factor of the variety, and the genetic mechanism of the fruit aroma is difficult to be revealed from the essence by using the traditional genetic research method. The linkage mapping based on the genealogy and the correlation analysis based on the natural population are the main methods to carry out the research on the quantitative characters of the plants nowadays. The study uses female parent '87-1.' and male parent '9-22 The molecular genetic map of the Eurasian grape was constructed by the combination of SSR and SRAP, and the main aroma components in the fruit of the grape were studied by using the combination of SSR and SRAP. On the other hand, on the other hand, 92 varieties of resource groups were used as the test materials, and based on the VvDXS and the P-TPS genes related to the synthesis of the wild type substances, the association analysis of the polymorphism of the gene in different resource groups and the aroma characteristics of the grape fruits was carried out, and the functional SNP sites were excavated. The main results of the test are as follows: 1. by comparing the hybridization population' 87-1 'to the hybrid population' 87-1 ', to make a difference' 9-22 'The composition and the variety group carry out the determination of the content of the aroma substances, and in the hybridization group, the aromatic camphor alcohol, the citronellol, the orange flower alcohol, the geraniol and the aromatic folic acid are found to have a wide super-affinity phenomenon, It is found that there is a great difference in the distribution of the different aroma substances, in which the content of the linalool is in the range of 0-5 & mu; g/ kg and the two extreme regions higher than 25 & mu; g/ kg; the citronellol and the orange flower alcohol are distributed more uniformly in the content intervals; and the alcitronol, the geraniol, the content of the aroma of the fragrant and folic acid is mainly in the interval of 0-5 & mu; g/ kg. in that hybridization group' 87-1, '%' 9-22 The genetic map was constructed for the mapping group, 156 male parent-specific sites, 142 male parent-specific sites, and 215 parent-specific sites, and 19 linkage groups were formed in both the parent and the two-parent genetic maps. wherein the female parent '87-1' The genetic map contains 308 markers, covering the general plot distance of 2017. 9cM, the average distance between the sites is 6.55 cM, and the male parent '9-22' The genetic map contains 295 markers, covering the general plot distance of 1803. 3cM, and the average distance between the sites is 6.11cM.3. Six kinds of aroma substance QTLs were located by using the MapQTL 5.0 location software. A total of 48 QTL sites associated with the aroma compounds were found in the constructed genetic map. Among them, two linkage groups of LG3 and LG5 have more QTL sites. Lo14-1, Lin15-1, L-Ter15-1, Cit14-1, Nere15-2 and Ger15-1 QTLs associated with the aroma substances are located between the linkage group LG3 marker VMC1A5 and the VMC2E7. By marking the physical location ratio pair, it was found that there was an HDR gene associated with the metabolic pathway of the opioid substance within the interval. On the other hand, five QTLs related to the aroma substance are located between the linkage group LG5-labeled VLG5-C-1 and the UDV 106, and five QTLs related to the aroma substance are located between the LG5-C-1 and the UDV 106. and it was found that there was a DXS gene in this interval associated with the metabolic pathway of the opioid. The relationship between the mutation of P852 and P1678 and the content of the aroma substance was obtained by the correlation analysis of 22 SNP sites in the VvDXS gene and 6 kinds of aroma substances. Among them, when the allele of the locus P852 is G/ T, the content of the aromatic compounds of the corresponding variety is higher than the content of the four kinds of aromatic substances. When the allele of the site P1678 is A/ A, the variety shows a strong flavor, and the content of the linalool, the alcitol, the citronellol, the citronellol, the geraniol and the aromatic folic acid is high. The corresponding variety of the A/ G genotype may not belong to the non-flavor type. The study found that the mutation of the S1556 site could significantly affect the content of the p-hydroxybenzene in the grape fruit of the corresponding genotype. The allele of the locus S1556 is C/ T, and the corresponding variety has a higher content of the high-yield-equivalent.
【学位授予单位】：沈阳农业大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：S663.1
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