葡萄果實香氣物質的QTL定位及關聯(lián)分析研究
發(fā)布時間：2018-11-22 08:05
【摘要】：葡萄果實中香氣物質是由多基因控制的復雜性狀,其不僅受到栽培技術、環(huán)境條件等因素影響,更與品種基因型和遺傳因素密切相關,利用傳統(tǒng)的遺傳研究方法較難從本質上揭示果實香氣的遺傳機制。而基于系譜的連鎖作圖和基于自然群體的關聯(lián)分析是現(xiàn)今開展植物數(shù)量性狀研究的主要方法。本研究以母本'87-1' 和父本'9-22'雜交獲得的149株雜交后代為作圖群體,采用SSR和SRAP兩種分子標記技術相結合,構建歐亞種葡萄的分子遺傳圖譜,并對葡萄果實中主要香氣成分進行了 QTL定位研究。另一方面,以92個品種資源群體為試材,基于與萜類物質合成相關的VvDXS和α-TPS基因,對基因在不同品種資源群體中的多態(tài)性與葡萄果實香氣特征進行關聯(lián)分析,發(fā)掘功能性SNP位點。試驗主要結果如下:1.通過對雜交群體'87-1' × '9-22'組合和品種群體進行香氣物質含量的測定,在雜交群體中發(fā)現(xiàn)芳樟醇、香茅醇、橙花醇、香葉醇、香葉酸幾種物質存在廣泛的超親現(xiàn)象,而αα-萜品醇較少出現(xiàn)超親現(xiàn)象;在品種群體香氣物質的檢測中,發(fā)現(xiàn)不同香氣物質的分布存在較大差異,其中芳樟醇含量集中在0-5μg/kg和高于25μg/kg的兩個極端區(qū)域;香茅醇、橙花醇在各含量區(qū)間分布較為均勻;αα-萜品醇、香葉醇、香葉酸香氣物質含量主要集中在0-5μg/kg的區(qū)間內。2.以雜交群體'87-1' × '9-22'為作圖群體,構建了遺傳圖譜,共獲得母本特有位點156個,父本特有位點142個,雙親共有位點215個,雙親遺傳圖譜均形成19個連鎖群。其中,母本'87-1'遺傳圖譜包含308個標記,覆蓋總圖距2017.9cM,位點間平均距離為6.55 cM;父本'9-22'遺傳圖譜包含295個標記,覆蓋總圖距1803.3 cM,位點間平均距離為6.11cM。3.應用MapQTL 5.0定位軟件進行六種香氣物質QTL定位。在所構建的遺傳圖譜中共計發(fā)現(xiàn)了與香氣物質相關的QTL位點48個。其中,LG3和LG5兩個連鎖群獲得了較多的QTL位點。位于連鎖群LG3標記VMC1A5與VMC2E7之間定位Lin14-1、Lin15-1、α-Ter15-1、Cit14-1、Ner15-2 和 Ger15-1 六個與香氣物質相關的 QTL 位點。通過標記物理位置比對,發(fā)現(xiàn)在該區(qū)間內存在與萜類物質代謝路徑相關的HDR基因。另一方面,位于連鎖群LG5標記VLG5-C-1與UDV106之間定位Lin14-3、α-Ter15-2、Ner15-3、Ger15-2和Geracid14-1五個與香氣物質相關的QTL位點。并且發(fā)現(xiàn)在該區(qū)間內存在與萜類物質代謝路徑相關的DXS基因。4.通過對VvDXS基因中22個SNP位點與6種香氣物質進行關聯(lián)分析,最終得到了位點P852和P1678變異與香氣物質含量存在顯著關聯(lián)。其中,位點P852等位基因為G/T雜合時,其對應品種的芳樟醇、αα-萜品醇、橙花醇、香葉酸四種香氣物質含量較高。位點P1678的等位基因為A/A時,該品種表現(xiàn)為濃香型,且芳樟醇、α-萜品醇、香茅醇、橙花醇、香葉醇、香葉酸含量較高。該位點為A/G基因型對應的品種極可能不屬于無香型品種。5.通過對α-TPS基因與αα-萜品醇之間的關聯(lián)分析,本研究發(fā)現(xiàn)了 S1556位點的突變會會顯著影響其所對應基因型的葡萄果實中α-萜品醇含量。位點S1556的等位基因為C/T時對應品種有較高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.
【學位授予單位】：沈陽農業(yè)大學
【學位級別】：博士
【學位授予年份】：2017
【分類號】：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.
【學位授予單位】：沈陽農業(yè)大學
【學位級別】：博士
【學位授予年份】：2017
【分類號】：S663.1
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