葡萄果實(shí)香氣物質(zhì)的QTL定位及關(guān)聯(lián)分析研究
發(fā)布時(shí)間：2018-11-22 08:05
【摘要】：葡萄果實(shí)中香氣物質(zhì)是由多基因控制的復(fù)雜性狀,其不僅受到栽培技術(shù)、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(guān),利用傳統(tǒng)的遺傳研究方法較難從本質(zhì)上揭示果實(shí)香氣的遺傳機(jī)制。而基于系譜的連鎖作圖和基于自然群體的關(guān)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(biāo)記技術(shù)相結(jié)合,構(gòu)建歐亞種葡萄的分子遺傳圖譜,并對葡萄果實(shí)中主要香氣成分進(jìn)行了 QTL定位研究。另一方面,以92個(gè)品種資源群體為試材,基于與萜類物質(zhì)合成相關(guān)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(shí)香氣特征進(jìn)行關(guān)聯(lián)分析,發(fā)掘功能性SNP位點(diǎn)。試驗(yàn)主要結(jié)果如下:1.通過對雜交群體'87-1' × '9-22'組合和品種群體進(jìn)行香氣物質(zhì)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(zhì)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(zhì)的檢測中,發(fā)現(xiàn)不同香氣物質(zhì)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(gè)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(zhì)含量主要集中在0-5μg/kg的區(qū)間內(nèi)。2.以雜交群體'87-1' × '9-22'為作圖群體,構(gòu)建了遺傳圖譜,共獲得母本特有位點(diǎn)156個(gè),父本特有位點(diǎn)142個(gè),雙親共有位點(diǎn)215個(gè),雙親遺傳圖譜均形成19個(gè)連鎖群。其中,母本'87-1'遺傳圖譜包含308個(gè)標(biāo)記,覆蓋總圖距2017.9cM,位點(diǎn)間平均距離為6.55 cM;父本'9-22'遺傳圖譜包含295個(gè)標(biāo)記,覆蓋總圖距1803.3 cM,位點(diǎn)間平均距離為6.11cM。3.應(yīng)用MapQTL 5.0定位軟件進(jìn)行六種香氣物質(zhì)QTL定位。在所構(gòu)建的遺傳圖譜中共計(jì)發(fā)現(xiàn)了與香氣物質(zhì)相關(guān)的QTL位點(diǎn)48個(gè)。其中,LG3和LG5兩個(gè)連鎖群獲得了較多的QTL位點(diǎn)。位于連鎖群LG3標(biāo)記VMC1A5與VMC2E7之間定位Lin14-1、Lin15-1、α-Ter15-1、Cit14-1、Ner15-2 和 Ger15-1 六個(gè)與香氣物質(zhì)相關(guān)的 QTL 位點(diǎn)。通過標(biāo)記物理位置比對,發(fā)現(xiàn)在該區(qū)間內(nèi)存在與萜類物質(zhì)代謝路徑相關(guān)的HDR基因。另一方面,位于連鎖群LG5標(biāo)記VLG5-C-1與UDV106之間定位Lin14-3、α-Ter15-2、Ner15-3、Ger15-2和Geracid14-1五個(gè)與香氣物質(zhì)相關(guān)的QTL位點(diǎn)。并且發(fā)現(xiàn)在該區(qū)間內(nèi)存在與萜類物質(zhì)代謝路徑相關(guān)的DXS基因。4.通過對VvDXS基因中22個(gè)SNP位點(diǎn)與6種香氣物質(zhì)進(jìn)行關(guān)聯(lián)分析,最終得到了位點(diǎn)P852和P1678變異與香氣物質(zhì)含量存在顯著關(guān)聯(lián)。其中,位點(diǎn)P852等位基因?yàn)镚/T雜合時(shí),其對應(yīng)品種的芳樟醇、αα-萜品醇、橙花醇、香葉酸四種香氣物質(zhì)含量較高。位點(diǎn)P1678的等位基因?yàn)锳/A時(shí),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(diǎn)為A/G基因型對應(yīng)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(guān)聯(lián)分析,本研究發(fā)現(xiàn)了 S1556位點(diǎn)的突變會(huì)會(huì)顯著影響其所對應(yīng)基因型的葡萄果實(shí)中α-萜品醇含量。位點(diǎn)S1556的等位基因?yàn)镃/T時(shí)對應(yīng)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(xué)位授予單位】：沈陽農(nóng)業(yè)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：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(xué)位授予單位】：沈陽農(nóng)業(yè)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：S663.1
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