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为挖掘我国特有的果实香气浓郁的梨资源,弄清其果实芳香成分及含量,并进而研究它们在梨果实中的代谢与调节机制。采用气相色谱质谱联用(GC-MS)技术,分析了我国部分秋子梨品种果实的香气成分,在基础上,以挖掘出的‘南果梨’为试材,采用RACE技术克隆了与其果实特异香气成分生物合成相关的α-法尼烯合成酶基因(PuFS),并采用半定量PCR(RT-PCR)技术分析了该基因在其果实不同发育时期和不同部位的时空表达规律。结果表明:‘荣香’、‘龙香’、‘香水’和‘晚香’的主要香气成分为醛类物质,‘南果’、‘大南果’和‘寒香’梨主要香气成分为酯类物质。在‘南果’梨中还检测出了高含量的α-法尼烯,而在其他品种中α-法尼烯相对含量较低或未检测到,这可能是‘南国’梨具有浓郁香气的主要原因;随着果实的成熟,PuFS基因在‘南果梨’果皮中的表达量呈逐渐下降的趋势,绿熟期表达量最高,商熟期表达量开始降低,完熟期表达量非常弱。而PuFS在3个时期果肉中的表达量均很弱,且差异不明显。总体上,PuFS基因在绿熟期的表达量高于商熟期和完熟期,在果皮中的表达量显著高于果肉。
In order to excavate the fragrant pear resources that are endemic to our country, find out the aromatic components and contents of their fruits, and then study their metabolism and regulation mechanism in pear fruit. Based on the GC-MS technique, the aroma components of some varieties of Actinidia species in China were analyzed. Based on the results of the excision of ’Nanguo pear’ by RACE technique, The biosynthesis-related α-farnesene synthase gene (PuFS) was analyzed by semi-quantitative PCR (RT-PCR) to analyze the temporal and spatial expression of the gene at different developmental stages and in different parts of its fruit. The main aroma components of ’Rongxiang’, ’Longxiang’, ’Perfume’ and ’Xiangxiang’ were aldehydes. The main aroma components of ’Nan Guo’, ’Da Nan Guo’ and ’Han Xiang’ Esters. A high content of alpha-farnesene was also detected in the ’Nanguo’ pear, whereas the relative content of a-farnesene was lower or undetectable in other varieties, which could be the result of the strong aroma of ’Nanguo’ pear The main reason was that the expression of PuFS gene in ’Nanguo pear’ peel decreased gradually with the ripening of fruit. The expression level of PuFS gene in green ripe stage was the highest, the expression in commercial ripe stage began to decrease, and that in mature stage was very weak. However, the expression of PuFS in the three stages of flesh was very weak, and the difference was not obvious. In general, the expression level of PuFS gene in green ripe stage was higher than that in commercial ripe stage and mature stage, and the expression level of PuFS gene in peel was significantly higher than that of pulp.