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中国精品科技期刊2020
吴晴阳, 周子维, 武清扬, 倪子鑫, 赖钟雄, 孙云. 乌龙茶加工过程中α-法呢烯的形成关键调控基因的筛选与表达分析[J]. 食品工业科技, 2020, 41(15): 135-142. DOI: 10.13386/j.issn1002-0306.2020.15.022
引用本文: 吴晴阳, 周子维, 武清扬, 倪子鑫, 赖钟雄, 孙云. 乌龙茶加工过程中α-法呢烯的形成关键调控基因的筛选与表达分析[J]. 食品工业科技, 2020, 41(15): 135-142. DOI: 10.13386/j.issn1002-0306.2020.15.022
WU Qing-yang, ZHOU ZI-wei, WU Qing-yang, NI Zi-xin, LAI Zhong-xiong, SUN Yun. Screening and Expression Analysis of Key Regulator Gene Associated with α-Farnesene Formation during Manufacturing Process of Oolong Tea[J]. Science and Technology of Food Industry, 2020, 41(15): 135-142. DOI: 10.13386/j.issn1002-0306.2020.15.022
Citation: WU Qing-yang, ZHOU ZI-wei, WU Qing-yang, NI Zi-xin, LAI Zhong-xiong, SUN Yun. Screening and Expression Analysis of Key Regulator Gene Associated with α-Farnesene Formation during Manufacturing Process of Oolong Tea[J]. Science and Technology of Food Industry, 2020, 41(15): 135-142. DOI: 10.13386/j.issn1002-0306.2020.15.022

乌龙茶加工过程中α-法呢烯的形成关键调控基因的筛选与表达分析

Screening and Expression Analysis of Key Regulator Gene Associated with α-Farnesene Formation during Manufacturing Process of Oolong Tea

  • 摘要: 为探讨乌龙茶加工过程中做青工序的机械力对香气成分中α-法呢烯的影响及形成机理,本实验以乌龙茶品种黄旦的鲜叶、萎凋叶、摇青叶为供试材料,未摇青等时间摊放叶为对照组,采用GC-TOF-MS 方法检测乌龙茶加工过程中α-法呢烯含量,结合转录组和茶树基因组数据库筛选α-法呢烯合成酶(AFS)关键基因,构建系统进化发育树,利用实时荧光定量PCR(Quantitative Real-time PCR,qRT-PCR)进行数据验证,并进行α-法呢烯含量与基因表达水平间的相关性分析。结果表明,α-法呢烯的含量在第一次摇青时有所下降,由于机械胁迫的累加,摇青叶中α-法呢烯的含量逐渐增加并在第三次摇青时达到峰值(0.9485%)。筛选获得的CL11384.Contig7_All转录本与茶树α-法呢烯合成酶基因(CsAFS,GenBank登录号:MH125264.1)高度相似,暂命名为CsAFS7基因,该基因表达模式与测序数据相一致。皮尔逊相关系数分析表明,乌龙茶加工过程中α-法呢烯含量与CsAFS7基因的相关表达水平之间存在极显著正相关(r=0.951,P<0.01)。因此,乌龙茶做青过程中α-法呢烯的累积可能受到CsAFS7基因的表达调控。

     

    Abstract: For purpose of understanding the influence of mechanical force during the turning-over process on the formation of α-farnesene, in this experiment, fresh tea leaves, solar withering tea leaves, turned-over tea leaves of the oolong tea cultivar Camellia sinensis cv.Huangdan as test materials and spread tea leaves (collected at the same time point) as control group were detected by headspace-gas chromatograpy-mass spectrometry (GC-TOF-MS).Thereafter, α-farnesene synthase (AFS) gene was selected based on Tea Tree Genemo Database and phylogenetic tree was constructed. The data were measured by quantitative real-time PCR and the relative correlation between α-farnesene content and relative gene expression level was also analyzed. The results indicated that although the α-farnesene content in first-time turned-over leaves decreased under the mechanical force, it still increased gradually with the treatments and reached the peak at the third-time turned-over point (0.9485%).The CL11384.Contig7_All gene was remarkably similar to CsAFS (GenBank accession no. MH125264.1), so CL11384.Contig7_All gene was named as CsAFS7 tentatively since the gene expression pattern was corresponding with the sequencing data. The correlation between α-farnesene content of manufacturing-process oolong tea and relative expression level of CsAFS7 gene were evaluated by Pearson correlation coefficient, and the results showed that there was a significant positive correlation (r=0.951, P<0.01). To sum up, the accumulation of α-farnesene content during manufacturing process might be regulated by CsAFS7 gene expression manipulation.

     

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