基于转录组测序和RT-qPCR技术的烟草糖酯合成基因挖掘  

Excavating tobacco sugar ester synthesis genes based on transcriptome sequencing and RT-qPCR

作　　者：黄申 闫茗熠 陈梦月 尚紫博 杨晨 张金林[3] 褚智国 毛多斌 HUANG Shen;Yan Mingyi;CHEN Mengyue;SHANG Zibo;YANG Chen;ZHANG Jinlin;CHU Zhiguo;MAO Duobin(College of Tobacco Science and Engineering,Zhengzhou University of Light Industry,Zhengzhou 450001,China;Jinan Cigarette Factory,China Tobacco Shandong Industrial Co.,Ltd.,Jinan 250000,China;Technology Center,China Tobacco Shandong Industrial Co.,Ltd.,Jinan 250000,China)

机构地区：[1]郑州轻工业大学烟草科学与工程学院,河南郑州450001 [2]山东中烟工业有限责任公司济南卷烟厂,山东济南250000 [3]山东中烟工业有限责任公司技术中心,山东济南250000

出　　处：《轻工学报》2023年第6期78-84,117,共8页Journal of Light Industry

基　　金：中国烟草总公司科技重点研发项目(110202002001,110202202006)。

摘　　要：为挖掘调控烟草糖酯合成的功能基因,采用转录组测序对烟草腺毛细胞和叶细胞的差异表达基因及其功能进行分析,并利用RT-qPCR对分析结果进行验证。结果表明:共获得11962个腺毛细胞和叶细胞的差异表达基因,其中5145个上调表达基因,6817个下调表达基因;上调表达基因主要包括生物学进程(2265个)、分子功能(1169个)、细胞成分(363个)3大类,可将基因序列定位到122条代谢通路中,其中与糖酯合成相关的糖酵解/糖异生、淀粉和蔗糖代谢、苯丙烷类生物合成被高度富集,腺毛细胞中酰基糖通路显著上调,有12个基因在功能上与酰基转移酶有关;在腺毛细胞中,分析得到的12个基因中有11个基因的表达量都高于叶细胞,其中gene63826、gene16194和gene37511表达分别上调195.79倍、166.23倍和132.65倍。上述11个基因可能参与调控腺毛细胞合成糖酯,为下一步验证基因功能提供依据。To study the genes regulating the synthesis of sugar esters in tobacco,transcriptome sequencing analysis was performed on tobacco glandular hair cells and bundle sheath cells,and RT-qPCR was used for verification.The results showed that 11962 differentially expressed genes were obtained,of which 5145 were up-regulated and 6817 were down-regulated.GO annotation of differentially expressed genes showed that the up-regulated genes mainly included biological processes(2265),molecular functions(1169),and cellular components(363),with a total of 3793 GO terms.Based on the assignment of KEGG pathway,the sequence was mapped to 122 metabolic pathways.Glycolysis/gluconeogenesis,starch and sucrose metabolism,phenylpropanoid biosynthesis related to glycoester synthesis were highly enriched,and acyl sugar pathway in glandular hairs was significantly up-regulated.By comparing with the tobacco reference genome,12 up-regulated acyltransferase genes were identified in glandular hair cells.RT-qPCR results showed that the expression of 11 genes was higher than that of the control.The expression of gene 63826,gene 16194 and gene 37511 increased by 195.79 times,166.23 times and 132.65 times,respectively.The above 11 genes may be involved in regulating the synthesis of sugar esters in glandular hair cells,providing a basis for further verification of gene function.

关 键 词：烟草糖酯 腺毛细胞 转录组 差异表达基因 RT-QPCR 

分 类 号：TS413[农业科学—烟草工业]