柚芽变株系类胡萝卜素代谢差异基因的转录组学分析  

Transcriptome Analysis of Carotenoid Metabolism Differential Genes in Pomelo Bud Strain

作　　者：潘鹤立 潘腾飞[1] 佘文琴[1] 徐世荣[1] 李小婷[1] 黄汉唐 陈源[2] 吴少华[1] 潘东明[1] PAN Heli;PAN Tengfei;SHE Wenqin;XU Shirong;LI Xiaoting;HUANG Hantang;CHEN Yuan;WU Shaohua;PAN Dongming(College of Horticulture,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China;Institute of Agricultural Engineering and Technology,Fujian Academy of Agricultural Sciences,Fuzhou,Fujian 350003,China)

机构地区：[1]福建农林大学园艺学院,福建福州350002 [2]福建省农业科学院农业工程技术研究所,福建福州350003

出　　处：《热带作物学报》2020年第11期2165-2175,共11页Chinese Journal of Tropical Crops

基　　金：2017年福建省科技厅区域发展项目(No.2017N3005);2018年福建省发改委农业“五新”工程项目(闽发改农业[2018]114号);福建省农业科学院“青年科技英才百人计划”项目(No.YC2015-10)。

摘　　要：为了研究‘琯溪蜜柚’果实红肉突变株系类胡萝卜素代谢和芽变的分子机制,本研究以‘琯溪蜜柚’及其芽变一代品种‘红肉蜜柚’和芽变二代品种‘三红蜜柚’果实为材料,使用RNA-seq技术研究3个品种3个发育时期果肉类胡萝卜素的代谢。经生物信息学分析,共筛选出参与类胡萝卜素生物合成显著差异表达的10个基因,分别为八氢番茄红素合成酶基因(PSY)、六氢番茄红素脱氢酶基因(ZDS)、类胡萝卜素异构酶基因(CRTISO1、CRTISO2)、紫黄质脱环氧酶基因(NPQ1)、9-顺式-环氧类胡萝卜素双加氧酶基因(NCED3、NCED5)、脱落醛氧化酶基因(AAO3)、ABA 8’-羟化酶基因(CYP707A1)、类胡萝卜素裂解双加氧酶基因(CCD4)。共表达分析揭示了7个家族转录因子的表达模式与类胡萝卜素合成基因高度相关,分别为MADS、bZIP、bHLH、MYB、AP2/ERF、NAC和WRKY。通过实时荧光定量PCR(qRT-PCR)验证了部分基因的表达模式,结果与RNA-seq结果高度一致。此外,经对‘红肉蜜柚’果肉转色期的初生和次生代谢通路DEGs富集分析,发现类胡萝卜素的积累过程中,细胞壁、脂质、类黄酮代谢差异基因大量富集。研究结果揭示,‘琯溪蜜柚’果实红肉突变品种‘红肉蜜柚’和‘三红蜜柚’果实类胡萝卜素的形成和积累,是由类胡萝卜素的代谢相关酶基因差异表达和转录因子调控引起的;在果肉红色芽变的同时,伴随着与汁胞粒化相关的基因突变。这对进一步开展柚子果实类胡萝卜素的形成与积累、汁胞粒化研究,为解析柚果实不同颜色突变、汁胞粒化的机制有重要参考价值。In order to study the molecular mechanism of carotenoid metabolism of‘Guanximiyou’pomelo red mutant line,‘Guanximiyou’pomelo(GX),its budding first generation‘Hongroumiyou’(HR)and budding second generation‘Sanhongmiyou’(SH)were used to study the carotenoid metabolism in the pulp of the three varieties at three development stages by the RNA SEQ technology.By bioinformatics analysis,10 differentially expressed genes were isolated to participate in the biosynthesis of carotenoids,namely,PSY,ZDS,CRTISO1/2,NPQ1,and 9-cis-epoxy carotenoids,the genes of oxygenase(NCED3/5),abscisic aldehyde oxidase(AAO3),ABA 8’-hydroxylase(CYP707A1),carotenoid cleavage dioxygenases gene(CCD4).Co expression analysis revealed that the expression patterns of seven family transcription factors were highly correlated with carotenoid synthesis genes,which were MADS,bZIP,bHLH,MYB,AP2/ERF,NAC and WRKY.The expression patterns of some genes were verified by real-time quantitative PCR(qRT-PCR),and the results were highly consistent with those of RNA-seq.In addition,DEGs enrichment analysis of primary and secondary metabolic pathways in‘red flesh honey pomelo’showed that during the accumulation of carotenoids,genes of cell wall,lipid and flavonoid metabolism were abundant.The results showed that the red flesh mutation of GX was caused by the differential expression of carotenoid metabolism related enzyme genes and the regulation of transcription factors;at the same time,the red bud mutation in flesh was accompanied by the mutation related to juice granulation.This may provide a valuable reference for further study on the formation and accumulation of carotenoids and juice granulation,and also provide a way to analyze the mechanism of different color mutation and juice granulation in pomelo fruit.

关 键 词：琯溪蜜柚 芽变 类胡萝卜素 转录组 基因 转录因子 

分 类 号：S666.3[农业科学—果树学]