西瓜果肉柠檬黄色的遗传分析和基因定位  

Genetic Analysis and Gene Mapping of Canary Yellow in Watermelon Flesh

作　　者：刁卫楠 袁平丽[1] 龚成胜 赵胜杰[1] 朱红菊[1] 路绪强[1] 何楠[1] 杨东东 刘文革[1] DIAO WeiNan;YUAN PingLi;GONG ChengSheng;ZHAO ShengJie;ZHU HongJu;LU XuQiang;HE Nan;YANG DongDong;LIU WenGe(Zhengzhou Fruit Research Institute,Chinese Academy of Agricultural Sciences,Zhengzhou 450009)

机构地区：[1]中国农业科学院郑州果树研究所,郑州450009

出　　处：《中国农业科学》2021年第18期3945-3958,共14页Scientia Agricultura Sinica

基　　金：国家重点研发计划(2018YFD0100704);中国农业科学院科技创新工程(CAAS-ASTIP-2016-ZPRI);现代产业技术体系建设专项(CARS-25-03);中国农业科学院基本科研业务费专项(1610192020103);国家自然科学基金(31672178,31471893)。

摘　　要：【目的】对西瓜白色和柠檬黄色果肉的色素成分、色素含量、遗传规律进行研究,通过BSA-seq进行基因定位,并预测与柠檬黄色果肉相关的候选基因,为深入研究西瓜柠檬黄色果肉的遗传与分子机制奠定理论基础。【方法】本研究选用‘冰糖脆’(ⅠP1,白色果肉)和‘喜华’(ⅠP2,柠檬黄色果肉),‘萨省奶油瓜’(ⅡP1,白色果肉)和‘新金兰选’(ⅡP2,柠檬黄色果肉)4份纯合自交系材料为亲本分别配置杂交组合,构建了两个六世代群体。利用高效液相色谱法(HPLC)对4个亲本材料4个不同发育时期的类胡萝卜素组分和含量进行测定。利用集群分离分析法(bulked segreant analysis,BSA)实现对两个BSA-seq群体(BSA-seqⅠ和BSA-seqⅡ)的初定位,然后根据西瓜参考基因组‘97103’V2注释信息挖掘候选基因,并通过实时荧光定量PCR(qRT-PCR)对候选基因进行验证。【结果】在西瓜果实发育过程中,紫黄质和叶黄素在双亲中差异性积累,其中紫黄质具有更高的含量,且在柠檬黄色果肉中的含量显著高于白色果肉。成熟期西瓜白色果肉中紫黄质含量为(10.96±4)μg·g^(-1) DW,柠檬黄果肉中紫黄质含量为(22.84±2)μg·g^(-1) DW;成熟期西瓜白色果肉中叶黄素含量为(2.23±1)μg·g^(-1) DW,柠檬黄果肉中叶黄素含量为(3.97±1)μg·g^(-1) DW。在构建的两组六世代分离群体中,ⅠF_(1)、ⅡF_(1)、ⅠBC_(1)P_(1)、ⅡBC_(1)P_(1)群体西瓜果肉颜色均为非柠檬黄色,F2群体中西瓜果肉非柠檬黄色与柠檬黄色的分离比符合3∶1的孟德尔分离比例,ⅠBC_(1)P_(2)、ⅡBC_(1)P_(2)回交群体果肉非柠檬黄色和柠檬黄色分离比符合1∶1,表明西瓜果肉柠檬黄色对白色为隐性性状。通过对BSA-seqⅠ和BSA-seqⅡ数据进行SNP和InDel关联分析,将控制西瓜果肉柠檬黄色的主效位点定位在6号染色体24.00-24.61 Mb的区域内,该区域内共有70个基因。结合西瓜参考基因组注释信息及【Objective】In order to find out the formation mechanism of watermelon canary yellow flesh color,the pigment composition,pigment contents,and the inheritance of white and canary yellow flesh watermelon color were studied in this study,and the candidate genes related to canary yellow flesh were predicted by BSA-seq.【Method】Two six generations populations were constructed by crossing four materials,including Bingtangcui(white flesh)and Xihua(canary yellow flesh),as well as Sashengnaiyougua(white flesh)and Xinjinlanxuan(canary yellow flesh).High performance liquid chromatography(HPLC)was used to determine the carotenoid composition and content in flesh of four parents at four different development stages.Two BSA-seq populations(BSA-seq I and BSA-seq II)were initially located by using the bulked sergeant analysis(BSA),and then the candidate genes were screened according to the annotation information of watermelon reference genome‘97103’V2,and verified by real-time quantitative polymerase chain reaction(qRT-PCR).【Result】During the development of watermelon fruit,violaxanthin and lutein accumulated differently in both parents,and the content of violaxanthin was higher in canary yellow flesh than in white flesh.The content of violaxanthin in white and canary yellow flesh watermelon fruits were(10.96±4)μg·g^(-1) DW and(22.84±2)μg·g^(-1) DW,respectively.The content of lutein in white and canary yellow flesh watermelon fruits were(2.23±1)μg·g^(-1) DW and(3.97±1)μg·g^(-1) DW,respectively.The content of violaxanthin was about 7 times of lutein.Two six generations segregation populations were analyzed,and the results showed that the flesh color of F_(1) andⅠBC_(1)P_(1) andⅡBC_(1)P_(1) populations were non-canary yellow,the segregation ratio of non-canary yellow and canary yellow in F2 population were consistent with the Mendelian segregation ratio of 3:1,and the segregation ratio of non-canary yellow and canary yellow inⅠF_(1),ⅡF_(1),ⅠBC_(1)P_(2) andⅡBC_(1)P_(2) backcross populations wer

关 键 词：西瓜 果肉 柠檬黄色 类胡萝卜素 基因定位 

分 类 号：S651[农业科学—果树学]