摘要
【目的】菜心口感脆嫩,营养价值高,是华南地区人们最喜爱的蔬菜之一,其中糖的种类和含量是决定菜心甜度和风味的主要因素。分析菜心不同器官糖含量及与糖代谢和转运相关基因的表达,以探明导致菜心不同器官糖含量差异的分子机制。【方法】采用高效液相色谱法测定菜心叶片、薹茎和花蕾中可溶性糖、蔗糖、葡萄糖和果糖的含量;通过转录组测序技术对菜心叶片、薹茎和花蕾基因的表达量进行全面系统的分析,并利用DESeq2软件鉴定3个不同器官间的差异表达基因,分析糖代谢通路及转运过程涉及的酶编码基因的表达,在基迪奥生信云平台将筛选到的与糖代谢转运相关的差异表达基因和糖含量进行相关性分析,同时利用实时荧光定量PCR(qRT-PCR)技术对部分基因的表达量进行验证。【结果】糖含量测定结果表明,菜心不同器官之间糖含量差异明显。可溶性糖、蔗糖、葡萄糖和果糖含量均呈现花蕾>薹茎>叶片的趋势。花蕾中葡萄糖含量分别为薹茎和叶片的1.3倍和1.6倍,果糖含量分别为薹茎和叶片的1.42倍和1.78倍。通过分析糖代谢关键酶和转运蛋白编码基因与糖含量变化趋势的一致性,共找出18个与糖含量变化趋势一致的基因。通过分析糖代谢转运中相关差异基因的表达与糖种类及含量的相关性和一致性,鉴定了14个与糖含量变化趋势一致的DEG。合并7个共有基因(Bra020096、Bra029914、Bra033419、Bra037980、Bra027398、Bra006129和Bra006130),共计25个基因与糖含量变化趋势一致。qRT-PCR结果分析表明筛选到的8个差异基因的表达量FPKM值与相对表达水平高度一致,说明测序结果准确可靠。【结论】菜心花蕾中的糖含量高于薹茎和叶片,是由合成相关酶编码基因的表达在薹茎和花蕾中高于叶片所致,研究结果为揭示菜心不同器官中糖代谢及转运的分子机制奠定了一定基础。
【Objective】Chinese flowering cabbage has become one of the most popular vegetables in southern China due to its crunchy taste and high nutritional value.The type and content of sugars are the main factors for determining the sweetness and flavour of Chinese flowering cabbage.Therefore,this paper analyzed the sugar content and the expression of genes related to sugar metabolism and transport in different organs,so as to investigate the molecular mechanisms responsible for the differences in sugar content in different organs of Chinese flowering cabbage.【Method】In this experiment,the content of soluble sugars,sucrose,glucose and fructose in leaves,stalks and flower buds were determined by high performance liquid chromatography(HPLC).The expression of genes in leaves,stalks and buds was analyzed comprehensively and systematically by transcriptome sequencing(RNA-seq),and the differentially expressed genes(DEGs)between three different organs were identified using DESeq2 software.In addition,the expression of genes encoding enzymes involved in the sugar metabolic pathway and transport process was analyzed,and the correlation with sugar content were analyzed on OmicShare.Moreover,the expression of some genes was verified using quantitative real-time fluorescence PCR(qRT-PCR).【Result】The sugar content varied significantly among the different organs of Chinese flowering cabbage.Soluble sugars,sucrose,glucose and fructose all showed a trend of buds>stalks>leaves.The glucose content in flower buds was 1.3 and 1.6 times of that in stalks and leaves;the fructose content was 1.42 and 1.78 times of that in stalks and leaves,respectively.By analyzing the relation and consistency between the expression of genes coding enzymes involving in sugar metabolism and transport and content of the sugar,a total of 18 genes were screened.By analyzing the correlation and consistency of the expression of DEGs among different organs with sugar species and content,14 DEGs were identified.By merging 7 common genes(Bra020096,Bra029914,Bra033419,Bra037980,Bra027398,Bra006129 and Bra006130),expression of 25 genes in total were consistent with the sugar content.qRT-PCR results showed that the relative expression of the eight DEGs were highly consistent with their FPKM values from RNA-seq,indicating that the sequencing results were accurate and reliable.【Conclusion】The sugar content in the flower buds of Chinese flowering cabbage was higher than that in stalks and leaves,because the expression of genes encoding sugar synthesis-related enzymes was higher in buds than in stalks and leaves.These results laid a certain foundation for revealing the molecular mechanism of sugar metabolism and transport in different organs of Chinese flowering cabbage.
作者
冯献君
王丽
王彤
侯雷平
李梅兰
FENG XianJun;WANG Li;WANG Tong;HOU LeiPing;LI MeiLan(College of Horticulture,Shanxi Agricultural University,Taigu 030801,Shanxi;Modern Agricultural Industry Development Service Center,Agricultural and Rural Bureau,Zuoquan County,Zuoquan 032600,Shanxi)
出处
《中国农业科学》
CAS
CSCD
北大核心
2023年第11期2158-2171,共14页
Scientia Agricultura Sinica
基金
山西省重点研发计划重点项目(201703D211001-04-01)
山西农业大学曲沃果蔬研究院项目(2021QWGS-3)。
关键词
菜心
器官
糖类
转录组
基因
Chinese flowering cabbage
organs
sugar
RNA-seq
gene
