摘要
【目的】黄酮类化合物具有抗炎、抗癌、抑菌等多种功效,是全缘叶绿绒蒿的主要药用成分之一。通过分析全缘叶绿绒蒿不同部位的空间代谢组与转录组信息,挖掘调控黄酮类化合物合成的关键基因,为研究全缘叶绿绒蒿类黄酮合成机制提供理论参考,并为提高类黄酮含量遗传育种奠定分子基础。【方法】以全缘叶绿绒蒿的根、茎、叶和花瓣为材料,对不同部位进行转录组测序,并通过空间代谢组数据分析黄酮类化合物在不同部位中的分布情况,利用加权基因共表达网络分析(WGCNA)鉴定出与黄酮类化合物合成密切相关的关键模块和关键基因。另外,挑选12个基因进行qRT-PCR,验证转录组数据的可靠性。【结果】全缘叶绿绒蒿中黄酮类化合物在不同部位呈现差异积累,花瓣是黄酮类化合物积累的主要部位,同时明确了8种主要黄酮类化合物。转录组测序共获得20085个表达基因,仅在花中表达的基因有286个,是仅在其他部位中表达基因数量的3.6—4.2倍。利用WGCNA对过滤后的高表达差异基因进行划分,共获得14个共表达模块,确定了关键模块MEturquoise和MEgreen与8个主要黄酮类化合物显著相关(P<0.05)。KEGG分析发现这两个模块基因主要在代谢相关的通路中富集,在黄酮类化合物合成相关通路上也有基因富集,两个模块分别包含了18个和6个与黄酮类化合物合成相关的基因,并从模块中筛选到14个核心结构基因(5个CHS、2个HIDH、2个CCoAOMT以及FLS、CYP75B1、CHI、HCT和CYP73A)和1个转录因子HB2,这些基因多在花瓣或是茎中高表达。qRT-PCR所测基因表达变化趋势与转录组基本一致,表明利用该转录组数据所得出的分析结果可信。【结论】全缘叶绿绒蒿黄酮类化合物积累和基因表达在不同器官间具有显著差异,联合分析筛选到与黄酮类化合物积累密切相关的14个核心结构基因和1个转录因子,这些基因可能在调控全缘叶绿绒蒿不同器官黄酮类化合物的合成和差异积累过程中起到关键作用。
【Objective】Flavonoids are known for their anti-inflammatory,anti-cancer,and antibacterial properties,and are one of the main medicinal components of Meconopsis integrifolia.By analyzing the spatial metabolome information and transcriptome data from various parts(roots,stems,leaves and petals)of M.integrifolia,the key genes regulating the flavonoid synthesis could be excavated.This research could provide valuable insights into the mechanism underlying flavonoids synthesis in M.integrifolia,paving the way for genetic breeding aimed at enhancing flavonoid content.【Method】Transcriptome sequencing was conducted on the root,stem,leave and petal of M.integrifolia to analyze the distribution of flavonoids across different parts using spatial metabolomic data.The weighted gene co-expression network analysis(WGCNA)was employed to identify key modules and key genes closely related to flavonoid synthesis.To validate the reliability of the transcriptome data,12 genes were selected for qRT-PCR analysis.【Result】Flavonoids accumulation in M.integrifolia varied across different parts,with petals being the primary site of accumulation,where 8 main flavonoids were identified.Transcriptome sequencing revealed a total of 20085 expressed genes,among which 286 genes expressed were exclusively expressed in flowers,showing 3.6-4.2 times more expression than that in other plant parts.Using WGCNA to categorize highly expressed differential genes,a total of 14 co-expression modules were identified,and the key modules,including MEturquoise and MEgreen,were significantly associated with 8 main flavonoids(P<0.05).KEGG analysis demonstrated that the genes within these two modules were primarily enriched in metabolism-related pathways,with some genes enriched in pathways related to flavonoid synthesis.MEturquoise and MEgreen comprised 18 and 6 genes related to flavonoid synthesis,respectively,and screened 14 core structural genes(5 CHSs,2 HIDHs,2 CCoAOMTs and FLS,CYP75B1,CHI,HCT,and CYP73A)and one transcription factor HB2,which were predominantly highly expressed in petals or stems.The consistent gene expression trends between qRT-PCR and transcriptome data were observed,which showed that the analysis results derived from the transcriptome data were reliable.【Conclusion】The accumulation of flavonoids and gene expression patterns in different organs of M.integrifolia varied significantly,and 14 core structural genes and one transcription factor were screened to be closely related to the accumulation of flavonoids across different organs.These genes might play a key role in regulating the synthesis and differential accumulation of flavonoids in different organs of M.integrifolia.
作者
陈晓涓
王海菊
王富敏
雍清青
黄顺满
屈燕
CHEN XiaoJuan;WANG HaiJu;WANG FuMin;YONG QingQing;HUANG ShunMan;QU Yan(College of Landscape Architecture and Horticulture,Southwest Forestry University,Kunming 650224;Southwest Engineering and Technology Research Center of Landscape Architecture,National Forestry and Grassland Administration,Kunming 650224;Yunnan Engineering Research Center for Functional Flower Resources and Industrialization,Kunming 650224)
出处
《中国农业科学》
CAS
CSCD
北大核心
2024年第15期3053-3070,共18页
Scientia Agricultura Sinica
基金
国家自然科学基金(32160404,31460218)
云南省“万人计划”青年拔尖人才项目(YNWR-QNBJ-2019-211)。
