Article(id=1198652630173057628, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1198652605778985059, articleNumber=null, orderNo=null, doi=10.16438/j.0513-4870.2023-0516, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1682438400000, receivedDateStr=2023-04-26, revisedDate=1688054400000, revisedDateStr=2023-06-30, acceptedDate=null, acceptedDateStr=null, onlineDate=1763710656921, onlineDateStr=2025-11-21, pubDate=1691769600000, pubDateStr=2023-08-12, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1763710656921, onlineIssueDateStr=2025-11-21, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1763710656921, creator=13701087609, updateTime=1763710656921, updator=13701087609, issue=Issue{id=1198652605778985059, tenantId=1146029695717560320, journalId=1189982191388893191, year='2023', volume='58', issue='8', pageStart='0', pageEnd='2540', issueExtLink='null', onlineDate='null', pubDate='1691769600000', pubDateStr='2023-08-12', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1763710651106, creator='13701087609', updateTime=1763710739504, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1198652976601596347, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1198652605778985059, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1198652976601596348, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1198652605778985059, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=2522, endPage=2531, ext={EN=ArticleExt(id=1198652630567322245, articleId=1198652630173057628, tenantId=1146029695717560320, journalId=1189982191388893191, language=EN, title=Systematic analysis of MYB transcription factors related to the geniposide biosynthesis in Gardenia jasminoides Ellis based on whole genome, columnId=null, journalTitle=Acta Pharmaceutica Sinica, columnName=null, runingTitle=null, highlight=null, articleAbstract=

MYB transcription factors are involved in the regulation of various secondary metabolites biosynthesis. Gardenia jasminoides Ellis is the commonly used Chinese herbal medicine, and its main active ingredient is geniposide. Here, leaves and flower buds at different developmental stages of G. jasminoides were used to explore MYB transcription factors related to geniposide biosynthesis based on genome and transcriptome analysis. Transcriptome data analysis showed that, different from the expression of the common pathway genes for terpenoid biosynthesis, the expression level of genes in the specific pathway of geniposide biosynthesis was significantly higher in flower buds than in leaves, which was the same as the organ accumulation pattern of this component. And the promoter regions of geraniol synthase, iridoid synthase and geniposidic acid methyltransferase involved in the specific pathway all contained multiple MYB-binding sites. A total of 105 MYB transcription factors were obtained by annotating the coding genes of G. jasminoides, which were divided into 68 1R-MYB, 33 R2R3-MYB, 3 3R-MYB and 1 atypical MYB transcription factor according to the number of conserved domain. Based on the analysis of phylogenetic tree and quantitative real-time PCR, three candidate MYB transcription factors related to geniposide biosynthesis were selected, including potential positive regulation factor GjMYB23 and negative regulation factors GjMYB31 and GjMYB73. The results of this study will lay a foundation for searching the regulation of geniposide biosynthesis and further analysis of the quality formation mechanism of G. jasminoides, so as to promote the breeding of excellent varieties of G. jasminoides.

, authors=null, authorsList=Wen-jie XU, Yuan-hao HUANG, Rong-rong HAN, Yan-qin LIU, Min CAO, Jing-yuan SONG, authorCompany=null, correspAuthors=Jing-yuan SONG, authorNote=null, correspAuthorsNote=null, copyrightStatement=Copyright ©2023 Acta Pharmaceutica Sinica. All rights reserved., copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1198652633511723865, articleId=1198652630173057628, tenantId=1146029695717560320, journalId=1189982191388893191, language=CN, title=基于全基因组的栀子苷生物合成相关MYB转录因子系统分析, columnId=1190335348896011050, journalTitle=药学学报, columnName=研究论文, runingTitle=null, highlight=null, articleAbstract=

MYB转录因子参与调控多种次生代谢产物的生物合成。栀子是我国大宗常用中药材, 主要活性成分为栀子苷。本研究以栀子不同发育时期的叶片和花蕾器官为研究材料, 借助基因组和转录组分析, 全面挖掘栀子苷生物合成相关MYB转录因子。转录组数据分析表明, 不同于萜类合成共有途径基因表达模式, 栀子苷合成特异途径基因在花蕾中的表达量显著高于叶片, 表达模式与该成分的器官积累规律相同, 且香叶醇合酶、环烯醚萜合酶和栀子苷酸甲基转移酶编码基因的启动子区域均含有多个MYB转录因子结合位点。对栀子基因组编码基因进行注释, 共获得105个MYB转录因子, 根据保守结构域数量将其划分为68个1R-MYB、33个R2R3-MYB、3个3R-MYB和1个非典型MYB转录因子。进一步结合系统发育树和实时荧光定量PCR分析, 筛选出3个可能调控栀子苷生物合成的候选MYB转录因子, 包括潜在正调控转录因子GjMYB23及负调控转录因子GjMYB31和GjMYB73。本研究结果将为栀子苷生物合成调控研究及进一步解析栀子品质形成机制奠定基础, 以推动栀子优良品种选育。

, authors=null, authorsList=许文杰, 黄远浩, 韩蓉蓉, 刘燕琴, 曹敏, 宋经元, authorCompany=null, correspAuthors=宋经元, authorNote=null, correspAuthorsNote=
*宋经元, Tel: 86-10-57833199, E-mail:
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Key Lab of Chinese Medicine Resources Conservation, State Administration of Traditional Chinese Medicine of the People's Republic of China, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China
3. Engineering Research Center of Chinese Medicine Resource, Ministry of Education, Beijing 100193, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1198960118181228833, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, authorId=1198960117862461695, language=CN, stringName=宋经元, firstName=经元, middleName=null, lastName=宋, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=1, 3, *, address=1.中国医学科学院北京协和医学院药用植物研究所, 国家中医药管理局中药资源保护重点研究室, 北京 100193
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Plant Physiol J (植物生理学报), 2022, 585: 904-918., articleTitle=Identification and analysis of 1R MYB transcription factor subfamily in tobacco, refAbstract=null), Reference(id=1198960126947324049, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, doi=10.3390/ijms20133195, pmid=null, pmcid=null, year=2019, volume=20, issue=null, pageStart=3195, pageEnd=null, url=null, language=null, rfNumber=[30], rfOrder=29, authorNames=null, journalName=Int J Mol Sci, refType=null, unstructuredReference=Yong Y, Zhang Y, Lyu Y. A MYB-related transcription factor from Lilium lancifolium L. (LlMYB3) is involved in anthocyanin biosynthesis pathway and enhances multiple abiotic stress tolerance in Arabidopsis thaliana[J]. Int J Mol Sci, 2019, 20: 3195., articleTitle=A MYB-related transcription factor from Lilium lancifolium L. (LlMYB3) is involved in anthocyanin biosynthesis pathway and enhances multiple abiotic stress tolerance in Arabidopsis thaliana, refAbstract=null)], funds=[Fund(id=1198960122472002244, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, awardId=82073966, language=CN, fundingSource=国家自然科学基金项目(82073966), fundOrder=null, country=null), Fund(id=1198960122627191506, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, awardId=cstc2020jscx-cylhX0008, language=CN, fundingSource=川渝道地药材绿色种植技术研究与应用(cstc2020jscx-cylhX0008), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1198960115094221771, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, xref=null, ext=[AuthorCompanyExt(id=1198960115110998989, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, companyId=1198960115094221771, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1. Key Lab of Chinese Medicine Resources Conservation, State Administration of Traditional Chinese Medicine of the People's Republic of China, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China), AuthorCompanyExt(id=1198960115127776207, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, companyId=1198960115094221771, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1.中国医学科学院北京协和医学院药用植物研究所, 国家中医药管理局中药资源保护重点研究室, 北京 100193)]), AuthorCompany(id=1198960115299742694, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, xref=null, ext=[AuthorCompanyExt(id=1198960115308131304, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, companyId=1198960115299742694, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2. 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Engineering Research Center of Chinese Medicine Resource, Ministry of Education, Beijing 100193, China), AuthorCompanyExt(id=1198960115442349049, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, companyId=1198960115421377525, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3.中药资源教育部工程研究中心, 北京 100193)])], figs=[ArticleFig(id=1198960120139968988, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, language=EN, label=null, caption=null, figureFileSmall=/cWO5dU60iGd6xdtrC30kg==, figureFileBig=InR3iuYYv+tRwc1IEZGzcA==, tableContent=null), ArticleFig(id=1198960120249020903, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, language=CN, label=Figure 1, caption= Genes involved in the biosynthetic pathway of geniposide and their expression in <i>Gardenia jasminoides</i>. A: Terpene biosynthetic pathway. G3P: Glyceraldehyde 3-phosphate; DXP: 1-Deoxy-<i>D</i>-xylulose-5-phosphate; MEP: 2-C-Methyl-<i>D</i>-erythritol-4-phosphate; CDP-ME: 4-(Cytidine 5′-diphospho)-2-C-methyl-<i>D</i>-erythritol; CDP-MEP: 2-Phospho-4-(cytidine 5′-diphospho)-2-C-methyl-<i>D</i>-erythritol; MECPP: 2-C-Methyl-<i>D</i>-erythritol-2, 4-cyclodiphosphate; HMBPP: 1-Hydroxy-2-methyl-2-butenyl-4-diphosphate; HMG-CoA: 3-Hydroxy-3-methylglutaryl CoA; MVA: Mevalonate; MVP: Mevalonate phosphate; MVPP: Mevalonate diphosphate; IPP: Isopentenyl pyrophosphate; DMAPP: Dimethylallyl pyrophosphate; B: Specific step in the biosynthetic pathway of iridoid glycoside in <i>Gardenia jasminoides.</i> Red arrows represent catalytic steps that have been elucidated in <i>Gardenia jasminoides</i>; C: Expression heatmap of the genes involved in the biosynthetic pathway of geniposide. YL: Young leaf; ML: Mature leaf; SFB: Small flower bud; LFB: Large flower bud; FPKM: Fragments per kilobase of exon per million; D: Distribution of MYB transcription factors binding sites in the promoter of the specific pathway genes for geniposide biosynthesis. The promoter sequence was 2 000 bp upstream of the transcription start site , figureFileSmall=/cWO5dU60iGd6xdtrC30kg==, figureFileBig=InR3iuYYv+tRwc1IEZGzcA==, tableContent=null), ArticleFig(id=1198960120387432950, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, language=EN, label=null, caption=null, figureFileSmall=1jMZQ+q6nH1xAixKcPBLng==, figureFileBig=D0YV21aQ4Su177PeQx+kog==, tableContent=null), ArticleFig(id=1198960120483901955, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, language=CN, label=Figure 2, caption= Chromosome location of MYB transcription factor genes in <i>Gardenia jasminoides</i> , figureFileSmall=1jMZQ+q6nH1xAixKcPBLng==, figureFileBig=D0YV21aQ4Su177PeQx+kog==, tableContent=null), ArticleFig(id=1198960120605536788, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, language=EN, label=null, caption=null, figureFileSmall=s5N9vcnHWZsqfKONpR3boQ==, figureFileBig=jpTGLS2wmPN50HnUSThkeA==, tableContent=null), ArticleFig(id=1198960120739754527, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, language=CN, label=Figure 3, caption= Sequence analysis of MYB transcription factors in <i>Gardenia jasminoides</i>. A: The phylogenetic tree of GjMYBs; B: The motif distribution of GjMYBs; C: Four conserved motifs of GjMYBs , figureFileSmall=s5N9vcnHWZsqfKONpR3boQ==, figureFileBig=jpTGLS2wmPN50HnUSThkeA==, tableContent=null), ArticleFig(id=1198960120903332401, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, language=EN, label=null, caption=null, figureFileSmall=Zj1o5y1bYiGiozSCi9ONXg==, figureFileBig=muOjTs17+nPemNEEzN3m/w==, tableContent=null), ArticleFig(id=1198960120999801397, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, language=CN, label=Figure 4, caption= Analysis of MYB transcription factors related to geniposide biosynthesis. A: Phylogenetic tree and expression pattern of expressed <i>GjMYB</i> genes; B: Tertiary structures of selected eight GjMYB transcription factors , figureFileSmall=Zj1o5y1bYiGiozSCi9ONXg==, figureFileBig=muOjTs17+nPemNEEzN3m/w==, tableContent=null), ArticleFig(id=1198960121134019139, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, language=EN, label=null, caption=null, figureFileSmall=BQAGqR7FiJWkRquskWsNGA==, figureFileBig=IXP5Ck8pY7yL8DPY212F1w==, tableContent=null), ArticleFig(id=1198960121247265360, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, language=CN, label=Figure 5, caption= Relative expression levels of selected MYB transcription factor genes related to geniposide biosynthesis in <i>Gardenia jasminoides</i> , figureFileSmall=BQAGqR7FiJWkRquskWsNGA==, figureFileBig=IXP5Ck8pY7yL8DPY212F1w==, tableContent=null), ArticleFig(id=1198960121389871711, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
Gene Gene name Accession ID/reference
DXS 1-Deoxy-D-xylulose-5-phosphate synthase Q38854
DXR 1-Deoxy-D-xylulose-5-phosphate reductoisomerase ARU08103
CMS 4-Diphosphocytidyl-2-C-methyl-D-erythritol synthase Q5N8G1
CMK 4-Diphosphocytidyl-2-C-methyl-D-erythritol kinase O81014
MCS 2-C-Methyl-D-erythritol-2, 4-cyclodiphosphate synthase Q9CAK8
HDS 4-Hydroxy-3-methylbut-2-enyl diphosphate synthase F4K0E8
HDR 4-Hydroxy-3-methylbut-2-enyl diphosphate reductase Q94B35
AACT Acetoacetyl-CoA thiolase AAM00280
HMGS 3-Hydroxy-3-methylglutaryl CoA synthase P54873
HMGR 3-Hydroxy-3-methylglutaryl CoA reductase P43256
MVK Mevalonate kinase P46086
PMK 5-Phosphomevalonate kinase Q9C6T1
MVD Mevalonate-5-diphosphate decarboxylase O23722
IS Iridoid synthase MN120556
IS2 Iridoid synthase 2 MN120557
IS4 Iridoid synthase 4 MN120558
GPPS Geranyl diphosphate synthase AHA82035
GES Geraniol synthase QCT83300
IO Iridoid oxidase AHK60833
8HGO 8-Hydroxygeraniol oxidoreductase AHK60836
G10H Geraniol 10-hydroxylase Q8VWZ7
GAMT1 Geniposidic acid methyltransferase 1 [9]
GAMT2 Geniposidic acid methyltransferase 2 [9]
), ArticleFig(id=1198960121494729323, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, language=CN, label=Table 1, caption=

Genes involved in the biosynthetic pathway of iridoid glycosides. Sequences of GAMT1 and GAMT2 genes were used with permission from reference[9], copyright (2021) Elsevier B. V.

, figureFileSmall=null, figureFileBig=null, tableContent=
Gene Gene name Accession ID/reference
DXS 1-Deoxy-D-xylulose-5-phosphate synthase Q38854
DXR 1-Deoxy-D-xylulose-5-phosphate reductoisomerase ARU08103
CMS 4-Diphosphocytidyl-2-C-methyl-D-erythritol synthase Q5N8G1
CMK 4-Diphosphocytidyl-2-C-methyl-D-erythritol kinase O81014
MCS 2-C-Methyl-D-erythritol-2, 4-cyclodiphosphate synthase Q9CAK8
HDS 4-Hydroxy-3-methylbut-2-enyl diphosphate synthase F4K0E8
HDR 4-Hydroxy-3-methylbut-2-enyl diphosphate reductase Q94B35
AACT Acetoacetyl-CoA thiolase AAM00280
HMGS 3-Hydroxy-3-methylglutaryl CoA synthase P54873
HMGR 3-Hydroxy-3-methylglutaryl CoA reductase P43256
MVK Mevalonate kinase P46086
PMK 5-Phosphomevalonate kinase Q9C6T1
MVD Mevalonate-5-diphosphate decarboxylase O23722
IS Iridoid synthase MN120556
IS2 Iridoid synthase 2 MN120557
IS4 Iridoid synthase 4 MN120558
GPPS Geranyl diphosphate synthase AHA82035
GES Geraniol synthase QCT83300
IO Iridoid oxidase AHK60833
8HGO 8-Hydroxygeraniol oxidoreductase AHK60836
G10H Geraniol 10-hydroxylase Q8VWZ7
GAMT1 Geniposidic acid methyltransferase 1 [9]
GAMT2 Geniposidic acid methyltransferase 2 [9]
), ArticleFig(id=1198960121599586940, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
Gene Forward primer (5′→3′) Reverse primer (5′→3′)
Actin TCCTCTTCCAGCCTTCTATC GCTCATACGGTCAGCAATAC
GjMYB9 TACTCGTACGAAGAGGTGAG GTTTACGACCTGGAATTCTCC
GjMYB16 GTGTTAGTGCAGGGTCTTAC TGCGTTTCCAGACTCTAATG
GjMYB19 CAGTTCAATGCCACCTCTAA AGGATGGCTACTGTTGATATTG
GjMYB23 GGACGAAAGTCAAGACTCTAAG TTGTGTTCCAATACTCCTCAAT
GjMYB31 CATGGAAGCAGCAAGTTTAAG GACTGTGGAAGTCCAATAGATAG
GjMYB73 GCCCAATACGATAAGGAGAC GTAGTTGGGATACGGAACAC
GjMYB76 CGAATGCCTCTTCCTTCTTC AGGCTAATGGATGGCTTTATAC
GjMYB101 CTCCTCATCATCGTCCATTTC CAGCTGGCTGCATTATTCT
), ArticleFig(id=1198960121742193292, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, language=CN, label=Table 2, caption=

Primers for qRT-PCR of MYB transcription factor genes in Gardenia jasminoides. MYB: v-myb avian myeloblastosis viral oncogene homolog

, figureFileSmall=null, figureFileBig=null, tableContent=
Gene Forward primer (5′→3′) Reverse primer (5′→3′)
Actin TCCTCTTCCAGCCTTCTATC GCTCATACGGTCAGCAATAC
GjMYB9 TACTCGTACGAAGAGGTGAG GTTTACGACCTGGAATTCTCC
GjMYB16 GTGTTAGTGCAGGGTCTTAC TGCGTTTCCAGACTCTAATG
GjMYB19 CAGTTCAATGCCACCTCTAA AGGATGGCTACTGTTGATATTG
GjMYB23 GGACGAAAGTCAAGACTCTAAG TTGTGTTCCAATACTCCTCAAT
GjMYB31 CATGGAAGCAGCAAGTTTAAG GACTGTGGAAGTCCAATAGATAG
GjMYB73 GCCCAATACGATAAGGAGAC GTAGTTGGGATACGGAACAC
GjMYB76 CGAATGCCTCTTCCTTCTTC AGGCTAATGGATGGCTTTATAC
GjMYB101 CTCCTCATCATCGTCCATTTC CAGCTGGCTGCATTATTCT
), ArticleFig(id=1198960121876411032, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
Gene name Gene ID Class AA size Gene name Gene ID Class AA size
GjMYB1 Gj1P4T24 1R 552 GjMYB54 Gj9A610T144 1R 124
GjMYB2 Gj1A24T33 1R 147 GjMYB55 Gj9X626T25 1R 314
GjMYB3 Gj1P524T10 1R 280 GjMYB56 Gj9A636T100 1R 243
GjMYB4 Gj1P528T16 1R 779 GjMYB57 Gj9A654T110 1R 204
GjMYB5 Gj2A357T40 1R 257 GjMYB58 Gj9A661T106 1R 239
GjMYB6 Gj2P379T15 R2R3 586 GjMYB59 Gj9X666T65 R2R3 369
GjMYB7 Gj2A386T122 1R 642 GjMYB60 Gj9A682T134 1R 112
GjMYB8 Gj2P388T19 1R 993 GjMYB61 Gj9P698T16 1R 176
GjMYB9 Gj2A393T78 1R 97 GjMYB62 Gj9A770T55 R2R3 307
GjMYB10 Gj2A400T31 1R 912 GjMYB63 Gj9A774T63 1R 265
GjMYB11 Gj2P448T7 1R 293 GjMYB64 Gj9A778T112 Atypical 1 099
GjMYB12 Gj3A273T7 1R 134 GjMYB65 Gj9A798T79 R2R3 267
GjMYB13 Gj4A31T63 1R 596 GjMYB66 Gj9P800T24 1R 551
GjMYB14 Gj4A284T28 R2R3 372 GjMYB67 Gj9A829T62 1R 323
GjMYB15 Gj5A22T109 1R 267 GjMYB68 Gj9A839T43 1R 191
GjMYB16 Gj5A229T11 1R 210 GjMYB69 Gj9A839T49 1R 172
GjMYB17 Gj6P85T6 1R 463 GjMYB70 Gj9P839T9 1R 943
GjMYB18 Gj6A273T22 R2R3 437 GjMYB71 Gj9A842T63 1R 614
GjMYB19 Gj6P295T8 1R 308 GjMYB72 Gj9A842T65 1R 183
GjMYB20 Gj7A210T21 R2R3 491 GjMYB73 Gj9X870T42 1R 91
GjMYB21 Gj7A210T23 R2R3 487 GjMYB74 Gj9X871T92 1R 88
GjMYB22 Gj7P308T5 1R 495 GjMYB75 Gj9A887T113 1R 301
GjMYB23 Gj7A335T61 1R 77 GjMYB76 Gj9P925T22 1R 410
GjMYB24 Gj7P355T9 1R 71 GjMYB77 Gj9X971T52 R2R3 267
GjMYB25 Gj7P357T7 1R 222 GjMYB78 Gj9X980T34 1R 94
GjMYB26 Gj7A357T62 R2R3 336 GjMYB79 Gj9X981T83 1R 93
GjMYB27 Gj7P357T10 R2R3 243 GjMYB80 Gj9X995T40 R2R3 334
GjMYB28 Gj7A405T102 1R 464 GjMYB81 Gj9P1015T8 R2R3 1 812
GjMYB29 Gj8A42T41 R2R3 335 GjMYB82 Gj9A1031T85 R2R3 232
GjMYB30 Gj8A84T56 1R 467 GjMYB83 Gj9A1036T88 3R 904
GjMYB31 Gj8A308T22 1R 204 GjMYB84 Gj9P1051T18 R2R3 518
GjMYB32 Gj8E331T0 1R 145 GjMYB85 Gj9A1052T114 R2R3 290
GjMYB33 Gj8A382T92 1R 88 GjMYB86 Gj9P1069T25 1R 309
GjMYB34 Gj8P396T16 1R 1 397 GjMYB87 Gj9A1083T89 R2R3 271
GjMYB35 Gj9A7T75 1R 280 GjMYB88 Gj9A1088T92 1R 336
GjMYB36 Gj9P11T26 1R 1 293 GjMYB89 Gj10A273T124 1R 497
GjMYB37 Gj9A69T86 R2R3 297 GjMYB90 Gj10P276T12 R2R3 400
GjMYB38 Gj9A76T57 3R 560 GjMYB91 Gj10A280T93 1R 293
GjMYB39 Gj9A81T123 1R 278 GjMYB92 Gj10P280T27 R2R3 223
GjMYB40 Gj9A87T89 1R 552 GjMYB93 Gj10P280T28 R2R3 203
GjMYB41 Gj9A130T84 R2R3 1 027 GjMYB94 Gj10X286T42 1R 528
GjMYB42 Gj9X141T53 1R 107 GjMYB95 Gj10X300T28 1R 86
GjMYB43 Gj9P154T10 1R 433 GjMYB96 Gj10X304T40 R2R3 354
GjMYB44 Gj9A202T95 1R 487 GjMYB97 Gj11A117T23 R2R3 295
GjMYB45 Gj9A212T108 R2R3 529 GjMYB98 Gj11A233T32 R2R3 491
GjMYB46 Gj9A213T96 R2R3 308 GjMYB99 Gj11A233T34 R2R3 487
GjMYB47 Gj9P220T9 1R 697 GjMYB100 Gj11X253T23 1R 118
GjMYB48 Gj9A225T92 1R 89 GjMYB101 Gj11A376T83 R2R3 316
GjMYB49 Gj9A286T43 1R 185 GjMYB102 Gj11A408T77 R2R3 348
GjMYB50 Gj9A312T88 3R 981 GjMYB103 Gj11A411T50 1R 454
GjMYB51 Gj9A564T94 R2R3 246 GjMYB104 Gj11A417T81 1R 333
GjMYB52 Gj9A584T54 R2R3 278 GjMYB105 Gj11A433T110 1R 205
GjMYB53 Gj9A610T142 1R 300
), ArticleFig(id=1198960122123874981, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198652630173057628, language=CN, label=Table 3, caption=

The classification of MYB transcription factor genes in Gardenia jasminoides. AA size: The number of amino acid

, figureFileSmall=null, figureFileBig=null, tableContent=
Gene name Gene ID Class AA size Gene name Gene ID Class AA size
GjMYB1 Gj1P4T24 1R 552 GjMYB54 Gj9A610T144 1R 124
GjMYB2 Gj1A24T33 1R 147 GjMYB55 Gj9X626T25 1R 314
GjMYB3 Gj1P524T10 1R 280 GjMYB56 Gj9A636T100 1R 243
GjMYB4 Gj1P528T16 1R 779 GjMYB57 Gj9A654T110 1R 204
GjMYB5 Gj2A357T40 1R 257 GjMYB58 Gj9A661T106 1R 239
GjMYB6 Gj2P379T15 R2R3 586 GjMYB59 Gj9X666T65 R2R3 369
GjMYB7 Gj2A386T122 1R 642 GjMYB60 Gj9A682T134 1R 112
GjMYB8 Gj2P388T19 1R 993 GjMYB61 Gj9P698T16 1R 176
GjMYB9 Gj2A393T78 1R 97 GjMYB62 Gj9A770T55 R2R3 307
GjMYB10 Gj2A400T31 1R 912 GjMYB63 Gj9A774T63 1R 265
GjMYB11 Gj2P448T7 1R 293 GjMYB64 Gj9A778T112 Atypical 1 099
GjMYB12 Gj3A273T7 1R 134 GjMYB65 Gj9A798T79 R2R3 267
GjMYB13 Gj4A31T63 1R 596 GjMYB66 Gj9P800T24 1R 551
GjMYB14 Gj4A284T28 R2R3 372 GjMYB67 Gj9A829T62 1R 323
GjMYB15 Gj5A22T109 1R 267 GjMYB68 Gj9A839T43 1R 191
GjMYB16 Gj5A229T11 1R 210 GjMYB69 Gj9A839T49 1R 172
GjMYB17 Gj6P85T6 1R 463 GjMYB70 Gj9P839T9 1R 943
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基于全基因组的栀子苷生物合成相关MYB转录因子系统分析
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许文杰 1 , 黄远浩 1 , 韩蓉蓉 2 , 刘燕琴 2 , 曹敏 2 , 宋经元 1, 3, *
药学学报 | 研究论文 2023,58(8): 2522-2531
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药学学报 |研究论文 2023 , 58 (8) : 2522 -2531
基于全基因组的栀子苷生物合成相关MYB转录因子系统分析
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许文杰1, 黄远浩1, 韩蓉蓉2, 刘燕琴2, 曹敏2, 宋经元1, 3, *
作者信息
  • 1.中国医学科学院北京协和医学院药用植物研究所, 国家中医药管理局中药资源保护重点研究室, 北京 100193
  • 2.重庆市药物种植研究所, 重庆 408435
  • 3.中药资源教育部工程研究中心, 北京 100193
通讯作者:
*宋经元, Tel: 86-10-57833199, E-mail:
Systematic analysis of MYB transcription factors related to the geniposide biosynthesis in Gardenia jasminoides Ellis based on whole genome
Wen-jie XU1, Yuan-hao HUANG1, Rong-rong HAN2, Yan-qin LIU2, Min CAO2, Jing-yuan SONG1, 3, *
Affiliations
  • 1. Key Lab of Chinese Medicine Resources Conservation, State Administration of Traditional Chinese Medicine of the People's Republic of China, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China
  • 2. Chongqing Institute of Medicinal Plant Cultivation, Chongqing 408435, China
  • 3. Engineering Research Center of Chinese Medicine Resource, Ministry of Education, Beijing 100193, China
出版时间: 2023-08-12 doi: 10.16438/j.0513-4870.2023-0516
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MYB转录因子参与调控多种次生代谢产物的生物合成。栀子是我国大宗常用中药材, 主要活性成分为栀子苷。本研究以栀子不同发育时期的叶片和花蕾器官为研究材料, 借助基因组和转录组分析, 全面挖掘栀子苷生物合成相关MYB转录因子。转录组数据分析表明, 不同于萜类合成共有途径基因表达模式, 栀子苷合成特异途径基因在花蕾中的表达量显著高于叶片, 表达模式与该成分的器官积累规律相同, 且香叶醇合酶、环烯醚萜合酶和栀子苷酸甲基转移酶编码基因的启动子区域均含有多个MYB转录因子结合位点。对栀子基因组编码基因进行注释, 共获得105个MYB转录因子, 根据保守结构域数量将其划分为68个1R-MYB、33个R2R3-MYB、3个3R-MYB和1个非典型MYB转录因子。进一步结合系统发育树和实时荧光定量PCR分析, 筛选出3个可能调控栀子苷生物合成的候选MYB转录因子, 包括潜在正调控转录因子GjMYB23及负调控转录因子GjMYB31和GjMYB73。本研究结果将为栀子苷生物合成调控研究及进一步解析栀子品质形成机制奠定基础, 以推动栀子优良品种选育。

栀子  /  栀子苷  /  MYB  /  转录因子  /  基因组

MYB transcription factors are involved in the regulation of various secondary metabolites biosynthesis. Gardenia jasminoides Ellis is the commonly used Chinese herbal medicine, and its main active ingredient is geniposide. Here, leaves and flower buds at different developmental stages of G. jasminoides were used to explore MYB transcription factors related to geniposide biosynthesis based on genome and transcriptome analysis. Transcriptome data analysis showed that, different from the expression of the common pathway genes for terpenoid biosynthesis, the expression level of genes in the specific pathway of geniposide biosynthesis was significantly higher in flower buds than in leaves, which was the same as the organ accumulation pattern of this component. And the promoter regions of geraniol synthase, iridoid synthase and geniposidic acid methyltransferase involved in the specific pathway all contained multiple MYB-binding sites. A total of 105 MYB transcription factors were obtained by annotating the coding genes of G. jasminoides, which were divided into 68 1R-MYB, 33 R2R3-MYB, 3 3R-MYB and 1 atypical MYB transcription factor according to the number of conserved domain. Based on the analysis of phylogenetic tree and quantitative real-time PCR, three candidate MYB transcription factors related to geniposide biosynthesis were selected, including potential positive regulation factor GjMYB23 and negative regulation factors GjMYB31 and GjMYB73. The results of this study will lay a foundation for searching the regulation of geniposide biosynthesis and further analysis of the quality formation mechanism of G. jasminoides, so as to promote the breeding of excellent varieties of G. jasminoides.

Gardenia jasminoides  /  geniposide  /  MYB  /  transcription factor  /  genome
许文杰, 黄远浩, 韩蓉蓉, 刘燕琴, 曹敏, 宋经元. 基于全基因组的栀子苷生物合成相关MYB转录因子系统分析. 药学学报, 2023 , 58 (8) : 2522 -2531 . DOI: 10.16438/j.0513-4870.2023-0516
Wen-jie XU, Yuan-hao HUANG, Rong-rong HAN, Yan-qin LIU, Min CAO, Jing-yuan SONG. Systematic analysis of MYB transcription factors related to the geniposide biosynthesis in Gardenia jasminoides Ellis based on whole genome[J]. Acta Pharmaceutica Sinica, 2023 , 58 (8) : 2522 -2531 . DOI: 10.16438/j.0513-4870.2023-0516
中药栀子来源于茜草科栀子属植物栀子(Gardenia jasminoides Ellis) 的干燥成熟果实, 味苦, 性寒, 具有泻火除烦、清热利湿和凉血解毒等功效。现代药理研究表明, 栀子在保肝利胆、保护神经、抗炎和抗血栓等方面具有一定活性, 是280余种中药制剂和2 330余种治疗疾病经典方剂配方[1], 如急救名药安宫牛黄丸和血脉清片等。栀子天然产物包括环烯醚萜、二萜、有机酸和黄酮等多种化合物, 主要活性成分环烯醚萜多以糖苷形式存在, 其中栀子苷(geniposide) 含量最高, 《中华人民共和国药典》(2020年版一部) 规定该成分是栀子的质量控制指标。近期多项研究发现, 栀子苷在阿尔茨海默症和抑郁症等疾病治疗中展现出多种生理活性, 是潜在的多种脑部疾病预防和治疗药物或先导化合物[2, 3]
栀子始记于《神农本草经》, 迄今已有千余年栽培历史, 主产于江西、四川、重庆和福建等长江以南多地[4], 具有较高遗传多样性[5]。栀子年需求量在6 000吨左右, 这促使人们对栀子进行盲目引种与大规模种植, 品种混杂导致药材质量参差不齐[6], 严重影响栀子疗效及产业化发展。栀子苷生源途径研究可以为栀子种质选育提供关键分子标志物, 广受关注。与众多萜类成分相同, 栀子苷的合成前体物是来源于甲基赤藓醇4-磷酸途径(MEP) 和甲羟戊酸途径(MVA) 的异戊烯基二磷酸(IPP) 和二甲基烯丙基二磷酸(DMAPP)。IPP和DMAPP在一系列酶的催化下合成环烯醚萜醇骨架, 其中香叶醇合酶(GES) 是环烯醚萜类成分合成特异途径的第一个酶[7], 环烯醚萜合酶(IS) 则催化关键的环化过程[8]。上述产物进一步在糖基转移酶、甲基转移酶和氧化还原酶的作用下合成栀子苷等环烯醚萜苷成分, 其中催化栀子苷合成最后关键步骤的栀子苷酸甲基转移酶(GAMT) 已被鉴定[9], 为栀子苷代谢调控研究提供了重要靶基因。
植物MYB (v-myb avian myeloblastosis viral oncogene homolog) 转录因子广泛参与胁迫响应、次生代谢调控和激素应答等过程[10], 与黄酮、萜类和木质素等天然产物合成密切相关[11]。根据保守结构域SANT的数量, MYB转录因子可划分为4种类型: 1R-MYB、R2R3-MYB、3R-MYB及非典型MYB (4个或5个MYB结构域)[12]。研究人员已在香雪兰(Freesia x hybrida)[13]、丹参(Salvia miltiorrhiza)[14]和黄花蒿(Artemisia annua)[15, 16]等中发现调控萜类天然产物合成的MYB转录因子, 但目前尚未见栀子苷生物合成调控相关MYB转录因子报道。前期转录组数据分析和靶向代谢物检测结果表明, 栀子叶片中的栀子苷含量较低, 而花蕾中该成分含量较高, 且栀子苷合成途径关键酶编码基因GAMT的表达模式与该成分的器官积累规律一致[9]。因此, 为探究栀子苷的生物合成是否受到MYB转录因子调控, 本研究同样以栀子不同发育时期叶片和花蕾器官为研究材料, 通过栀子基因组及转录组数据系统分析, 全面挖掘调控栀子苷生物合成特异途径基因表达的MYB转录因子, 为栀子优良种质选育及代谢工程研究提供潜在遗传资源。
转录组数据分析  栀子转录组数据为实验室前期积累数据[9], 包括幼嫩叶片(young leaf, YL)、成熟叶片(mature leaf, ML)、小花蕾(small flower bud, SFB, 长约2 cm) 和大花蕾(large flower bud, LFB, 长约3 cm), 每个器官三个生物学重复。基于栀子参考基因组(NCBI: ASM1310374v1), 借助Hisat2 (v. 2.1.0) 和Cufflinks (v. 2.1.1) 软件分别进行序列比对及基因表达值FPKM (fragments per kilobase of exon per million reads mapped) 计算。从NCBI数据库(https://www.ncbi.nlm.nih.gov/) 下载拟南芥(Arabidopsis thaliana)、长春花(Catharanthus roseus) 和栀子中环烯醚萜苷合成途径基因的蛋白序列(表 1[9]), 以其为参考序列, 使用本地版Blastp (v. 2.10.0) 对栀子相同功能蛋白进行注释。提取栀子中已验证功能基因及与其他物种高度同源基因的4个器官表达值, 使用TBtools (v. 1.108) 软件, 以Log10 (FPKM+1) 绘制表达热图。
MYB转录因子结合位点预测  根据栀子参考基因组, 提取栀子中已表征功能的栀子苷合成特异途径基因上游2 000 bp为启动子序列, 包括GESIS和2个GAMT。使用New PLACE在线工具(https://www.dna.affrc.go.jp/PLACE) 预测上述基因启动子区域的顺式元件结合位点, 使用TBtools软件进行可视化。
MYB转录因子家族分析  基于本课题组已有的栀子基因组注释编码基因序列, 使用plantTFDB数据库(http://planttfdb.gao-lab.org/index.php) 进行转录因子预测。使用SMART在线工具(http://smart.embl-heidelberg.de) 预测MYB转录因子的SANT保守结构域, 过滤无完整结构域基因。使用MEME在线工具(https://meme-suite.org/meme/tools/meme) 预测MYB转录因子保守基序, 数量设置为20。使用TBtools软件对MYB转录因子系统发育树和保守基序进行可视化。分别使用SOPMA在线工具(https://npsa-prabi.ibcp.fr/cgi-bin/npsa_automat.pl?page=npsa%20_sopma.html) 和Swiss-Model (https://swissmodel.expasy.org/) 预测MYB转录因子的二级结构和三级结构。
候选MYB转录因子挖掘  收集与萜类合成相关的MYB转录因子蛋白序列[13-15, 17-21], 提取在栀子任一器官表达MYB基因(FPKM > 10) 的蛋白序列, 分别使用MAFFT和IQ-TREE2软件对其进行多序列比对和系统发育树构建, 自动筛选最优建树模型。根据栀子4个器官的基因表达值FPKM, 使用R软件(v. 3.4.3) Cor函数计算基因间表达相关系数, 提取与栀子苷合成特异途径基因表达模式呈正、负相关的MYB转录因子基因。最终选取与已知萜类代谢相关MYB转录因子亲缘关系较近、编码基因与栀子苷合成特异途径基因表达模式相关的MYB转录因子作为栀子苷生物合成调控候选转录因子。
候选MYB基因qRT-PCR分析  候选GjMYB基因实时荧光定量PCR (qRT-PCR) 验证所用样本包括YL、ML、SFB和LFB, 均为2021年5月采集于重庆市南山区, 实验室-80 ℃冰箱保存。该材料与转录组测序样本为同批次采集, 但为不同样本。使用多糖多酚植物总RNA提取试剂盒(TIANGEN, DP441) 提取栀子样品总RNA。使用PrimerQuest在线工具(https://sg.idtdna.com/Primerquest) 设计MYB基因的qRT-PCR引物(表 2)。扩增片段长度约100 bp, Tm值为60 ℃左右。以栀子YL、ML、SFB和LFB样本质量合格的反转录cDNA为模板, 每个器官2个生物学重复, 每个基因3个技术重复, 使用SYBR Green I染料(CWBIO, CW0955M) 进行MYB基因表达相对定量分析。以栀子Actin为内参基因, 以YL器官为对照组, 根据2–ΔΔCt方法计算候选MYB基因相对表达量。
根据栀子基因组注释文件, 计算4个器官35 967个编码基因的表达水平, 并提取已鉴定栀子苷生源途径基因及环烯醚萜苷合成途径同源基因的表达值。结果显示, 与萜类合成共有途径(图 1A) 基因相比, 除GPPS外的环烯醚萜醇骨架形成及修饰基因均在不同发育阶段的花蕾器官中特异性高表达(P < 0.05), 而在不同发育时期叶片中表达量较低, 尤其是ML样本(图 1BC)。该结果说明栀子苷合成特异途径基因的表达均具有组织特异性, 且表达模式与栀子苷在不同器官中的积累规律一致。
由于栀子苷合成特异途径基因均为器官特异性表达, 提取已验证功能的栀子苷合成特异途径基因上游2 000 bp序列, 包括GESISGAMT1GAMT2 (图 1D)。New PLACE顺式作用元件分析结果显示, GESIS和2个GAMT基因启动子区域均含有16~18个MYB转录因子结合位点, 如MYB1AT、MYBST1、MYBCORE和MYB2CONSENSUSAT等, 提示栀子苷生物合成基因的器官特异性表达可能受到MYB转录因子调控。
通过PlantTFDB数据库对栀子编码基因进行转录因子预测, 共获得119个MYB转录因子。进一步使用SMART数据库筛选含有完整SANT结构域的MYB, 最终确定105个MYB转录因子, 根据其编码基因的染色体位置排序命名为GjMYB1~GjMYB105。基于SANT结构域数量将104个MYB转录因子划分为3类, 包括68个1R-MYB、33个R2R3-MYB和3个3R-MYB (表 3)。此外, GjMYB64含有5个SANT结构域, 为非典型MYB转录因子。
对105个MYB转录因子进行序列分析发现, 其平均蛋白长度为395个氨基酸。所有MYB转录因子基因不平均分布于栀子的11条染色体, 其中9号染色体包含54个MYB转录因子编码基因, 平均MYB基因数量最多(图 2)。MEME预测结果显示, 所有MYB转录因子的保守motif为1~12个(图 3)。数量最多的1R-MYB均含有motif 1、motif 2、motif 3和motif 4中至少1个motif, 除GjMYB81外的32个R2R3-MYB注释到motif 1、motif 2、motif 3和motif 4中至少2个motif, 3个3R-MYB均包含motif 1、motif 2和motif 3。该结果说明, motif 1、motif 2、motif 3和motif 4可能是MYB转录因子保守序列。
提取表达GjMYB基因(FPKM > 10) 的蛋白序列, 将53个GjMYB转录因子与已鉴定参与萜类合成调控的MYB转录因子构建系统发育树。结果显示, 拟南芥、留兰香(Mentha spicata)、香雪兰、黄花蒿、丹参、白桦(Betula platyphylla) 和番茄(Solanum lycopersicum) 中的10个萜类合成调控MYB转录因子聚在一支, 7个GjMYB与其亲缘关系较近, 包括GjMYB9、GjMYB16、GjMYB19、GjMYB23、GjMYB31、GjMYB76和GjMYB101 (图 4A)。在7个GjMYB基因中, GjMYB9GjMYB23GjMYB101表达量较高, GjMYB23GjMYB31GESISGAMT1GAMT2的表达模式相关性较高, 表达相关系数分别大于0.91和小于-0.91。此外, 通过对53个GjMYB基因的表达模式分析发现, GjMYB73基因表达量极高, 且其与上述功能基因的表达模式呈现高度负相关(相关系数小于-0.90)。因此, 将上述8个GjMYB作为调控栀子苷生物合成的候选MYB转录因子。转录因子二维结构分析显示, 筛选的8个GjMYB转录因子均包含α-螺旋、β-转角、β-折叠和无规则卷曲构型, 且以α-螺旋和无规则卷曲为主, 与三维结构同源建模结果相近(图 4B)。
根据基因注释CDS序列对筛选的8个GjMYB基因进行qRT-PCR验证。结果显示, 除GjMYB19GjMYB101外, 其余6个GjMYB基因的表达模式与转录组数据分析结果相近(图 5), 即GjMYB23在花蕾器官中表达量最高, GjMYB31GjMYB73在叶片中的表达量高于花蕾器官, 且成熟叶片中表达量最高。GjMYB9GjMYB16GjMYB76与栀子苷合成特异途径基因的表达模式相关性相对较低, 且与栀子苷的器官积累规律无显著关联。据此, 推测GjMYB23是正向调控栀子苷生物合成的MYB转录因子, 而GjMYB31和GjMYB73可能参与该成分合成的负调控过程。
栀子是临床常用中药之一, 代表性活性成分为环烯醚萜苷, 研究人员已从栀子中分离和鉴定出48种该类化合物[1], 指标成分栀子苷是多种环烯醚萜苷的进一步修饰产物及合成底物。由于次生代谢产物合成基因及其表达调控因子是优良种质选育的重要遗传信息, 研究人员先后鉴定了栀子苷生物合成特异途径基因GESISGAMT1GAMT2, 然而缺乏相应的基因表达调控机制研究。前期研究发现, 栀子的叶片和花蕾器官中栀子苷含量存在较大差异, 活性成分积累呈现器官特异性分布。在此基础上, 本文进一步对栀子苷生源途径基因的表达模式进行分析发现, 所有栀子苷合成特异途径基因均为器官特异性表达, 且表达模式均与栀子苷积累规律相同。此外, GESIS及2个GAMT基因启动子区域包含多个MYB转录因子结合位点, 推测由于MYB转录因子调控栀子苷生物合成基因的器官特异性表达导致活性成分在特定部位合成。
多组学数据联合分析可全面且精准挖掘次生代谢产物合成基因及调控转录因子[22-24], 栀子染色体水平基因组[25]为系统分析栀子苷生物合成相关MYB转录因子提供良好的数据支撑。本研究通过转录组测序、多种生物信息学分析及qRT-PCR验证, 共筛选到3个潜在的栀子苷生物合成相关MYB转录因子。首先, 通过系统发育树分析发现, 候选转录因子GjMYB23和GjMYB31与萜类成分合成相关MYB转录因子聚在一支, 提示已知天然产物合成调控MYB转录因子序列存在一定保守性。qRT-PCR实验证实GjMYB23基因在花蕾中高表达, 而GjMYB31基因在叶片中表达量相对较高, 即两个基因的表达模式分别与栀子苷的器官积累规律呈现正相关与负相关, 推测二者分别是栀子苷生物合成正调控和负调控转录因子。其次, 鉴于栀子苷合成特异途径基因的表达及该成分的积累均呈现器官特异性, 根据基因表达定量分析筛选出高表达的候选基因GjMYB73, 其表达模式与栀子苷合成特异途径基因ISGESGAMTs的表达趋势及栀子苷含量分布为负相关, 可能参与栀子苷生物合成负调控过程。此外, 需要特别说明的是, GjMYB73蛋白序列与MYB转录因子家族中的RAD蛋白同源性较高, 这导致其与萜类合成相关MYB转录因子亲缘关系较远。研究表明, MYB-RAD类蛋白参与对称花型发育过程[26, 27], 但GjMYB73基因在花蕾中表达量极低, 可能具有新的调控功能。
本研究筛选出的3个潜在栀子苷生物合成调控MYB转录因子均为1R-MYB转录因子。然而, 多数研究发现R2R3-MYB型转录因子在植物天然产物合成中发挥重要作用[28], 其不仅可以直接调控靶基因表达, 还可以与光和激素等协同参与次生代谢产物合成。Reddy等[17]在留兰香中发现了能直接与香叶基焦磷酸合酶(GPPS) 编码基因的顺式作用元件结合, 通过抑制GPPS表达负调控单萜合成的R2R3-MYB转录因子。Liu等[16]发现只有当黄花蒿中的R2R3型转录因子AaMYB108存在时, 光和茉莉酸才能促进青蒿素合成。1R-MYB转录因子研究基础相对薄弱[29], 仅有少量研究发现其参与天然产物合成[30]。因此, 本研究结果可能将进一步拓展1R-MYB转录因子的调控功能, 下一步计划通过酵母单杂交或转基因技术等验证候选GjMYB转录因子是否参与栀子苷的生物合成调控过程。
作者贡献: 许文杰负责实验设计、数据分析、基因表达定量实验及文章撰写; 黄远浩参与基因表达定量实验; 韩蓉蓉、刘燕琴及曹敏负责栀子样品采集; 宋经元负责总体实验设计及稿件修改等。
利益冲突: 本文无任何利益冲突。
  • 国家自然科学基金项目(82073966)
  • 川渝道地药材绿色种植技术研究与应用(cstc2020jscx-cylhX0008)
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2023年第58卷第8期
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doi: 10.16438/j.0513-4870.2023-0516
  • 接收时间:2023-04-26
  • 首发时间:2025-11-21
  • 出版时间:2023-08-12
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  • 收稿日期:2023-04-26
  • 修回日期:2023-06-30
基金
国家自然科学基金项目(82073966)
川渝道地药材绿色种植技术研究与应用(cstc2020jscx-cylhX0008)
作者信息
    1.中国医学科学院北京协和医学院药用植物研究所, 国家中医药管理局中药资源保护重点研究室, 北京 100193
    2.重庆市药物种植研究所, 重庆 408435
    3.中药资源教育部工程研究中心, 北京 100193

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*宋经元, Tel: 86-10-57833199, E-mail:
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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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