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Methods Transcriptome sequencing and analysis was performed on the three tissues of the flowerstalk, rhizome and umbel of B. subcupularis, then searching the candidate enzyme genes involved in triterpene ester biosynthesis by PfamScan, constructing the yeast expression vector of BsubOSCs gene which was introduced into the Saccharomyces cerevisiae lanosterol synthase gene(erg7)-deficient mutant strain GIL77 for functional characterization. Results A total of 87 candidate genes involved in pentacyclic triterpene esters biosynthesis were identified from the transcriptome of B. subcupularis, most of them were highly expressed in umbel, suggesting that umbel may be the main site of synthesis of pentacyclic triterpene esters. The enzyme function of two BsubOSC genes were characterized, which was found that BsubOSC2 could catalyze 2, 3-oxidosqualene to form β-amyrin, but no catalytic product of BsubOSC1 was detected. Conclusion Based on transcriptome analysis, the key enzyme genes involved in pentacyclic triterpene esters biosynthesis were found in B. subcupularis, and BsubOSC2 is a β-amyrin synthetase gene, and is one of the key enzyme genes for the biosynthesis of balanophorin A. This study provides gene resources for further analysis of the biosynthesis pathway of triterpene ester compounds such as balanophorins, and facilitate the research on molecular breeding of B. subcupularis., authors=LI Cuixia, WANG Yina, ZHAO Xiu, XIANG Guisheng, ZHANG Guanghui, YANG Shengchao, authorsList=LI Cuixia, WANG Yina, ZHAO Xiu, XIANG Guisheng, ZHANG Guanghui, YANG Shengchao, authorCompany=null, correspAuthors=null, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, 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=1304406837219054119, articleId=1304406836967395878, tenantId=1146029695717560320, journalId=1302319053441957962, language=CN, title=转录组分析揭示杯茎蛇菰三萜酯生物合成途径的关键基因, columnId=1304140203967410414, journalTitle=中草药, columnName=药材与资源, runingTitle=null, highlight=null, articleAbstract=目的 探究杯茎蛇菰Balanophora subcupularis中五环三萜酯生物合成途径的关键酶基因,为其生物合成机制及蛇菰属药用植物资源开发利用奠定了一定的研究思路和理论基础。方法 对杯茎蛇菰花柄、根茎和花序3个组织进行转录组测序与分析,通过PfamScan搜索三萜酯生物合成候选酶基因,并构建了BsubOSCs基因的酵母表达载体,将其导入酿酒酵母Saccharomyces cerevisiae羊毛甾醇合酶基因(erg7)缺陷突变株GIL77中以表征其功能。结果 从杯茎蛇菰转录组中挖掘到可能参与五环三萜酯生物合成的相关酶基因共87条,其中大多数基因在花序中表达量最高,说明花序可能是五环三萜酯的主要合成部位。2条BsubOSCs基因的酶功能表征发现BsubOSC2能够催化2,3-氧化鲨烯生成β-香树脂醇,并没有检测到BsubOSC1的催化产物。结论 通过转录组分析发现了杯茎蛇菰五环三萜酯的生物合成关键酶基因,其中BsubOSC2为β-香树脂醇合成酶基因,是蛇菰素A生物合成的关键酶基因之一。为进一步解析蛇菰素等三萜酯化合物的生物合成途径提供了基因资源,促进了杯茎蛇菰分子育种的研究。, authors=李翠霞1,2,3,4, 王益娜1,2,3,4, 赵秀1,2,3,4, 向贵生2,3,4, 张广辉1,2,3,4, 杨生超1,2,3,4,5, authorsList=李翠霞, 王益娜, 赵秀, 向贵生, 张广辉, 杨生超, authorCompany=1 云南农业大学农学与生物技术学院, 云南 昆明 650201;
2 云南农业大学 西南中药材种质创新与利用国家地方联合工程研究中心, 云南 昆明 650201;
3 云南农业大学 云南省药用植物生物学重点实验室, 云南 昆明 650201;
4 云南特色植物提取实验室, 云南 昆明 650106;
5 红河学院生物科学与农学学院, 云南 蒙自 661199, correspAuthors=杨生超, authorNote=李翠霞: 李翠霞,硕士研究生,研究方向为药用植物合成生物学。Tel:18487183726 E-mail:2082554237@qq.com, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=YOgnM+TYJiMiADD63Gj56Q==, pdfFileSize=2831888, 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=云南特色植物提取实验室自主研究项目基金 (2022YKZY004))}, authors=null, keywords=[Keyword(id=1304406837453935145, tenantId=1146029695717560320, journalId=1302319053441957962, articleId=1304406836967395878, language=CN, orderNo=1, keyword=杯茎蛇菰), Keyword(id=1304406837542015530, tenantId=1146029695717560320, journalId=1302319053441957962, articleId=1304406836967395878, language=CN, orderNo=2, keyword=转录组), Keyword(id=1304406839198765611, tenantId=1146029695717560320, journalId=1302319053441957962, articleId=1304406836967395878, language=CN, orderNo=3, keyword=三萜酯生物合成), Keyword(id=1304406839303623212, tenantId=1146029695717560320, journalId=1302319053441957962, articleId=1304406836967395878, language=CN, orderNo=4, keyword=氧化鲨烯环化酶), Keyword(id=1304406839366537773, tenantId=1146029695717560320, journalId=1302319053441957962, articleId=1304406836967395878, language=CN, orderNo=5, keyword=酰基转移酶), Keyword(id=1304406839454618158, tenantId=1146029695717560320, journalId=1302319053441957962, articleId=1304406836967395878, language=EN, orderNo=1, keyword=Balanophora subcupularis P. 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Prakash Chaturvedula V S, Schilling J K, Miller J S, et al.Two new triterpene esters from the twigs of Brachylaena ramiflora from the Madagascar rainforest[J]. J Nat Prod,2002, 65(8):1222-1224.
Dai T T, Li L C, Qi W, et al. Balanophorin B inhibited glycolysis with the involvement of HIF-1α[J]. Life Sci,2021, 267:118910.
Lin C N, Tome W P. Antihepatotoxic principles of Sambucus formosana[J]. Planta Med, 1988, 54(3):223-224.
Liu J, Yin X, Kou C X, et al. Classification, biosynthesis,and biological functions of triterpene esters in plants[J]. Plant Commun, 2024, 5(4):100845.
Xu R, Fazio G C, Matsuda S P T. On the origins of triterpenoid skeletal diversity[J]. Phytochemistry, 2004,65(3):261-291.
Abe I. Enzymatic synthesis of cyclic triterpenes[J]. Nat Prod Rep, 2007, 24(6):1311-1331.
Bontpart T, Cheynier V, Ageorges A, et al. BAHD or SCPL acyltransferase? What a dilemma for acylation in the world of plant phenolic compounds[J]. New Phytol, 2015,208(3):695-707.
D’Auria J C. Acyltransferases in plants:A good time to be BAHD[J]. Curr Opin Plant Biol, 2006, 9(3):331-340.
Hofmann K. A superfamily of membrane-bound Oacyltransferases with implications for Wnt signaling[J]. Trends Biochem Sci, 2000, 25(3):111-112.
Zhou Y, Ma Y S, Zeng J G, et al. Convergence and divergence of bitterness biosynthesis and regulation in Cucurbitaceae[J]. Nat Plants, 2016, 2:16183.
Shang Y, Ma Y S, Zhou Y, et al. Plant science.Biosynthesis, regulation, and domestication of bitterness in cucumber[J]. Science, 2014, 346(6213):1084-1088.
Kim Y C, Choi D, Cha A, et al. Critical enzymes for biosynthesis of cucurbitacin derivatives in watermelon and their biological significance[J]. Commun Biol, 2020, 3(1):444.
Sohrabi R, Ali T, Harinantenaina Rakotondraibe L, et al.Formation and exudation of non-volatile products of the Arabidiol triterpenoid degradation pathway in Arabidopsis roots[J]. Plant Signal Behav, 2017, 12(1):e1265722.
Huang A C, Jiang T, Liu Y X, et al. A specialized metabolic network selectively modulates Arabidopsis root microbiota[J]. Science, 2019, 364(6440):eaau6389.
Mugford S T, Qi X Q, Bakht S, et al. A serine carboxypeptidase-like acyltransferase is required for synthesis of antimicrobial compounds and disease resistance in oats[J]. Plant Cell, 2009, 21(8):2473-2484.
Mugford S T, Louveau T, Melton R, et al. Modularity of plant metabolic gene clusters:A trio of linked genes that are collectively required for acylation of triterpenes in oat[J]. Plant Cell, 2013, 25(3):1078-1092.
Owatworakit A, Townsend B, Louveau T, et al.Glycosyltransferases from oat(Avena)implicated in the acylation of avenacins[J]. J Biol Chem, 2013, 288(6):3696-3704.
Choi H S, Han J Y, Cheong E J, et al. Characterization of a pentacyclic triterpene acetyltransferase involved in the biosynthesis of taraxasterol andψ-taraxasterol acetates in lettuce[J]. Front Plant Sci, 2022, 12:788356.
Chen Q L, Steinhauer L, Hammerlindl J, et al.Biosynthesis of phytosterol esters:Identification of a sterol O-acyltransferase in Arabidopsis[J]. Plant Physiol, 2007,145(3):974-984.
Chen X L, Fang D M, Xu Y X, et al. Balanophora genomes display massively convergent evolution with other extreme holoparasites and provide novel insights into parasite-host interactions[J]. Nat Plants, 2023, 9(10):1627-1642.
Bolger A M, Lohse M, Usadel B. Trimmomatic:A flexible trimmer for Illumina sequence data[J]. Bioinformatics,2014, 30(15):2114-2120.
Deng Z L, Münch P C, Mreches R, et al. Rapid and accurate identification of ribosomal RNA sequences via deep learning[J]. Nucleic Acids Res, 2022, 50(10):e60.
Patro R, Duggal G, Love M I, et al. Salmon provides fast and bias-aware quantification of transcript expression[J]. Nat Methods, 2017, 14(4):417-419.
Buchfink B, Reuter K, Drost H G. Sensitive protein alignments at tree-of-life scale using DIAMOND[J]. Nat Methods, 2021, 18(4):366-368.
Ito R, Masukawa Y, Hoshino T. Purification, kinetics,inhibitors and CD for recombinant β-amyrin synthase from Euphorbia tirucalli L and functional analysis of the DCTA motif, which is highly conserved among oxidosqualene cyclases[J]. FEBS J, 2013, 280(5):1267-1280.
李传旺,张贺,饶攀,等.植物五环三萜类化合物生物合成途径研究进展[J]. 中草药, 2021, 52(11):3436-3452.
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转录组分析揭示杯茎蛇菰三萜酯生物合成途径的关键基因
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中草药 | 药材与资源 2026,57(1): 282-294
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中草药 |药材与资源 2026 , 57 (1) : 282 -294
转录组分析揭示杯茎蛇菰三萜酯生物合成途径的关键基因
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李翠霞1,2,3,4, 王益娜1,2,3,4, 赵秀1,2,3,4, 向贵生2,3,4, 张广辉1,2,3,4, 杨生超1,2,3,4,5
作者信息
    1 云南农业大学农学与生物技术学院, 云南 昆明 650201;
    2 云南农业大学 西南中药材种质创新与利用国家地方联合工程研究中心, 云南 昆明 650201;
    3 云南农业大学 云南省药用植物生物学重点实验室, 云南 昆明 650201;
    4 云南特色植物提取实验室, 云南 昆明 650106;
    5 红河学院生物科学与农学学院, 云南 蒙自 661199
通讯作者:
杨生超
作者简介:
李翠霞: 李翠霞,硕士研究生,研究方向为药用植物合成生物学。Tel:18487183726 E-mail:2082554237@qq.com
Transcriptome analysis revealing key genes of triterpene ester biosynthetic pathway in Balanophora subcupularis
  • LI Cuixia, WANG Yina, ZHAO Xiu, XIANG Guisheng, ZHANG Guanghui, YANG Shengchao
  • Affiliations
    doi: 10.7501/j.issn.0253-2670.2026.01.025
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    目的 探究杯茎蛇菰Balanophora subcupularis中五环三萜酯生物合成途径的关键酶基因,为其生物合成机制及蛇菰属药用植物资源开发利用奠定了一定的研究思路和理论基础。方法 对杯茎蛇菰花柄、根茎和花序3个组织进行转录组测序与分析,通过PfamScan搜索三萜酯生物合成候选酶基因,并构建了BsubOSCs基因的酵母表达载体,将其导入酿酒酵母Saccharomyces cerevisiae羊毛甾醇合酶基因(erg7)缺陷突变株GIL77中以表征其功能。结果 从杯茎蛇菰转录组中挖掘到可能参与五环三萜酯生物合成的相关酶基因共87条,其中大多数基因在花序中表达量最高,说明花序可能是五环三萜酯的主要合成部位。2条BsubOSCs基因的酶功能表征发现BsubOSC2能够催化2,3-氧化鲨烯生成β-香树脂醇,并没有检测到BsubOSC1的催化产物。结论 通过转录组分析发现了杯茎蛇菰五环三萜酯的生物合成关键酶基因,其中BsubOSC2为β-香树脂醇合成酶基因,是蛇菰素A生物合成的关键酶基因之一。为进一步解析蛇菰素等三萜酯化合物的生物合成途径提供了基因资源,促进了杯茎蛇菰分子育种的研究。
    杯茎蛇菰  /  转录组  /  三萜酯生物合成  /  氧化鲨烯环化酶  /  酰基转移酶
    Objective To explore the key enzyme genes of the pentacyclic triterpene esters biosynthesis pathway in Balanophora subcupularis, which lays a certain research idea and theoretical foundation for its biosynthesis mechanism and the development and utilization of medicinal plant resources in the genus Balanophora. Methods Transcriptome sequencing and analysis was performed on the three tissues of the flowerstalk, rhizome and umbel of B. subcupularis, then searching the candidate enzyme genes involved in triterpene ester biosynthesis by PfamScan, constructing the yeast expression vector of BsubOSCs gene which was introduced into the Saccharomyces cerevisiae lanosterol synthase gene(erg7)-deficient mutant strain GIL77 for functional characterization. Results A total of 87 candidate genes involved in pentacyclic triterpene esters biosynthesis were identified from the transcriptome of B. subcupularis, most of them were highly expressed in umbel, suggesting that umbel may be the main site of synthesis of pentacyclic triterpene esters. The enzyme function of two BsubOSC genes were characterized, which was found that BsubOSC2 could catalyze 2, 3-oxidosqualene to form β-amyrin, but no catalytic product of BsubOSC1 was detected. Conclusion Based on transcriptome analysis, the key enzyme genes involved in pentacyclic triterpene esters biosynthesis were found in B. subcupularis, and BsubOSC2 is a β-amyrin synthetase gene, and is one of the key enzyme genes for the biosynthesis of balanophorin A. This study provides gene resources for further analysis of the biosynthesis pathway of triterpene ester compounds such as balanophorins, and facilitate the research on molecular breeding of B. subcupularis.
    Balanophora subcupularis P. C. Tam  /  transcriptome  /  triterpene ester biosynthesis  /  oxidosqualene cyclase  /  acyltransferase
    李翠霞, 王益娜, 赵秀, 向贵生, 张广辉, 杨生超. 转录组分析揭示杯茎蛇菰三萜酯生物合成途径的关键基因. 中草药, 2026 , 57 (1) : 282 -294 . DOI: 10.7501/j.issn.0253-2670.2026.01.025
    LI Cuixia, WANG Yina, ZHAO Xiu, XIANG Guisheng, ZHANG Guanghui, YANG Shengchao. Transcriptome analysis revealing key genes of triterpene ester biosynthetic pathway in Balanophora subcupularis[J]. Chinese Traditional and Herbal Drugs, 2026 , 57 (1) : 282 -294 . DOI: 10.7501/j.issn.0253-2670.2026.01.025

      云南特色植物提取实验室自主研究项目基金 (2022YKZY004)

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    Buchfink B, Reuter K, Drost H G. Sensitive protein alignments at tree-of-life scale using DIAMOND[J]. Nat Methods, 2021, 18(4):366-368.
    Ito R, Masukawa Y, Hoshino T. Purification, kinetics,inhibitors and CD for recombinant β-amyrin synthase from Euphorbia tirucalli L and functional analysis of the DCTA motif, which is highly conserved among oxidosqualene cyclases[J]. FEBS J, 2013, 280(5):1267-1280.
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    Kushiro T, Shibuya M, Masuda K, et al. Mutational studies on triterpene synthases:Engineering lupeol synthase intoβ-amyrin synthase[J]. J Am Chem Soc, 2000, 122(29):6816-6824.
    Hans J, Hause B, Strack D, et al. Cloning, characterization,and immunolocalization of a mycorrhiza-inducible 1-deoxy-D-xylulose 5-phosphate reductoisomerase in arbuscule-containing cells of maize[J]. Plant Physiol,2004, 134(2):614-624.
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    2026年第57卷第1期
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    doi: 10.7501/j.issn.0253-2670.2026.01.025
    • 接收时间:2025-08-06
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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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