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该实验以金花葵叶片为实验材料,通过分子克隆技术成功获得了金花葵类黄酮生物合成途径中的关键基因AmCHI的编码区序列(coding sequences,CDS)及起始密码子ATG前1111bp的启动子区序列,运用生物信息学方法对其序列进行分析。结果表明:在金花葵叶片中克隆得到的AmCHI基因包含882bp的CDS,编码293个氨基酸残基,预测其相对分子质量为31.65kDa,等电点为4.78,通过对AmCHI进行系统进化树分析发现AmCHI与木槿HsCHI的同源性最高。在AmCHI启动子区域包含转录起始的基本元件、响应激素和非生物胁迫以及与组织特异性表达相关的顺式作用元件。该研究为阐明AmCHI基因在金花葵黄酮类化合物合成中的功能及调控机制奠定了基础,也为通过分子育种手段提高金花葵黄酮含量提供了理论依据。
Abstract：In this study, using Abelmoschus manihot L.leaves as experimental materials, the coding sequence(CDS) of the key gene AmCHI in the flavonoid biosynthesis pathway of A.manihot and the 1111bp promoter sequence before the start codon ATG were successfully obtained through molecular cloning technology, and the sequences were analyzed using bioinformatics methods.The results showed that the AmCHI gene cloned from A.manihot leaves contains an 882bp CDS, encoding 293 amino acid residues, with a predicted relative molecular weight of 31.65kDa and an isoelectric point of 4.78.Phylogenetic tree analysis of AmCHI revealed that it has the highest homology with HsCHI from Hibiscus syriacus.The promoter region of AmCHI ontains basic elements for transcription initiation, cis-acting elements responsive to hormones and abiotic stresses, and those related to tissuespecific expression.This study lays a foundation for elucidating the function and regulatory mechanism of the AmCHI gene in flavonoid synthesis in A.manihot, and also provides a theoretical basis for improving flavonoid content in A.manihot through molecular breeding.
[1]杨美娟,黄坤艳,李玲,等.金花葵化学成分及其生物活性研究进展[J].安徽农业科学,2019,47(17):5-7.
[2]程丽敏,董爽,梁文明,等.金花葵茎可溶性糖的提取及抗氧化性研究[J].食品研究与开发,2016,37(22):59-63.
[3]楚洪涛,刘仲旭,麻明杰,等.金花葵的主要研究简述[J].现代农业研究,2024,30(07):6-12.
[4]ZOUY H,XU D,CHEN Y Z,et al.Combined metabolome and transcriptome analyses reveal the flavonoids changes and biosynthesis mechanisms in different organs of Hibiscus manihot L.[J].Frontiersin plant science,2022,13:817378.
[5]HOUJ Q,ZOU H,REN L P,et al.Transcriptome and metabolome analysis reveal the flavonoid biosynthesis mechanism of Abelmoschus manihot L.at different anthesis stages[J].Metabolites,2023,13(02):216.
[6]刘琦,辛秀兰,兰蓉,等.金花葵花中总黄酮提取工艺的研究[J].食品研究与开发,2011,32(07):19-21.
[7]周宇涵,徐小迪,高君,等.响应面法优化金花葵花总黄酮提取工艺及抗氧化活性分析[J].分子植物育种,2023,21(06):2024-2031.
[8]宋琳琳,黄蓓蓓,张朝晖.金花葵花中总黄酮的超声波提取工艺[J].贵州农业科学,2016,44(02):70-72.
[9]周宇涵,徐小迪,高君,等.金花葵不同组织的转录组测序及分析[J].分子植物育种,2023,21(09):2890-2899.
[10]MEHDYM C,LAMB J.Chalcone isomerasec DNA cloning and m RNA induction by fungal elicitor,wounding and infection[J].The EMBO journal,1987,6(06):1527-1533.
[11]RA LSTON L,SU BRAMANIAN S,MATSUNO M,et al.Partial reconstruction of flavonoid andi soflavonoid biosynthesisin yeast usingsoybean type and type II chalcone isomerases[J].Plant physiology,2005,137(04):1375-1388.
[12]YIN C,ZHANG X D,GAO Z Q,et al.The research progress of chalcone isomerase (CHI) in plants[J].Molecular biotechnology,2019,61(01):32-52.
[13]SHIMADAN,AOKI T,SATO S,et al.A cluster of genes encodes the two types of chalcone isomerase involved in the biosynthesis of general flavonoids and legume-specific 5-deoxy(iso)flavonoids in Lotus japonicus[J].Plant physiology,2003,131(03):941-951.
[14]CHENGA X,ZHANG X,HAN J,et al.Identification of chalconei somerasei n the basal land plants reveals an ancient evolutionof enzymatic cyclization activity for synthesis of flavonoids[J].The new phytologist,2018,217(02):909-924.
[15]RYANK G,SWINNY E,WINEFIELD C,et al.Flavonoids and UV photoprotection in Arabidopsis mutants[J].C-Ajournalof biosciences,2001,56(09 10):745-754.
[16]NGAKI N,LOUIE G V,PHILIPPE R N,et al.Evolution of the chalcone-isomerase fold from fatty-acid binding to stereospecific catalysis[J].Nature,2012,485(7399):530-533.
[17]JIANGW,YIN Q,WU R,et al.Role of chalconei somerase-like proteini n flavonoid biosynthesisi n Arabidopsist haliana[J].Journa l of experimental botany,2015,66(22):7165-7179.
[18]MOUSTAFAE,WONG E.Purification and properties of chalcone-flavanone isomerase from soya bean seed[J].Phytochemistry,1967,6(05):625-632.
[19]张忠润,韩曙.紫荚豌豆查尔酮异构酶PsCHI基因的克隆及表达分析[J].贵州农业科学,2025,53(02):1-8.
[20]吴彦庆,赵大球,王静,等.芍药查尔酮异构酶基因(CHI)克隆、密码子偏好性分析以及蛋白结构功能预测[J].华北农学报,2016,31(02):71-80.
[21]王爱华,刘文欣,马超,等.火龙果HpCML18基因的克隆、定位及启动子分析[J/OL].分子植物育种,2025,1-16[2025-07-21].https://link.cnki.net/urlid/46.1068.S.20250717.1736.002.
[22]徐朝阳,张苛苛,王晨晨,等.生菜查尔酮异构酶基因克隆及体外酶活测定[J].植物生理学报,2023,59(11):2135-2143.
[23]张苛苛,孙术富,谭宇萍,等.菘蓝查尔酮异构酶基因的克隆与体外酶活鉴定[J].中国中药杂志,2023,48(06):1510-1517.
[24]吴冰,祝钦泷,郭余龙,等.查尔酮异构酶基因的分子特征及其在基因工程中的应用[J].植物生理学通讯,2008(01):175-181.
[25]MORITA Y,TAKAGI K,FUKUCHI-MIZUTANI M,et al.A chalcone isomerase-like protein enhances flavonoid productionand flower pigmentation[J].The plant journal,2014,78(02):294-304.
[26]MUIRS R,COLLINS J,ROBINSON S,et al.Overexpression of petunia chalcone isomerase in tomato results in fruit containing increased levels of flavonols[J].Nature biotechnology,2001,19(05):470-474.
[27]翟妞,郑庆霞,刘萍萍,等.烟草查尔酮异构酶基因2(NtCHI2)功能分析[J].烟草科技,2020,53(09):1-5.
[28]GRACE ML,CHANDRASEKHARAN B,HALL T C,et al.Sequence and spacing of TATA box elements are criticalfor accurate initiation from the beta-phaseolin promoter[J].The journal of biological chemistry,2004,279(09):8102-8110.
[29]张小辉,祁艳霞.真核生物启动子TATA-box·GC-box和CAAT-box的分析[J].安徽农业科学,2008(04):1380-1381.
[30]郭依萍,石晶静,周美琪,等.白桦BpbZIP1基因抗旱耐盐分析及ABRE元件结合鉴定[J].林业科学研究,2020,33(05):68-76.
[31]高玉姣,夏伟羿,戴梦清,等.小麦MYB转录因子挖掘及功能研究进展[J/OL].麦类作物学报,2025,1-13[2025-07-21].https://link.cnki.net/urlid/61.1359.s.20250709.0933.002.
[32]陈乃钰,张国香,张力爽,等.ABF转录因子在植物响应非生物胁迫中的作用[J].植物遗传资源学报,2021,22(04):930-938.
[33]SONGX,JIN L,GUAN X,et al.Comprehensive multi-omics analysis reveals regulatory mechanisms of red and blue lightcombination on phenolic metabolite accumulation in larch[J].Industrial crops products,2025,225:120526.
[34]WANGF,WANG R,GU H X,et al.Integrative transcriptome and metabolome analysis reveals the mechanism of exogenousmelatonin alleviating drought stress in maize roots[J].Plant physiology and biochemistry,2023,199:107723.
基本信息:
DOI：10.13478/j.cnki.jasyu.2025.03.002
中图分类号:S681.9
引用信息:
[1]楚洪涛,张婷,李钰莹,等.金花葵AmCHI基因的克隆及生物信息学分析[J].延边大学农学学报,2025,47(03):10-17.DOI:10.13478/j.cnki.jasyu.2025.03.002.
基金信息:
国家自然科学基金项目（31960619）; 吉林省教育厅项目（JJKH20180887KJ）