LIU Hui,ZHANG Heng,YE Lihong,et al.Construction of Overexpression Vector for DchCHS1 and Its Transient Expression in Detached Floral Buds of Dianthus chinensis[J].Northern Horticulture,2023,(20):47-53.[doi:doi：10.11937/bfyy.20230838]
DchCHS1过表达载体构建及其在中国石竹离体花苞中瞬时表达分析
- Title:
- Construction of Overexpression Vector for DchCHS1 and Its Transient Expression in Detached Floral Buds of Dianthus chinensis
- 文章编号:
- 1001-0009(2023)20-0047-07
- 分类号:
- S 681.5
- 文献标志码:
- A
- 摘要:
- 以中国石竹为试材，将构建的重组载体pBI121-DchCHS1-GUS转化到农杆菌GV3101中，采用农杆菌侵染的方法，研究了DchCHS1（KX893854）对中国石竹花色的影响，以期建立适用于中国石竹基因功能的快速鉴定体系。结果表明：DchCHS1过表达后，引起查尔酮合成酶（CHS）活性提高，黄酮含量显著增加，但会导致花瓣花色变浅甚至有些部位褪色的现象，同时花瓣中花色素苷含量显著降低。综上，查尔酮合成酶是植物类黄酮物质合成途径和花色素苷合成途径的关键酶，筛选、鉴定CHS基因，有利于花色改良及代谢工程的研究。
- Abstract:
- The recombinant vector pBI121-DchCHS1-GUS was transformed into agrobacterium GV3101，and the influence of DchCHS1(KX893854) on the color of D.chinensis was studied by agrobacterium infection method,in order to establish a rapid identification system for gene function of D.chinensis.The results showed that after DchCHS1 overexpression，CHS activity and flavonoid content increased significantly，but the flower color became lighter and some parts faded，and the content of anthocyanin in petals decreased significantly.In conclusion，chalcone synthase (CHS) was a key enzyme in plant flavonoid synthesis and anthocyanin synthesis pathway.Screening and identification of CHS gene was conducive to the study of flower color improvement and metabolic engineering.
参考文献/References:
［1］CHEN S，PAN X，LI Y，et al.Identification and characterization of chalcone synthase gene family members in Nicotiana tabacum［J］.Journal of Plant Growth Regulation，2017,36(2):374-384.［2］谢修志，陈兆平，王小菁.非洲菊查尔酮合酶基因的克隆序列分析及在大肠杆菌中的表达［J］.热带亚热带植物学报，2004,12(5):431-434.［3］赵风治，马钰婕，韦韬,等.花椰菜查尔酮合酶基因的克隆及功能分析［J］.南开大学学报(自然科学版)，2016,49(4):61-66.［4］王旭，韩春乐，周亚楠，等.黄秋葵查尔酮合成酶基因 AeCHS 的克隆与表达分析［J］.植物遗传资源学报，2014，15(3)：561-567．［5］孙燕,高德民.牡丹花色性状的研究进展［J］.寒旱农业科学,2023,2(1):17-22.［6］关淑文，王毅，郝佳波,等.竹叶花椒查尔酮合成酶基因克隆与表达［J］.分子植物育种，2020,18(16):5300-5305.［7］OHNO S，HOSOKAWA M，KOJIMA M，et al.Simultaneous post-transcriptional gene silencing of two different chalcone synthase genes resulting in pure white flowers in the octoploid dahlia［J］.Planta，2011,234(5)：945-958.［8］周兴文，李纪元，朱宇林.金花茶查尔酮合成酶基因CnCHS的克隆及遗传转化研究［J］.植物研究，2015,35(3):327-332.［9］陈洁，安利清，王涛,等.百合查尔酮合成酶基因克隆及其转化烟草的花色表达分析［J］.西北植物学报，2012,32(8):1511-1517.［10］王仲伟.浙江红山茶花青素和脂肪酸合成相关基因克隆与表达分析［D］.南京：南京农林大学，2013.［11］夏玉凤，耿晓娜，王万双,等.转查尔酮合酶基因对烟草花色及花器官的影响［J］.生物技术通报，2010,1(12):93-98.［12］李辛雷,陈发棣.菊花种质资源与遗传改良研究进展［J］.植物学通报,2004(4):392-401.［13］张衡,贺学勤.农杆菌介导的中国石竹基因瞬时过表达体系建立［J］.北方园艺,2022(23):68-73.［14］LIU X J，CHUANG Y N，CHIOU C Y，et al.Methylation effect on chalcone synthase gene expression determines anthocyanin pigmentation in floral tissues of two Oncidium orchid cultivars［J］.Planta，2012,236(2)：401-409.［15］吴榕，田再民，郑国华,等.白三叶 CHS 基因的过表达提高烟草类黄酮的含量［J］.草业科学，2020,37(2)：305-313.［16］LIU J，HAO X L，HE X Q.Characterization of three chalcone synthase-like genes in Dianthus chinensis［J］.Plant Cell Tissue and Organ Culture,2021,146:1-10.［17］LIVAK K J，SCHMITTGEN T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2-△△CT method［J］.Methods，2001,25(4):402-408.［18］LI F，JIN Z，QU W，et al.Cloning of a cDNA encoding the Saussurea medusa chalcone isomerase and its expression in transgenic tobacco［J］.Plant Physiology Biochemity，2006,44:455-461.［19］李颖畅.植物花色苷［M］.北京:化学工业出版社，2013.［20］刘建新，丁华侨，葛亚英,等.擎天凤梨花色素合成关键基因〖STBX〗CHS、F3′H和DFR〖STBZ〗的克隆及表达分析［J］.分子植物育种，2017,15(3):805-813.［21］TUTEJA J H.Tissue-specific gene silencing mediated by a naturally occurring chalcone synthase gene cluster in glycine max［J］.Plant Cell，2004,16(4):819-835.［22］CHEN J C，JIANG C Z，GOOKIN T，et al.Chalcone synthase as a reporter in virus-induced gene silencing studies of flower senescence［J］.Plant Molecular Biology，2004，55(4):521-530.［23］WANG C K，CHEN P Y，WANG S M，et al.Cosuppression of tobacco chalcone synthase using Petunia chalcone synthase constructs results in white flowers［J］.Botanical Studies，2006，47(1):71-82.［24］于晓南，张启翔.观赏植物的花色素苷与花色［J］.林业科学，2002(3)：147-153.［25］SHANG Y，SCHWINN K E，BENNETT M J，et al.Methods for transient assay of gene function in floral tissues［J］.Plant Methods，2007,3(1):1-12.［26］侯泽豪,王书平,魏淑东,等.植物花青素生物合成与调控的研究进展［J］.广西植物,2017,37(12):1603-1613.［27］ZHANG L，DING R，CHAI Y，et al.Engineering tropane biosynthetic pathway in Hyoscyamus niger hairy root cultures［J］.Proceedings of the National Academy of Sciences，2004,101(17)：6786-6791.
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备注/Memo
第一作者简介：刘慧（1993-），女，硕士研究生，研究方向为观赏植物逆境栽培生理。E-mail：liuu0729@163.com.责任作者：贺学勤（1970-），女，博士，教授，现主要从事观赏植物栽培生理等研究工作。E-mail：xueqinhe2013@163.com.基金项目：内蒙古自然科学基金资助项目（2021MS03041）。收稿日期：2023-03-17