首页|广藿香PcGA2ox1基因克隆、VIGS载体构建和表达分析

广藿香PcGA2ox1基因克隆、VIGS载体构建和表达分析

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  • 国家科技期刊平台
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  • NSTL
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赤霉素2-氧化酶(gibberellin 2-oxidases，GA2ox)是赤霉素(GAs)代谢途径关键酶，调节活性GAs降解，在植物株型调控中发挥重要作用。为探寻GA2ox基因在广藿香中的功能，本研究对广藿香PcGA2ox1 基因进行克隆，得到 561 bp的CDS序列。生物信息学分析结果表明，PcGA2ox1 蛋白编码 186 个氨基酸，为亲水性蛋白，分子量为 20。98 kDa，等电点为 5。81，不含信号肽，无跨膜结构域，具有GA2ox超基因家族的保守结构域，主要定位在细胞核。系统发育分析结果显示，PcGA2ox1 基因与丹参GA2ox7 的亲缘关系最为密切。qRT-PCR结果显示，PcGA2ox1 基因在广藿香中的表达具有组织特异性，叶中表达量最高，根中表达量最低。用病毒诱导基因沉默(VIGS)技术侵染广藿香的结果显示，侵染两周后，PcGA2ox1 基因表达量比阴性对照下降 83%，比空白对照下降 71%，沉默效果明显。本研究结果可为深入了解PcGA2ox1 结构、功能和作用机理奠定基础，并为广藿香优良品种选育提供参考。
Cloning,VIGS Vector construction and Expression Analysis of PcGA2ox1 Gene in Pogostemon cablin(Blanco)Benth
Gibberellin 2-oxidases(GA2ox)is a key enzyme in metabolic pathway of gibberellins(GAs),which regulates the degradation of active GAs and plays important roles in plant type regulation.In order to explore the function of GA2ox gene in Pogostemon cablin,we cloned PcGA2ox1 gene of P.cablin and obtained 561 bp of CDS sequence.The results of bioinformatics analysis showed that PcGA2ox1 protein enco-ded 186 amino acids,was a hydrophilic protein with the molecular weight as 20.98 kDa and the isoelectric point as 5.81,contained neither signal peptide nor transmembrane domain,had the conserved domain of GA2ox supergene family,and was mainly located in nucleus.Phylogenetic analysis showed that PcGA2ox1 was most closely related to Salvia miltiorrhiza GA2ox7.The qRT-PCR results showed that PcGA2ox1 gene had tis-sue-specific expression in P.cablin with the highest level in leaves and the lowest level in roots.Infestation of P.cablin by virus-induced gene silencing(VIGS)technology showed that PcGA2ox1 gene expression decreased by 83%compared with negative control and 71%compared with blank control two weeks later,and the silen-cing effect was obvious.The results of this study could lay a foundation for an in-depth understanding of the structure,function and mechanism of PcGA2ox1,and provide a reference for the selection and breeding of ex-cellent P.cablin varieties.

Pogostemon cablinGibberellin 2-oxidases(GA2ox)Gene cloningVirus-induced gene si-lencing(VIGS)Expression analysis

严雅玲、曾晴、严寒静、何梦玲、张宏意

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广东药科大学中药学院,广东 广州 510006

广藿香 赤霉素2-氧化酶(GA2ox) 基因克隆 病毒诱导基因沉默(VIGS) 表达分析

广东省中医药局科研项目广东省重点领域研发计划广东省普通高等学校重点领域专项广州市科技计划

202411772020B0202210022023ZDZX4025SL2023B03J00748

2024

10.14083/j.issn.1001-4942.2024.05.003

山东农业科学
山东省农业科学院,山东农学会,山东农业大学

山东农业科学

CSTPCD北大核心
影响因子：0.578
ISSN：1001-4942
年,卷(期)：2024.56(5)
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