首页|红豆草OvIF7GT基因克隆与表达分析

红豆草OvIF7GT基因克隆与表达分析

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异黄酮是豆科植物中一类典型的次级代谢产物,其化学结构多样性对于植物生长发育及抵御环境胁迫具有重要功能。本研究以高寒地区豆科牧草红豆草(Onobrychis viciifolia)为研究材料,根据转录组测序数据,从红豆草叶片中克隆获得一个异黄酮7-O-糖基转移酶(Isoflavone 7-O-glucosyltransferase,OvIF7GT)基因,并对该基因进行生物信息学、亚细胞定位和干旱胁迫下表达水平的分析。结果显示:1)本研究克隆得到的OvIF7GT大小为1 512 bp,编码503个氨基酸,蛋白质相对分子质量为56 774。36 Da;其编码蛋白是一种亲水性蛋白,具有两个跨膜结构域,62个磷酸化结合位点,理论等电点为5。51。2)保守结构域分析推测,OvIF7GT蛋白属于异黄酮糖基转移酶家族,主要由α-螺旋(41。75%)和无规则卷曲(35。79%)构成;系统发育进化关系分析表明该蛋白与蒺藜苜蓿(Medicago truncatula)和鹰嘴豆(Cicer arietinum)中的IF7GT亲缘关系较近;此外,亚细胞定位分析显示OvIF7GT主要存在于细胞质中,在细胞核中也有分布。3)实时荧光定量PCR(qRT-PCR)结果表明,在干旱胁迫72h后,红豆草幼苗中OvIF7GT显著受到干旱胁迫的诱导表达,且在叶中比根中的表达量更高。该研究结果表明,OvIF7GT参与了红豆草干旱胁迫的响应,可能在红豆草的抗旱中具有潜在功能。
Gene cloning and expression analysis of OvIF7GT in Onobrychis viciifolia
Isoflavones are a class of typical secondary metabolites in legumes.The diversity of their chemical structure aids plant growth and development and resistance to environmental stress.In the present study,a gene encoding isoflavone 7-O-glucosyltransferase(OvIF7GT)was cloned from the leaf of Onobrychis viciifolia with specific primers designed from the transcriptome data,and the gene was subjected to bioinformatics analysis,subcellular localization,and relative expression analysis under drought stress.The results revealed that the gene OvIF7GT had the sequence size of 1 512 bp and encodes a protein with 503 amino acids and molecular weight of 56 774.36 Da.The protein was proposed to be hydrophilic and had two transmembrane domains,62 phosphorylation binding sites,and a theoretical isoelectric point of 5.51.Its conserved domain analysis showed that the protein OvIF7GT contained a-helix(41.75%)and random coils(35.79%);hence,it is proposed to be belonged to the isoflavone glucosyltransferase family.Its phylogenetic analysis showed that the protein OvIF7GT has the closest relationship with the Medicago truncatula and Cicer arietinum homologues.Subcellular localization analysis showed that OvIF7GT was mainly present in the cytoplasm and also distributed in the nucleus.The real-time fluorescence quantification(qRT-PCR)result revealed that the relative expression level of OvIF7GT gene was significantly induced by drought stress,the highest level was at 72 h under drought stress,and the expression level in the leaves was higher than that in roots.Therefore,the gene OvIF7GT probably plays a role in drought stress response and resistance of O.viciifolia.

Onobrychis viciifoliaIsoflavone 7-O-glucosyltransferasegene cloningsubcellular localizationreal-time fluorescence quantitative PCRdrought stressexpression analysis

李珍珍、张正芳、马晓兰、王文颖、邱庆辉、王小梅、周华坤、殷恒霞

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青海大学省部共建三江源生态与高原农牧业国家重点实验室,青海西宁 810016

青海大学生态环境工程学院,青海西宁 810016

青海师范大学生命科学学院/青藏高原生物多样性形成机制与综合利用省级重点实验室,青海西宁 810008

中国科学院西北高原生物研究所/青海省寒区恢复生态学重点实验室,青海西宁 810008

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红豆草 异黄酮7-O-糖基转移酶 基因克隆 亚细胞定位 实时荧光定量PCR 干旱胁迫 表达分析

2024

草业科学
中国草原学会 兰州大学草地农业科技学院

草业科学

CSTPCD北大核心
影响因子:0.854
ISSN:1001-0629
年,卷(期):2024.41(12)