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马尾松冷休克蛋白(CSP)基因克隆及表达分析OA

Cloning and Expression Analysis of Cold Shock Protein(CSP)Gene in Pinus massoniana

中文摘要英文摘要

冷休克蛋白(cold shock protein,CSP)是进化保守的DNA/RNA结合蛋白,调控植物抗寒能力建成与生长发育.为解析马尾松(Pinus massoniana)CSP基因家族特征,本研究基于马尾松全长转录组鉴定PmCSPs,并分析其系统进化、结构功能及幼苗低温胁迫响应下的表达规律.鉴定并克隆获得2个无内含子PmCSP基因.其中PmCSP1开放阅读框共642 bp,编码213个氨基酸残基;PmCSP2开放阅读框共603 bp,编码200个氨基酸残基.编码的多肽链均含有冷休克结构域和CCHC锌指结构域,但锌指结构域的数量有差异,PmCSP1有3个,而PmCSP2仅有2个.系统进化分析表明,PmCSPs与油松的亲缘关系更近.原生质体瞬时表达分析表明,PmCSPs蛋白主要定位于细胞核和细胞质.PmCSPs在马尾松幼苗叶和顶芽中的表达量高于其他组织.低温胁迫下,PmCSP1、PmCSP2分别在全初生叶幼苗、全次生叶幼苗中上调表达,处理8h时PmCSPs的表达量达到最大值,表明PmCSPs是马尾松幼苗应对低温胁迫的早期响应基因,且在不同叶型幼苗中可能参与特异性调控.本研究为深入揭示PmCSP基因功能以及针叶树异生叶幼苗抗寒能力建成的调控机制提供理论依据.

Cold shock protein(CSP)is an evolutionarily conserved DNA/RNA binding protein that plays a key role in the establishment of cold tolerance and growth,as well as in the development of plants.In order to understand the family characteristics of CSP gene(PmCSP)in Pinus massoniana,this study used full-length transcriptome data to identify the PmCSP gene family and to analyse its phylogeny,structure and functions,as well as the expression patterns of seedlings in response to cold stress.The results showed that 2 PmCSP gene family members were identified and cloned,both were intronless.The open reading frame of PmCSP1 was 642 bp in length,encoding 213 amino acids residues.The open reading frame of PmCSP2 was 603 bp in length,encoding 200 amino acids residues.The encoded polypeptide chains contained both cold shock domains and CCHC zinc finger domains,but differed in the number of zinc finger domains.PmCSP1 contained 3 domains,while PmCSP2 only contained 2 domains.Phylogenetic analysis showed that PmCSPs were more closely related to P.tabuliformis.Protoplast transient expression analysis showed that PmCSPs proteins were mainly located in the nucleus and cytoplasm.The expression of PmCSPs in the leaves and terminal buds of P.massoniana seedlings were higher than those in other tissues.Under cold stress,PmCSP1 and PmCSP2 were upregulated in the primary needle and secondary needle seedlings,respectively,with PmCSP expression peaking at 8 h,which indicated that PmCSPs were early response genes for P.massoniana seedlings in response to cold stress and may be involved in specific molecular regulation in heteroblastic foliage seedlings.This study provides a theoretical basis for further revealing the functions of the PmCSP gene family and the regulatory mechanisms established for the cold resistance of conifer heteroblastic foliage seedlings.

李雪颖;徐盈盈;赵渊祥;吴峰

贵州大学贵州省森林资源与环境研究中心/贵州省高原山地林木培育全省重点实验室,贵阳 550025||贵州大学林学院,贵阳 550025贵州大学贵州省森林资源与环境研究中心/贵州省高原山地林木培育全省重点实验室,贵阳 550025||贵州大学林学院,贵阳 550025贵州大学贵州省森林资源与环境研究中心/贵州省高原山地林木培育全省重点实验室,贵阳 550025||贵州大学林学院,贵阳 550025贵州大学贵州省森林资源与环境研究中心/贵州省高原山地林木培育全省重点实验室,贵阳 550025||贵州大学林学院,贵阳 550025

农业科技

马尾松幼苗低温胁迫异生叶表达模式CCHC锌指结构域

Pinus massoniana seedlingsCold stressHeteroblastic foliageExpression patternCCHC zinc finger domains

《农业生物技术学报》 2026 (3)

658-671,14

贵州省重大科技专项(黔科合重大专项字[2024]010)国家自然科学基金(32260406)

10.3969/j.issn.1674-7968.2026.03.017

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