林业科学研究2025，Vol.38Issue(2)：48-57,10.DOI:10.12403/j.1001-1498.20240209
落叶松LkTCTP启动子的克隆及表达分析
Isolation and Characterization of a Larix Promoter Derived from the Translationally Control Tumor Protein(TCTP)Gene
摘要
Abstract
[Objective]To reveal LkTCTP function during somatic embryogenesis and evaluate its feasibil-ity as a constitutive promoter,based on its expression patterns and promoter activity analysis.[Method]The promoter region of LkTCTP was cloned using an improved genome walking technique.The expres-sion patterns of LkTCTP under different temperature and hormone treatments were examined by qRT-PCR.Promoter activity and tissue localization were analyzed in model plants and Larix by constructing an expression vector for the fusion of LkTCTP promoter and GUS.[Result]A～3.0 kb fragment of the LkTCTP promoter was obtained.In this region we identified many important potential cis-acting elements related to hormones and stress resistance predicted by PLANTCARE,such as AuxRR core/TGA element,TATC box,TCA element,LTR,MBS/TC rich repeats.The expression of LkTCTP could be induced and up-regulated by high temperature(42℃,37℃)and hormone treatments(75 μmol·L-1 ABA、50 μmol·L-1 GA、50 μmol·L-1 IAA、50 μmol·L-1 SA).The 2.0 kb promoter of LkTCTP could drive constitutive expres-sion of GUS in Arabidopsis and could also be used for subcellular localization of transient expressed in to-bacco.Notably,significant GUS staining was observed in Larix embryonic cells following a brief 6-day in-fection,particularly when the P19 protein,which suppresses transcription silencing and enhances exogen-ous gene expression,was introduced.GUS staining was also evident in transgenic somatic embryo seed-lings,with expression levels at the base of the cotyledons is significantly higher than at the leaf tips,indic-ating tissue-specific expression in Larix.[Conclusion]The prediction of cis-acting elements in the LkTCTP promoter and its response to temperature and hormones,suggests that it may be closely related to stress resistance responsiveness.The expression patterns in model plants and Larix indicate its feasibility as a constitutive promoter.However,further validation using promoters of different lengths is still needed to provide a theoretical basis for the development of genetic engineering elements suitable for Larix.关键词
落叶松/TCTP/启动子克隆/体细胞胚/表达分析Key words
Larix/TCTP/promoter cloning/somatic embryogenesis/expression analysis分类
农业科技引用本文复制引用
黄雨芹,张如鑫,张馨元,王俊臣,齐力旺,张立峰..落叶松LkTCTP启动子的克隆及表达分析[J].林业科学研究,2025,38(2):48-57,10.基金项目
农业生物育种国家科技重大专项(2023ZD04058) （2023ZD04058）
国家自然基金项目(32171811) （32171811）
中央级公益性科研院所基本科研业务费专项资金项目(CAFYBB2019SY011) （CAFYBB2019SY011）
