西南林业大学学报2025，Vol.45Issue(1)：55-67,13.DOI:10.11929/j.swfu.202402027
云南松GA20氧化酶基因的克隆与表达分析
Cloning and Expression Analysis of GA20 Oxidase Gene in Pinus yunnanensis
摘要
Abstract
To explore the effects of exogenous gibberellins on the growth and development of Pinus yunnan-ensis,and to analyze the expression of the key synthase gene GA20ox,P.yunnanensis seedlings were selected as materials and treated with exogenous spraying of gibberellin and the gibberellin inhibitor Paclobutrazol at concen-trations of 100,200,and 300 mg/L,respectively,to observe the changes in seedling height and ground diameter.At the same time,by screening and comparing the transcriptome data of P.yunnanensis,a homologous sequence of GA20ox gene was obtained.RT-PCR technology was used for gene cloning,and bioinformatics analysis,codon preference analysis,and gene expression analysis were performed.The results showed that under different con-centration gradients,exogenous 200 mg/L gibberellin significantly promoted the height growth of P.yunnanensis seedlings,while 100 mg/L gibberellin significantly promoted their ground diameter growth.The GenBank login number of P.yunnanensis GA20ox(PyGA20ox)obtained through cloning is OR651279,with a coding region length of 1107 bp,encoding 368 amino acids,with a theoretical relative molecular weight of 41.7 kDa.Codon preference analysis showed that the gene prefers to use codons ending in A/U,Saccharomyces cerevisiae was suit-able as a microbial heterologous expression receptor for this gene,and both Nicotiana benthamiana and Arabidop-sis thaliana were suitable as receptor materials for genetic transformation and functional studies of this gene.The results of fluorescence quantitative PCR showed that PyGA20ox was expressed in roots,stems,leaves,and stem tips,and was significantly expressed in stems.However,the expression pattern of this gene varied under different hormone treatments.This experiment determined the significant promoting effect of gibberellin on the growth of P.yunnanensis through exogenous hormone treatment,with a suitable spraying concentration of 200 mg/L.The key enzyme PyGA20oxin the gibberellin biosynthesis pathway was cloned and expressed.关键词
云南松/植物生长调节剂处理/基因克隆/密码子偏好性/表达分析Key words
Pinus yunnanensis/hormone treatment/gene clone/codon usage bias/expression analysis分类
林学引用本文复制引用
李瑞连,王玉倩,母德锦,徐骏飞,蔡年辉,许玉兰,陈林..云南松GA20氧化酶基因的克隆与表达分析[J].西南林业大学学报,2025,45(1):55-67,13.基金项目
国家自然科学基金项目(32360381)资助 （32360381）
云南省高层次人才培养支持计划青年拔尖人才项目(YNWR-QNBJ-2019-075)资助 （YNWR-QNBJ-2019-075）
云南省农业联合专项重点项目(202301BD070001-152)资助 （202301BD070001-152）
云南省农业联合专项面上项目(202301BD070001-035)资助 （202301BD070001-035）
云南省研究生导师团队建设项目(2022-97)资助. （2022-97）
