山东农业科学2025，Vol.57Issue(8)：15-23,9.DOI:10.14083/j.issn.1001-4942.2025.08.003
花烟草(Nicotiana alata)叶绿素酶基因NalaCLH2的克隆与表达模式分析
Cloning and Expression Pattern of Chlorophyllase Gene NalaCLH2 in Nicotiana alata
摘要
Abstract
Chlorophyllase facilitates the first step of chlorophyll degradation and play key roles in the se-nescence of leaves,flowering and fruit ripening.To elucidate the function of chlorophyllase2(CLH2)gene in Nicotiana alata,the homologous cloning method was used to clone CLH2 gene from Nicotiana alata and the bioinformatics analysis was conducted.Furthermore,real-time fluorescence quantitative PCR was used to ana-lyze the tissue-specific expression of this gene and the expression characteristics of Nicotiana alata seedlings treated with abiotic stresses and different hormones.The results showed that the open reading frame(ORF)of NalaCLH2 was 960 bp(GenBank accession number as PP445034)encoding 319 amino acids.The NalaCLH2 protein had the molecular weight as 34.59 kDa and the theoretical isoelectric point as 6.75,and it was a stable non-transmembrane non-secretory protein mainly located in chloroplasts.NalaCLH2 was a hydrophilic protein containing chlorophyllase2 conservative domain but without signal peptide and transmembrane structure.Tis-sue-specific expression analysis showed that NalaCLH2 gene was expressed in the roots(taproots and lateral roots),stems(upper,middle and lower stems),leaves(upper and lower leaves)and flowers(calyces and petals)of Nicotiana alata,but its expression level was the highest in calyx and the lowest in petal.NalaCLH2 gene was upregulated by low temperature,abscisic acid(ABA),gibberellic acid(GA)and methyl jasmonate(MeJA)induction,while NaCl,drought and brassinolide(BR)inhibited its expression.These results of this study could provide a basis for further analysis of the function of NalaCLH2 gene in abiotic stresses and hor-mone signal transduction pathway of Nicotiana alata.关键词
花烟草/叶绿素酶2/基因克隆/表达模式分析Key words
Nicotiana alata/Chlorophyllase2/Gene cloning/Expression pattern analysis分类
农业科技引用本文复制引用
侯佩,张君旗,蒋彪,李猛,赵晨颖..花烟草(Nicotiana alata)叶绿素酶基因NalaCLH2的克隆与表达模式分析[J].山东农业科学,2025,57(8):15-23,9.基金项目
郑州轻工业大学博士科研基金项目(2015BSJJ020) （2015BSJJ020）
河南省科技特派员项目(SPY20180015) （SPY20180015）
