亚热带农业研究2025，Vol.21Issue(1)：1-10,10.DOI:10.13321/j.cnki.subtrop.agric.res.2025.01.001
杉木硝态氮转运蛋白基因ClNRT1.1的克隆与表达
Cloning and expression analysis of nitrate transporter gene ClNRT1.1 in Cunninghamia lanceolata
摘要
Abstract
[Purpose]This study aimed to elucidate the bioinformatic characteristics and expression patterns of the nitrate transport-er gene ClNRT1.1in Cunninghamia lanceolata under varying nitrogen(N)and phosphorus(P)conditions,providing insights for breeding Cunninghamia lanceolata varieties with enhanced N and P utilization efficiency.[Method]ClNRT1.1 was cloned from Cunninghamia lanceolata seedlings grown under normal N and P conditions by rapid amplification of cDNA ends(RACE).Bioinfor-matics tools were employed to analyze the features of ClNRT1.1 and the physicochemical properties and structure of the protein.Transcriptomic data and real-time quantitative PCR were used to investigate the expression differences of ClNRT1.1 under the treat-ments of nitrate nitrogen addition and low P.[Result](1)Bioinformatic analysis revealed that ClNRT1.1 encoded a 331-amino-acid protein with notable hydrophobicity and stability.The protein harbored a conserved major facilitator superfamily(MFS)domain(PLN00028),9 transmembrane helical structures,and 9 hydrophobic regions.Its secondary structure was dominated by α-helices,with serine as the primary phosphorylation site.Multiple cis-acting elements responsive to plant growth,hormones,light,and stress were identified in the promoter region.Phylogenetic analysis indicated highly conserved evolution of ClNRT1.1,which showed the highest homology to the corresponding protein in Cryptomeria japonica.(2)Subcellular localization confirmed the membrane localiza-tion of ClNRT1.1,suggesting a nitrate transport function.(3)Transcriptomic data demonstrated that ClNRT1.1 expression was sig-nificantly upregulated under low-P and low-N conditions but suppressed under high-N treatment.Notably,high-P combined with low-N strongly induced the expression of ClNRT1.1,which implied its role in nitrate uptake and N-P crosstalk regulation.(4)Real-time quantitative PCR results revealed that low-P stress induced significant upregulation of ClNRT1.1 across all families,particularly in Family 24,where the expression was upregulated by 3-folds compared with that under normal-P conditions.Family-specific ex-pression patterns of ClNRT1.1 in leaves and stems under low-P stress were observed:significant upregulation in Families 10 and 24,but downregulation in Family 65.[Conclusion]ClNRT1.1 plays a critical role in low-P adaptation of Cunninghamia lanceolata through transmembrane nitrate transport and response to N-P synergistic signaling,serving as a new target for genetic improvement of Cunninghamia lanceolata regarding N and P utilization efficiency.关键词
杉木/硝态氮转运蛋白/基因克隆/氮、磷协同/低磷胁迫Key words
Cunninghamia lanceolata/nitrate transporter/gene cloning/N-P synergy/low phosphorus stress分类
林学引用本文复制引用
陈凡,曹睿倪,张娅静,赵铭臻,李明..杉木硝态氮转运蛋白基因ClNRT1.1的克隆与表达[J].亚热带农业研究,2025,21(1):1-10,10.基金项目
国家自然科学基金面上项目"基于特异种质资源精准鉴定和GWAS关联分析的杉木磷效率性状遗传解析与良种选育"(32471906). （32471906）
