种子2025，Vol.44Issue(8)：74-84,93,12.DOI:10.16590/j.cnki.1001-4705.2025.08.074
不同花色忍冬LjDFR2基因的克隆及表达模式分析
Analysis on Cloning and Expression Patterns of LjDFR2 Gene in Lonicera japonica Thunb.with Different Colors
摘要
Abstract
To explore the function of the DFR gene in the flower color formation of Lonicera japonica Thunb.and Lonicera japonica var.chinensis(Wats.)Bak.,the DFR2 gene was obtained from Loni-cera japonica Thunb.and Lonicera japonica var.chinensis(Wats.)Bak.by qRT-PCR technology.The full-length cDNA of this gene in Lonicera japonica Thunb.and Lonicera japonica var.chinensis(Wats.)Bak.was obtained,both of which sequences were 1 176 bp.There was a difference of one base in the nucleotide sequence,while there was no difference in the amino acid sequence.It was named LjDFR2.The LjDFR2 gene encoded 391 amino acids,with 6 exons and 5 introns.Its secondary structure was mainly composed of α-helices and random curls.It was a stable hydrophilic protein without signal pep-tides and had one transmembrane domain,which was located in the cell membrane and the nucleus.Multiple alignment of amino acid sequences and phylogenetic analysis indicated that LjDFR2,like other plant DFR pro-teins,contained highly conserved NADPH and the substrate binding domain was most closely related to the LcDFR of Lonicera caerulea,which also belonged to a large branch with Chrysanthemum ×morifolium and Saussurea involucrata,and they contained similar motifs.The closer the relationship,the more similar the motifs.The promoter of the LjDFR2 gene contained a large number of stress response elements.The qRT-PCR results indicated that the LjDFR2 gene could respond to drought-salt stress,and its expression showed a trend of first increasing and then decreasing under drought stress,and a trend of first decreasing and then increasing under salt stress.In addition,the expression level of the LjDFR2 gene was the highest in flowers,the lowest in the stems of Lonicera japonica Thunb.,and the lowest in the roots of Lonicera japonica var.chinensis(Wats.)Bak.,The expression level was the highest in the S1 stage of flower development and the lowest in the S5 stage.Overall,the expression level of LjDFR2 in each tissue of the darker Lonicera japoni-ca var.chinensis(Wats.)Bak.was significantly higher than that of Lonicera japonica Thunb.,and the ex-pression level in the darker pre-flowering S1,S2 and S3 was also significantly higher than that of Lonicera japonica Thunb.,Therefore,it was speculated that LjDFR2 could be directly related to the accumulation of anthocyanins.关键词
忍冬/DFR/基因克隆/胁迫处理/表达分析Key words
Lonicera japonica Thunb./DFR/gene cloning/stress treatment/expression analysis分类
农业科技引用本文复制引用
鲁丹丹,余永亮,乔美玉,梁慧珍,郭水柱,谭政委,苏小雨,孙瑶,曹艺雯,王利娜,李磊,李春明..不同花色忍冬LjDFR2基因的克隆及表达模式分析[J].种子,2025,44(8):74-84,93,12.基金项目
河南省重点研发专项(241111310200、231111110800、251111310600、251111111800) （241111310200、231111110800、251111310600、251111111800）
河南省农科院新兴学科发展专项(2024XK01) （2024XK01）
财政部和农业农村部:国家现代农业产业技术体系资助(CARS-21) （CARS-21）
河南省外籍科学家工作室(GZS2024025) （GZS2024025）
中央引导地方科技发展资金(Z20241471131) （Z20241471131）
河南省农科院自主创新项目(2025ZC45、2025ZC44) （2025ZC45、2025ZC44）
河南省科技攻关项目(252102111079) （252102111079）
河南省农科院优秀青年科技基金(2024YQ15、2024YQ16) （2024YQ15、2024YQ16）
