褪黑素诱导西伯利亚百合的转录组分析及LoERF96基因克隆  

Transcriptome Analysis of Lilium Siberian Induced by Melatonin and LoERF96 Gene Cloning

作　　者：刘烨 孙亮 赵国林 彭梦群 贾文杰[3] 李想 张小平[1] 王有国[1] LIU Ye;SUN Liang;ZHAO Guolin;PENG Mengqun;JIA Wenjie;LI Xiang;ZHANG Xiaoping;WANG Youguo(College of Horticulture and Landscape,Yunnan Agricultural University,Kunming 650201,China;School of Agriculture,Jinhua University of Vocational Technology,Jinhua 321017,China;Flower Research Institute,Yunnan Academy of Agricultural Sciences,Kunming 650205,China)

机构地区：[1]云南农业大学园林园艺学院,云南昆明650201 [2]金华职业技术大学农学院,浙江金华321017 [3]云南省农业科学院花卉研究所,云南昆明650205

出　　处：《云南农业大学学报（自然科学版）》2024年第4期119-130,共12页Journal of Yunnan Agricultural University:Natural Science

基　　金：国家自然科学基金项目(32060690)。

摘　　要：【目的】研究西伯利亚百合萌发阶段的分子调控机制,筛选响应褪黑素、促进百合鳞茎萌发的潜在乙烯响应因子(ethylene response factor,ERF)。【方法】对褪黑素处理下的西伯利亚百合小鳞茎关键时期样本进行转录组测序分析,并对差异表达基因进行功能注释,筛选响应褪黑素、促进百合鳞茎萌发的ERF潜在基因。对筛选的关键基因LoERF96进行克隆和生物信息学分析,通过实时荧光定量分析小鳞茎4个不同生长发育时期中LoERF96基因的表达量。【结果】根据转录组数据筛选得到780个差异表达基因,其中上调表达基因392个,下调表达基因388个。GO富集分析结果显示:富集的差异基因主要生物学过程包括代谢过程、细胞过程、生物调节、刺激响应、信号、生殖过程等。KEGG代谢途径分析结果显示:差异基因的代谢途径包括植物激素信号转导、植物MAPK信号途径、苯丙烷生物合成、淀粉和蔗糖代谢、光合作用、碳代谢、内质网中的蛋白质加工等。通过同源克隆得到LoERF96基因,该基因序列全长435 bp,编码144个氨基酸,有1个AP2保守结构域,蛋白相对分子质量约为16.32 ku,理论等电点为4.94,是没有跨膜结构的亲水性蛋白,预测亚细胞定位于细胞核中。RT-qPCR表达结果显示:在百合4个不同生长发育过程中,LoERF96基因的相对表达量总体呈先上升后下降的趋势。【结论】西伯利亚百合中的乙烯响应因子LoERF96受褪黑素诱导表达,这一结果为探究百合AP2/ERF转录因子萌发阶段的生物学功能提供了理论基础和参考。[Purpose]To study the molecular regulatory mechanism of the germination stage of Lilium Siberian,and to screen the potential ethylene response factor(ERF)that responds to melatonin to promote the germination of Lilium Siberian.[Methods]Transcriptome sequencing analysis was performed on the samples of Lilium Siberian bulblets under melatonin treatment during the critical period,and the differentially expressed genes were functionally annotated,screening the potential ERF genes that respond to melatonin to promote the germination of lily bulblets.The screened key gene LoERF96 was cloned and bioinformatically analyzed,and the expression of LoERF96 gene was quantified by RT-qPCR in four different growth and development periods of small bulbs.[Results]According to the transcriptome data,780 differential expressed genes were obtained,including 392 up-regulated genes and 388 down-regulated genes.The results of GO enrichment analysis showed that:the biological processes of enriched differential genes included metabolic processes,cellular processes,biological regulation,stimulus response,signaling,reproductive process,and so on.KEGG metabolic pathway analysis showed that:the metabolic pathways of the differential genes included plant hormone signal transduction,plant MAPK signaling pathway,phenylpropanoid biosynthesis,starch and sucrose metabolism,photosynthesis,carbon metabolism,protein processing in endoplasmic reticulum,and so on.The LoERF96 gene was obtained by homologous cloning,the fulllength sequence of the gene was 435 bp,encoding 144 amino acids,with one AP2 conserved structural domain,the relative molecular weight of the protein was about 16.32 ku,and the theoretical isoelectric point was 4.94;it was a hydrophilic protein with no transmembrane structure,and the subcellular localization was predicted to be in nucleus.RT-qPCR results showed that:in the four different growth and development periods of lily,the relative expression level of LoERF96 gene showed a general trend of first increase and then decrease.[Conc

关 键 词：百合 LoERF96 基因克隆 转录组 生物信息学分析 

分 类 号：S682.265.01[农业科学—观赏园艺]