卷丹百合LlSHMT基因的克隆、表达及功能  

Cloning,Expression and Function of LlSHMT Gene in Lilium lancifolium

作　　者：郝俊夷 李佳敏 张智萱 梁佳惠 潘文强 孙明[1] 杜运鹏 Hao Junyi;Li Jiamin;Zhang Zhixuan;Liang Jiahui;Pan Wenqiang;Sun Ming;Du Yunpeng(Beijing Forestry University,Beijing 100083,P.R.China;Beijing Academy of Agriculture and Forestry Sciences)

机构地区：[1]北京林业大学,北京100083 [2]北京市农林科学院

出　　处：《东北林业大学学报》2025年第5期58-65,共8页Journal of Northeast Forestry University

基　　金：国家自然科学基金项目(32371954、32302599、32471936);北京市农林科学院优秀青年基金项目(YXQN202303);北京市农林科学院国际科技合作平台建设项目(2024-13);雄安新区科技创新专项项目(2022XAGG0100)。

摘　　要：为研究卷丹百合(Lilium lancifolium)珠芽发生的内在机制,以卷丹和大花卷丹(L.leichtlinii var.maximowiczii)为试验材料,在珠芽S1时期(珠芽小白点时期),取卷丹和大花卷丹上下部叶腋处进行转录组测序,通过转录组分析筛选到植物一碳代谢与光呼吸的关键酶——丝氨酸羟甲基转移酶(SHMT)。利用生物信息学相关的软件对其生化特性进行预测;通过实时荧光定量PCR检测LlSHMT在不同百合、不同时期的表达量,及对卷丹进行生长素吲哚-3-乙酸(IAA)、生长素极性运输抑制剂(NPA)以及打顶处理后的基因表达情况;利用基因沉默技术验证LlSHMT在珠芽发生过程中的作用。结果表明:LlSHMT序列全长为909 bp,编码302个氨基酸;LlSHMT在卷丹S1时期上部叶腋处表达量显著高于卷丹S1时期下部叶腋处和大花卷丹上下部叶腋;沉默LlSHMT基因导致卷丹珠芽发生数量显著减少,证明了LlSHMT在珠芽发生过程中的正调控作用。综上,LlSHMT在卷丹珠芽发生过程中发挥促进作用。To study the internal mechanism of bulbil formation in Lilium lancifolium,L.lancifolium and L.leichtlinii var.maximowiczii were used as experimental materials.During the S1 stage of bulbil(the stage of small white bulbil spots),the upper and lower leaf axils of L.lancifolium and L.leichtlinii var.maximowiczii were sampled for transcriptome sequencing.Through transcriptome analysis,serine hydroxymethyltransferase(SHMT),a key enzyme in plant one-carbon metabolism and photorespiration,was screened out.Bioinformatics-related software was used to predict its biochemical characteristics.Real-time fluorescence quantitative PCR was used to detect the expression levels of LlSHMT in different lilies and at different stages,as well as the gene expression of L.lancifolium after treatment with indole-3-acetic acid(IAA),auxin polar transport inhibitor(NPA),and topping.Gene silencing technology was used to verify the role of LlSHMT in the process of bulbil formation.The results showed that the full-length sequence of LlSHMT was 909 bp,encoding 302 amino acids.The expression level of LlSHMT in the upper leaf axils of L.lancifolium during the S1 stage was significantly higher than that in the lower leaf axils of L.lancifolium during the S1 stage and in the upper and lower leaf axils of L.leichtlinii var.maximowiczii.Silencing the LlSHMT gene led to a significant decrease in the number of bulbil formed in L.lancifolium,which proved the positive regulatory role of LlSHMT in the process of bulbil formation.In conclusion,LlSHMT plays a promoting role in the process of bulbil formation in L.lancifolium.

关 键 词：卷丹 珠芽 LlSHMT 表达分析 基因沉默 

分 类 号：S682.29[农业科学—观赏园艺]