摘要
以云南松一年生幼苗为研究材料,基于转录组数据,筛选到2个可能参与木质素单体糖基化修饰的候选基因(PyUGT1,PyUGT2)。利用RT-PCR技术克隆它们的全长开放阅读框序列,对这两个基因编码的蛋白质的理化性质、结构进行生信分析,检测其在不同组织中的表达水平,并和木质素通路小分子作相关性分析。结果显示:(1)PyUGT1的ORF长度为1458 bp,编码485个氨基酸;PyUGT2的ORF长为1476 bp,编码491个氨基酸。(2)PyUGT1和PyUGT2蛋白的结构均主要以α螺旋和无规则卷曲为主,是一种不稳定蛋白。(3)系统进化分析表明PyUGT1和PyUGT2单独聚为一支,与杨树等植物的UGT72B亚家族的亲缘关系最为接近。(4)表达分析表明PyUGT1在云南松的嫩茎和针叶中表达量最高且差异显著,PyUGT2在5个组织中的相对表达量无显著差异。(5)相关性分析表明PyUGT1和PyUGT2与部分木质素通路小分子含量有一定的相关性。研究结果表明,PyUGT1和PyUGT2在云南松木质素单体的糖基化修饰中发挥重要的作用,其中PyUGT1可能参与了云南松嫩茎和针叶中木质素单体的糖基化修饰,PyUGT2可能在不同的组织中都参与了木质素单体的糖基化修饰。该研究为进一步挖掘云南松中具有糖基化修饰功能的UGT基因提供科学依据。
Based on the transcriptome of Pinus yunnanensis,two glycosyltransferase candidate genes(PyUGT1,PyUGT2)involved in lignin monomerization were screened.Their full-length open-reading frame sequences were cloned by RT-PCR,and the biological and chemical properties and structures of the proteins encoded by these genes were analyzed.The correlation analysis was made with small molecules of lignin pathway.Result showed:(1)The ORF of PyUGT1 was 1458 bp,485 amino acids,and that of PyUGT2 was 1476 bp,491 amino acids.(2)The structure of PyUGT1 and PyUGT2 proteins are mainlyα-helices and irregular coiling,and they are unstable proteins.(3)Phylogenetic analysis showed that PyUGT1 and PyUGT2 were clustered into a single branch,which was most closely related to the UGT72B subfamily of poplar and other plants.(4)Expression analysis showed that PyUGT1 had the highest expression level in tender stems and needles of P.yunnanensis,and the relative expression level of PyUGT2 had no significant difference among the five tissues.(5)Correlation analysis showed that PyUGT1 and PyUGT2 were correlated with the secondary metabolite of the lignin pathway.These results suggest that PyUGT1 and PyUGT2 play an important role in the glycosylation modification of lignin monomers in P.yunnanensis,in which PyUGT1 may be involved in the glycosylation modification of lignin monomers in the stems and needles of P.yunnanensis,PyUGT2 may be involved in glycosylation modification of lignin monomers in different tissues.This study provides a basis for further studies on the functions of PyUGT1 and PyUGT2 in P.yunnanensis.
作者
母德锦
陈林
陈诗
许玉兰
蔡年辉
冯玲
唐军荣
MU Dejin;CHEN Lin;CHEN Shi;XU Yulan;CAI Nianhui;FENG Ling;TANG Junrong(Key Laboratory of National Forestry and Grassland Administration on Biodiversity Conservation in Southwest China,Southwest Forestry University,Kunming Yunnan 650224,P.R.China)
出处
《西部林业科学》
CAS
北大核心
2023年第3期109-117,127,共10页
Journal of West China Forestry Science
基金
国家自然科学基金项目(32260386)
云南省“兴滇英才支持计划”青年人才专项(XDYC-QNRC-2022-0203)
西南林业大学博士启动基金资助项目(112116)。
关键词
云南松
基因克隆
糖基转移酶
表达分析
Pinus yunnanensis
gene cloning
glycosyltransferase
expression analysis
