马尾松大小径材miRNA的差异表达分析  

Differential expression analysis of miRNA between large diameter wood and small diameter wood of Pinus massoniana

作　　者：周紫晶 范付华 尚先文[1,2] 覃慧娟 王聪慧 丁贵杰 谭健晖 ZHOU Zijing;FAN Fuhua;SHANG Xianwen;QIN Huijuan;WANG Conghui;DING Guijie;TAN Jianhui(Research Center for Forest Resources and Environment of Guizhou Province/Key Laboratory of Forest Cultivation in Plateau Mountain of Guizhou Province,Guizhou University,Guiyang 550025,China;College of Forestry,Guizhou University,Guiyang 550025,China;Guangxi Forestry Research Institute,Nanning 530002,China)

机构地区：[1]贵州大学贵州省森林资源与环境研究中心/贵州省高原山地林木培育重点实验室,贵阳550025 [2]贵州大学林学院,贵阳550025 [3]广西壮族自治区林业科学研究院,南宁530002

出　　处：《植物生理学报》2020年第10期2111-2120,共10页Plant Physiology Journal

基　　金：国家重点研发计划(2017YFD0600302);广西创新驱动发展专项(桂科AA17204087-4);国家重点研发计划后补助(黔科合平台人才[2018]5261号);贵州省一流学科建设项目(GNYL[2017]007)。

摘　　要：本研究采用Illumina HiSeq 2500高通量测序技术对马尾松大径材和小径材进行小RNA分子测序,构建了6个小RNA文库,共检测到1.42×10^7~1.59×10^7条序列。鉴定得到1567个miRNA,其中已知保守miRNA 783个,新miRNA 784个;差异表达miRNA 67个,其中31个在大径材马尾松中显著上调,36个显著下调。最终预测得到28044个靶基因,67个差异表达miRNA对应592个靶基因,GO分析表明这些差异表达的miRNA主要参与了细胞组分、分子功能和生物过程中共计27个类别,KEGG分析表明它们参与了44种不同的途径。利用实时荧光定量PCR验证了4个差异表达miRNA的表达趋势与小RNA测序结果相似,且对其靶基因可能起负调控作用。结果表明某些miRNA(如miR171、miR397、novel-m0462-5p、novel-m0101-3p、novel-m0504-3p、novel-m0279-3p等)可能参与马尾松生长发育过程,对马尾松的次生生长、维管形成层发育过程、木质素生物合成和植物激素信号转导过程等多方面产生作用,从而造成马尾松大径材和小径材的差异。本研究为今后马尾松大径材的培育工作提供参考。In this study, Illumina HiSeq 2500 high throughput sequencing technique was used for small RNA molecular sequencing of large diameter wood and small diameter wood of Pinus massoniana. Six small RNA libraries were constructed, and a total of 1.42×10^7–1.59×10^7 sequences were detected. 1 567 miRNAs were identified, of which 783 were known conservative miRNAs, 784 were new miRNAs. There were 67 differentially expressed miRNAs, of which 31 were significantly up-regulated and 36 were significantly down-regulated in P. massoniana. Finally, 28 044 target genes were predicted, among which 592 target genes of 67 differentially expressed miRNA were identified. GO analysis showed that these differentially expressed miRNA target genes were mainly involved in 27 categories in cellular components, molecular function and biological processes. KEGG analysis showed that they were involved in 44 different pathways. Through real-time fluorescence quantitative PCR, it was verified that the expression trends of the 4 differentially expressed miRNA were similar to the sequencing results of small RNA, and may have a negative regulatory effect on its target genes. Results show that some miRNAs(such as miR171, miR397, novel-m0462-5 p, novel-m0101-3 p, novel-m0504-3 p, novel-m0279-3 p, etc.) may be involved in the growth and development of P. massoniana. And they play an important role in secondary growth, vascular cambium development, lignin biosynthesis and plant hormone signal transduction of P. massoniana, resulting in the difference between large diameter and small diameter material. This study will provide reference for the cultivation of large diameter material of P. massoniana in the future.

关 键 词：马尾松 大径材 小径材 MIRNA 生长发育 

分 类 号：S791.248[农业科学—林木遗传育种]