杉木施肥引发基因表达响应机制的转录组分析  

A transcriptome analysis of the gene expression responses induced by fertilization treatments in Chinese fir

作　　者：齐明[1] 何贵平[1] 王海蓉[2] 邱勇斌 程亚平 徐肇友 QI Ming;HE Guiping;WANG Hairong;QIU Yongbin;CHENG Yaping;XU Zhaoyou(Research Institute of Subtropical Forestry of CAF,Fuyang 311400,Zhejiang,China;Zhejiang Provincial Key of Suichang County,Zhejiang province,Suichang 323300,Zhejiang,China;Forestry Farm of Kaihua County Zhejiang Province,Kaihua 324300,Zhejiang,China;Qingyuan Forest farm of Qingyuan County,Zhejiang province,Qingyuan 323805,Zhejiang,China;Foresty Academy of Longquan City,Zhejiang Province,Longquan 323700,Zhejiang,China)

机构地区：[1]中国林业科学研究院亚热带林业研究所,浙江富阳311400 [2]浙江省遂昌县林业技术推广总站,浙江遂昌323300 [3]浙江省开化县林场,浙江开化324300 [4]浙江省庆元县庆元林场,浙江庆元323805 [5]浙江省龙泉市林科院,浙江龙泉323700

出　　处：《中南林业科技大学学报》2020年第4期101-110,共10页Journal of Central South University of Forestry & Technology

基　　金：中国林科院中央级公益性科研院所基本科研业务费专项资金资助子项(CAFYBB2017 ZA001-1-2);浙江省“十三五”林木新品种育种专项(2016C02056-5)。

摘　　要：【目的】从基因表达水平上,研究施肥引起杉木基因表达变化规律,为杉木施肥理论与MAS育种实践,提供科学依据。【方法】以开化1年生的杉木无性系开6扦插苗为研究对象,4种施肥处理,每处理三个重复,采用Illumina HiSeq4000高通量测序技术进行测序,对测序结果进行无参转录组分析。【结果】1)转录组测序共产生clean reads5.0E+07 nt到7.1E+07 nt,总拼接长度724341090 nt,通过Trinity组装,获得了74288个unigenes(基因),然后将组装序列在6个数据库(Nr,Swiss-prot,eggNOG,KEGG,Pfam,GO)进行BLASTX分析,获取功能注释信息;2)韦恩图揭示基因对不同施肥处理的响应是:在-P-N1-VS-WT比较组中,未施肥处理,有4650个特异表达基因,其中包含了若干耐性基因,其它比较组也获得了相应的结果。杉木施肥诱导一些基因的开启和上调,同时也导致一些基因表达的关闭或下调;不同处理间的表达差异基因数在不同施肥组间变化很大,显著差异表达基因从施磷与对照比较组的429个到施磷与施氮比较组的4942;3)差异表达极显著基因的热聚类分析结果,揭示同一比较组,不同处理间基因表达是互补的关系:同一基因要么高表达,要么低表达;4)GO富集与分类显示施肥对杉木影响显著的基因主要定位在叶绿体、细胞骨架、细胞膜、细胞核、细胞质上;5)GO富集分析的结果表明施氮(磷)肥,不仅可以促进氮(磷)的吸收与代谢,而且也能促进磷(氮)的吸收与代谢;6)对施磷肥与氨酰基-tRNA大分子化合物合成的生理生化过程进行了分析。基于以上研究结果:今后可对转录组分析获得的重要基因进行研究,其结果可用于指导杉木施肥和杉木营养遗传育种MAS选择。【结论】RNA-seq高通量测序技术,对于研究非模式植物(包括基因组比较大的针叶树)施肥分子机制,是一种快速而简便的方法。研究结果表明,施肥不仅激活了与N、P的吸收、转运和利用有【Objective】The change rule of Chinese fir gene expression caused by fertilization was studied,which provided knowledge for Chinese fir fertilization theory and MAS breeding practice.【Method】The test material was the 1-year clone K6,four fertilization treatments,three repeats for each treatment.Illumina HiSeq4000 high-throughput sequencing technology was used for sequencing,non-reference transcriptome analysis was performed on the sequencing data.【Result】1)In 12 samples,the transcriptome sequencing produced clean reads from 5.0E+07 nt to 7.1E+07 nt,and the total splicing length was 724,341,090 nt.Through Trinity assembly,74,288 unigene genes were obtained.and the assembled sequences were analyzed by BLASTX in six databases(Nr,swiss-prot,eggNOG,KEGG,Pfam,GO)to obtain functional annotation information;2)Venn diagram revealed the response of genes to different fertilization treatments:in the–P1-N1-VS-WT comparison group,there were 4,650 specifically expressed genes without fertilization group,including several tolerance genes,other comparison groups also obtained corresponding results;3)Chinese fir fertilization induces the opening and up-regulation of some genes,and also leads to the closing or down-regulation of some genes;The number of differentially expressed genes in different treatments varied greatly among different fertilization groups,The significantly differentially expressed genes ranged from 429 of the group3_VS_group4 to 4,942 of the group3_VS_group2;4)The results of the heat-map clustering graph of significant differentially expressed genes of the six comparison groups were as follows:the up-regulation and down-regulation of gene expression in different treatments of the same comparison group were complementary:either high expression or low expression.5)Furthermore,the GO enrichment and classification revealed GO terms mainly located in chloroplasts,cytoskeleton,cell membrane,nucleus and cytoplasm,etc,which indicated fertilization had significant influence on Chinese fir.The analytical res

关 键 词：杉木 转录组测序 氮肥 磷肥 差异表达基因 

分 类 号：S718.46[农业科学—林学]