梁山慈竹高通量转录组测序及差异表达基因分析

摘要

分析梁山慈竹及其体细胞突变体的转录组，挖掘功能基因并对其差异表达基因进行筛选和分析，为梁山慈竹遗传改良提供理论依据。利用 RNA-Seq 技术进行转录组测序，对测序结果进行 de novo 拼接和功能注释；并对差异表达基因进行筛选及 COG、GO、KEGG 数据库中进行比对注释，此外，基于 Swiss-Prot 功能注释结果，分析纤维素和木质素相关功能基因的表达量差异。测序结果表明，共获得86575631条 reads，de novo 组装得到84741条 unigenes，共有49829条被 Nr、COG、GO、KEGG、Swiss-Prot 注释。从梁山慈竹实生植株（对照）和体细胞突变体 No．30这2个测序样本中，筛选出3572条差异表达 unigenes，757条差异表达 unigenes 在 COG 分类体系中具有详细的蛋白功能释义，2213条差异表达 unigenes 在 GO 数据库具有功能定义，385条 unigenes 被注释到94条 KEGG Pathways 中。纤维素合成相关纤维素合酶、过氧化物酶、泛素连接酶和热休克蛋白在梁山慈竹体细胞突变体 No．30中表达量升高，木质素合成相关 MYB4、4-香豆酸 CoA 连接酶、肉桂醇脱氢酶、肉桂酰-CoA 还原酶和漆酶在突变体中表达量降低。提供了全面的梁山慈竹转录组信息，获得了一批在梁山慈竹纤维素和木质素生物合成过程中有重要功能的基因序列。%The transcriptome from the shoots of Dendrocalamus farinosus and its somatic mutant was sequenced using the RNA-Seq technology to elucidate its functional gene and analyze its differential expressed gene,providing a theoretical basis for its genetic improvement.The sequencing data was assembled by de novo assembly and the dif-ferential expressed gene screened was annotated in COG,GO and KEGG database.In addition,the differential ex-pression of gene related to cellulose and lignin biosynthesis was analyzed,basing on the Swiss-Prot function annota-tion.The sequencing results showed that a total of 86 575 631 reads were produced and assembled into a total of 84 741 unigenes by de novo,among which 49 829 were annotated in Nonredundant protein,Cluster of Orthologous Groups of proteins,Gene Ontology,Kyoto Encyclopedia of Genes and Genomes and Swiss-Prot database.Besides,a total of 3 572 differential expressed unigenes were identified from the plant of seeds (CK)and the somatic mutant No.30.Of these differential expressed genes,757 unigenes had a detailed protein functions in the COG classification system,2 213 unigenes had the function definition in the GO database,385 unigenes were annotated to 94 KEGG Pathways.The expression of genes encoding CesA,Prx,Ubiquitin-conjugating enzyme and Heat shock proteins in-creased in somatic mutant No.30,while,the expression of genes encoding MYB4,4CL,CAD,CCR and LAC de-creased.Our data provide the most comprehensive transcriptomic resource for Dendrocalamus farinosus and the tran-script sequences with important function related to cellulose and lignin biosynthesis were found,which provide the most precious information resources for the further research on bamboo.