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基于Illumina HiSeq~(TM)2000平台,对健康与感病杧果顶芽的转录组进行测序,采用BLAST软件将获得的Unigene与NCBI-nr、Swiss-Prot、KEGG和COG数据库进行比对,根据基因功能注释后分析杧果病健组织的差异表达基因(DEGs),并对DEGs进行GO和Pathway富集分析。结果表明:2个样品共获得119 815条Unigene,N50为1 546 bp,平均片段长度为880 bp;鉴定了29 878个DEGs,对随机挑选的11个差异表达基因进行了qRT-PCR验证,结果与转录组一致。以corrected P-value≤0.05为阈值的代谢途径有22条,其中大多数代谢途径与植物的抗逆响应密切相关;153个DEGs参与了淀粉和蔗糖代谢途径,DEGs主要是编码糖类异构酶、水解酶和转移酶等,参与葡萄糖水解、细胞碳水化合物、丙酮酸盐和核苷酸代谢等生物进程;在抗氧化生物过程中,编码活性氧代谢相关酶且log_2Ratio>10或<-10以上的DEGs有20个,14个属上调表达,表明活性氧代谢在杧果与病原互作过程中起到重要的调解作用;F.mangiferae侵染杧果后,以log_2Ratio>10或<-10为筛选条件,共获得酚类代谢相关差异表达基因53个,其中40个DEGs上调表达,推测杧果可能是通过合成加固细胞壁的木质素或生成抑菌作用的酚类化合物来提高对病原菌的抵抗能力;杧果与F.mangiferae互作过程中,钙信号传导、SA信号途径和丝裂原活化蛋白信号传导途径相关基因表达下调,造成了下游植物抗病基因RPM1的表达受到抑制,这可能是杧果畸形病的主要成因之一。
Based on the Illumina HiSeq 2000 platform, the transcriptome of healthy and susceptible buds was sequenced. The obtained Unigene was compared with NCBI-nr, Swiss-Prot, KEGG and COG databases by using BLAST software. Differential gene expression profiles (DEGs) were analyzed after annotation of gene function, and DEGs were analyzed by GO and Pathway enrichment. The results showed that a total of 119 815 Unigene were obtained in two samples, with an N50 of 1 546 bp and an average fragment length of 880 bp. A total of 29 878 DEGs were identified and 11 differentially expressed genes randomly selected were verified by qRT-PCR. The results Consistent with transcriptome. There are 22 metabolic pathways with the corrected P-value≤0.05 as the threshold, most of which are closely related to plant stress response. 153 DEGs are involved in the metabolic pathway of starch and sucrose, and DEGs mainly encode carbohydrate isomerase , Hydrolases and transferases, involved in biological processes such as glucose hydrolysis, cellular carbohydrates, pyruvate and nucleotide metabolism; in the antioxidant biological process, encoding enzymes related to active oxygen metabolism and log_2Ratio> 10 or <-10 above 20 of DEGs and 14 of them were upregulated, indicating that reactive oxygen metabolism played an important mediating role in the interaction between ginkgo biloba and pathogen. After F. mangiferae infected ginkgo, log_2Ratio> 10 or <-10 Screening conditions, a total of 53 differentially expressed genes related to phenolic metabolism were obtained, of which 40 DEGs were up-regulated. It is speculated that the fruit may enhance the resistance to pathogenic bacteria through the synthesis of lignin that reinforces the cell wall or phenolic compounds that produce antibacterial activity ; In the process of interaction between F. mangiferae and F. mangiferae, the expression of calcium signaling, SA signaling and mitogen-activated protein signal transduction related genes were down-regulated, resulting in the expression of the downstream plant resistance gene RPM1 To inhibition, this may be one of the main causes of congenital malformations.