苹果褐斑病菌侵染苹果属山定子的转录组学分析

Abstract
Figure/Table
References
Related Citation (15)

Download: PDF (2900 KB) HTML (1 KB)
Export: BibTeX | EndNote (RIS)

Abstract Marssonina leaf blotch of apple caused by Diplocarpon mali, is one of the main apple (Malus domestica) leaf defoliation diseases in apple production areas, which can result in severe etiolation and abscission of leaves during the growing season. In this study, a new generation of high-throughput sequencing technology Illumina Hiseq2500 transcriptome sequencing was used for transcriptome sequencing in M. baccata infected with Diplocarpon mali, and then bioinformatic methods were used for gene expression profile and gene function prediction. The results showed that 46.86 Gb sequence information was abtained. Through filtering, splicing, assembling and going redundancy, 48 868 unigenes were abtained. After assessing the length distribution, GC content, expression level and other aspects of the unigenes, the sequencing data showed good quality and high reliability. Homology of the 48 868 unigenes were predicted by common databases NR (Non-Redundant), Swiss-port Protein Sequence Database, Eukaryotic Orthologous Groups of Proteins (KOG), Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG), and 25 797 unigenes were directly aligned onto the genes of apple (M. domestica). Compared with KOG database, the unigenes could be divided into 25 categories according to their function, including general function prediction for 19.45%, signal transduction mechanisms for 11.84%, and the cell motility was the least. Taking KEGG database as a reference, the unigenes were located to 145 branched metabolic pathways, including antibiotic biosynthesis for 842, purine metabolism for 740, starch and sucrose metabolism for 437, T cell receptors signaling pathways for 292. Then RSEM software was used to calculate the differential expression, and edgeR software was used for differential expression analysis. By setting the threshold false discovery rate (FDR) ＜0.05, log2(fold change (FC))＞ 2 or log2(FC)＜-2 to screen differentially expressed genes, all 1 230 upregulated genes and 1 869 downregulated genes were identified. The enrichment results of KEGG Pathway showed that 94 differential genes participated in tyrosine metabolism, plant-pathogen interaction, plant hormone signal transduction and biosynthesis of amino acids, etc.. Expression of some regulated genes validated by qRT-PCR showed consistence with transcriptome analysis. In this study, the second generation of high-throughput transcriptome sequencing technology was used to research the related genes of the Malus baccata in the resistence to Diplocarpon mali, which provides theoretical foundation for clearing the resistence mechanism.

Key words： Malus baccata Marssonina leaf blotch of apple Transcriptome High-throughput sequencing

Received: 19 August 2016 Published: 24 December 2016

	Service

	E-mail this article
	Add to my bookshelf
	Add to citation manager
	E-mail Alert
	RSS
	Articles by authors
	LI Hai-Lu
	SONG Yan-Yan
	FENG Gao
	HUANG Li-Li
	QIANG Xiao-Yu
	HAN Jing-Mei

Cite this article:

LI Hai-Lu,SONG Yan-Yan,FENG Gao, et al. Transcriptome Analysis of the Malus baccata Infected with Diplocarpon mali[J]. , 2017, 25(1): 32-42.

URL:

http://journal05.magtech.org.cn/Jwk_ny/EN/ OR http://journal05.magtech.org.cn/Jwk_ny/EN/Y2017/V25/I1/32

黄锐, 黄富, 张晋,等. 2008. 杂交稻及其亲本抗瘟性的生化机制[J]. 西南农业学报, 21(1):80-83.（Huang R, Huang F, Zhang J et al.2008. Biochemical mechanism of resistance of hybrid rice sand their parents to rice blast[J].Southwest China Journal of Agricultural Sciences, 21(1):80-83.）
任斌. 2015. 山定子抗苹果褐斑病过程中活性氧迸发和内质网应激相关基因表达研究[D]. 硕士学位论文,西北农林科技大学,导师:黄丽丽.pp.17-18.(Ren B.2015.Studies on oxidative burst and expression of end op las mic reticulum stress related Gene sin resistance of Malus baccata to Diplocarpon Mall[D].Thesis for M.S.,Northwest A&F University, Supervisor:Huang L L,pp.17-18.)
田伟, 田义轲, 王彩虹, 等. 2010. 苹果组织总RNA提取方法的比较研究[J]. 青岛农业大学学报:自然科学版, 27(2):122-125.（Tian W, Tian Y K, Wang C H et al. 2010. Comparison of methons for toal RNA extraction apple tissues[J]. Editorial Dept.of Journal of QAU, 27(2):122-125.）
王洁, 赵华, 苏苏, 等. 2012. 苹果属不同种对褐斑病菌生长发育的影响[J]. 西北农业学报, 21(5):60-64, 83.（Wang J, Zhao H, Su S et al.2012. Effect of different malus species on growth and development of diplocarpon mali[J]. Acta Agriculturae Boreali-occidentalis Sinica, 21(5):60-64, 83.）
Asselbergh B, De V D, H?fte M. 2008. Global switches and fine-tuning-ABA modulates plant pathogen defense[J]. Molecular plant-microbe interactions, 21(6):709-719.
Dahl C C V, Baldwin I T.2007. Deciphering the Role of Ethylene in Plant–Herbivore Interactions[J]. Journal of Plant Growth Regulation, 26(2):201-209.
Grabherr M G, Haas B J, Yassour M, et al. 2013. Trinity: reconstructing a full-length transcriptome without a genome from RNA-Seq data[J]. Nature Biotechnology, 29(7):644-652.
Howe G. 2004. Jasmonates as signals in the wound response[J]. Journal of Plant Growth Regulation, 23(3):223-237.
Kim SJ, Maeda T, Sarker MZ, et al. 2007. Identification of anthocyanins in the sprouts of buckwheat[J]. Journal of Agricultural and Food Chemistry, 55(15):6314-6318.
Konishi T, Ohnishi O, Konishi T, et al. 2006. A linkage map for common buckwheat based on microsatellite and AFLP markers[J]. Fagopyrum, 23.
Li, Dewey N. 2011. RSEM: accurate transcript quantification from RNA-Seq data with or without a reference genome[J]. Bmc Bioinformatics, 12(1):93-99.
Livak K J, Schmittgen T D. 2001. Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2 ?ΔΔ C T, Method[J]. Methods, 25(4):402-408.
Mortazavi A, Williams B A, Mccue K, et al. 2008. Mapping and quantifying mammalian transcriptomes by RNA-Seq[J]. Nature Methods, 5(7):621-628.
Navarro L, Bari R, Achard P, et al. 2008. DELLAs control plant immune responses by modulating the balance of jasmonic acid and salicylic acid signaling[J]. Current Biology, 18(9):650-5.
Pieterse C M J, Leon-Reyes A, Van der Ent S, Van Wees S C M. 2009. Networking by small-molecule hormones in plant immunity[J]. Nature Chemical Biology, 5(5):308-316.
Pozo M J, Loon L C V, Pieterse C M J. 2004. Jasmonates - Signals in Plant-Microbe Interactions[J]. Journal of Plant Growth Regulation, 23(3):211-222.
Robinson M D, Mccarthy D J, Smyth G K. 2009. edgeR: a Bioconductor package for differential expression analysis of digital gene expression dataSMotn[J]. Bioinformatics, 26(1):139–140.
Shan L, He P, Li J, et al. 2008. Bacterial effectors target the common signaling partner BAK1 to disrupt multiple MAMP receptor-signaling complexes and impede plant immunity[J]. Cell Host & Microbe, 4(1):17-27.
Siemens J, Keller I J, Kunz S, et al. 2006. Transcriptome analysis of Arabidopsis clubroots indicate a key role for cytokinins in disease development[J]. Molecular plant-microbe interactions, 19(5):480-94.
Van Loon L C, Geraats B P J, Linthorst H J M. 2006. Ethylene as a modulator of disease resistance in plants[J]. Trends in Plant Science, 11(4):184-91.
Wang D, Pajerowska-Mukhtar K, Culler A H, et al. 2007. Salicylic acid inhibits pathogen growth in plants through repression of the auxin signaling pathway[J]. Current Biology, 17(20):1784-1790.
Zhao H, Han Q, Wang J, et al. 2013. Cytology of infection of apple leaves by Diplocarpon mali[J]. European Journal of Plant Pathology, 136(1):41-49.
Yin L, Zou Y, Ke X, et al. 2013. Phenolic responses of resistant and susceptible Malus, plants induced by Diplocarpon mali[J]. Scientia Horticulturae, 164(164):17-23.