Scientia Agricultura Sinica ›› 2017, Vol. 50 ›› Issue (17): 3323-3336.doi: 10.3864/j.issn.0578-1752.2017.17.007

• PLANT PROTECTION • Previous Articles     Next Articles

Transcriptome Analysis of Creeping Bentgrass (Agrostis stolonifera) Infected with Rhizoctonia solani

SHI Yi, NIU KuiJu, MA HuiLing   

  1. College of Pratacultural Science, Gansu Agricultural University/Key Laboratory of Grassland Ecosystem, Ministry of Education/Sino-U.S. Center for Grazingland Ecosystem Sustainability, Lanzhou 730070
  • Received:2017-03-06 Online:2017-09-01 Published:2017-09-01

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Abstract: 【Objective】The objective of this study is to reveal the gene expression pattern of creeping bentgrass (Agrostis stolonifera) after been inoculated with Rhizoctonia solani, by comparing the gene expression level between plant with disease and without disease during gene transcription, and to identify the key genes of turfgrass responding to pathogen infection. 【Method】A. stolonifera which was grown for 14 days, was inoculated with R. solani. Leaf samples of plant with or without disease were collected after 3 days. High-throughput sequencing technology and Trinity software were used for obtaining the transcriptome. Different expression genes were screened with |log2 (fold change)| ＞1, q-value＜0.005 as threshold and the assembly was used as reference. Bioinformatics software was used to analyze the transcriptome difference. iTAK was used for transcription factors. All the DEGs were blasted with plant resistance gene database to find the R protein. Mapman software was used to analyze the signaling pathway. 【Result】 Using the high-throughput transcriptome sequencing, 125 253 092 were obtained. 466 761 transcripts were assembled by Trinity software. More than half of them were longer than 700 bp and N50=1 100 bp. CD-HIT was used to choose 334 212 transcripts as Unigene with the average length of 573 bp and N50=791 bp. By comparing the A. stolonifera transcripts with disease and without disease, 7 937 up-regulated genes and 1 570 down-regulated genes were obtained. Among the up-regulated genes, 296 of them are transcription factors (TFs), while 142 of down-regulated genes can be defined as TFs. Those TFs were classified as 58 families, including 54 zinc-finger protein containing TFs C2H2, which was the most, then 22 C3H TFs. A total of 451 of different expression genes (DEGs) can be annotated as plant R-protein, which can be classified into 33 families. Among these families, NBS-LRR containing resistance protein, LRR-like receptor protein kinase, ABC-2 type transporter, U-box domain containing protein kinase, and heat shock protein gene, these 5 families showed the most variation. A bunch of up-regulated DEGs can be mapped in plant biotic response pathway and enriched in pathogen recognition, ROS eliminate, signaling transport, programmed cell death, pathogenesis-related protein and so on, while the down-regulated genes were enriched in plant growth and development pathways. The expression level of random selected DEGs was tested with qRT-PCR, and they were consistent with the result of RNA-seq analysis. The random selected genes included 12 C2H2 TFs, 10 C3H TFs and 12 R-protein genes.【Conclusion】The inoculation of pathogen caused great expression difference of A. stolonifera transcriptome. Most of the TFs, R-protein and defense-related genes were up-regulated to suppress pathogen growth. The inherent resistance of A. stolonifera to R. solani has been generated with the co-action of all these genes.

Key words: Agrostis stolonifera, Rhizoctonia solani, transcriptome, different expression genes, disease resistance mechanism

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