Transcriptome Differential Expression Analysis of Mycocentrospora acerina Under Antagonism by Brevibacillus laterosporus S2-31

doi:10.13560/j.cnki.biotech.bull.1985.2020-0545

Abstract

Abstract:

This work aims to explore the molecular mechanism of antagonistic action of Brevibacillus laterosporus against leaf blight pathogen Mycocentrospora acerina by analyzing the transcriptome characteristics of pathogen antagonized by Brevibacillus laterosporus and studying the enrichment of differentially expressed genes（DEGs）and metabolic pathway. First,the antagonistic effect of S2-31 on leaf blight pathogen was observed on confrontation culture. Then the differences of gene expression level of pathogen under antagonistic and normal growth conditions were conducted by RNA sequencing,and verified by RT-qPCR. Finally,DEGs was annotated and enriched by relevant databases. M. acerina mycelium appeared shrank and malformed after confrontation cultivation. A total of 3 681 DEGs were obtained after sequencing,among which 2 224 were up-regulated and 1 457 were down-regulated. GO enrichment showed that all DEGs were annotated into 8 subpopulations of biological process,8 subpopulations of cell components and 4 subpopulations of molecular functions. KEGG enrichment analysis showed that 732 unigenes were located at 115 biological pathways,and amino sugar,nucleoside sugar metabolism,glycosylphosphatidylinositol（GPI）anchor biosynthesis,and peroxisomes were the most pathways enriched by DEGs. There were a relatively large number of DEGs in starch and sucrose metabolism,spliceosome and endocytosis pathways. The up-regulated DEGs were mainly enriched in the metabolic pathways,while the down-regulated DEGs were mainly enriched in the genetic information processing and cell process pathways. Furthermore,DEGs related to mycelial growth were screened from the significantly enriched pathways,and the results revealed that B. laterosporus mainly inhibited the formation of peroxisomes,GPI anchors,and the process of endocytosis of pathogenic mycelial. The results of RT-qPCR on the selected DEGs were consistent with those of RNA sequencing. The antagonism of B. laterosporus S2-31 inhibited the growth of leaf blight pathogen in vitro and induced significant changes in its transcriptome characteristics,which were mainly related to the pathways of metabolism,cell process and genetic information processing.

Key words: Brevibacillus laterosporus, Asarum leaf blight, Mycocentrospora acerina, biological control, transcriptomics

LIU Ya-ling, YU Ying, LU Hai-kun, LEI Hui-xia, SUI Xin, GUO Jing. Transcriptome Differential Expression Analysis of Mycocentrospora acerina Under Antagonism by Brevibacillus laterosporus S2-31[J]. Biotechnology Bulletin, 2021, 37(2): 111-121.

Figures/Tables 11

References 32

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Sample	Clean reads	Error rate/%	Q30/%	GC content/%
CK-1	41 693 598	0.0279	91.52	53.76
CK-2	41 493 354	0.0267	92.53	53.31
CK-3	42 041 596	0.0267	92.59	54.02
S-1	40 579 598	0.027	92.26	53.33
S-2	42 027 532	0.0276	91.78	53.2
S-3	41 786 006	0.027	92.28	52.81

Sample	Clean reads	Error rate/%	Q30/%	GC content/%
CK-1	41 693 598	0.0279	91.52	53.76
CK-2	41 493 354	0.0267	92.53	53.31
CK-3	42 041 596	0.0267	92.59	54.02
S-1	40 579 598	0.027	92.26	53.33
S-2	42 027 532	0.0276	91.78	53.2
S-3	41 786 006	0.027	92.28	52.81

Pathway	Gene ID	Gene description	FC（S/CK）	P value
Peroxisome	TRINITY_DN5187_c2_g3	PEX19	0.148	6.239E-04
	TRINITY_DN4909_c1_g15	alpha/beta-hydrolase	7.412	1.452E-06
	TRINITY_DN2942_c0_g1	PEX16	0.317	2.685E-03
Endocytosis	TRINITY_DN5265_c0_g11	Arf GTPase activating protein	0.165	1.759E-05
	TRINITY_DN5050_c1_g5	vacuolar-sorting protein-like protein snf7	0.159	1.382E-04
	TRINITY_DN4927_c2_g3	GTP binding	0.214	9.932E-04
	TRINITY_DN5015_c8_g6	f-actin-capping protein subunit alpha	0.304	3.159E-06
	TRINITY_DN5245_c3_g6	actin cytoskeleton organization	0.463	0.0098
GPI-anchor biosynthesis	TRINITY_DN5325_c0_g1	GPI：protein transamidase	0.291	1.858E-03
	TRINITY_DN4901_c0_g4	PIG-A	0.298	0.00012
	TRINITY_DN4400_c0_g1	PIG-C	0.385	0.0005
	TRINITY_DN4854_c1_g10	PIG-H	0.418	0.0012
	TRINITY_DN5334_c1_g6	PIG-L	0.446	0.0021
Spliceosome	TRINITY_DN4979_c1_g13	ATP binding	0.156	1.290E-10
	TRINITY_DN5195_c2_g7	pre-mRNA splicing factor	0.282	1.466E-05
	TRINITY_DN5299_c2_g2	splicing factor 3b	0.390	1.489E-04
	TRINITY_DN5211_c1_g1	spliceosomal tri-snRNP complex assembly	0.468	0.015

Pathway	Gene ID	Gene description	FC（S/CK）	P value
Peroxisome	TRINITY_DN5187_c2_g3	PEX19	0.148	6.239E-04
	TRINITY_DN4909_c1_g15	alpha/beta-hydrolase	7.412	1.452E-06
	TRINITY_DN2942_c0_g1	PEX16	0.317	2.685E-03
Endocytosis	TRINITY_DN5265_c0_g11	Arf GTPase activating protein	0.165	1.759E-05
	TRINITY_DN5050_c1_g5	vacuolar-sorting protein-like protein snf7	0.159	1.382E-04
	TRINITY_DN4927_c2_g3	GTP binding	0.214	9.932E-04
	TRINITY_DN5015_c8_g6	f-actin-capping protein subunit alpha	0.304	3.159E-06
	TRINITY_DN5245_c3_g6	actin cytoskeleton organization	0.463	0.0098
GPI-anchor biosynthesis	TRINITY_DN5325_c0_g1	GPI：protein transamidase	0.291	1.858E-03
	TRINITY_DN4901_c0_g4	PIG-A	0.298	0.00012
	TRINITY_DN4400_c0_g1	PIG-C	0.385	0.0005
	TRINITY_DN4854_c1_g10	PIG-H	0.418	0.0012
	TRINITY_DN5334_c1_g6	PIG-L	0.446	0.0021
Spliceosome	TRINITY_DN4979_c1_g13	ATP binding	0.156	1.290E-10
	TRINITY_DN5195_c2_g7	pre-mRNA splicing factor	0.282	1.466E-05
	TRINITY_DN5299_c2_g2	splicing factor 3b	0.390	1.489E-04
	TRINITY_DN5211_c1_g1	spliceosomal tri-snRNP complex assembly	0.468	0.015

Gene ID	Gene description	Primer sequences（5'-3'）
TRINITY_DN4979_c1_g13	ATP binding	CGAACGAACCACCAAACAT	GGAACCAGTAGGGGCACAG
TRINITY_DN5195_c2_g7	pre-mRNA splicing factor	CGAAACATCCCTTACCATT	AGTACGACCAGCACGACCG
TRINITY_DN5265_c0_g11	Arf GTPase activating protein	CGTGAAGAAGCGGAGCGTA	GCGACAGGTGTGGGGGATA
TRINITY_DN5050_c1_g5	vacuolar-sorting protein-like protein snf7	CCCGTCTCAAACCATCTACAAC	ATTATTTGGGAGCTGGTCTGTT
TRINITY_DN4873_c1_g11	chitin binding	TGGCAGGTGCGGTAAGGAGT	TGTGGTGGTGGAAGGTGGTC
TRINITY_DN4909_c1_g15	alpha/beta-hydrolase	CTCCGCACCAAGCAACAAAC	AGCCCCAGTCATGCCCAATC
TRINITY_DN5416_c0_g1	gamma-glutamyltranspeptidase-like protein	CTCTTCCCTGGTCCACTGTT	TGTTGCGTTCTGATTTGCTG