Combined Transcriptomic and Metabolomic Analysis of Virulence Differences among Three Lecanicillium lecanii Strains

doi:10.13560/j.cnki.biotech.bull.1985.2024-1281

Abstract

Abstract:

Objective To study the virulence differences among various strains of Lecanicillium lecanii, and to identify key differentially expressed genes (DEGs) and differential secondary metabolites (DEMs) associated with strain virulence through integrated transcriptomic and metabolomic data analyses. Method RNA-seq and LC-MS/MS techniques were used to detect the gene expression and secondary metabolites of three L. lecanii strains cultured for 8 d and liquid fermentation for 8 d, and the differential genes were verified by RT-qPCR. Result Indoor virulence results showed that J-1 secondary metabolites were the most virulent against Myzus persicae, with highly significant differences from J-2 and V-1. Transcriptome and metabolome results: 225 DEGs and 59 DEMs were obtained for J-1_vs_J-2, 2464 DEGs and 75 DEMs were obtained for J-1_vs_V-1. Differential genes and differential metabolites were enriched by KEGG analysis, mainly in phenylalanine metabolism, tyrosine metabolism and ABC transporter protein, 79 DEGs and 19 DEMs, salicylic acid, 3-hydroxyphenylacetic acid, phenyllactic acid, hippuric acid, and methyl beta-D-galactoside were significantly up-regulated and norepinephrine was highly significantly down-regulated between high and low virulence strains. c75905.graph_c0,c78027.graph_c0,c77968.graph_c0, c78586.graph_c1, c74779.graph_c0, and c78871.graph_c0 were significantly up-regulated. RT-qPCR results showed that the key differential genes were consistent with the expression trends in the transcriptome. Conclusion Six key genes and six significantly enriched differential metabolites that may be involved in regulating the virulence of L. lecanii strains are found by combined transcriptome and metabolome analysis, which may provide a basis for the construction of highly virulent strains.

Key words: Lecanicillium lecanii, transcriptomic, metabolomic, virulence

CHAI Jun-fa, HONG Bo, JIA Yan-xia. Combined Transcriptomic and Metabolomic Analysis of Virulence Differences among Three Lecanicillium lecanii Strains[J]. Biotechnology Bulletin, 2025, 41(8): 311-321.

Figures/Tables 10

References 37

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名称 Name	CAS	纯度 Purity	品牌 Brands
甲醇	67-56-1	≥99.0%	Thermo
乙腈	75-05-8	≥99.9%	Thermo
甲酸	64-18-6	LC-MS grade	TCI
甲酸铵	540-69-2	≥99.9%	Sigma
H₂O			Millipore
Succinic acid-2，2，3，3，-d4	14493-42-6		sigma
Cholic acid-2，2，3，4，4，-d5	53007-09-3		sigma
DL-Tryptophan-2，3，3-d3	340257-61-6		CDN
DL-Methionine-3，3，4，4-d4	93709-61-6		CDN
L-PHENYLALANINE （RING-D5）	63-91-2	98%	CIL
CHOLINE CHLORIDE（TRIMETHYL-D9）	/	98%	CIL

名称 Name	CAS	纯度 Purity	品牌 Brands
甲醇	67-56-1	≥99.0%	Thermo
乙腈	75-05-8	≥99.9%	Thermo
甲酸	64-18-6	LC-MS grade	TCI
甲酸铵	540-69-2	≥99.9%	Sigma
H₂O			Millipore
Succinic acid-2，2，3，3，-d4	14493-42-6		sigma
Cholic acid-2，2，3，4，4，-d5	53007-09-3		sigma
DL-Tryptophan-2，3，3-d3	340257-61-6		CDN
DL-Methionine-3，3，4，4-d4	93709-61-6		CDN
L-PHENYLALANINE （RING-D5）	63-91-2	98%	CIL
CHOLINE CHLORIDE（TRIMETHYL-D9）	/	98%	CIL

基因ID Gene ID	正向引物Forward primer （5′-3′）	反向引物 Reverse primer （5′-3′）
GAPDH	F： CCCCAAACACCTCATCCTCA	R： ATGCCAACCTTGACGGGAG
c78027.graph_c0	F： CAATCCGGCCCGACTACTTCT	R： TTACATGGCAACAACTGGTGAGG
c77500.graph_c2	F： TCTGAGAGTGTCCCGTCGAT	R： GCGAGACTCGATCCAGAGTG
c78385.graph_c0	F： CGCAGTTCAACTTGTGGTGG	R： AACGAGACGGCAACATGAGT
c74339.graph_c0	F： CATGGGCGGTTCGATACTCA	R： ATAGGTCCAGCGGTCAGAGT
c77841.graph_c0	F： ATCACGCTTCCACATTCACTCAC	R： CTTGAAATCCTGGAGGTTGATGTAG
c80667.graph_c0	F： CGTGGTGATAGTAATGAGAACGGAC	R： GGCGTGTCATGCTACGTTCATAT
c71547.graph_c0	F： GGCCAATCAATGGGACGAAA	R： CACATCGCCGACCAACCAA

基因ID Gene ID	正向引物Forward primer （5′-3′）	反向引物 Reverse primer （5′-3′）
GAPDH	F： CCCCAAACACCTCATCCTCA	R： ATGCCAACCTTGACGGGAG
c78027.graph_c0	F： CAATCCGGCCCGACTACTTCT	R： TTACATGGCAACAACTGGTGAGG
c77500.graph_c2	F： TCTGAGAGTGTCCCGTCGAT	R： GCGAGACTCGATCCAGAGTG
c78385.graph_c0	F： CGCAGTTCAACTTGTGGTGG	R： AACGAGACGGCAACATGAGT
c74339.graph_c0	F： CATGGGCGGTTCGATACTCA	R： ATAGGTCCAGCGGTCAGAGT
c77841.graph_c0	F： ATCACGCTTCCACATTCACTCAC	R： CTTGAAATCCTGGAGGTTGATGTAG
c80667.graph_c0	F： CGTGGTGATAGTAATGAGAACGGAC	R： GGCGTGTCATGCTACGTTCATAT
c71547.graph_c0	F： GGCCAATCAATGGGACGAAA	R： CACATCGCCGACCAACCAA

样本名称 Sample	Read number	Clean bases	碱基质量值≥Q30 Base quality value（%）	GC含量 GC content（%）
J-1-1	19 650 825	5 859 575 818	94.76	55.46
J-1-2	22 287 859	6 648 086 524	95.63	55.51
J-1-3	21 026 399	6 275 725 238	95.40	55.51
J-2-1	22 649 343	6 759 978 410	95.32	55.33
J-2-2	19 611 279	5 792 883 170	96.13	54.98
J-2-3	20 383 277	6 077 510 742	95.28	55.53
V-1-1	20 089 720	5 971 239 342	94.51	55.26
V-1-2	19 968 516	5 946 912 808	95.82	55.62
V-1-3	24 919 447	7 407 074 958	95.32	54.73