Molecular Mechanism of the Generation of Asexual Spores of the Mango Fungal Pathogen (Colletotrichum gloeosporioides) Induced by Mechanical Injuries

doi:10.11983/CBB20026

Abstract

Abstract: Colletotrichum gloeosporioides is a prevalent pathogen that causes anthracnose in mango (Mangifera indica). Mycelium of C. gloeosporioides will accumulate a large number of conidia in 24-48 hours after mechanical injuries. However, it often accumulates none or few conidia during indoor culture, and the gene regulatory networks of the response to injury for a short-time (ST), or the key metabolic pathways involved in the response has not been explored. In this study, RNA-seq was carried out on RNA samples obtained at 5 time points within 2 hours after mechanical injuries. The differentially expressed genes were enriched by GO enrichment and KEGG metabolic pathway. The expression dynamics of mycelia in response to ST injury stress was analyzed. Based on a nonlinear ordinary differential equation model coupled with variable selection techniques, inter-module networks were constructed. The results showed that 417 differentially expressed genes were obtained, which belong to 12 clustered modules. KEGG enrichment analysis of differentially expressed genes was enriched in the process of pyruvate metabolism, sulfur metabolism, aflatoxin biosynthesis, diterpenoid biosynthesis. Combined with functional annotation, 12 core genes were identified that significantly correlated with ST injury-induced expression. These results provide valuable references for further research on asexual development and pathogenicity in C. gloeosporioides.

Key words: mango, Colletotrichum gloeosporioides, injury stress, RNA sequencing, expression dynamics

Liyan Wang, Mengyao Lu, Yue Tong, Xiangbin Xu, Zhengke Zhang, Lanhuan Meng, Xuequn Shi, Haichao Song. Molecular Mechanism of the Generation of Asexual Spores of the Mango Fungal Pathogen (Colletotrichum gloeosporioides) Induced by Mechanical Injuries[J]. Chinese Bulletin of Botany, 2020, 55(5): 551-563.

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URL: https://www.chinbullbotany.com/EN/10.11983/CBB20026

https://www.chinbullbotany.com/EN/Y2020/V55/I5/551

Figures/Tables 8

Table 1 Primers for real-time quantitative PCR

Transcript	Upstream primer sequence (5°?3°)	Downstream primer sequence (5°?3°)
ELA27200	CCTCAAACTTCGCAGATG	GGCTGTCGTGTAGAACTG
ELA31518	ACTGCGAATCACATCATCTCA	TAAGAGGTCGGCGTCAGA
ELA26856	CAGCAGATGGTCAAGAGG	GAGGAGACGAAGGAAGGA
ELA23533	TCAGCCTTGTTGGACCTTAG	CGTAGCATCGGACCTTGT
ELA34567	GAGCAGGTATCAGAAGACA	CCAGTTCGTAAGCCAGAT
Actin	ACGCTTCTCATCTCCAAGATCCGT	AGAGAGCCTCGTTGTCAATGCAGA

Figure 1 Gene expression data and GO enrichment for differentially expressed genes under different treatments (A) Gene expression in different samples; (B) Correlation between different samples; (C) GO enrichment for differentially expressed genes. MF: Molecular function; BP: Biological process; CC: Cell components

Table 2 KEGG enrichment result for differentially expressed genes

Description	Gene ratio	Bg ratio	P-value	Gene ID	Count
Aflatoxin biosynthesis	6/113	46/2826	0.009071	CGGC5_14023/CGGC5_13120/CGGC5_13638/ CGGC5_13456/CGGC5_10453/CGGC5_9572	6
Pyruvate metabolism	7/113	72/2826	0.023327	CGGC5_14045/CGGC5_7569/CGGC5_3411/ CGGC5_10319/CGGC5_11829/CGGC5_3455/ CGGC5_5764	7
Sulfur metabolism	3/113	19/2826	0.037811	CGGC5_5794/CGGC5_12200/CGGC5_15291	3
Diterpenoid biosynthesis	3/113	21/2826	0.048998	CGGC5_1921/CGGC5_15293/CGGC5_11451	3

Figure 2 Expression profiles for each cluster of differentially expressed genes

Figure 3 Enrichment analysis for each gene cluster of differentially expressed genes (A) GO enrichment analysis for each cluster; (B) KEGG enrichment analysis for each cluster. The x-axis represents the different cluster and the y-axis is the name of enrichment pathway. Different P-value are represented by dots with different colors, while the number of differentially expressed genes in each pathway are displayed for the size of the dots. P-value ranges from 0-0.15. The closer the value to 0, the more significant the enrichment is.

Table 3 Twelve core genes identified in 12 cluster modules of differentially expressed genes

Module	Gene numbers	Transcript	Gene description
1	1	ELA23757	Pep1
1	6	ELA25282	Cytochrome p450
1	16	ELA23240	Nad-dependent epimerase dehydratase
2	20	EFCGT00000013177	5.8S_rRNA
2	25	ELA35632	Polyketide synthase
2	44	ELA25574	Integral membrane protein
2	49	ELA24193	Short chain dehydrogenase reductase family
2	57	ELA35820	Ankyrin repeat-containing protein
2	189	ELA25228	Ankyrin repeat protein
2	26	ELA33919	Laccase
6	76	ELA24504	Major facilitator superfamily transporter
11	232	ELA37190	Catalase

Figure 4 Inter-module regulatory network reconstruction of differentially expressed genes Different colors correspond to different clustered modules. The numbers in the boxes represent the interacting gene numbers in core modules. The 12 core genes are included.

Figure 5 The dynamic expression patterns and data correlation of five core genes Different lowercase letters indicate significant differences among treatments (P<0.05).

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