Interactions of the Fungal Community in the Complex Patho-System of Esca, a Grapevine Trunk Disease
Abstract
:1. Introduction
- (i)
- To elucidate differences in the fungal community inhabiting little degraded (healthy + black necrosis) or highly degraded (white rot) wood of grapevines showing esca foliar symptoms (symptomatic, Sy) or not (asymptomatic, As).
- (ii)
- To determine the most informative point for detection of fungal infection within a vine. To this end, a comparison of the fungal population at four different parts (above the graft, A, strain cross, B, and right arm, C and left arm, D) of the vine was completed.
- (iii)
- To compare both culture-dependent and culture-independent methods to identify and quantify the fungal species which colonized the grapevine wood. We characterized the wood mycobiome composition by ITS1 sequencing on an Illumina MiSeq instrument. Moreover, quantification of the fungus Phaeomoniella chlamydospora was achieved by using TaqMan qPCR. Molecular results were compared with the cultivable community of fungi to establish the most efficient methodology.
2. Results
2.1. Culture-Dependent Microbiological Analysis
2.2. Culture-Independent Fungal Community Analysis
2.2.1. Illumina® NGS (Next Generation Sequencing) Results
2.2.2. Results of P. chlamydospora Quantification by q-PCR
2.3. Biological Conclusions
3. Discussion
3.1. Comparison of Both Culture-Dependent and Culture-Independent Methods
3.2. Pathogenetic Role of P. chlamydospora and Botryosphaeriaceae, Fungi Associated to GTDs
3.3. Basidiomycota and Hymenochataceae Family as Key of Pathotrophs
3.4. New Insights on Biocontrol and Microbiome Interaction
4. Materials and Methods
4.1. Vineyard
4.2. Wood Sampling
4.3. Culture-Dependent Microbiological Analysis
4.4. Culture-Independent Fungal Community Analysis (qPCR and Illumina Technology)
4.4.1. DNA Extraction
4.4.2. Amplification, Library, Illumina Sequencing and Bioinformatics Processing
4.4.3. Quantification of the Esca Causal Agent Phaeomoniella chlamydospora by qPCR
4.5. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
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Symptoms | S | H’ | J’ |
---|---|---|---|
As-BH | 213.25 ± 32.99 b | 2.33 ± 0.35 ab | 0.45 ± 0.06 ab |
As-WR | 138.00 ± 12.88 b | 1.82 ± 0.12 b | 0.37 ± 0.02 b |
Sy-BH | 405.09 ± 57.92 a | 3.41 ± 0.32 a | 0.57 ± 0.05 a |
Sy-WR | 202.50 ± 39.12 b | 2.67 ± 0.30 ab | 0.52 ± 0.06 ab |
ANOVA (p-values) | 8.64 (0.0001) | 5.40 (0.0030) | 3.71 (0.0182) |
Symptoms | C (%) | N (%) |
---|---|---|
As-BH | 27.20 ± 1.99 | 0.17 ± 0.06 |
As-WR | 36.03 ± 5.46 | 0.23 ± 0.03 |
Sy-BH | 27.33 ± 1.30 | 0.19 ± 0.02 |
Sy-WR | 28.17 ± 2.69 | 0.20 ± 0.03 |
ANOVA (p-values) | 1.70 (0.244) | 0.42 (0.747) |
Symptoms Category | Foliar Esca Symptoms | Wood Symptoms Observed in Trunk | GC | SP |
---|---|---|---|---|
As-BH | Asymptomatic | Healthy wood and black necrosis | C04 | A, B, C, D |
C26 | A, B, C, D | |||
D86 | A, B, C, D | |||
As-WR | Asymptomatic | White rot | D43 | A, B, C, D |
E08 | A, B, C, D | |||
E60 | A, B, C, D | |||
Sy-BH | Symptomatic | Healthy wood and black necrosis | D96 | A, B, C, D |
D80 | A, B, C, D | |||
C01 | A, B, C, D | |||
Sy-WR | Symptomatic | White rot | C40 | A, B, C, D |
C16 | A, B, C, D | |||
C41 | A, B, C, D |
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Martín, L.; García-García, B.; Alguacil, M.d.M. Interactions of the Fungal Community in the Complex Patho-System of Esca, a Grapevine Trunk Disease. Int. J. Mol. Sci. 2022, 23, 14726. https://doi.org/10.3390/ijms232314726
Martín L, García-García B, Alguacil MdM. Interactions of the Fungal Community in the Complex Patho-System of Esca, a Grapevine Trunk Disease. International Journal of Molecular Sciences. 2022; 23(23):14726. https://doi.org/10.3390/ijms232314726
Chicago/Turabian StyleMartín, Laura, Blanca García-García, and María del Mar Alguacil. 2022. "Interactions of the Fungal Community in the Complex Patho-System of Esca, a Grapevine Trunk Disease" International Journal of Molecular Sciences 23, no. 23: 14726. https://doi.org/10.3390/ijms232314726