IS FUNGAL COMMUNITY OF CHESTNUT ORCHARD SOILS

Transcription

1 9 SEPTEMBER, 2015 IS FUNGAL COMMUNITY OF CHESTNUT ORCHARD SOILS RESILIENT TO FUNGAL INOCULATION? ÂNGELA SOARES, FRANCISCA REIS, ERIC PEREIRA, PAULA BAPTISTA, TERESA LINO-NETO

2 The relationships between plant-microbe and microbe-microbe can be complex

3 Application of innoculants is an attractive agricultural and forestry practice Biofertilizers (such as nitrogen-fixing microorganisms or mycorrhyzae) Phytostimulators (microbes expressing phytohormones, such as Azospirillum) Biological control agents (such as Trichoderma, Penicillium) that protect plants against phytopathogens.

4 SAPROTROPHIC FUNGUS: HYPHOLOMA FASCICULARE High occurrence in healthy chestnut groves Strong antagonist activity (Phytophtora spp.) Used as a biocontrol agent against Armillaria spp. 2) Is Hypholoma fasciculare introduction in chestnut groves able to reshape fungal community? 1) 2) Comparison between a well-adapted community (native soil) and a recently disturbed community

5 Chestnut and soil sampling 100-year-old chestnut orchard Terroso region, Parque Natural de Montesinho, Bragança Low abundance of H. fasciculare between Set Dec

6 Chestnut and soil sampling Experimental design Germination of seeds in sterile sand Pruning of radicle tips Plantlets transference into plastic cuvettes (vermiculite mixture) Plants transference to chestnut grove soil

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13 Chestnut and soil sampling Experimental design DNA extraction Ultra Clean Soil DNA Isolation kit (MO BIO Laboratories) Three soil samples/pot were mixed together Sample preparation and cell lysis Column washing DNA elution

14 Chestnut and soil sampling Experimental design DNA extraction DNA amplification (fungal ITS1 region) ITS1-F ITS2 Adapted from Bugni and Ireland (2004). Nat. Prod. Rep. 21:

15 Chestnut and soil sampling Experimental design DNA extraction DNA amplification (fungal ITS1 region) 454 pyrosequencing Sequence processing and data analysis

16 Sequence processing and data analysis Sterile (S) Non-sterile (NS) Wo/Hf +Hf Wo/Hf +Hf T0 T6M T6M T1Y T0 T6M T6M T1Y (S,T0) (S,T6M) (S,T6M+Hf) (S,T1Y+Hf) (NS,T0) (NS,T6M) (NS,T6M+Hf) (NS,T1Y+Hf) Total Raw reads 17,634 23,581 27,070 19,996 19,556 24,333 27,476 16, ,469 Post-filtering 13,602 15,793 19,795 11,793 9,815 12,289 14,687 12, ,076 Fungal reads 8,101 10,559 6,919 8,668 8,684 10,427 12,886 11,785 78,029 Number of OTUs

17 Phyla richness (OTUs nr) 0.2% 2.0% 38.9% 58.9% Ascomycota Basidiomycota Zygomycota Unclassified Phyla abundance (reads nr) 0.0% 1.8% 40.9% 57.4% Higher number of Ascomycota OTUs Higher abundance of basideomycetes

18 OTUs number ABUNDANCE > 50 READS HYPHOLOMA FASCICULARE IMPACT ON SOIL FUNGAL COMMUNITY S NS RICHNESS a a a a T0 T6M NS soils were richer than S soils Mostly parasitic species During chestnut growth there was an increase in the number of OTUs

19 OTUs number ABUNDANCE > 50 READS HYPHOLOMA FASCICULARE IMPACT ON SOIL FUNGAL COMMUNITY S NS a a a RICHNESS a T0 T6M T6M T1Y Hf Hf a a a a Does Hypholoma fasciculare disturbs fungal community of S soils? Is NS (native) soils more resilient to fungal inoculation? Abundance of Cortinarius limonius (M) and Paxillus involutus (M) species. Abundance of Auricularia cornea (PS) and Hebeloma mesophaeum (M) species.

20 HYPHOLOMA FASCICULARE IMPACT ON SOIL FUNGAL COMMUNITY CLUSTERING ACCORDING TO SIMILARITY Fungal community of NS soils more homogenous NS soils seems to present a buffering-like effect

21 HYPHOLOMA FASCICULARE IMPACT ON THE FUNCTIONAL GROUPS ABUNDANCE > 50 READS Mycorrhizal (M) NS soils Parasite (P) ABUNDANCE Parasite/Saprotrophic (PS) Saprotrophic (S)

22 CONCLUSIONS Well-adapted fungal community is more resistant to H. fasciculare inoculation than recently-stablished fungal community. H. fasciculare application increases the abundance of mycorrhizal fungi and reduces parasites abundance. This is the first environmental impact of Hypholoma fasciculare application for biological control purposes

23 ACKNOWLEDGMENTS University of Minho (UM) Polytechnic Institute of Bragança (IPB) Teresa Lino-Neto Paula Baptista Francisca Reis Eric Pereira Ângela Soares PTDC/AGR-AAM/099556/2008