Molecular Detection and Identification of Diatrypaceous Airborne Spores in Australian Vineyards Revealed High Species Diversity Between Regions

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2023 Jun 2

PMID 37267392

Authors

Regina Billones-Baaijens

Meifang Liu

Mark R Sosnowski

Matthew R Ayres

Sandra Savocchia

Affiliations

Soon will be listed here.

Abstract

The grapevine trunk disease, Eutypa dieback (ED), causes significant vine decline and yield reduction. For many years, the fungus Eutypa lata was considered the main pathogen causing ED of grapevines in Australia. Recent studies showed other Diatrypaceous fungi were also associated with vines exhibiting dieback symptoms but there is limited information on how these fungal pathogens spread in vineyards. Thus, information on the spore dispersal patterns of Diatrypaceous fungi in different wine regions will assist in identifying high-risk infection periods in vineyards. Using more than 6800 DNA samples from airborne spores collected from eight wine regions in south-eastern Australia over 8 years using a Burkard spore trap, this study investigated the diversity and abundance of Diatrypaceous species, using multi-faceted molecular tools. A multi-target quantitative PCR (qPCR) assay successfully detected and quantified Diatrypaceous spores from 30% of the total samples with spore numbers and frequency of detection varying between regions and years. The high-resolution melting analysis (HRMA) coupled with DNA sequencing identified seven species, with E. lata being present in seven regions and the most prevalent species in the Adelaide Hills, Barossa Valley and McLaren Vale. Cryptovalsa ampelina and Diatrype stigma were the predominant species in the Clare Valley and Coonawarra, respectively while Eutypella citricola and Eu. microtheca dominated in the Hunter Valley and the Riverina regions. This study represents the first report of D. stigma and Cryptosphaeria multicontinentalis in Australian vineyards. This study further showed rainfall as a primary factor that triggers spore release, however, other weather factors that may influence the spore release in different climatic regions of Australia still requires further investigation.

References

Benight A, Wartell R, Howell D . Theory agrees with experimental thermal denaturation of short DNA restriction fragments. Nature. 1981; 289(5794):203-5. DOI: 10.1038/289203a0. View

Morales-Cruz A, Amrine K, Blanco-Ulate B, Lawrence D, Travadon R, Rolshausen P . Distinctive expansion of gene families associated with plant cell wall degradation, secondary metabolism, and nutrient uptake in the genomes of grapevine trunk pathogens. BMC Genomics. 2015; 16:469. PMC: 4472170. DOI: 10.1186/s12864-015-1624-z. View

Molyneux R, Mahoney N, Bayman P, Wong R, Meyer K, Irelan N . Eutypa dieback in grapevines: differential production of acetylenic phenol metabolites by strains of Eutypa lata. J Agric Food Chem. 2002; 50(6):1393-9. DOI: 10.1021/jf011215a. View

Garganese F, Ippolito A, di Rienzo V, Lotti C, Montemurro C, Sanzani S . A new high-resolution melting assay for genotyping Alternaria species causing citrus brown spot. J Sci Food Agric. 2018; 98(12):4578-4583. DOI: 10.1002/jsfa.8986. View

Ririe K, Rasmussen R, Wittwer C . Product differentiation by analysis of DNA melting curves during the polymerase chain reaction. Anal Biochem. 1997; 245(2):154-60. DOI: 10.1006/abio.1996.9916. View

Gonzalez-Dominguez E, Berlanas C, Gramaje D, Armengol J, Rossi V, Berbegal M . Temporal Dispersal Patterns of , Causal Agent of Petri Disease and Esca, in Vineyards. Phytopathology. 2020; 110(6):1216-1225. DOI: 10.1094/PHYTO-10-19-0400-R. View

Xanthopoulou A, Ganopoulos I, Tryfinopoulou P, Panagou E, Osanthanunkul M, Madesis P . Rapid and accurate identification of black aspergilli from grapes using high-resolution melting (HRM) analysis. J Sci Food Agric. 2018; 99(1):309-314. DOI: 10.1002/jsfa.9189. View

Bustin S, Benes V, Garson J, Hellemans J, Huggett J, Kubista M . The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem. 2009; 55(4):611-22. DOI: 10.1373/clinchem.2008.112797. View

Vossen R, Aten E, Roos A, den Dunnen J . High-resolution melting analysis (HRMA): more than just sequence variant screening. Hum Mutat. 2009; 30(6):860-6. DOI: 10.1002/humu.21019. View

10.

Li W, Xi B, Yang W, Hawkins M, Schubart U . Complex DNA melting profiles of small PCR products revealed using SYBR Green I. Biotechniques. 2003; 35(4):702-4, 706. DOI: 10.2144/03354bm07. View

11.

Crous P, Slippers B, Wingfield M, Rheeder J, Marasas W, Philips A . Phylogenetic lineages in the Botryosphaeriaceae. Stud Mycol. 2008; 55:235-53. PMC: 2104729. DOI: 10.3114/sim.55.1.235. View

12.

Onetto C, Sosnowski M, Van Den Heuvel S, Borneman A . Population genomics of the grapevine pathogen Eutypa lata reveals evidence for population expansion and intraspecific differences in secondary metabolite gene clusters. PLoS Genet. 2022; 18(4):e1010153. PMC: 9007359. DOI: 10.1371/journal.pgen.1010153. View

13.

Billones-Baaijens R, Urbez-Torres J, Liu M, Ayres M, Sosnowski M, Savocchia S . Molecular Methods to Detect and Quantify Botryosphaeriaceae Inocula Associated With Grapevine Dieback in Australia. Plant Dis. 2019; 102(8):1489-1499. DOI: 10.1094/PDIS-11-17-1854-RE. View

14.

Diguta C, Rousseaux S, Weidmann S, Bretin N, Vincent B, Guilloux-Benatier M . Development of a qPCR assay for specific quantification of Botrytis cinerea on grapes. FEMS Microbiol Lett. 2010; 313(1):81-7. DOI: 10.1111/j.1574-6968.2010.02127.x. View

15.

Andolfi A, Mugnai L, Luque J, Surico G, Cimmino A, Evidente A . Phytotoxins produced by fungi associated with grapevine trunk diseases. Toxins (Basel). 2012; 3(12):1569-605. PMC: 3268457. DOI: 10.3390/toxins3121569. View

16.

Sosnowski M, Lardner R, Wicks T, Scott E . The Influence of Grapevine Cultivar and Isolate of Eutypa lata on Wood and Foliar Symptoms. Plant Dis. 2019; 91(8):924-931. DOI: 10.1094/PDIS-91-8-0924. View

17.

Moisy C, Berger G, Flutre T, Le Cunff L, Peros J . Quantitative Assessment of Grapevine Wood Colonization by the Dieback Fungus Eutypa lata. J Fungi (Basel). 2018; 3(2). PMC: 5715921. DOI: 10.3390/jof3020021. View

18.

Pouzoulet J, Mailhac N, Couderc C, Besson X, Dayde J, Lummerzheim M . A method to detect and quantify Phaeomoniella chlamydospora and Phaeoacremonium aleophilum DNA in grapevine-wood samples. Appl Microbiol Biotechnol. 2013; 97(23):10163-75. DOI: 10.1007/s00253-013-5299-6. View

19.

Urbez-Torres J, Haag P, Bowen P, Lowery T, OGorman D . Development of a DNA Macroarray for the Detection and Identification of Fungal Pathogens Causing Decline of Young Grapevines. Phytopathology. 2015; 105(10):1373-88. DOI: 10.1094/PHYTO-03-15-0069-R. View

20.

Pouzoulet J, Rolshausen P, Schiavon M, Bol S, Travadon R, Lawrence D . A Method to Detect and Quantify Eutypa lata and Diplodia seriata-Complex DNA in Grapevine Pruning Wounds. Plant Dis. 2019; 101(8):1470-1480. DOI: 10.1094/PDIS-03-17-0362-RE. View