Fig 1.
Symptoms of Eutypa dieback of grapevines caused by Eutypa lata.
(A) Cross-section of the cordon with wedge-shaped wood necrosis (solid arrow) and stunted shoots (dashed arrows); (B) vine cordon with dieback (bracket) and chlorotic, stunted shoots (dashed circle).
Table 1.
Multi-target primers designed for quantitative PCR (qPCR) and High Resolution Melting Analysis (HRMA) used in this study.
Table 2.
Fungal isolates used for the development of molecular tools to detect, quantify and identify Diatrypaceous species.
Table 3.
Fungal isolates of non-target species used for testing the specificity of primers DITS-1F and DITS-1R and DIA-17F and DIA-122R using conventional PCR, quantitative PCR (qPCR) and High Resolution Melting Analysis (HRMA).
Table 4.
Determination of the limit of detection (LOD) of the qPCR using different concentrations of Diat-5S gBlocks®.
Table 5.
Determination of the limit of detection (LOD) of the qPCR using different concentrations of genomic DNA of Eutypa lata, Cryptovalsa ampelina, Eutypella citricola and E. leptoplaca.
Table 6.
The calculated number of copies of the ribosomal RNA (rRNA) gene in one haploid genome of the four representative Diatrypaceae species.
Fig 2.
High resolution melting analysis (HRMA) for the identification of Diatrypaceous species in Australian vineyards.
(A) melt curve profiles; (B) normalised graph; (C) difference graph with Eutypella citricola as the core genotype which was converted to a horizontal line.
Fig 3.
Detection of Diatrypaceous spores from 2-day samples from different wine growing regions in South Australia (SA) and New South Wales (NSW) during the 2014–2016 (A) and 2017–2021 (B) spore surveillance studies. Black bars (■) are the total samples that tested positive to Diatrypaceous spores and grey bars (■) are the proportion of the positive samples that were trapped during or within 48 hours of a rain event.
Fig 4.
Average number of Diatrypaceous spores detected in all 2-day samples for each month in different wine growing regions in Australia from 2014–2016.
Differences in the number of samples (n) between years and regions were due to the missing data (red bars) associated with spore trap deployment dates or spore trap malfunctions.
Fig 5.
Average number of Diatrypaceous spores detected in all 2-day samples for each month in different wine growing regions in Australia from 2017–2021.
Differences in the number of samples (n) between years and regions were due to the missing data (red bars) associated with spore trap delopyment dates or spore trap malfunctions.
Fig 6.
Species diversity of Diatrypaceous airborne spores from different wine growing regions in Australia.
Airborne spores were collected using a Burkard volumetric spore trap and DNA extracted from 2-day spore tape sections were identified by High Resolution Melting Analysis (HRMA, ■ ■) and DNA sequencing (■) using Diatrypaceous multi-target primers DITS-1F and DITS-1R that can amplify 305–350 bp of the ribosomal DNA ITS region. Bars in light orange (■) are the DNA samples with similar HRMA curves as Eutypa lata and identified as variants. Csp. is the abbreviation for the genus Cryptosphaeria.