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Evidence that Eutypa lata and other diatrypaceous species
occur in New South Wales vineyards
W. M. Pitt
A,C
, R. Huang
A
, F. P. Trouillas
B
, C. C. Steel
A
and S. Savocchia
A
A
National Wine and Grape Industry Centre, Charles Sturt University, Locked Bag 588, Wagga Wagga,
NSW 2678, Australia.
B
Department of Plant Pathology, University of California, Davis, California 95616, USA.
C
Corresponding author. Email: wpitt@csu.edu.au
Abstract. Eutypa dieback, caused by the fungus Eutypa lata is a serious disease of grapevines that affects vineyard
productivity and longevity. Grapevines displaying foliar symptoms typical of Eutypa dieback or evidence of dead spurs,
cankers, or discoloured vascular tissue, were surveyed from 77 vineyards throughout New South Wales (NSW), Australia.
Fungal cultures were tentatively identified based on cultural morphology, before further identification using sequence
analysis of rDNA internal transcribed spacer regions. E. lata and several other species from the Diatrypaceae including
Cryptovalsa ampelina, and species of Eutypella and Diatrypella were isolated from diseased grapevines. Eutypa dieback was
found to be more widespread in NSW than first thought, with confirmation that the disease is present both in the Central
Ranges and southern NSW districts, regions recognised for their cooler climates and higher annual rainfall, both of which
favour the growth of E. lata.
Introduction
The ascomycete fungus Eutypa lata (=E. armeniaceae), first
described from apricot, is a significant pathogen of grapevines,
both in Australia and abroad, where it is responsible for the
vascular disease Eutypa dieback (Carter 1957; Carter et al. 1983).
While both the anamorph (Libertella blepharis) and teleomorph
are produced on dead wood, the pathogen is disseminated solely
by ascospores that are released from mature perithecia for up to
36 h after rainfall (Dubos 1987). Infection commences when
ascospores germinate on fresh pruning wounds or other areas of
the vine that have been damaged by vineyard operations
(Sosnowski et al. 2004). E. lata subsequently colonises the
xylem tissue, cambium and phloem and eventually girdles the
vine as a result of canker formation around infected wounds
(Munkvold and Marois 1995). Wounds are most susceptible to
infection in early winter, becoming less so as the temperature rises
later in the year, owing to increased competition from other
natural wound colonisers and the initiation of sap flow as the
growth cycle commences in spring (Dubos 1987; Chapuis et al.
1998). Foliar symptoms, which are caused by toxins produced
by the fungus, include stunted shoots with shortened internodes
and small distorted, chlorotic leaves with cupped or tattered
margins (Mahoney et al. 2005). A characteristic wedge-shaped
zone of dead wood is also common in the trunks and cordons
of infected vines (Moller and Kasimatis 1981). Without
intervention, vines may die within several years, thereby
reducing vineyard productivity, longevity and economic
sustainability (Wicks and Davies 1999; Siebert 2001).
Diagnosis of the disease can be difficult as infection is
followed by a period of latency. This delay in symptom
expression as the fungus colonises the vascular tissue may be as
brief as 1 year (Tey-Rulh et al. 1991), but generally occurs over a
period of 3–8 years following infection (Carter 1978). As a result,
infected vines may appear asymptomatic (Loschiavo et al. 2007).
Similarly, the expression of foliage symptoms may fluctuate from
season to season (Sosnowski et al.2007b), can be influenced by
grapevine cultivar and fungal isolate (Sosnowski et al. 2007a)
and can be confused with other diseases or disorders or masked
by the growth of healthy foliage from neighbouring vines
(Lardner et al. 2005). In culture, where diagnosis is based on
the anamorph, the situation is equally difficult as hyphae lack
diagnostic characters; conidial production may be inconsistent
and morphological features of the anamorph are insufficient to
distinguish E. lata from other ascomycetes, especially other
genera of the Diatrypaceae possessing Libertella anamorphs and
including Eutypella,Diatrypella,Diatrype,Cryptosphaeria and
Cryptovalsa (Glawe and Rogers 1984; Acero et al. 2004; Mostert
et al. 2004). Furthermore, the fungus is often overgrown by
faster growing species cohabitating the same piece of wood
(Rolshausen et al. 2004).
Eutypa dieback is widespread throughout many of the
premium wine-growing areas of Australia (Sosnowski et al.
2007b), and has been reported to affect more than 60% of
vines in some South Australian vineyards (Highet and Wicks
1998). A recent survey showed that Eutypa dieback is widespread
in the Adelaide Hills region, despite the majority of vines being
planted within the past decade (Loschiavo et al. 2007). This
suggests that the fungus can establish rapidly in new grapevines
planted under favourable conditions. With premium wines now
being produced from cooler climate regions of New South Wales
CSIRO PUBLISHING
www.publish.csiro.au/journals/app Australasian Plant Pathology, 2010, 39,97–106
Australasian Plant Pathology Society 2010 10.1071/AP09051 0815-3191/10/010097
(NSW) having climatic conditions not unlike those found
in the Adelaide Hills, the status of Eutypa dieback in NSW is
of increasing concern. This study aimed to determine the
incidence and distribution of Eutypa dieback throughout
NSW. Other diatrypaceous fungi isolated from diseased
grapevines were also documented as some of these species
have previously been implicated in the death and decline of
grapevines (Mostert et al. 2004; Luque et al. 2006).
Materials and methods
Isolation and morphological identification
Between November 2006 and April 2008 field surveys were
conducted from 77 vineyards across NSW, encompassing
seven major grape-growing regions (Table 1), viz. Big Rivers
(growing season spatial mean average temperature from
1 October 1971 to 30 April 2000, ~21.1C), Central Ranges
(~18.9C), Hunter Valley (~20.7C), Southern NSW
(~18.3C), South Coast (~17.8C), Northern Rivers (~20.8C),
Northern Slopes (~18.6C). A total of 1846 wood samples
were collected from the cordons or trunks of grapevines with
evidence of dieback, including dead spurs or cordons, cankers or
bleached and discoloured tissue. Five 2-mm
2
portions of wood
from each sample, were excised from the margin of diseased and
healthy tissue, surface sterilised in 8% bleach (active ingredient
1.0% w/v sodium hypochlorite; LabServ by Biolab Australia
Ltd, Clayton, Vic., Australia) for 2 min and transferred to potato
dextrose agar (PDA; Oxoid Ltd, Basingstoke, Hampshire,
England). Cultures were incubated in the dark at 25C for
5–7 days before being transferred onto fresh PDA to create
pure cultures. Foliar symptoms typical of Eutypa dieback were
encountered infrequently during the surveys, but in such cases
wood samples were extracted and prepared for fungal isolation
as described previously. Diatrypaceous species were tentatively
identified based on gross cultural morphology (Glawe and
Rogers 1984; Glawe and Jacobs 1987; Carter 1991; Mostert
et al. 2004). In December 2008, additional surveys and
collections were conducted from grapevines both in the Hunter
Valley and Tumbarumba (NSW). Foliage symptoms of Eutypa
dieback were not observed, at either location, and in contrast to
the previous collections, isolations were made from fruiting
bodies collected on dead wood from aged vines or debris from
the vineyard floor. Isolations of diatrypaceous fungi from these
samples were made directly from ascospores as described by
Trouillas and Gubler (2004), with species tentatively identified
based on morphology of the teleomorph (Glawe and Rogers
1984).
DNA extraction, amplification and sequencing
Before extraction of DNA, selected Diatrypaceae isolates
were pure cultured by hyphal tip, before being transferred by
colonised agar plug, to 50-mL Falcon tubes containing 20 mL
of potato dextrose broth (Oxoid). Broth cultures were incubated
on a Sartorius Certomat BS-1 (Goettingen, Germany) orbital
shaker at 90 rpm (=0.4779g, or approximately 0.5g; the orbital
mechanism has a rotational radius of 5 cm) for 7 days at
25C. Mycelia were harvested by filtration, lyophilised and
DNA extracted using the Qiagen Plant Mini Kit according to
the manufacturer’s instructions (Qiagen Pty Ltd, Clifton hills,
Vic., Australia). Molecular identification of E. lata and other
diatrypaceous species was achieved via amplification and
comparison of rDNA internal transcribed spacer (ITS) regions
(ITS1, 5.8S and ITS2) using the oligonucleotide primers ITS1
and ITS4 (White et al. 1990).
Each PCR reaction contained 0.1 volume of 10buffer
(containing 15 mM MgCl
2
, Qiagen), 200 mM each of dNTP,
0.15 mM of each primer, 1 unit of HotStart Taq DNA polymerase
(Qiagen), ~50 ng of DNA template, and were adjusted with sterile
nanopure water to a total volume of 50 mL. PCR reactions
were performed using an Eppendorf Master Thermocycler
(Hamburg, Germany). Amplification was achieved by an
initial step of 15 min at 95C, followed by 40 cycles of 30 s at
94C, 45 s at 55C, and 1.5 min at 72C, with a final extension of
5 min at 72C. PCR products were separated by electrophoresis
on 1% agarose containing 0.5Tris-borate-EDTA buffer, and
photographed under UV light after staining with ethidium
bromide (0.5 mg/L).
For sequencing, PCR products were purified using the
QIAquick PCR purification kit (Qiagen). ITS regions were
sequenced in both directions by the Australian Genome
Research Facility (University of Queensland, St Lucia, Qld,
Australia), and identification of diatrypaceous species
confirmed by comparison of ITS sequences of our isolates
with those available in GenBank. Individual sequences were
compiled in BioEdit sequence alignment editor (Hall 1999)
and aligned for comparison using ClustalX (Thompson et al.
1997).
Isolates of E. lata,Cryptovalsa ampelina,Eutypella and
Diatrypella used in this study are maintained on PDA agar
slopes at 4C in the collection at the National Wine and Grape
Industry Centre (Charles Sturt University, Wagga Wagga, NSW,
Australia), and representative isolates of each species were
deposited in the Australian Scientific Collections Unit (NSW
Department of Primary Industries, Orange, NSW, Australia).
DNA sequences of representative isolates used in this study
were submitted to GenBank (Table 1).
Results
A total of 73 isolates developing white, cottony mycelium
and conforming to gross morphological descriptions of
the Diatrypaceae (Glawe and Rogers 1984; Carter 1991;
Luque et al. 2006) were isolated from diseased grapevines.
E. lata was isolated from ~0.65% of grapevines surveyed
(12 isolates), with the fungus being reported for the first time
both in the Central Ranges and southern NSW regions (Fig. 1).
In each case, foliar symptoms typical of Eutypa dieback were
present including stunted shoots with shortened internodes and
cupped leaves with necrotic margins, but the teleomorph was not
observed and identification of the anamorph was accomplished
via sequencing and comparison of rDNA ITS sequences. Other
diatrypaceous species isolated from initial field collections
included 14 isolates of Eutypella (0.76% of samples, Fig. 2),
23 isolates of Diatrypella (1.25%, Fig. 3), and 21 isolates of
C. ampelina (1.14%, Fig. 4), which were tentatively identified
according to morphological descriptions (Glawe and Rogers
1984). In all cases specimens were isolated from cankers from
diseased trunks or cordons. Further isolations from fruiting bodies
98 Australasian Plant Pathology W. M. Pitt et al.
Table 1. Diatrypaceous species isolated from grapevines throughout New South Wales
Region (# vines sampled),
location
Cultivar Age Species (# isolated) Accession numbers
A
Central Ranges (246)
Orange Chardonnay 9 Eutypella sp. (1) –
Grenache 10 Diatrypella sp. (3) –
Cryptovalsa ampelina (1) –
Sauvignon blanc 14 Eutypa lata (2) EU835160, DAR79045
C. ampelina (2) –
Cabernet sauvignon 14 Diatrypella sp. (2) –
C. ampelina (1) –
Canowindra Verdello 14 Diatrypella sp. (1) –
C. ampelina (4) –
Chardonnay 13 E. lata (2) EU835166, DAR79048
EU835167, DAR79049
Cowra Chardonnay 15 C. ampelina (2) –
Mudgee Chardonnay 8 C. ampelina (1) –
Northern Slopes (300)
Bendemeer Shiraz 12 Eutypella sp. (1) –
Armidale Pinot grigio 8 Eutypella sp. (1) –
Deepwater Semillon 12 Eutypella sp. (3) –
Diatrypella sp. (2) –
Inverell Shiraz 13 Diatrypella sp. (2) –
Southern New South Wales (525)
Murrumbateman Shiraz 37 E. lata (2) EU835162, DAR79046
EU835163, DAR79047
C. ampelina (4) EU835150, DAR79050
EU835151, DAR79051
EU835152, DAR79052
Cabernet sauvignon 36 Eutypella sp. (1) –
29 E. lata (1) –
Young Chardonnay 19 C. ampelina (2) –
Diatrypella sp. (1) –
Eutypella sp. (1) –
Berridale Riesling 24 Diatrypella sp. (1) –
Tumbarumba Pinot noir 28 Diatrypella sp. (2) –
16 Diatrypella sp. (3) –
20 Diatrypella sp. (1) –
Chardonnay –Diatrypella sp. (2) –
Eutypa lata (1) EU835164, DAR79128
Sauvignon blanc –C. ampelina (2) –
South Coast (250)
––– –
Big Rivers (427)
Wagga Wagga Shiraz 11 Diatrypella sp. (1) –
Book Book Shiraz 12 C. ampelina (1) EU835154, DAR79054
Griffith Shiraz 39 E. lata (3) EU835157, DAR79040
Traminer 27 C. ampelina (1) –
Ruby Cabernet 33 E. lata (1) EU835156, DAR79043
Chardonnay 41 Eutypella sp. (6) –
Northern Rivers (62)
Port Macquarie Pinot noir 22 C. ampelina (1) –
Hunter Valley (36)
Pokolbin Semillon 22 Diatrypella sp. (2) –
C. ampelina (1) FJ800509, DAR79966
Eutypella sp. (4) FJ800513, DAR79970
FJ800514, DAR79971
FJ800515, DAR79972
FJ800520, DAR79977
A
GenBank number and DAR (Australian Scientific Collections Unit, NSW Department of Primary Industries, Orange, NSW,
Australia) herbarium number, respectively.
Diatrypaceae in New South Wales Australasian Plant Pathology 99
collected in the Hunter Valley tentatively identified an additional
four isolates of Eutypella and one isolate of C. ampelina based on
features of the stromata and teleomorph, but no foliar symptoms
of Eutypa dieback were observed, nor was E. lata isolated
from the samples collected. Molecular identification of isolates
from this latter survey subsequently confirmed the identity of
C. ampelina and revealed the presence of several different species
of Eutypella, but neither these isolates nor isolates of Diatrypella
could be identified at the species level based on available data
from GenBank. No diatrypaceous species were isolated from the
South Coast region of NSW.
Discussion
The generic concept in the Diatrypaceae is principally based
on stromatic characters, such as the degree of stromatal
development, configuration of perithecial necks and type of
host tissue in which stromata occur (Glawe and Rogers 1984).
In general, diatrypaceous asci are clavate to spindle-shaped, long
stipitate with a truncate or blunt apex, often containing
cytoplasmatic strands in the apex and frequently possess a
thicker walled region above the ascospores (Carmaran et al.
2006). Differentiation of the various genera and species within
the Diatrypaceae is difficult, as many are indistinguishable,
possessing few if any unique taxonomic features. Apart from
Cryptovalsa and Diatrypella, which can be clearly separated on
the basis of their polysporous asci, the number of ascospores
being the only aspect of the ascus regularly used for diagnostic
purposes (Carmaran et al. 2006), the other common species,
viz. Cryptosphaeria,Diatrype,Eutypa and Eutypella are
indistinguishable, and all comprise octosporous asci, with
differentiation instead relying on the use of molecular tools
(Rolshausen et al. 2004).
Several genera of the Diatrypaceae are known to occur on
grapevines throughout the world (Farr et al. 1989). Until recently
E. lata was thought to be responsible for dead and declining
Growing season temperature
<13°
°
C
13–15°
°
C
15–17°
°
C
17–19°
°
C
19–21°
°
C
21–24°
°
C
>24°
°
C
Northern Rivers
Northern Slopes
South Coast
Central Ranges
Southern NSW
Big Rivers
Hunter Valley
Sample Sites
N
0125 250 500 km
Fig. 1. Growing season mean average temperature distribution bands across New South Wales (1 October 1971–30 April 2000) and the major grape-growing
regions, including sites of isolation of Eutypa lata. Data obtained from the Australian Government Bureau of Meteorology (2005), and map produced
following protocol of Hall and Jones (2009).
100 Australasian Plant Pathology W. M. Pitt et al.
vines routinely observed in the Hunter Valley region of
NSW. However, foliar symptoms of the disease have not been
observed, nor has the fungus been isolated from material collected
during extensive surveys of the region (Castillo-Pando et al.
2001; Creaser et al. 2003; Qiu et al. 2006; Savocchia et al. 2007).
Recently, E. lata was reported in the Riverina region of NSW
for the first time (Pitt et al. 2007). Although prominent in South
Australian vineyards, Eutypa dieback had not been reported
north of Wentworth (latitude 3420S, longitude 14150E) in
NSW, which is on the Victorian border (T. Wicks, pers.
comm.), and until now, no definitive records of the fungus
exist from the wine-growing regions of south-eastern NSW.
Extensive surveys conducted throughout NSW have now
shown that E. lata is more widespread than first thought, with
new isolations from Canowindra and Orange in the Central
Ranges and Murrumbateman and Tumbarumba in southern
(a)
(b)(c)
Fig. 2. Eutypella sp. from Vitis vinifera,(a) poorly developed stromata embedded in bark with perithecial
necks erumpent in groups (inset: close-up of perithecia embedded in mixture of fungal and host tissue),
(b) squash mount of asci, and (c) close-up of clavate to spindle-shaped, long stipitate, octosporous ascus
comprising eight allantoid to moderately curved, subhyaline to subolivaceous ascospores (bar = 50 mm).
Diatrypaceae in New South Wales Australasian Plant Pathology 101
NSW (Fig. 1). While Edwards and Pascoe (2004) reported
the presence of Eutypa from grapevine samples received
from Tumbarumba during a routine diagnostic survey, cultures
appeared not to have been identified to species, nor characterised
at a molecular level. Regardless, these results suggest that
E. lata may be well suited to the cooler climate regions of
NSW where low temperatures and high rainfall favour the
growth of the fungus. Certainly, the isolation of E. lata from
diseased grapevines in Orange (latitude 33150S, longitude
14910E) represents a small but measurable expansion in the
geographic range of the pathogen and the most northerly
occurrence of E. lata reported in Australia.
While the epidemiology (Petzoldt et al. 1981; Carter 1991;
Munkvold and Marois 1995) and management (Munkvold and
(a)
(b)(c)
Fig. 3. Diatrypella sp. from Vitis vinifera,(a) well developed stromata, erumpent through bark (inset:
close-up of perithecia, surrounded by pseudoparenchymatous tissue), (b) squash mount of asci, and
(c) close-up of spindle-shaped, long stipitate, polysporous ascus comprising ~128 allantoid to moderately
curved, subhyaline to subolivaceous ascospores (bar = 50 mm).
102 Australasian Plant Pathology W. M. Pitt et al.
Marois 1993; Weber et al. 2007; Sosnowski et al. 2008) of E. lata,
has been studied extensively, little is known about the other
members of the Diatrypaceae, many of which are known to occur
not only on grapevines but on other hosts including, apples,
cherries, pears, olives and poplars (Farr et al. 1989). Several
species of Eutypella and Diatrypella, as well as C. ampelina,a
pathogen in its own right (Mostert et al. 2004; Luque et al. 2006),
were isolated from grapevines from many regions throughout
NSW. Interestingly, all three of these genera were present in the
Hunter Valley region, and clearly possess a far greater geographic
range than E. lata. In contrast, E. lata was present only in the
cooler climate regions of southern NSW, and once again the
fungus was not recovered from samples collected from the Hunter
Valley, nor was it or any other diatrypaceous species isolated from
the South Coast region of NSW. In the latter case, the reasons for
this are unknown; however, our failure to isolate E. lata from the
(a)
(b)(c)
Fig. 4. Cryptovalsa ampelina from Vitis vinifera,(a) poorly developed stromata embedded in
decorticated wood (inset: close-up of perithecia, singularly erumpent), (b) squash mount of asci and,
(c) close-up of spindle-shaped, long stipitate, mulitsporous ascus comprising ~32 allantoid, subolivaceous
ascospores (bar = 50 mm).
Diatrypaceae in New South Wales Australasian Plant Pathology 103
Hunter Valley and other regions throughout NSW in no way
suggests that these regions are free from the disease. But, it is
possible that other genera within the Diatrypaceae are
contributing to the dieback phenomenon generally attributed to
E. lata (Trouillas et al. 2001).
Why species such as C. ampelina,Eutypella and Diatrypella
are prominent in the Hunter Valley, seemingly in the absence
of E. lata remains a mystery. Perhaps the poorly developed stroma
of C. ampelina, which is embedded in the bark or decorticated
wood, as opposed to well developed and erumpent through bark
as with E. lata (Glawe and Rogers 1984), affords the fungus
some measure of protection from the elements, thereby enabling
the fungus to survive under a greater range of environmental
conditions. In all diatrypaceous genera, stromata are thought to
aid in conserving moisture, but in some genera, their function is to
aid discharge of ascospores by rupturing the host’s bark, thereby
exposing perithecial ostioles (Glawe and Rogers 1984). If
the reduced stroma composed of both fungal and host tissue
preserves less moisture or fails to expose ostioles to the
environment, effectively limiting ascospore discharge, fruiting
bodies may be better preserved, their survival enhanced,
their ability to withstand adverse conditions heightened and
geography improved as a function of age. Equally the reverse
could be true, or maybe the host ranges of C. ampelina and the
other more prominent diatrypaceous species are simply larger
than that of E. lata; as species with larger host ranges often have
greater geographic distributions (Glawe and Rogers 1984).
Other explanations for the geographic disparity among the
different members of the Diatrypaceae are speculative at best,
although clearly the environmental conditions in the northern
regions of NSW, including the Hunter Valley, are less favourable
to E. lata than to some of the other species in the Diatrypaceae,
likely due to stricter rainfall and temperature requirements of
E. lata (Sosnowski et al. 2005, 2007b). While the Mediterranean
climate common in South Australia where Eutypa dieback is
extensive contrasts well with the cooler climate regions of NSW,
the subtropical climate of the Hunter Valley region, with higher
average daily temperatures appears unfavourable for the
establishment of Eutypa dieback, despite the fact that rainfall
in the region is suitable for the disease. Notably, in a recent report
concerning the seasonal variation in Eutypa dieback symptoms,
Sosnowski et al. (2005) reported a positive relationship between
disease incidence and spring temperatures; the higher the
temperature, the lower the incidence of disease, as determined
by expression of foliage symptoms. Glawe and Rogers (1984)
reported that the teleomorph of the fungus fails to form in
regions receiving less than 330 mm of annual rainfall.
However, plants infected with E. lata have been found in
such locations, with long-distance and airborne dispersal of
ascospores known to accompany the onset of rainfall (Carter
1957; Ramos et al. 1975; Glawe and Rogers 1984). With an
average annual rainfall greater than 600 mm (Australian
Government Bureau of Meteorology 2008; data for Cessnock),
understandably, researchers continue to be baffled by the absence
of Eutypa dieback in the Hunter Valley, a disease that for all
intensive purposes should be present there. Incidentally, US
researchers have also noted large geographical differences in
the distribution of E. lata with respect to climate (Urbez-Torres
et al. 2006).
To date, the pathogenicity of E. lata (Carter 1991) and
C. ampelina (Mostert et al. 2004; Luque et al. 2006) to
grapevines has been confirmed, but investigations on the
virulence of many of the other diatrypaceous species towards
grapevines are limited (Trouillas et al. 2001). Eutypella vitis has
been reported to cause xylem necrosis and foliar symptoms
similar to those caused by E. lata (Wolf 2006), for which it
was suggested as an ulterior cause of Eutypa dieback
(Myers 2008). Furthermore, two reports from research
conducted in California show that E. leptoplaca and
Diatrypella are regularly isolated from diseased grapevines
and that inoculation of fresh pruning wounds with these
species can cause disease (Rolshausen et al. 2004; Trouillas
and Gubler 2004).
As cultures of diatrypaceous fungi are often indistinguishable
from one another, the co-occurrence of multiple diatrypaceous
fungi in diseased wood of grapevines could also lead to
misidentification of the correct agents of disease, especially
where multiple species are isolated from the same infection.
While this occurred in only two instances in the present study,
whereby C. ampelina and Diatrypella were cultured from the
same piece of wood (Central Ranges), more than half of the
C. ampelina isolates collected in the survey co-occurred with
species of the Botryosphaeriaceae, which are also well known
trunk disease pathogens of grapevines (van Niekerk et al. 2004;
Savocchia et al. 2007). The frequent occurrence of C. ampelina in
conjunction with other known grapevine pathogens was thought
by Luque et al. (2006) to represent a synergistic association by a
facultative pathogen, which he then used as an explanation for the
moderate virulence of the fungus. However, in recent years,
C. ampelina has been isolated repeatedly from grapevines both
in Australia and overseas, and under suitable conditions has been
shown to be a pathogen in its own right (Mostert et al. 2004;
Luque et al. 2006). Nevertheless, competition from other trunk
disease pathogens like the Botryosphaeriaceae may play a role in
reducing the incidence of the Diatrypaceae. While unlikely
with respect to C. ampelina, interactions of this nature may
contribute to the low incidence of E. lata throughout the state,
or alternatively to its absence in some regions. To date, no reports
of this nature have been published.
This study has shown that many of the other diatrypaceous
species are more widespread and abundant than E. lata in
NSW. While the exact species causing trunk diseases in the
vineyard may be inconsequential to many grapegrowers, due to
increasing knowledge that the recommended management
strategies pertain to many of the grapevine canker causing
fungi (van Niekerk et al. 2002; van Niekerk et al.2006),correct
diagnoses of the causal agent may be essential to predict the
severity of the disease and hence the urgency of management. In
a major study of the pathogenicity of nine Botryosphaeriaceae
species isolated from grapevines in California, Urbez-Torres and
Gubler (2009) showed that four species within this family were
equivalent to, or greater in virulence than E. lata. Other researchers
have reported considerable variation in the pathogenicity of
isolates of E. lata both to grapevines and other hosts, suggesting
the existence of two pathotypes, vastly different in virulence
(Carter et al. 1985). This variation in pathogenicity of individual
isolates of E. lata,aswellastheinfluence of grapevine cultivar
(Sosnowski et al.2007a), and the effect of climate on seasonal
104 Australasian Plant Pathology W. M. Pitt et al.
disease expression (Petzoldt et al. 1981; Sosnowski et al.2007b),
can greatlyinfluence the severityand perceived importance of such
canker diseases. Because it is impossible to discriminate wood
symptoms caused by the different trunk disease fungi, the
aggressiveness of the pathogen and its identification is of the
utmost importance for effective management.
As few of the diatrypaceous species have been studied in detail
in Australia, their incidence, distribution and pathogenicity
towards grapevines and other cultivated crops requires further
research. In the interim, vigilant monitoring, avoiding pruning
during and directly after rainfall, protection of pruning wounds,
and removal and incineration of dead infected wood from the
vineyard remain the best methods of managing Eutypa dieback
and other infections that may be caused by the Diatrypaceae.
Acknowledgements
Thiswork was supportedby the WinegrowingFutures Program,a joint initiative
of the Grape and Wine Research and Development Corporation and the
National Wine and Grape Industry Centre (NWGIC). The authors wish to
thank Mark Sosnowski and Adrian Loschiavo (South Australian Research
and Development Institute) for advice and assistance in identifying and
collecting diatrypaceous species. Many thanks also to Andrew Hall
(NWGIC) for assistance with the preparation of maps and the grapegrowers
and winemakers of NSW who provided access to their properties and
participated in this study.
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