Technical article

Global strategy for the control

of Grapevine Trunk Diseases
This project has received funding from
the European Unions Horizon 2020
research and innovation programme
under grant agreement No 652601

Grapevine trunk diseases (GTDs), caused by a complex of pathogenic fungi, have increased especially following the pro-
hibition of chemical products as sodium arsenite (for Esca) first and the Benzimidazoles Benomyl and carbendazim,
later. Their eradication is not simple: currently, no efficient treatment exists. For this reason and in order to avoid GTDs
spread, prophylactic methods should be used and implemented both in nursery and vineyard, since its planting

Symptomatology of Grapevine Trunk Diseases (GTDs)

GTDs, caused by a complex of pathogenic fungi, affect the
perennial organs of the vine, causing their death in the long
term. Two forms, a chronic and a severe called apoplectic, are
found in the vineyard. The chronic form causes vigor loss and
leads to the death of a part of vine and then to its totality. Fo-
liar symptoms are characterized by necrotic areas (staining or
tabby appearance) or stunting. The inflorescences and fruits
may dry out or take on a pumpkins and peas appearance.
GTDs cause necrosis and degradation in the wood by bloc-
king the vessels or forming cankers. This article focuses pri-
marily on the following three trunk diseases: Esca, Botryos-
phaeria dieback and Eutypa dieback.

Symptoms and causal agents of Esca

Esca is a complex disease caused by a various pathogenic
fungi such as Phaeomoniella chlamydospora and Phaeoacre-
monium aleophilum. Fomitiporia mediterranea is responsible
for amadou. This disease has an apoplectic form and a chro-
nic form. The rapid form is characterized by rapid drying of
a part or the entire vine. The chronic form is characterized
by an inter-nervous staining, yellow on white grape varieties Pictures : Characteristic symptoms of Esca : foliar symptoms on Cv Caber-
and red on black grape varieties with a yellow border. Foliar net. Sauvignon (1), Sauvignon B. (2) central necrosis (3) and symptoms on
necrosis then evolves towards a leaves drying. Two types of fruits (4) .
necrosis can be observed in the wood. A central necrosis
with a clear, tender zone in the center surrounded by a black Symptoms and causal agents of Botryosphaeria dieback
border and a sectorial necrosis with a clear and tender zone Botryosphaeria dieback has long been mistaken for Esca, as
surrounded by a hard brown/black zone are characteristic of the symptoms may be similar. This disease can be observed
Esca. Fruits may dry out resulting in crop losses. in its chronic or apoplectic form. Botryosphaeria dieback is
caused by fungi of the genus Botryosphaeria (B. obtusa, B.
parva and B. stevensii) as well as by Diplodia seriata and Neo-

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 Global strategy for the control of Grapevine Trunk
Diseases
fusicoccum parvum. Between the veins, leaves are staining in
red for red varieties and in yellow/orange for white varieties
and then become necrotic. After removing the bark, a brown
band surrounded by a yellow/orange zone or a brown/black
sectorial necrosis, which can go from the shoot to the graft
point, is visible. The severe form is characterized by rapid
desiccation of the strain and destruction of the inflorescences
and fruits.
Pictures : Characteristic symptoms of Botryopshaeria dieback : foliar symp-
toms on black (1) variety, dessication of a shoot (2) and wood necrosis
(3-4).
Symptoms and causal agent of Eutypa dieback
The fungus Eutypa lata is responsible for this disease. The
internodes of the diseased strains are shortened and the
branches are stunted. The leaves are deformed and chlorotic
and sagging or millerandage can be observed on the berries.
On wood, the symptoms are in the form of a brown and hard
sectorial necrosis.
Pictures : Characteristic symptoms of Eutypa dieback : stunting (1) and sec-
toral necrosis (2) .
Factors influencing the expression of GTDs symptoms
Biotic or abiotic factors could act in the GTDs development
in field and on symptoms expression. Climatic conditions
have an important role in the pathogen development cycles.
Dry periods, causing water stress in the plant are favorable
to pathogens development (Stamp, 2001). On the contrary,
the presence of other microorganisms could interfere with
pathogens, either limiting or even preventing their develop-
ment.
Foliar symptoms could not appear or differ from year to year
according to different factors. Vineyard age and cultivar, trai-
ning and pruning system could also influence GTDs foliar
symptoms expression (Grosman & Doublet 2012).
The prevalence of Esca/BDA is maximal between 12 and 18
years and then gradually decreases until 40 years whereas it is
25 - 40 years for Eutypiosis (Grosman & Doublet, 2012).
Preventive methods to avoid GTDs during the esta-
blishment of the vineyard
The quality of grafted plants, the cultivar and planting condi-
tions are important elements to be considered if one wants
to manage the GTDs since the vineyard implant. It has been
pointed out that during the production of plant materials, se-
veral stages may predispose young plants to the development
of GTDs parasites (Gramaje & Armengol, 2011, Lecomte et al.,
2008, Stamp 2001).In general, it is better to chose good quality
plants, with a well developed root system and with a com-
plete heal of the grafting point. Even if not compulsory, some
nurseries have started to apply physical (Hot Water Treat-
ment - HWT), chemical (fungicides) or biological methods
(Trichoderma) to prevent GTDs infections during the plant
production process. Treatments with hot water during 30 min
can limit populations of P. chlamydospora and some strains of
Botryosphaeriaceae (Larignon et al., 2009, Vigues et al., 2009,
Elena et al., 2015). Acquiring high-standart, health and cer-
tified plants is therefore important to assure a good sanitary
quality of the vineyard in the next years.
Some grapes varieties or rootstocks appear to be more sen-
sitive to GTDS than others so that choice should then be
oriented towards the most resistant. Cv. Pinot Noir, Syrah and
Cot tend to be less sensitive to Esca /BDA than cv. Sauvignon,
Ugni B, Gewurztraminer or Trousseau. Rupestris or Riparia
Gloire of Montpellier rootstocks are more resistant to Esca
whereas 101-14, 3309 and Gravesac tend to be more sensitive,
but this may differ depending on climatic conditions (Roby et
al., 2015).
Planting in a decompressed soil may be carried out between
late autumn and early summer, during the vines dormant
period. Prior to planting, treatment with Trichoderma, cypro-
dinil and fludioxonil or metiram and pyraclostrobin products
may be performed by root dipping . Attention should be paid
to the root system, leaving it undamaged and able to spread
properly. During the vineyard implant watering of the vines
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 Global strategy for the control of Grapevine Trunk
Diseases

is important to avoid water stress. It is advisable to tutor the The training system seems to play a role in the impact of GTDs,
plant in order to promote vertical growth of the trunk to be less but the information is divergent. It is preferable to opt for a
sensitive to the damage related to the weeding tools. Thereafter, less severe pruning, generating small wounds and fewer dis-
the choice of the vine formation system must be considered and turbances for the vessels development. Long pruning should
allow a physiologically rational size, facilitating sap flows. be preferred and many studies show that goblets and Guyot
systems should be preferred. In Bordeaux, a study showed that
Soil management to prevent GTDs symptoms expression was increased in Guyot (short canes).
Eutypa dieback foliar symptoms were higher in short pruning
Plant stress caused by unbalanced supply of nutrients, bad drai-
than in long one, but the mortality was lower in the long pru-
nage or soil compaction plays an important role in the develop-
ned vines. The latter, such as those in Guyot, are damaged by
ment of GTDs and their symptoms. Soil is the main source of
numerous wounds localized in the upper part of the trunk; on
inoculum and GTDs inoculum mainly spread through rain to
the contrary, short pruned vines (Cordon or goblet) had larger
pruning wounds. Direct infection into roots is possible so pro-
total area of wounds.
per soil management can be a way to prevent GTDs. Generally,
The Guyot-Poussard pruning method, recently proposed to
a good soil management, where air and water circulate easily
vine growers to limit GTDs, preserve the sap flux since pru-
and where water does not stagnate, could avoid stress in GTDs
ning wounds are located on the upper part of the cordon. The
affected vines so they dont show foliar symptoms. Unreaso-
wounds are smaller, less numerous and located on young wood
nable nitrogen fertilization should be avoided (low C/N ratio)
that would be more resistant to infections than old one. This
as this promotes excessive vigor that reduces the polyphenols
kind of training system could limit GTDs but it still needs to be
production having a role in pathogen resilience. In this direc-
scientifically evaluated.
tion, the establishment of permanent or non-permanent vege-
tation cover crop in the inter-row could balance the availability
of nutrients also improving soil structure. Deep-rooted species
such as Medicago sativa improves soil structure and air circu-
lation.

Pruning system, an important reach

Pruning wounds are the main entry point for pathogens cau-
sing GTDs. The wider and more numerous the wounds are, the
larger the area where spores of GTDS pathogens can infect is.
In addition, pruning wound may be correlated with the un-
derlying necrosis (also known as desiccation or drying cone).
These necrosis could be infected by other wood pathogens and/
or deteriorate the sap flux, increasing physiological stress and
thus the harmful effects of chronic and severe GTDs forms. To
avoid these necrosis and their effects, the residual length of the
pruned spur should be at least 1.5 times its diameter. Therefore,
the training system, the pruning time and the wound protection
are elements to be taken into account. Even if scientifically proved, the contamination trough pru-
Late pruning during the dormant season and close to budbreak ning scissors is quite irrelevant compared to those by pruning
is recommended, since wounds heal better with mild tempe- wounds. Consequently, it is not strictly necessary to disinfect
ratures. Infection of pruning wounds tend to be lower in au- these tools to limit Esca and Eutypa dieback (Larignon, 2007).
tumn than in winter however, late pruning can be made in vine
growing areas with dry spring . Wound susceptibility is mostly Because pruning wounds are the gateway to GTDs pathogens,
influenced by the relative humidity and rainfall (Luque et al., it is recommended to protect them preventively by using fungi-
2014). Precipitation and temperature have a direct effect on the cides (Phytopast-V for Eutypa dieback), bio-control products or
pathogens distribution. It is therefore recommended to prune mastics. These products have to be applied locally on the wounds
vines during dry periods since available inoculum is signifi- and as soon as possible after pruning. Wounds can be protected
cantly lower. Double pruning or mechanical pre-pruning is against Eutypa dieback by bush application or spraying with a
done to speed up final pruning and to reduce GTDs incidence solution of carbendazim and fluzilazole (Lecomte et al., 2004).
in spurpruned vineyards. Infections by D. seriata, Inocutis sp. and P. chlamydospora can
 Global strategy for the control of Grapevine Trunk
Diseases
be significantly reduced using a mixture of benomyl, pyraclos-
trobin, tebuconazole and thiophanate-methyl (Daz y La Torre,
2013). Spraying is more convenient, faster and less expensive,
but can easily be washed away by rain. Trichoderma-based pro-
ducts (Di Marco et al., 2000) or chitosan (Bertsch et al., 2013)
can also be used to protect wounds, as well as mastics that act
as physical barrier.
Destruction of inoculum sources
The GTDs inoculum is found on diseased and dead plants
(leaves, grapes, and necrotic wood and old wood). Pruning de-
bris, symptomatic and dead vines are a potential source of new
infections in the vineyard. Pruning debris, for example, can
bring viable inoculum of Botryosphaeria dieback pathogens up
to 42 months. Thus, it is necessary to try as much as possible to
get rid of all sources as quickly as possible (VITI 2/2006 reso-
lution). To destroy potential inoculum, pruning woods may be
crushed and buried in the soil, burned (if permitted by law),
crushed and compacted (40 to 50 C for 6 months) or remove
them from the parcel.
Trunk renewal to restore vines
To rejuvenate the trunk of GTDs affected vines, the trunk must
be cut above the rootstock and below necrosis. The vine will
then produce sucker that will have to be tutored in order to
obtain a new trunk. This simple technique has good results
with vigorous varieties. This technique has been shown to be
effective in controlling Eutypa dieback (BNIC, Chamber of
Agriculture, 1989, BNIC, INRA Bordeaux, 1989, Mette et al.,
2004, Sosnowski et al., 2011). It seems that it is also could be
also useful for Esca and BDA, but depending on the sanitary
status of the remaining part ( if the trunk below the cut has
GTDs infections, vines could express GTDs symptoms again in
the following years (Calzarano et al., 2004), (Larignon & Yobre-
gat, 2016.) It is possible to anticipate the trunk renewal by kee-
ping a sucker several years in advance. Therefore, when cutting
the vine, the new one is alrea-
dy formed. If the renewal is
done during the winter, it
is preferable to protect the
wound in order to limit infec-
tions by GTDS. Smart (2015)
established a guideline (The
Timely Trunk Renewal pro-
tocol) to do the renewal. As
part of the Winetwork pro-
ject, a survey carried out in
southwestern France estima-
ted the cost of this operation
between 225 and 275 / ha for
a plot planted at 4500 vines/ha with a proportion of 250 vines.
Practices to restaure diseased vines
Re-grafting to limit repercussions of GTDs on vines
An alternative method to manage GTDs diseased vines is the re-
grafting. The operation consists in grafting again diseased vines
starting from its healthy rootstock. It seems that slotted grafting
is the most appropriate method for regenerating diseased vines
(SICAVAC). This grafting method can be carried out in spring
or autumn, when the sap is in circulation but not very active,
and during dry periods. It is necessary to cut the vine and to
verify that at the cuts location it is healthy (free from decay or
the graft will also be diseased). Sur-grafted vines produce a half-
harvest the year after the graft and harvest return to normal two
years later. The advantage is that the root system and therefore
the age of the original vine is preserved, giving a production
with the same quality of the healty ones. This technique is quite
complex but the recovery rate could reach up to 80 - 90%.
Re-grafting can be carried out in service provision or by the
winegrower himself. The cost of the service can range from 1.35
to 2.05 HT per vine, depending on the surface. In other words,
the cost for the purchase of supplies, for the use of equipment,
workforce and fixed charges for the first two years is estimated
at 12 000/ha (Source Chambre dAgriculture Loir-et-Cher). It
is necessary to contract an employee for 3 months full-time to
sustain grafted-vines for 4500 vines (Source IFV).
Trunk cleaning to save vines
Trunk cleaning is a kind of clea-
ning surgery, which could be
effective to limit Esca and BDA.
It consists in removing the rot-
ten white wood from the trunk,
as a dentist could do for caries.
Trunk cleaning can be perfor-
med as soon as the first symp-
toms of GTDs occur and when
the presence of rotten wood is
observed. If done early in the
season, the harvest of the year
can be preserved. To clean, it is
necessary to locate the pieces of
rotten and spongy wood that are affected. Healthy wood and
sap flux should be preserved.
In Sancerre, a winegrower observes that 99% of the cured di-
seased vines are now free from Esca (Source SICAVAC). Simi-
larly, a trial carried out in Alsace shows the non-cured vines,
compared to cured, express more and fatally symptoms of
GTDs. The same test indicates that it takes an average of 5 mi-
nutes to cure diseased vines, but this depends on the reached
level of vines and winegrowers experience (Source IFV). These
studies need to be extended to determine how long the vines are
free from GTDs.
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 Global strategy for the control of Grapevine Trunk
Diseases

Use of phytosanitary for limiting GTDs symptoms Copper nails: This technique consists in planting a copper nail
in a diseased trunk. Copper is supposed to diffuse from the nail
Since the prohibition of sodium arsenite and Escudo, no fun-
into the trunk through the sap and exerts a fungicidal effect on
gicide is allowed against GTDs.
GTDs fungi. This practice has been tested by a winegrower for
Systemic fungicides such as N-carboxymethyl-3-cyano-4-
3 years in Germany but for the moment no scientific validation
(2,3-dichlorophenyl) pyrrole can circulate in the phloem (Chol-
is available.
let et al., 2004, Jousse, 2004) and have an effect on Eutypa lata.
Other studies have been conducted with Fenpiclonil (Jousse,
2004) to control Esca. GTDs could be controlled by preventive H2O2: Injecting H2O2 using a syringe is an innovative tech-
products like tebuconazol + synthetic resins or Esquive WP (T. nique applied in the Douro vineyards in Portugal, Galicia in
atroviride I-1237), Folicur (tebuconazole), Shirlan (fluazinam) Spain and Southern France in vines with symptoms of Esca or
or Cabrio (pyraclostrobin). An application of Bion (acibenzo- BDA. It would appear that the results are positive for GTDs, but
lar-S-methyl) + Cuprocol (Cu oxiclorure) then of Bion + Score have not been scientifically evaluated. However, H2O2is not
(difeconazole) seems to reduce Botryosphaeria dieback inci- always accepted by law in some countries. Plants, when sub-
dence and severity. Boron is able to reduce infection by E. lata jected to biotic attack, produce H2O2. Reactive oxygen species
(Sosnowski et al., 2008; Rolshausen et al., 2010). Several trials (ROS) exert a direct toxic activity on microorganisms and often
have also been conducted with natural defectives, Trichoderma inhibit its development. It is recognized that ROS have a role
or fertilizers, as preventive or curative but no results are signifi- in the gene expression of the hypersensitive response (Lamb &
cant. Foliar application of a mixture of calcium chloride, magne- Dixon, 1997), in parietal reinforcements and in plant resistance,
sium nitrate and fucal algae extract for several years resulted in allowing the expression of defense genes and accumulation of
a significant reduction in symptoms in the treated vine. Pathogenesis-Related proteins (Van Breusegem & Dat, 2006).
Injection of H2O2 using
a syringe is an innovative
New methods that could avoid harmful effects of GTDs
technique applied in the
Douro vineyards in Por-
Layering: this can be done to renew the missing vine trunk
tugal, Galicia in Spain and
and maintain the production level of a plot. This consists in
Southern France in vines
burying a branch from the mother plant and bringing it out of
with symptoms of Esca
the ground at the end. In this way, the buried part takes root
or BDA. It would appear
and creates a plant with the same characteristics as the mother
that the results are posi-
plant. The mother plant must be healthy and the marcot must
tive for GTDs, but have not
remain linked to the mother plant due to the risk of Phylloxera.
been scientifically evalua-
ted. However, H2O2is not
Copper nanoparticles
always accepted by law in
The injection of NP of Cu using a syringe is an innovative prac-
some countries.
tice carried out in the vineyard of Galicia in Spain to fight Esca.
Two injection and 4 to 5 foliar treatments of a mixture of Cu NP
Soaking in fungicides: Before planting, the grafted-welded
and a porcine blood amino acid elicitor are carried out. For the
plants are soaked in a fungicidal solution for 50 minutes. This
moment, no results can be pres-
technique is not scientifically evaluated. This innovative tech-
ented as to the effect of this tech-
nique has been carried out in Portugal.
nique on the Esca. Copper has a
broad spectrum of action against
Conlusion
bacteria and fungi and has been
Overcome wood diseases is not an easy task as symptoms are
used longtime in agriculture as a
influenced by plenty of factors, such as climate effect on the
pesticide. Recently, copper nano-
fungal development in vineyards and its expression of symp-
particles (particles between 10-7
toms. Indigenous microflora can interact with GTDs fungi by
and 10-9 meters) have become
limiting or preventing their development. The growing condi-
very popular due to their physi-
tions and the pruning system and can be relevant factors to
cal, chemical and antimicrobial
limit GTDs. A better knowledge of GTDs would help to better
properties, as well as their abun-
understand the action mode of these diseases. Many methods,
dance (Betancourt et al., 2013).
already used in Europe vineyards, seem to be promising but still
The Cu NPs are effective in vitro
need to be validating scientifically. The application of a single
on pathogenic fungi and yeasts (Ren et al., 2009, Rupareli et al.,
control method to manage GTDs is only partially effective. It
2009, Ramyadevi et al., 2012).
seems important to try to apply different preventive techniques
to effectively manage GTDs.
Page 5
 Global strategy for the control of Grapevine Trunk
Diseases
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