SITY OF PANNONIA

GEORGIKON FACULTY, KESZTHELY

DOCTORAL SCHOOL OF CROP PRODUCTION AND

HORTICULTURAL SCIENCES

THESIS OF DOCTORAL (PhD)DISSERTATION

Leading professor: Dr. habil. László Kocsis

University professor

Associate professor: Dr. habil Géza Fischl

University professor

THE COMPARATIVE EXAMINATION OF VITIS SPECIES AND

VARIETIES AGAINST VINE DECLINE PATHOGENS, AND THE

POSSIBILITIES OF CONTROL

WRITTEN BY:

Zoltán Varga

Keszthely

2009

UNIVER

2

Introduction The economical and biological aging of a vineyard is not in a

same time. Economic aging is, when an area is not

economically worthwhile to maintain to continue to work on

the plantation, as the expenses exceed the revenues. A number

of factors can cause grapevine decline. In addition to abiotic

stress, biotic factors may play an important role. Grapevine

dieback caused by fungal pathogens is an extremely complex

problem.

The research of the problem is established for more than a 130

years, and there is still no effective or permanent solution for

the protection against these disease causing pathogens.

The research at University of Pannonia, Georgikon faculty

were primarily focused on the comparison of the susceptibility

of varieties.During the pathogenicity experiments the Pinot

variety group were examined against two pathogens, and the

susceptibility of common cultivated rootstocks were compared

to the experimental varieties grown in Cserszegtomaj. In the

United states were The testing of Vitis species. The

susceptibility of Vitis species were examined against fungal

pathogens causing vine-decline.

3

The testing of potential control methods of these pathogens

were the determined as the next aim of our study. The

definition of the time of the control, and the research of

effective fungicides were the primer aims.

The aims of our experiments 1. The testing of the susceptibility of Vitis species and

varieties, of fungal pathogens causing dieback.

2. To observe the sporulation of vine decline pathogens to

asses a possible date of effective defense.

3. Laboratory and field testing of plant protection

products.

4

Materials and methods

The experiments were took place in the University of

Pannonia, Faculty of Georgikon Department of Horticulture

and the University of California Davis. The determination of

the species was in the Plant pathology Department at

University of California Davis. The checking of the species

was according to the sequenal database of NCBI

(http://www.ncbi.nlm.nih.gov).

The monitoring of vine decline pathogens in the plantation 20 spore traps were put out in a plot of Vitis vinifera cv.

’Grüner Veltelini’. The spore trapping studies were conducted

from 8.th November 2007 to 15.th of April 2008. The traps were

transported to the laboratory every 14 days.

The testing of fungicides in laboratory and in field trials Two pathogens were tested during the experiment - Diplodia

seriata (Schwein.), Phomopsis viticola (Sacc.). The tests were

5

carried out in three different methods. Four replicates were

used in all occasions.

Two different dose of fungicides were granted on the samples,

or the medium. The dose in each case was determined

according top plant protection materials database (Szabadi,

publ. 2008) (hereinafter referred as the approved dose). The

treatments were applied with the approved dose and twice as

much.

1. method: The direct treatment of the mycelia

The fact of the method was that the mycelia sample from the

fungus is in direct contact with the fungicide. The diameter of

the cultures were measured from the second day of the

treatment daily.

2. method: The method of fungicide treated media

In the liquid medium 10 ml of fungicide was injected. After the

medias stupefaction, the sample from the fungus was put on the

surface. The diameter of the cultures were measured from the

second day of the treatment daily.

3. method: The surface of the media treated with the product

In the third method the product was spread on the surface of

the media, and the drop conidia suspension (30000 conidia/ml)

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of the pathogen was dropped on the surface. The diameter of

the cultures were measured from the second day of the

treatment daily.

The description of the field trials

The investigation happened in a plot of Vitis vinifera cv.

’Nektár’ planted in 1996. Two pathogens were used for the

experiments Diplodia seriata (Schwein.) and Phomopsis

viticola (Sacc.). The treatments were with 3 products in 4

replicas (Bordóilé®, Dithane DG Neotech®, Equation

contact®).

The treatments were done with two different methods:

1., First the conidia suspension was dropped on the pruning

wound than the fungicide product.

2., In the second case the treatments were switched.

Pathogenicity tests

The susceptibility species of the species of the Vitis genus

(Vitis berlandieri, Vitis champinii, Vitis rupestris, Vitis riparia)

was tested with vine decline pathogens (Phaeomoniella

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chlamydospora, Phaeoacremonium aelophilum, Verticillium

dahlie, Cylindrocarpon destructans).

During the other experiment the susceptibility of rootstock

varieties were inoculated with the fungus Phomopsis viticola

(Sacc.).

From the species Vitis vinifera the variety group of ’Pinot’ was

tested with two pathogens Diplodia seriata and Phomopsis

viticola

Results

The results of the monitoring of the pathogens During the survey the amount of Botrytis cinerea (Pers.),

Diplodia seriata (Schwein.) és a Phomopsis viticola(Sacc.)

colonies were counted.

On Figure 1. the average of the colonies, the tempereature and

precipitation data is shown.

0

5

10

15

20

25

2007

.11.08

.

2007

.11.22

.

2007

.12

Botrytis cinerea

Figure 1: Th

The resultsThe summary

Table 3.: The

colonies from

A kórokozók telepeinek megjelenése

8

.06.

2007

.12.20

.

2008

.01.17

.

2008

.01.31

.

2008

.02.14

.

2008

.02.28

.

2008

.03.14

.

2008

.03.28

.

2008

.04.10

.

2008

.04.24

.

-10

0

10

20

30

40

50

60

Temperatur and preceptitatio

n

Botryosphaeria obtusa Phomopsis viticola Átlag hőmérséklet (°C) Csapadék (mm)

e appearence of the colonies during the period of

the experiment

of the fungicide trials of the results is in Table 3-4

results of the diameter of the Diplodia seriata

the 2nd day.

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The average diameter of the colonies of Diplodia seriata (mm)

The name of the product (Dose) 2nd day 4th day 7th day 10th day 11th day Talendo® (0,25 l/ha) 16 48 62 81 90 Curzate R® (3kg/ha) 22 56 90 90 90 Equation contact ® (0,8 kg/ha) 0 0 0 0 0 Discus DF® (0,2 kg/ha) 20 33 75 85 90 Tiosol® (8-10%) 19 47 84 90 90 Thiovit jet® (3kg/ha) 30 52 79 90 90 Tanos 50DF® (0,4kg/ha) 20 75 90 90 90 Eclair 49WG® (0,5kg/ha) 11 36 73 82 90 Dithane DG Neotech® (1,2kg/ha) 0 10 24 68 90 Bordóilé FW® (6 l/ha) 22 62 90 90 90

Table 4.: The results of the diameter of the Phomopsis viticola

colonies from the 2nd day.

The average diemater of the colonies of Phomopsis viticola (mm)

The name of the product (Dose) 2nd day 4th day 7th day 10th day 11th day Talendo® (0,5 l/ha) 14 37 60 85 90 Curzate R® (6kg/ha) 23 64 90 90 90 Equation contact ® (1,6 kg/ha) 0 0 0 0 0 Discus DF® (0,4 kg/ha) 19 31 62 80 90 Tiosol® (16%) 19 46 54 79 90 Thiovit jet® (6kg/ha) 18 56 81 90 90 Tanos 50DF® (0,8kg/ha) 24 63 90 90 90 Eclair 49WG® (1kg/ha) 19 56 90 90 90 Dithane DG Neotech® (2,4 kg/ha) 0 0 0 0 0 Bordóilé FW® 12 l/ha) 24 77 90 90 90

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The results showed similarity in the three methods. In case of

four products the diameter of the treated colonies only reached

90 mm on the 11.th day. In case of the product called Dithane

DG Neotech the development of the colonies were the slowest.

In the occasion when double dose was used it destroyed both

fungus colonies. In case of Equation contact none of the

colonies development were observed already with the

recommended dose.

During the field trials the effect of Dithane DG Neotech® and

Equation contact® were tested.

The product Dithane DG Neotech® has an effect on the

inoculated fungi the distance of the tissue discoloration was

less than in the case of Bordóilé®. Both Phomopsis viticola

(Figure 2.), and Diplodia seriata (Figure 3.) were isolated from

the inoculated vines.

11

158

63

0

177 177 177

0

20

40

60

80

100

120

140

160

180

200

Phomopsis-Copper sulfate Phomopsis-Mankoceb Phomopsis-Famoxadon+Mankoceb

(mm)

Figure 2. The amount of tissue discoloration caused by

Phomopsis viticola(Control marked with green)

169

69

0

198 198 198

0

50

100

150

200

250

Diplodia-Copper sulfate Diplodia-Mankoceb Diplodia-Famoxadon+Mankoceb

(mm)

Figure 3.: The amount of tissue discoloration in case of

Diplodia seriata (Control marked with green)

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Pathogenicity tests

The testing of the susceptibility of Vitis species The pathogenicity tests were conducted in California and the

data were sent to by the researchers of the University of

California. The results of the tested species are summoned on

Figure 4-5. The figures are showing the average results both of

the tissue discoloration and the shoot growth.

0

5

10

15

20

25

30

35

Vitis riparia Vitis rupestris Vitis berlandieri Vitis champinii Vitis vinifera cv.Chardonnay

The length of the tissue discoloration

(mm)

Phaeomoniella chlamydospora Phaeoacremonium aelophilum Verticillium dahlie

Cylindrocarpon destructans Kontrol

Figure 4.: the length of the tissue discoloration On Figure 4. it is revealed that the longest tissue discoloration

was caused by Phaeoacremonium aelophilum in every

occasion. It can be seen as well that the length of the tissue

discoloration were the biggest int he tissues of Vitis vinifera.

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The susceptibility of Vitis champinii was the lowest in case of

Verticilliun dahlie. In case of the Control plants no tissue

discoloration was observed and no fungus could be isolated

from the wood.

0

5

10

15

20

25

30

Vitis riparia Vitis rupestris Vitis berlandieri Vitis champinii Vitis vinifera cv.Chardonnay

hajtáhosszúság (cm

)

Phaeomoniella chlamydospora Phaeoacremonium aelophilum Verticillium dahlie

Cylindrocarpon destructans Kontrol

Figure 5.: The length of the shoots at the end of the experiments

On Figure 5. it can be seen that the average length of the shoots

were longer in the control vines (two exception Vitis rupestris,

Vitis champinii). The growth of Vitis berlandieri was shorter,

compared to the other species, and in case of Verticillium

dahlie inoculation this data was even less. The squarest shoot

growing was observed in case of the Vitis vinifera.

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The susceptibility of rootstock varieties A certain degree of necrosis was examined in all of the

varieties, hybrids. However a considerable difference was

revealed between them. The less tissue discoloration was

measured by the widespread variety ’Fercal’. The experimental

varieties the G121 and the G212 showed less average

discoloration in the tissues. On Figure 6. the differences

between each varieties are summoned. At the end of the

experiments the average shoot length of the inoculated vines

were measured as well, and they were compared to the control

vines (Figure 7.). On the figure it can be seen that there were

no significant differences between the inoculated and control

vines.

Figure 6.: The amount of tissue discoloration in each variety

15

0

50

100

150

200

250

Teleki

5C

Teleki

5BB

Georg

ikon 2

8

G 121

GK 40

Ferca

l

Börne

r

Rugge

ri 140

G204

G212

G222

G225

G228

G235

G236

G239

G243

G246

G248

G251

(mm) Inoculated

Control

Figure 7. The average length of the shoots in case of the

inoculated and control vines

The results of the ’Pinot’ variety group testing The lengths of the tissue discoloration were longer in case of

the inoculation with Diplodia seriata than with Phomopsis

viticola. Except one occasion the ’Pinot gris’ variety. Out of

the four experimented varieties the ’Sauvignon blanc’ had the

highest data of Tissue discoloration in both of the fungi. The

variety ’Pinot blanc’ had the smallest amount of tissue

discoloration. The inoculated fungi were reisolated from the

vines. In case of the control a small tissue discoloration was

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measured and it was because of the method not because of a

fungus

Figure 8.: The length of tissue discoloration int he varieties

The biggest difference between the inoculated and control

vines in case of the shoot length were observed by the varieties

’Pinot noir’ and ’Sauvignon blanc’(Figure 9.). These

differences in case of ’Pinot blanc’ and ’Pinot gris’ were lower.

Figure 9.: The average length of the shoots by each variety

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Summary

The testing of fungicide products are started in laboratory

circumstances and ended on the field. According to our results,

the product containing famoxadone+mancozeb was the most

effective on the tested pathogens.

During our experiments there were no Vitis species from the

tested ones which had resistance against the tested pathogens,

but in case of susceptibility significant differences were found.

In case of ’Pinot’ variety group it was observed that these

differences can be seen between varieties as well. The variety

called ’Pinot blanc’ was the least susceptible to the tested

pathogens.

During the grafting process the susceptibility of the rootstocks

have a great importance. In case of sensitive varieties the

possibility of an early infection is always bigger. Our hope is

that our results can be useful for further experiments.

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New scientific results

1. During our experiments significant differences of

susceptibility were found in the varieties of Vitis vinifera. The

testing of the variety group ’Pinot’ significant differences were

observed between the varieties. The variety ’Pinot blanc’ was

the least susceptible on the tested pathogens (Diplodia seriata,

Phomopsis viticola).

2. In case the testing of the Vitis genus no resistant genotypes

were found so they are not good for resistance source.

3. The rootstock varieties widely used were susceptible to the

tested pathogens. There were differences between the varieties

but no resistance was found.

4. The highest amount of fungal spores causing vine decline is

presented int he air of a plantation during the pruning season,

so the time of the fungicide control should be in this period.

5. With the combination of famoxadon and mancozeb the

control of Diplodia seriata and Phomopsis viticola was

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succesful. The using of both agents protected the vines from

the infection of the pathogens. So the treatment with this

product during th time of the critical period could be a succeful

preventive method against these fungi.

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The summary of publications

Publications in hungarian connected to the

dissertation

Varga, Z., Kocsis, L. (2005): A szőlőszaporítóanyag-előállítás és a tőkeelhalást okozó kórokozók terjedésének összefüggései. LOV tudományos ülésszak, Budapest 2005. 334-335. Varga Z. (2006): A tőkeelhalás felmérése a cserszegtomaji kísérleti ültetvényben XVI. Növényvédelmi Fórum 2007 2006. január 26-27. Varga, Z., Tarczal, E., PodmanickyP., (2007): A korai tőkeelhalás felmérése a cserszegtomaji kísérleti ültetvényben XVII. Növényvédelmi fórum 2007 Keszthely Varga, Z., Kocsis, L., (2007): Érdemes megfontolni. Kertészet és szőlészet 56. 51-52, 26-27. Varga, Z., Tarczal, E., Kocsis, L. (2008): Szőlőtőke elhalás felmérése a Dunántúl egyes borvidékein 2007. évben. XVIII. Keszthelyi Növényvédelmi Fórum 2008. Keszthely január 30.-február 1. Varga, Z., Kocsis, L., Fischl G. (2009):A Szőlő korai tőkepusztulását okozó Phomopsis viticola (Sacc.) elleni növényvédőszeres védekezés lehetőségei (megjelenés alatt)

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Publications in foreign languages connected to the

dissertation

Varga Z., Fischl G., Tátrai B., Kocsis L. (2007): The evaluation of grapevine decline pathogens int he experimental field of the Georgikon Faculty of Agriculture in Cserszegtomaj. International journal of Horticulture Science 2007, 13, 19-22. Kocsis L., Fischl G., Varga Z., Kocsisné M. G., Tátrai B., (2006): Vine decline in grapevine rootstock plantation. 6th International cool climate symposium for viticulture and oenology New Zeland. 175 Varga Z., Kocsis L. (2008): The monitoring of vine decline pathogens ont he experimental field of Cserszegtomaj. 50th Georgikon Scientific Conference Keszthely 2008. ISBN 978-963-9639-32-4 Varga Z., Tarczal E. (2008): The effect of Phomopsis viticola (Sacc.) on the callusing formation during the grafting process 50th Georgikon Scientific Conference Keszthely 2008 ISBN 978-963-9639-32-4 Varga Z., Kocsis L., Fischl G., Tarczal E. (2009): Evaluation of the susceptibility of rootstock varieties against Phomopsis viticola (Sacc.). 60th Annual ASEV meeting technial abstracts, Napa CA 2009. 117.

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Publication in foreign languages

Kocsis L., Varga Z., Podmanicky P. (2006): Lime tolerance of grapevine rootstocks. 6th International cool climate symposium for viticulture and oenology New Zeland 2006. 174 Kocsis L., Varga Z., Pernesz Gy., (2006): Introduction of lime and drought tolerant rootstock variety. 9th International Conference on Grape Genetics and Breeding. Udine 2006 July Janousek C., Lorber J., Varga Z., Herche R., Wunderlich L. (2007): Fungicide control of scab in Red Delicious apple in the Sierra foothills, California, 2007 Plant Diesease Management Report RN: PF051 Podmanicky P., Kocsis L., Varga Z., Tarczal E., (2007): Effect of double Maturation Raisonée Technology and Cluster Thnning on Quality of harvest and wine, Am. J. Enol. Vitic., Sep 2007; 58: 410-419 Podmanicky P., Kocsis L., Varga Z., Tarczal E. (2007): The effect of canopy and crop management on the quality of grapejuice and wine. XXX. th OIV World congress Budapest 2007. Kocsis L., Varga Z., Podmanicky P., Varga Z., Győrffyné Janke G., Barát S., Császár A., Májer J. (2008): Developing of integrated viticulture int he upper part of the hill Somló. VII. Congrés International des terroirs viticoles 2008.

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Tarczal E., Varga Z., Kocsis L. (2008): Grapevine Rootstock Use: Physiological and Cultivation Technology. Hungarian Agricultural Research, 2008, 2-3. 31-36. Kocsis, L., Varga, Z., Pernesz, G. 2009. Introduction of a lime and drought tolerant rootstock variety. Proceedings of the IX. th International Conference on Grape Genetics and Breeding. Acta Horticulturae 827: 465-469. Tarczal E., Kállay M., Varga Z., Taksonyi P., Kocsis L. (2009): Rootstock-scion interactions and Correlations between Vine Vigor and Grape Quality, 60th Annual ASEV meeting technial abstracts, Napa CA 2009. 116.