UNIVERSITY OF PANNONIA GEORGIKON FACULTY, KESZTHELY DOCTORAL SCHOOL OF CROP PRODUCTION AND HORTICULTURAL SCIENCES

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1 UNIVERSITY OF PANNONIA GEORGIKON FACULTY, KESZTHELY DOCTORAL SCHOOL OF CROP PRODUCTION AND HORTICULTURAL SCIENCES Head of School: Dr. habil. Kocsis László, DSc University Professor Supervisors: Dr. habil. Kocsis László, DSc University Professor and Dr. Füzi István, PhD The Resistance of Erysiphe necator against QoI fungicides in Vineyards Doctor of Philosophy (PhD) Thesis Written by: Taksonyi Péter Keszthely

2 Table of Content 1. Introduction and Antecedents Objectives Material and Methods Comparative Research of QoI Active Ingredients and Fungicides Applicable Against Erysiphe necator (Shwein) in the Vineyard An in vitro Study of QoI Fungicides Used in Open Field Experiments Place of experiment The Research Material Method of Analysis An in vitro Analysis of QoI Resistance in Powdery Mildew Populations Collected in Different Vineyards in Hungary Place of Experiment The Research Material Method of Analysis Results and Inferences The Comparative Open Field Analysis of QoI Active Ingredients and Fungicides Used Against Erysiphe necator (Shwein) The Molecular Genetic Background of QoI Resistance in Powdery Mildew New Scientific Results Bibliography List of Publications

3 1. Introduction and Antecedents The powdery mildew, caused by Erysiphe necator (Schwein.) Burr, is one of the best known and the most frequent diseases in the vineyards of the world. In case of its appearance, photosynthesis decreases which has a direct effect on the amount and the quality of the crop. Since its first appearance in Europe in 1845, viticulturists have attempted to repel the disease in order to save the crop. First, based on their observation, they used natural materials for protection. Their first one was sulphur which was used already in 1802 against fungicides, however, not in vineyard. Nowadays, sulphur still has an important role in protection. As the disease was spreading, the number of possible chemicals which can be used for protection was also rose. Although, fungicides were already used 200 years ago, the question of resistance became significantly important only when systemic products were appeared. In crop production and horticulture, the use of contact fungicides did not provide longterm protection against pathogens because the weather conditions influenced their duration of operation. There was a need for chemicals which eliminate such problems to support the process of growing. Systemic fungicides provided solution for the above mentioned problems. After the use of systemic fungicides, soon appeared the population of the pathogens which were resistant to the applied active ingredients because of several specific modes of action of the systemic fungicides. At the beginning, this resulted in the reduction of efficiency and later, it resulted in the lack of efficiency. The main reason for the unsuccessful and ineffective protection was hidden in the one-sided use of active ingredients. Fungicide resistance is a phenomenon which endangers successful plant protection. Resistant population of wide ranges of different pathogenic fungi develops against systemic fungicides with specific mode of action as a result of their regular use for a shorter or longer period. Fungicide resistance is hardly mentioned by any literature in the 1960s. Most of the new systemic fungicides were appeared at that time. Since the 1970s, the resistance has been regular and nowadays, it has become a real danger in crop protection. As a result of the regulation of the European Union, the number of active ingredients has decreased from a hundreds to a few. The aim of this regulation was to eliminate those active ingredients from the production which were extremely harmful to the human body. 3

4 Such steps led companies to develop chemical compounds according to ones that can be found in nature. Thus, the group of QoI fungicides was created. QoI fungicides inhibit mitochondrial respiration. That is why they are called as QoI which is the abbreviation for Quinone outside Inhibitors. The Qoi fungicides are single site action fungicides and by this reason, there is a high chance for the development of resistance. The resistance is the result of natural mutation. The increasing ratio of the resistant mutants shows the decreasing efficiency of the fungicides. During our investigation, we attempted to apply in wide range all the fungicides used in grape growing and the recommended practices of the manufacturers. 4

5 2. Objectives The aim of the research, which was started at Georgikon Faculty of University of Pannonia, was to survey the effectiveness of QoI fungicides against powdery mildew and to work out rational use of these fungicides and fit them into a plant protection technology. We proposed to study the development of the resistance, the consequences of the one-sided use of chemicals and to identify QoI resistance at the genetic level. Our scientific objectives were the following: -researching of the effectiveness of QoI fungicides against powdery mildew in a vineyard where the population of Erysiphe necator is sensitive to the active ingredients, -following the development of the resistance of Erysiphe necator (Shwein) against QoI fungicides, -proofing for the resistance of E. necator against QoI fungicides at the genetic level, -surveying the spread of the QoI fungicide resistant powdery mildew fungus population in Hungary -working effective technological advices out for more effective protection against powdery mildew 5

6 3. Material and Methods 3.1. Comparative Research of QoI Active Ingredients and Fungicides Applicable Against Erysiphe necator (Shwein) in the Vineyard A vineyard of University of Pannonia Georgikon Public Benefit Organisation was designated for research. The vineyard is located in the Dunántúl Mountains, in its Bakony Region, on the south-west border of Keszthely Plateau in the Keszthely Mountains north-west of Cserszegtomaj. For the exact localization, the GPS coordinates are the following: 1. experiment (Olasz rizling GK 1): , ; 2. experiment (Merlot CI 181): , The area slopes towards north-east, every second row is covered by grass and the bottoms of the rows were treated with herbicide. The first chart represents the position of the two blocks of vine. The first block Olasz rizling GK 1 lies at right angle to the prevailing north wind while the second block Merlot CI 181 lies parallel to the prevailing north wind. Picture 1. The location of the examined area and the blocks at Cserszegtomaj. During the experiments, we made four experimental groups in each block. In the first group (see Table 1), we have only used QoI fungicide active ingredients. In the second group (see Table 1), we have used those groups of QoI fungicides which are used with other groups of active ingredients in practice and which are effective against Plasmopara viticola. The third group (see Table 2) is similar to the second one but in this case, contact active ingredients against Erysiphe necator have been added to the second group. In the fourth group 6

7 (see Table 3), we have sprayed according to the proposed protection programs of BASF, Bayer Cropscience and Syngenta, manufacturers of plant protection companies, from to. During the spraying, we have used the chemicals of the above mentioned companies applying their recommended orders and doses. We have divided some parcels as control groups in the examined area. The control groups and the parcels were on the same place in each year. During spraying, we used SOLO 450 motorized backpack sprayer with the capacity of 10 litres. We applied the adequate doses of active ingredients and chemicals per hectare in where we used mass concentration to express the amount of doses. During the application of the manufacturer s technology, we measured the amount of the spray per hectare. In 2009, we used those active ingredients which we had already used earlier but we decreased the territory of the experimental field in order to reduce the economic losses. In 2009, we doubled the dose of chemicals per hectare and mass concentration of solution of the active ingredients in the program carried out according to the recommendation of producing companies. The reason why we doubled the amount of the doses was to further prove the presence of the resistance/tolerance and to isolate it exactly in open field conditions. In, we applied the similar amount of active ingredients and chemicals like we had done it in 2009 and we also applied the same mass concentration. The formula of mass concentration is the following: 7

8 Table 1 The group of QoI active ingredients applied alone (I.) and the group of QoI active ingredients combined with active ingredients against downy mildew (II.) from to. Number of protection Group Treatment Year of experiment ρ B (g/l) Active ingredient I. QoI active ingredients 0,8 a 2009 Piraklostrobin 1,6 1,2 b 2009 Azoxistrobin 2,4 0,3 c 2009 Trifloxistrobin 0,6 II. a b c d QoI active ingredients with complementary active ingredients against P. viticola Piraklostrobin + metiram Azoxistrobin + folpet Trifloxistrobin + cimoxanil Cimoxanil + famoxadon ,5 1 0,4 0,8 Stock / plot 33/7 31/7 32/7 n=96+ 28/7 30/7 23/7 29/6 n=110+

9 Table 2 The combination of QoI active ingredients with active ingredients effective against downy mildew and powdery mildew (III.) from to. Group Treatment III. a b c Year of experiment Number of protection ρ B (g/l) Hatóanyag QoI active ingredients with complementary active ingredients against P. viticola E. necator (2) + 4 (Piraklostrobin + metiram) + sulphur (4) +8 (Trifloxistrobin + cimoxanil) + sulphur (Azoxistrobin + folpet) + penkonazol (0,5) + 4 (1) +8 (2) + 3 (4) +6 Stock / Plot 29/7 35/7 33/7 n=97+ 9

10 Table 3 The complex protection programmes of the manufacturers (IV.) from to. Number of protections Group IV. Treatment Year of experiment Dose (ha) Active ingredients Complex technology from the manufacturer /10 l /5l Stock / plot Kumulus S 5 kg/ha + Delan 700 WG 0,5 kg/ha Vivando 0,25 + Acrobat MZ 2,0 kg/ha Cabrio Top 2,0 kg/ha + Flamenco 0,5 Vivando 0,25 + Forum R 3,0 kg/ha Cabrio Top 2,0 kg/ha + Kumulus S 5,0 kg/ha Vivando 0,25 + Forum RézoxikloridWP 3,0 kg/ha Kumulus S 5 kg/ha Rézoxiklorid WP 3,0 kg/ha Mythos 30 SC 2,5 + Kumulus S 5 kg/ha Rézoxiklorid WP 3,0 kg/ha a (BASF) b (BAYER) c (SYNGENTA) 2009 Kumulus S 5 kg/ha + Delan 700 WG 0,5 kg/ha Kumulus S 5 kg/ha + Delan 700 WG 0,5 kg/ha Kén 5kg/ha 2009 Kén 5kg/ha Kén 5kg/ha 2009 Thiovit Jet 10,0 kg/ha Topas 100 EC 0,3 + Ridomil Gold MZ 68 WG 2,5kg/ha Vivando 0,25 + Acrobat MZ 2,0 kg/ha Vivando 0,25 + Acrobat MZ 2,0 kg/ha Falcon 0,3 + Melody Compact 1,5 kg/ha Falcon 0,3 + Melody Compact 1,5 kg/ha Falcon 0,3 + Melody Compact 1,5 kg/ha Bravo 500 2,5 + Topas 100 EC 0,3 Quadris Max 2,0 Cabrio Top 2,0 kg/ha + Flamenco 0,5 Cabrio Top 2,0 kg/ha + Collis 0,4 Eclair 0,5 kg/ha Eclair 0,5 kg/ha Eclair 0,5 kg/ha Ridomil Gold MZ 68 WG 2,5 kg/ha + Topas 100 EC 0,3 Topas 100 EC 0,3 + Pergado F 2,5kg/ha Vivando 0,25 + Forum R 3,0 kg/ha Vivando 0,25 + Forum R 3,0 kg/ha Falcon 0,3 + Melody Compact 1,5 kg/ha Falcon 0,3 + Melody Compact 1,5 kg/ha Falcon 0,3 + Melody Compact 1,5 kg/ha Quadris Max 2,0 Quadris Max 2,0 Cabrio Top 2,0 kg/ha + Kumulus S 5,0 kg/ha Cabrio Top 2,0 kg/ha + Collis 0,4 Eclair 0,5 kg/ha Eclair 0,5 kg/ha Eclair 0,5 kg/ha Quadris Max 2,0 Quadris Max 2,0 Vivando 0,25 + Forum RézoxikloridWP 3,0 kg/ha Vivando 0,25 + Forum+ Rézoxiklorid WP 3,0 kg/ha Falcon 0,3 + Melody Compact 1,5 kg/ha Falcon 0,3 + Melody Compact 1,5 kg/ha Falcon 0,3 + Melody Compact 1,5 kg/ha Quadris Max 2,0 Topas 100 EC 0,3 + Pergado F 2,5kg/ha X Kumulus S 5,0 kg/ha + Rézoxiklorid WP 3,0 kg/ha X X Kén 5,0 kg/ha + Rézoxiklorid WP 3,0 kg/ha Ridomil Gold MZ 68 WG 2,5 kg/ha + Topas 100 EC 0,3 + Chorus 75 WG 0,6 kg/ha X Cantus 1,0 kg/ha + Rézoxiklorid WP 3,0 kg/ha Cantus 1,0 kg/ha + Rézoxiklorid WP 3,0 kg/ha Kén 5,0 kg/ha + Rézoxiklorid WP 3,0 kg/ha + Teldor 1,0 Kén 5,0 kg/ha + Rézoxiklorid WP 3,0 kg/ha + Teldor 1,0 Kén 5,0 kg/ha + Rézoxiklorid WP 3,0 kg/ha + Teldor 1,0 Thiovit Jet 10,0 kg/ha + Ridomil Gold MZ 68 WG 2,5 kg/ha + Chorus 75 WG 0,6 kg/ha Thiovit Jet 10,0 kg/ha + Ridomil Plus 4,0 kg/ha + Chorus 75 WG 0,6 kg/ha 26/7 31/7 30/7 Topas 100 EC 0,3 + Ridomil Gold MZ 68 WG 2,5kg/ha Quadris Max 2,0 + Thiovit Jet 8,0 kg/ha Topas 100 EC 0,3 + Pergado F 2,5kg/ha Quadris Max 2,0 + Thiovit Jet 8,0 kg/ha Quadris Max 2,0 + Thiovit Jet 8,0 kg/ha Topas 100 EC 0,3 + Pergado F 2,5kg/ha Thiovit Jet 10,0 kg/ha + Ridomil Plus 4,0 kg/ha Thiovit Jet 10,0 kg/ha + Ridomil Plus 4,0 kg/ha + Chorus 75 WG 0,6 kg/ha n=87+ 10

11 During the collection, we ranked the infected leaves according to their powdery mildew covering from 0 to 5. Then, we determined the level of infection according to the method of Townsend and Heuberger (Gartner 1971). We examined 50 leaves on each parcel. The level of infection was also expressed in percentages. Level 0 or 0% reflect healthy leaves. Level 1 refers to 0-5% infection rate, level 2 is equal to 5-25% infection rate, level 3 shows 25-50% infection rate, level 4 represents 50-75% infection rate and level 5 indicated % infection rate. The data about the infection were averaged in each year, per treatment and groups of fungicides. The average of fungicide groups were analysed by the method of two-way analysis of variance with the help of SPSS 13.0 statistical software. We computed 95% confidence interval and the significance level between the groups of fungicides. We also determined the 95% confidence level and the significance level of infection in the case of each examined grape varieties. We analysed the correlation between grape varieties and treatments An in vitro Study of QoI Fungicides Used in Open Field Experiments Place of experiment We collected leaves from the parcel where only QoI active ingredients were sprayed on the experimental farm in Cserszegtomaj, between and and analysed them in vitro. These analyses were carried out at Georgikon Faculty of University of Pannonia, at the Research Group of Biotechnology at the Department of Horticulture The Research Material We collected samples from both varieties ( Olasz rizling GK 1 and Merlot CI 181 ) at the end of the growing season after the final infection level had developed on the foliage. The samples were collected from the middle fruiting cane of the middle stock of the parcels. It was important to exclude the possibility of faulty sampling as much as possible, which can be resulted by spay drift, while determining the place of sampling. Unlike the open field research, here we did collected infected clusters. 11

12 Method of Analysis We cut the infected leaves into small pieces. The leaves had been collected from 3 different levels of the foliage of the shoots. During the cutting, the used shears were sterilised by burning 80% ethanol on the surface of the shears. This process was repeated between cutting samples from different shoots and parcels of different treatment groups. Then, the infected pieces of the leaves were taken into 2 ml Eppendorf tubes with the help of sterilised forceps. The closed and catalogued samples were frozen in liquid nitrogen and stored on -80 C. Then, DNA was purified from the samples which were collected in. We had chosen samples from this year according to the results of the experiments of the active ingredients from Cserszegtomaj. The purification was carried out with the help of etyltrimethylammonium bromide (CTAB) method (Doyle and Doyle 1990) which was further optimised for obtaining good quality DNA. We applied PCR technology and used fungalspecific ITS1-ITS4 primers in order to monitor the adequate quality and amount of fungal DNA in the samples. We had chosen samples of good quality and detecting for the molecular analysis of the presence of QoI resistance. We designed primer sequences to detect resistance. The starting point was the detection of the point of mutation which can be found at nucleotide position 329 of the cytochrome b gene. Also, this point of mutation is responsible for QoI resistance against fungicides in other species. In order to detect the point of mutation, we designed a so-called inner primers onto the two sides of this point and a so-called outer primers for both directions a few hundred base pairs away. In PCR, the fragment of cytochrome bc1 gene, which is 653 base pairs (bp) in length and contains the point mutation, and DNA fragments 216 base pairs (bp) in length, which typical of resistant ones, were multiplied with the simultaneous use of BluGra124 (F) (5 - GGATGATTAATACGTTACATACA-3 ), CBR3 (R) (5 - CCTAATAATTTATTAGGTATAGATCTTA-3 ), and Enec1 R (5 - TAGTAATAACTGTTGCTG-3 ) primers. The PCR products were documented by GeneGenius Bio-Imaging System (Syngen, UK). 12

13 3.3. An in vitro Analysis of QoI Resistance in Powdery Mildew Populations Collected in Different Vineyards in Hungary Place of Experiment To obtain fungal spores, we made single spore isolation at Faculty of Sciences of University of Pécs, at the Department of General and Environmental Microbiology of the Institute of Biology. Then, the samples were prepared for storage at Georgikon Faculty of University of Pannonia, at the Department of Horticulture. The in vitro analyses were carried out at Georgikon Faculty of University of Pannonia, at the Research Group of Biotechnology at the Department of Horticulture The Research Material We collected samples in the middle of the growing season shortly after the powdery mildew infection. We did not succeed to collect samples from 2 wine regions out of 22. We found samples from 9 wine regions unsuitable, so we continued to analyse those which were the most suitable and could be accessible in large numbers. We also did collected samples from two vineyards, one from Szekszárd region and one from Eger region, where powdery mildew resistance against QoI fungicides had been first discovered (Füzi 2007). We collected 4 shoots, each one with 4-6 leaves. The samples were stored in humid environment in order to prevent damaging. They were taken into a plastic bag of appropriate size (30x50cm) half-filled with air next to a wet cotton wool ball. The plastic bags were stored in a cooler bag during the transportation and processing. Before the single spore isolation, we checked whether the mould of the fungus is suitable for further examination with the help of a binocular microscope Method of Analysis See

14 4. Results and Inferences 4.1. The Comparative Open Field Analysis of QoI Active Ingredients and Fungicides Used Against Erysiphe necator (Shwein) We tested different QoI active ingredients and chemicals against Erysiphe necator during the three-year open field experiment. During these years, potential QoI resistant lines appeared on the experimental field as a result of the chemical pressure. The development of resistance against the tested groups of active ingredients was much faster than the development of resistance against the group of SDHI fungicides despite the fact that both groups belong to the group of single site fungicides (Scheinpflug 1988; Tiemann et al. 1997). In most cases, resistance appeared when the same chemicals had been used through years (Steva and Clerjeau, 1990; Redl and Steinkeller, 1996; Halleen and Holz, 2001; Miller and Gubler, 2004; Wong and Wilcox, 2002). Our results showed that the effect of 2-3 years QoI active ingredients block treatment, using the widest possible range of registered active ingredients in Hungary, can be enough to the resistant phylum to appear. Most of the publications about the appearance of Qoi resistant E. necator proceed from the statement that resistant phyla have been found (Fernandez-Ortuno et al., ; Ishii et al., 2001). Our open field examinations showed what leads to the development of the resistance. The combination of QoI active ingredients with penconazole and sulphur can slow down the development of resistance (Group II). We did not experienced Steva s antagonistic observations between triadimenol and sulphur in the case of QoI active ingredients. The combinations against Plasmopara viticola (Group II) did not reduced the possibility of the development of resistance. The technologies offered by the manufacturers provided adequate protection against powdery mildew in all the three experimental years. The chemicals and active ingredients were used in combination and in rotation in the technological offers, so despite the presence of the resistant phyla, the chemicals and active ingredients provided high efficiency. According to the above mentioned, it is considered appropriate to follow the recommendation of the manufacturer, that is, QoI fungicides it is worth to be used in maximum 25-30% between sprayings. We agree with the statement of FRAC (1997), according to which the QoI treatments must follow the treatments of fungicides with other mode of action, but let us complete this statement with the application of sulphur. 14

15 4.2. The Molecular Genetic Background of QoI Resistance in Powdery Mildew 23 pathogens were defined as QoI resistant (Fraaije et al. 2005; Kim et al. 2003) from which four of them are powdery mildew: the Blumeria graminis f. sp. tritici (Fraaije et al. 2002), the B. graminis f. sp. hordei, the S. fuliginea and the Erysiphe necator. With the help of the primers, which were designed to the sequence of cythocrome bc 1 gene in B. graminis containing point mutation, we could detect the fragment 653 basepair in length containing point mutation in E. necator. Then, according to the results of sequencing, we could also detect that QoI resistance in E. necator is caused by the G C point mutation. We developed a fast PCR method by designing PASA primers for the isolation of individuals with resistant and sensitive alleles. We could isolate individuals with resistant alleles from individuals with sensitive alleles. We could also detect those fragments in resistant individuals which are typical for sensitive individuals. According to this, we can infer that the cythocrome bc 1 gene is present in heteroplasma in resistant individuals. QoI resistance can be detected quickly, even in samples of large number, with the help of the above mentioned PASA primers. Furthermore, resistant and sensitive individuals can be isolated clearly with the same method. Systemic active ingredients are playing an increasing role in the protection programs of large manufacturers. Considering cost-efficiency, the active ingredients have to compensate the lack of workforce by providing protection for longer period of time between sprayings. Because of the longer spaying cycles, the appearance of resistance can cause a great amount of crop loss. We can make protection more effective and safer in large-scale vineyards with the help of our quick resistance test because we can detect the possibly appearing mutants carrying resistance even in the vegetation period (between spraying cycles). The growth of the number of resistant individuals can be eliminated in vineyards with the help of the quick test because the further combinations of treatments of the spraying plan can be immediately changed after the detection of resistance. Thus, QoI active ingredients are excluded from further treatments and instead of them, systemic chemicals with different mode of action are applied supplemented by great amount of sulphur as we recommended earlier. Considering the test results, expenditure on fungicides spent during the vegetation period can be optimised and financial loss deriving from inefficiency can be eliminated both in case of grape production and current assets (fungicides). The results warn us that ignoring technological recommendations can leads to the increase of QoI resistance in Hungary. 15

16 5. New Scientific Results 1. We worked out a model system for studying resistance mechanism which develops as a result of one-sided use of QoI active ingredients. 2. Development of molecular genetic quick test for proving the presence of powdery mildew phyla resistant for QoI fungicides and similar ones in open field conditions. 3. Application of the quick test for proving the presence of resistant powdery mildew phyla in open field conditions. 4. We proved the presence of Erysiphe necator phyla resistant to QoI active ingredients in Cserszegtomaj vine region, Szekszárd and Eger vine regions, where signs of QoI fungicide resistance were detected earlier in the vineyards, in Hungary. As a result of this, we proved the usability of the new method. 5. Establishment of preventive vine protection technologies of resistance developing as a result of one-sided use, presentation of its background. 6. Proving efficiency differences between the groups of registered active ingredients used against E. necator. Explaining the process of the development of resistance. 16

17 6. Bibliography Doyle, J. J., and Doyle, J. L. (1990): A rapid total DNA preparation procedure for fresh plant tissue. Focus 12, Gartner, H. (1971): Versuche zur Bekämpfung von Botrytis cinerea (Grauschimmel) als Traubenfäule. Mitteilungen Klosterneuburg, 21: Fernández-Ortuno, D., Torés,J. A., De Vicente, A. and Pérez-Garcia, A. (): Field resistance to QoI fungicides in Podosphaera fusca is not supported by typical mutations in the mitochondrial cytochrome b gene. Pest Management Science, 64: Fraaije, B. A., Butters, J. A., Coelho, J. M., Jones, D. R. and Hollomon, D. W. (2002): Following the dinamics of strobilurin resistance in Blummeria graminis f. sp. tritici using quantitative allele-specific real-time PCR measurements with the fluorescent dye SYBR Green I. Plant Pathology, 51: Fraaije, B. A., Cools, H. J., Funtaine, J., Lovell, D. J., Motteram, J., West, J. S. and Lucas, J. A. (2005): Role of ascospores in further spread of QoI-resistant cytochrome b alleles (G143A) in field populations of Mycosphaerella graminicola. Phytopathology, 95: FRAC (1997): Status Report and Recommended Fungicide Resistance Management Guidelines. Global Crop Protection Federation, Belgium. Füzi I. (2007): A biztonság mindenek előtt. Növényvédelmi Tippek (BASF), 1-2 : Halleen, F. and Holz, G. (2001): An Overview of the Biology, Epidemiology and Control of Uncinula necator (Powdery Mildew) on Grapevine, with Reference to South Africa. South African Journal for Enolology and Viticulture, 22, 2: Ishii, H., Fraaije, B. A., Sugiyama, T., Noguchi, K., Nishimura, K., Takeda, T. Amano, T. and Hollomon D. W. (2001): Occurrence and molecular characterization of strobilurin resistance in cucumber powdery mildew and downy mildew. Phytophatology, 91:

18 Kim, Y. S., Dikson, E. W., Vincelli, P. and Farman, M. L. (2003): Field resistance to strobilurin (Q(o)I) fungicides in Pyricularia grisea caused by mutations in the mitochondrial cytochrome b gene. Phytopathology, 93: Miller, T. C. and Gubler, W. D. (2004): Sensitivity of California isolates of Uncinula necator to trifloxystrobin and spiroxamine, and update on triadimefon sensitivity. Plant Diseases, 88: Redl, H. and Steinkellner, S., (1996) Nachweiss einer sensitivitatsverminderung von Oidium gegenüber DMI-fungiziden im Österreichischen weinbau. Mitteilungen Klosterneuburg, 46: Scheinpflug, H., (1988): History of DMI fungicides and monitoring for resistance. In: Delph, C.J. (Eds.), Fungicide Resistance In North America. The American Phytophatological Society Press, Minnesota Steva, H., (1992): Resistance de l Oidium de la Vigne (Uncinula necator) aux fongicides inhibiteurs de la biosynthese des sterols. These de doctorat Universite Bordeaux II. Steva, H. and Clerjeau, M., (1990) Cross resistance to sterol biosynthesis inhibitor fungicides in strains of Uncinula necator isolated in France and Portugal. Meded. Fac. Landbouwwet. Rijksuniv. Gent 55: Tiemann, R., Berg, D., Kramer, W. and Pontzen, R. (1997): Biochemistry of the new fungicide KWG 4168 (Spiroxamine). Pflanzeschutz Nachrichten. Bayer, 50: Wong, F. P. and Wilcox, W. F. (2002): Sensitivity to azoxystrobin among isolates of Uncinula necator: Baseline distribution and relationship to myclobutanil sensitivity. Plant Diseases, 86:

19 7. List of Publications Publication Related to the Doctor of Philosophy (PhD) Thesis Scientific Articles Taksonyi, P., Kocsis, L., Mátyás, K. and Taller, J. (2012): The effect of Quinone outside Inhibitor fungicides on powdery mildew in grape vineyards in Hungary. Scientia Horticulture. Bírálat alatt. Taksonyi P., Maráczi K. és Kocsis L. (2011): Qoi-fungicidek alkalmazásának hatása a lisztharmat fertőzés kialakulására Kertgazdaság: Taksonyi, P., Tarczal, E., Maráczi, K., Holb, I. és Kocsis, L. (): Powdery mildew infection dependent on weather factors in vineyards near Keszthely in. International Journal of Horticultural Science, 16 (2): 55-59p. Taksonyi P., Füzi I. és Kocsis L. (2009): A szőlő egyes kórokozóinak QoIfungicidekkel szembeni rezisztenciájának kialakulása Magyarországon. Növényvédelem 45 (7): Scientific Presentations Taksonyi P., Kocsis, L. (): The powdery mildew infection of a vineyard at Lake Balaton in term of climate conditions. 28th International Horticultural Congress, Lisbon Congress Center aug 22-27,, p Taksonyi, P., Kocsis, L. (): The powdery mildew infection of a vineyard at Lake Balaton in term of climate conditions in. IHC Lisboa august Abstracts Volume II S16.306, 736. Taksonyi P., Maráczi K., Traczal E.(2009): Változó időjárási tényezők hatása a Keszthely környéki szőlőültetvények lisztharmat fertőzöttségére. 1 Szőlő és klíma konferencia, Kőszeg,2009. április

20 Taksonyi P. Kocsis L.(2009): Miért érdemes tudni, hogy mi az a QoI-rezisztencia? Metszési Bemutató Szakmai Nap, Keszthely, március. 20. Taksonyi P., Kocsis L. (2009): QoI-fungicidek alkalmazása Cserszegtomajon. Növényvédelmi fórum, Keszthely, február. Taksonyi P., Kadlicskó S.,Kocsis L., Gabi G. (): A szőlő előrejelzésre alapozott környezetkímélő növényvédelme a szekszárdi borvidéken. Növényvédelmi fórum, Keszthely,. január 30-február1. Taksonyi P., Kocsis L. (): A szőlő növényvédelmében alkalmazott természetes eredetű fungicidek és azokból származó mesterséges vegyületek történeti áttekintése. 50. Jubileumi Georgikon Napok, Keszthely,. szeptember p Publication Do Not Related to the Doctor of Philosophy (PhD) Thesis Kocsis, L., Taksonyi, P., Tarczal, E. (): The impact of climate change on grape production. Hungarian Agricultural Research 19(2): Taksonyi P. (2009): A szőlő ültetésének módjai és folyamata. Kertészet és Szőlészet, 2009.február. E. Tarczal, M. Kállay, Z. Varga, P. Taksonyi and L. Kocsis, (2009): Rootstock-scion interactions and correlations between vine vigor and grape quality. American Journal of Enology and Viticulture, 60(3): 408A-408A. L. Kocsis, É Baracsi Horváthné, K. Maráczi, P. Taksonyi, E. Tarczal, (2009): The impact of climate change in grape and ornimental production, Climate change: challange for training of applied plant scientists, Symposium and Training Course III: Impact of Climate Change on Corp Production, Martonvásár,