Fungicide resistance is one of the most important problems endangering the effectivity of practical plant protection today. The frequent and subsequent usage of specific fungicides results the emergence of resistant fungal populations. This threatens is especially high in case of Botrytis cinerea Pers.:Fr. being an endemic pathogen with frequent infection. Nowadays the main method of protection as against Botrytis cinerea is the application of chemical fungicides chemicals. Therefore, a better knowledge of local populations is necessary for the planning of the protection procedures.
Based on the results of our examinations we may establish that the growth of the examined samples showed a significant difference under in vitro circumstances, which shows a great deal of variability of the Botrytis cinerea populations in Hungary. Twenty-five Botrytis cinerea samples from different hosts were analyzed in this study. High resistance was found towards azoxistrobin in seven cases, and low resistance in eight cases.
It was also proved, that the B. cinerea is able to bypass the inhibition site of the azoxistrobin via the alternative oxidase. The presence of this altermative mitocondrial electrotransport route considerably reduces the effectivity of the chemical.
Botrytis cinerea (teleomorph Botryotinia fuckeliana (de Bary) Whetzel) is able to attack several economically important plants causing gray rot. Botrytis cinerea species complex includes two cryptic species (B. cinerea and B. pseudocinerea) that tolerate fungicides differently. On the basis of classical taxonomic markers, the two related species are very difficult to be distinguished; therefore, their separation is usually performed using molecular methods based on the time-consuming molecular analysis of several markers. Our goal was to find markers, which are suitable for the differentiation. Testing the nucleotide sequences of the alternative oxidase encoding gene, B. cinerea and B. pseudocinerea strains were clearly differentiated. Moreover, the analysis of the protein sequences of the enzyme with the maximum likelihood method reflected well the taxonomic relationships of the different fungi.
Botrytis cinerea has been reported as a species complex containing two cryptic species, groups I (Botrytis pseudocinerea) and II (B. cinerea sensu stricto). In order to compare the pathogenicity of group I and group II of B. cinerea, we have selected 4 strains of group I and 4 strains of group II. The results demonstrated that competitive infection of group II was more on grape, cucumber and paprika leaves, than group I. However the results on bean leaves did not correlate the applied B. cinerea group.
Botrytis cinerea causes gray mold on a high number of crop plants. Information about the populations of plant pathogen fungi may help to develop new strategies for the effective and economic crop protection with reduced fungicide usage. Hungarian B. cinerea populations were characterized with using different molecular genetic parameters. B. cinerea group I strains, characterized with high rate of fenhexamid resistance, could be detected only in restricted number. The Hungarian B. cinerea populations were characterized with high genetic diversity, and the regular occurrence of sexual reproduction. These results highlight the importance of rotating different type of fungicide in the plant protection technology against grey mould.
The research was carried out in a Gyenes Flower gardening between 2016 and 2017 in Kecskemét. The gardening was founded in 1978. Initially, the main plants were gerbera (Gerbera) and yucca (Yucca), later replaced by the geranium (Pelargonium) cultivation as a result of market demand. In horticulture, there are about than 80 variety geranium of the standing, running, semi-trailer types and English gnawing. The Pelargonium had different sizes and colors. The study was set up in 1,000–1,000 pieces of geraniums each year. The following pathogens have damaged the geranium stock: Botrytis cinerea, Pythium debaryanum, with a rare occurrence of Alternaria porri, Phytophthora cryptogea. The greatest destruction was caused by botrytis (Botrytis cinerea). In the first experimental year, 42% of the 1,000 geraniums tested were infected with fungal diseases (30% B. cinerea, 8% P. debaryanum, 4% other fungi). In 2017, fungal infections were detected on 380 geraniums in the 1,000 tested geraniums (290 Botrytis cinerea, 70 Pythium and 20 other fungal diseases). In addition to the use of fungicides, we increased the spatial position of geraniums, early irrigation and frequent ventilation to ensure successful control. By 2017, we were able to reduce the damage caused by pathogens by 4 percent.
In the mitochondrion of eukaryotes, cytochrome b is a component of respiratory chain complex III. Cytochrome b is encoded by the
cytochrome b (CYTB) gene located in the mitochondrial genome. The fungicidal activity of QoIs relies on their ability to inhibit mitochondrial respiration by binding at the so-called Qo site (the outer quinol-oxidation site) of the complex III. Since their introduction, QoIs (like azoxystrobin) have become essential components of plant disease control programs because of their wide-ranging efficacy against many agriculturally important fungal diseases like grey mould on various crops. QoI resistance primarily arises from a target-site-based mechanism involving mutations in the mitochondrial CYTB. As the management of grey mould is often dependent on chemicals, the rational design of control programs requires the information about the diversity of genes connected with resistance in field populations of the pathogen.
Monospore B. cinerea field isolates has been collected during 2008-2009 from different hosts in Hungary. PCR fragment length analysis
indicated the high frequency presence of type large intron in the isolates while in a few strains G143A substitution could also be detected.
These results indicated the heterogeneity of CYTB in the Hungarian B. cinerea populations, which possibly involve the heteroplasmy of this
mitochondrial gene, moreover indicates the existence op azoxystrobin resistant populations in Hungary.
This work was supported by NKFP-A2-2006/0017 grant. Erzsébet Fekete is a grantee of the János Bolyai Scholarship (BO/00519/09/8).
The genes being in the mitochondrial DNA primarily encode the enzymes of cellular respiration. Fungicides belonging to the family of quinol oxidase inhibitors (QoIs) play on important role in the protection against several plant diseases caused by fungi. These fungicides bind to the cytochrome bc1 complex so they block electron transport between cytochrome b and cytochrome c1. This way these fungicides inhibit the ATP synthesis consequently they inhibit the mitochondrial respiration. The QoI resistance has two mechanisms. One of them is the point mutation of the cytochrome b gene (CYTB), e.g. the substitution of a single glycine by alanine at position 143 results in high-resistance. The other is the cyanide-resistant alternative respiration sustained by the alternative oxidase.
In a cell there are several mitochondria. The phenomenon when the genomes of all mitochondria in the cell are identical is called homoplazmy. If in the cell there is wild and mutant mitochondrial DNA this is called heteroplasmy. Whether the mutation in the mitochondria causes fenotypical diversity or does not depend on the dose, i.e. it depends on the percentage of the changed mitochondrials. During our work we investigated Botrytis cinerea single spore isolates which have been collected in 2008-2009 on different host plants. Our goal was to decide whether heteroplasmy influences the level of resistance. We managed to detect the change of the level of heteroplasmy, so the change the level of the resistance due to the treatment with fungicide.