A subject of the diploma thesis was the optimization of a procedure of detecting a phytoplasma Candidatus Phytoplasma mali using woody indicator Golden Delicious in greenhouse conditions as an alternative method to the phytoplasma presence testing by biological indexing in field conditions. To control the presence of phytoplasma in the experimental plants, molecular method of real-time PCR was used. The experiment was set up on the demonstration area of the Department of Breeding and Propagation of Horticultural Plants in the grounds of Mendel University in Brno. Three different ways of transferring of the phytoplasma infection were compared: method of parallel grafting of the tested variety and of the indicator on the seedling in the period of dormancy (method number 1), method of summer budding of the tested variety on indicator grafted on the seedling in the period of dormancy (method number 2), and method of double summer budding of the indicator and of the tested variety on the seedling (method number 3). On a woody indicator Golden Delicious, varieties Virgin Czech, Canadian rennet, Gascoigne, Ribston and Boskoop were tested, 10 plants of each variety. The experiment lasted 10 months. A total of 150 tested plants were assessed by a visual inspection of the symptoms presence. During the monitoring period, none of the tested plants showed typical symptoms of the apple proliferation. Presence of the Candidatus Phytoplasma mali was verified using real-time PCR method in 60 tested (indicator) plants. The infection was confirmed in 12 of the indicator plants. The detected quantities of the phytoplasma in the tissues however reached very low values. The highest number of the infected plants was found in tests of the Gascoigne variety, established by the method of double summer budding of the indicator and of the tested variety on the seedling (method number 3). This method provided the most successful inoculation of the indicator by the phytoplasma. Based on the results obtained during the presented study, it can be declared that biological testing of the phytoplasma presence using short-term greenhouse tests is not reliable. When using testing with the indicator ´Golden Delicious´, it is recommended to realize the experiments for at least two growing seasons, as it is in the field tests. Due to the fact that during the time-limited realization period of the experiment in this study it was found that the phytoplasma was not sufficiently propagated, the final results of the comparison of the indicator plants inoculation methods will be available at the end of autumn of 2017.
| Identifer | oai:union.ndltd.org:nusl.cz/oai:invenio.nusl.cz:363118 |
| Date | January 2016 |
| Creators | Blahová, Kamila |
| Source Sets | Czech ETDs |
| Language | Czech |
| Detected Language | English |
| Type | info:eu-repo/semantics/masterThesis |
| Rights | info:eu-repo/semantics/restrictedAccess |
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