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BAHŞIEV, Aighiuni; MITIN, Valentin; MITINA, Irina; ZAMORZAEVA, Irina. Assessment of the load of tomato plants by phytoplasma. In: International Congress of Geneticists and Breeders from the Republic of Moldova. Ediția 11, 15-16 iunie 2021, Chişinău. Chișinău, Republica Moldova: Centrul Editorial-Poligrafic al Universităţii de Stat din Moldova, 2021, p. 18. ISBN 978-9975-933-56-8.
|International Congress of Geneticists and Breeders from the Republic of Moldova
Ediția 11, 2021
Congresul "International Congress of Geneticists and Breeders from the Republic of Moldova" |
Chişinău, Moldova, 15-16 iunie 2021
'Candidatus Phytoplasma solani' is a pathogen that colonizes the phloem of many crops causing disturbances in the normal growth and development of plants. The pathogen is endemic throughout the European Union and widely distributed in the world. Control of phytoplasma disease is difficult due to the systemic circulation in the host organism, being spread by vector insects, weeds and incorrect agronomic practices. Classical methods such as electron microscopy, in vitro cultivation and serological diagnosis are laborious and sometimes do not allow specific determination of the pathogen. Thus, molecular methods, especially based on PCR techniques, allow reliable identification of the phytoplasma species. Moreover, some of these methods enable the quantification of phytoplasma in plants. The purpose of the research was to determine the loading plants of two local tomato varieties with 'Ca. P. solani'. DNA was extracted from fruits of the Elvira and Mary Gratefully tomato varieties (10 plants in each variety) using DNAzol kit. Fruits for the analysis were collected in August, at the period of mass fruit ripening. The analysis of 'Ca. P. solani' loading of tomato plants was based on a number of consecutive dilutions of extracted DNA. Each dilution step was fivefold. The obtained solutions were used as template DNA in the first round of the nested-PCR with chaperonin primers ps2 and ps5. Second round was carried out with primer pair ps3-ps4.The result showed that the end point of phytoplasma detection was at eighth dilution (2.6x10-6) for the Elvira variety. For the Mary Gratefully variety the limit of 'Ca. P. solani' detection was at ninth dilution (5.1x10-7). So, the amount of phytoplasma DNA in the Elvira sample is a little less than in the Mary Gratefully sample. This result was confirmed in our further study of plant loading by phytoplasma in the same two tomato varieties by real-time PCR, which is mainly used for DNA quantification. Thus, we can conclude that the PCR assay by serial DNA dilutions can be used like alternative method for the quantification of 'Ca. P. solani' in tomato plants. We also believe that this method is an effective and relatively cheap approach, especially if mass analysis is required.