Primena savremenih biotehnologija u proizvodnji biljaka roda Impatiens / Application of modern biotechnology in the production plants of the genus Impatiens
2013
Milošević, Snežana
In this monograph are presented two important horticultural species – Impatiens walleriana L. And Impatiens hawkerii Bull. Their origin, distribution and morphological characteristics are shown. Based on the fact that these plants are natural hosts of Tomato spotted wilt virus (TSWV) the aim of this study was to determine the presenc eof this virus in greenhouses production of Impatiens at one locality in Serbia. The paper describes the presence of TSWV in Serbia and in the world, its economic importance, natural and experimantal hosts, morphology, genome, organization and variability, symptoms of infection, sources of infection, vectors and ways of expansion, as well as the detection and characterization. In the monograph are offered solutions to problems that result from viral infections by using modern biotechnological methods: in vitro culture meristem to eliminate the virus and genetic transformation using Agrobacterium tumefaciens who carries the gene for pac 1 for tolerance to viruses. In the paper attention is paid to the detection and identification of viruses in plant material, which are made by DAS-ELISA assay, by mechanical inoculation test plants, RT-PCR and sequencing RdRp gene from the L RNA viral genome. The author indicates that the application of biotechnological methods is necessary to establish an in vitro system for the regeneration of shoots with the addition of different concentrations of cytokinins (CPPU and TDZ). The paper confirmed that the regenerative potential depends on the applied concentration of cytokinins in MS medium. I. Walleriana L. Successfully regeneraed from shoots in vitro and I. Hawkerii Bull. From meristem with the elimination of TSWV. Sensitivitiy of the genusImpatiens to Agrobacterium was investigated with genetic transformetion I. Hawkerii Bull. By A. Rhizogenes A4M70GUS, because the development of adventives roots was quick and obvious evidence, and the success of the genetic transformation was confirmed by GUS and PCR essays.Obtaining a transformed plant in this paper for the first time confirmed the sensitivity of Impatiens plants to infection by Agrobacterium, which opens wide possibilities for further modification of this important horticultural species. I. hawkerii Bull. And I. walleriana L. Was transformed with A. tumefaciens C58C1pac1, the success of transformation was confirmed by PCR and RT-PCR. I. hawkerii Bull. Lines did not survive selection on antibiotic and obtained transformed lines I. walleriana L. Carrying the active pac construct and due to have the resistance to TSWV. The results presented in this work contribute to the production of plants tolerant to RNA viruses, thus reducing the cost of commercial production of planting material, which can be used in horticulture and other fields of agriculture.
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