Investigation of Causer of Tomato Pith Necrosis - Pseudomonas Mediterranea Cattara Et Al. 2002, In Republic Of Macedonia

Kovacevik, Biljana (2010) Investigation of Causer of Tomato Pith Necrosis - Pseudomonas Mediterranea Cattara Et Al. 2002, In Republic Of Macedonia. Masters thesis, University Goce Delcev- Stip.

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Abstract

Last several years from 2005 – 2009 the health condition of tomato plants is monitored in the region of Strumica. During this period, symptoms of Tomato Pith Necrosis – TPN, are observed for the first time at tomatoes grown in plasticgreen houses. A ubiquitous bacterium Pseudomonas mediterranea is isolated by standard procedure from the symptomatic tomato plants. Several years later, in 2009 the same bacterium is isolated from the soil, too. A total number of 22 isolates are investigated: nine strains isolated from the symptomatic plants (Ds-4, Ds-4/1, Ds-4/2, Ds-12, Ds-12/1, Ds-12/2, Ds-13, Ds-13/1 and Ds-13/2); four isolates from soil (S-2.1, S-2.2, S-2.3 и S-2.4); one positive control from Italy and one from Turkey (IPVCT 9.1, P.m); one control of P. corrugata (IPVCT 10.3) from Italy, and several negative controls obtained from the Belgium collection of microorganisms: Clavibacter mishiganensis subsp. michiganensis (LMG 2891), Pantoea agglomerans (LMG 2595), Pseudomonas syringae pv. syringae (LMG 5085), Pseudomonas viridiflava (LMG 5396), Pseudomonas fluorescence (LMG 5396) and Pseudomonas marginalis (LMG 5850). In in vitro conditions, are examined: pathogenicity of the isolates, hypersensitive reaction of tobacco, gram reaction, growth characteristics on different media, bacterial growth at 4 and 41˚C, tolerance of 5% and 7% NaCl, starch hydrolysis, gelatin lignifications, ammonia formation, formation of H2S from peptones, indol, nitrate reduction, aesculine hydrolysis and accumulation of poly-β-hydroxybuturate. Examined biochemical-physiological characteristics are: pectolitic activity on potato slices, oxidase activity, catalase activity, oxidative-fermentative test, dehidrolisis of arginie, urease activity, sensitivity to CuSO4 and streptomicine sulphate. The utilization of 96 different carbon dioxides and acids is investigated by BIOLOG tests. Genetic analysis, and final determination is obtained with multiplex PCR and two types of primers, according to Cattara et al., 2000 (Type I (PC5/1 and PC 5/2) and Type II (PC1/1 and PC 1/2). The result show the presence of new bacterial variety, not identified before in the Republic of Macedonia. All thirteen isolates from Macedonia, belongs to the variety of Pseudomonas mediterranea. A total number of 124 characteristics from 22 strains are used for the numerical taxonomy analysis (NTSYSpc version 2r, software package, Exeter Software, NY, USA): biochemical and physiological characteristics, resistence to copper and antibiotics, results obtained using BIOLOG plates, and introduction of HR in tobacco. The distance matrix is calculated by using the DICE coefficient and the cluster analysis is performed using the UPGMA (Rohlf, 1994). Final result show that all investigated isolates from the variety of Pseudomonas mediterranea and Pseudomonas corrugatа are grouped in one cluster with index of similarity of 0.937, in which the isolates are divided in three groups: A, B, and C. In the group A with the similarity index of 0.958, belongs control isolates IPVCT 9.1, IPVCT 10.3, and P.m. In the group B with the similarity index of 0.976, are Ds-4, Ds-4/1, Ds-4/2, S-2.1, S-2.2, S-2.3 and S-2.4. In the group C, with the similarity index of од 0.982, belongs Ds-12, Ds-12/1, Ds-12/2, Ds-13, Ds-13/1 and Ds-13/2.

Item Type: Thesis (Masters)
Subjects: Agricultural Sciences > Other agricultural sciences
Divisions: Faculty of Agriculture
Depositing User: Katerina Hadzi-Vasileva
Date Deposited: 07 Nov 2012 18:42
Last Modified: 12 Sep 2013 09:22
URI: http://eprints.ugd.edu.mk/id/eprint/598

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