Submitted by (lucia.rodrigues@ufrpe.br) on 2017-02-16T14:20:21Z No. of bitstreams: 1 Aldenir de Oliveira.pdf: 717930 bytes, checksum: eb22d1cac453b579b74b66da5fd0bc67 (MD5) / Made available in DSpace on 2017-02-16T14:20:21Z (GMT). No. of bitstreams: 1 Aldenir de Oliveira.pdf: 717930 bytes, checksum: eb22d1cac453b579b74b66da5fd0bc67 (MD5) Previous issue date: 2008-02-29 / This dissertation aimed to study: colonization of Acidovorax avenae subsp. citrulli in melons after inoculation of the first pair of true leaves, seeds and hermaphrodite flowers; bacterial survival in fruit and leaf residues incorporated to the soil at 0, 5, 10 and 15 cm depth and in soils without the host plant, under the influence of different soil type, temperature (10, 15, 20, 25, 30 and 35ºC) and humidity (50 and 100% of field capacity). In all studies a mutant resistant to 100 ppm of rifampicin (Aac1Rif) was utilized. Bacterial colonization was detected until 30 days after inoculation in the 10th pair of true leaves, with populations of 3.1 log UFC g-1 of leaf. In the same period, the shoot segment between the 10th and 11th leaf pair showed population of 3.52 log UFC g-1 of shoot. After seed inoculation the pathogen colonized the hypocotyl, roots, cotyledonary leaves, true leaves and shoots, until reach undetectable levels at 27 days after inoculation. Flower colonization by the bacteria was not verified. Aac1Rif was found in fruit and leaf residues at 0, 5 and 10 cm during 21 days and at 15 cm during 14 days. Highest population relative extinction rates (TERP)were presented by fruits on soil surface [0.1464 log (UFC) day -1] and on leaves at 10cm [0.084 log (UFC) day -1]. Aac1Rif survived on seven soil types only during three days and the soil C showed the highest TERP [0.9062 log (UFC) day-1]. Higher concentrations of Na+ and silt as well as higher populations of actinomycetes and Trichoderma correlated to faster extinction of Aac1Rif populations in soil. Generally for all soils the lowers TERP were found at 10 or 15ºC and the higher, at 30 or 35ºC. There was no significant (P=0.05) interaction between soil and humidity, however the T test showed significant difference (P=0.05) between the TERP at 100% [0.6685 log (UFC) day -1] and 50% [0.504591 log (UFC) day -1] of field capacity. Independent of temperature and humidity, Aac1Rif also survived in soil only during three days. / Esta dissertação teve como objetivos estudar: colonização de A. avenae subsp. citrulli em meloeiro a partir da inoculação no primeiro par de folhas verdadeiras, sementes e flores hermafroditas; sobrevivência da bactéria em restos de folhas e frutos incorporados ao solo a diferentes profundidades (0, 5, 10 e 15 cm) e em solos na ausência da planta hospedeira, sob a influência de diferentes tipos de solo (sete solos), temperaturas (10, 15, 20, 25, 30 e 35ºC) e umidades (50 e 100% da capacidade de campo). Para os estudos foi utilizado um mutante resistente a 100 ppm de rifampicina (Aac1Rif). Quando a bactéria foi inoculada nas folhas, a colonização foi detectada até os 30 dias, no 10º par de folhas verdadeiras, com população de 3,1 log UFC g-1 de folha. Nesse mesmo período, observou-se a colonização no segmento de ramo compreendido entre o 10º e 11º par de folhas com população de 3,52 log UFC g-1 de ramo. A partir de sementes, a bactéria colonizou o hipocótilo, raízes, folhas cotiledonares, folhas verdadeiras e ramos, até atingir níveis não detectáveis aos 27 dias após a inoculação. Não foi verificada colonização das flores pela bactéria. Aac1Rif foi encontrada em restosde frutos e folhas de meloeiro a 0, 5 e 10 cm durante 21 dias e a 15 cm por 14 dias. As maiores taxas de extinção relativa da população (TERP) ocorreram nos frutos na superfície do solo [0,1464 log (UFC) dia-1] e nas folhas a 10 cm [0,084 log (UFC) dia-1]. Aac1Rif sobreviveu nos sete tipos de solo apenas durante três dias e o solo C apresentou a maior TERP [0,9062 log (UFC) dia-1]. Maiores concentrações de Na+ e silte bem como maiores populações de actinomicetos e Trichoderma estiveram correlacionadas a mais rápida extinção da população de Aac1Rif no solo. Para a maioria dos solos, as menores TERP foram atingidas a 10 ou 15ºC e as maiores, a 30 ou 35ºC. Não houve interação significativa (P=0,05) entre solos e umidade, contudo o teste de T evidenciou diferença significativa (P=0,1) entre as TERP a 100% [0,6685 log (UFC) dia-1] e 50% [0,504591 log (UFC) dia-1] da capacidade de campo. Independente da temperatura e umidade, Aac1Rif também sobreviveu nos solos apenas por três dias.

Melon (Cucumis melo L.) and watermelon (Citrullus lanatus (Thunb.) Matsum and Nakai) belong to the family Cucurbitaceae. Bacterial fruit blotch (BFB) disease of cucurbits is an economically devastating plant disease that has caused an estimated loss of up to $450M on watermelon crops and $75M (worldwide) to the seed and transplant industries since 1996. Disease symptoms include water-soaked cotyledons, leaf necrosis, and internal fruit rot. Current commercial management strategies are very limited and include: seed production field sanitation, greenhouse transplant sanitation, copper-based bactericide sprays, crop rotation, disease-free healthy seeds, isolating diseased plants, and peroxyacetic acid seed treatments. The seedborne disease is usually spread by contaminated seeds, and there is a zero-tolerance policy in the seed industry for infected seeds. No nondestructive assays are commercially available to detect BFB in seeds. This research investigated several different aspects of BFB disease such as non-destructive seed detection, green tea seed treatment, candidate NB-LRR genes for disease resistance, and optimization of virus induced gene silencing for melon and watermelon crops. The potential application of attenuated total reflectance (ATR) Fourier transform infrared spectroscopy (ATR-FTIR) and high-resolution X-ray analysis methods for detection of BFB on seeds were evaluated. It was possible to detect BFB in seeds that were pistil inoculated via x-ray imaging and pericarp inoculated via ATR FT-IR. In vitro and in vivo experiments evaluated the potential of tea (Camellia sinensis) and tea polyphenols as seed treatments to sanitize seeds infected with A. citrulli. Green tea unlike black tea inhibited growth of A. citrulli because of polyphenols. Eighty one melon and forty four watermelon NB-LRR genes were reidentified, and genes that have potential resistance against A. citrulli on melon plants were screened based on host selectivity of the pathogen. Finally, the virus-induced, gene-silencing method was optimized for melon and watermelon for further analysis of potential disease resistance genes. BFB can be nondestructively identified in seeds and green tea may be an effective seed treatment with further development. Promising candidate R genes were identified that might confer stable resistance in the right genetic background. / Doctor of Philosophy / Melon and watermelon crops both belong to the gourd family. Bacterial fruit blotch (BFB) disease of cucurbits is an economically devastating plant disease that has caused an estimated loss of up to $450M on watermelon crops and $75M (worldwide) to the seed and transplant industries since 1996. Disease symptoms include water-soaked cotyledons, leaf necrosis, and internal fruit rot. Current commercial management strategies and detection methods are very limited. The seedborne disease is usually spread by contaminated seeds, and there is a zero-tolerance policy in the seed industry for infected seeds. This research investigated several different aspects of BFB disease such as non-destructive seed detection, green tea seed treatment, candidate disease resistance genes, and optimization of virus induced gene silencing methodology for melon and watermelon crops. There are currently no nondestructive assays available to detect BFB in seeds. We evaluated the potential application of attenuated total reflectance (ATR) Fourier transform infrared spectroscopy (ATR-FTIR) and high-resolution X-ray analysis methods for detection of BFB on seeds. It was possible to detect BFB inside layers of seeds that were naturally inoculated through the flowers via x-ray imaging and seedcoat inoculated via ATR FT-IR. In vitro and in vivo experiments evaluated the potential of tea and tea constituents as seed treatments to sanitize seeds infected with BFB. Green tea unlike black tea inhibited growth of BFB. Eighty one melon and forty four watermelon disease resistance genes were reidentified and genes that have potential resistance against BFB on melon plants were screened based on host selectivity of the pathogen. Finally, the virus induced gene silencing method was optimized for melon and watermelon plants for further analysis of potential disease resistance genes. BFB can be nondestructively identified in seeds and green tea may be an effective seed treatment with further development. Promising candidate resistance genes were identified that might confer stable resistance in the right genetic background.

Um dos principais fatores limitantes da produção do melão é a ocorrência de doenças. Entre os patógenos mais importantes estão as bactérias, que causam perdas significativas na produção. Causada pela bactéria Acidovorax avenae subsp. citrulli (Aac), a mancha-aquosa-do-melão, também conhecida como mancha-bacteriana-dofruto, é uma doença grave, que tem preocupado produtores do nordeste, sendo que todos os tipos de melão apresentam suscetibilidade ao patógeno. A principal fonte de inóculo para esta bactéria é a semente infectada. Este trabalho teve como objetivo o desenvolvimento de um meio semi-seletivo para detecção e identificação de Acidovorax avenae subsp. citrulli em sementes de melão, para testes de rotina em laboratórios de patologia de sementes. Por meio de testes de fungitoxicidade, antibiogramas qualitativos e quantitativos e testes bioquímicos foi desenvolvido o seguinte meio semiseletivo para a detecção de Aac em sementes de melão: dextrose (5 g/L), NaCl (5 g/L), peptona (5 g/L), KH2PO4 (2 g/L), MgSO4.7H2O (0,2 g/L), vermelho de fenol (0,012 g/L), uréia (25 g/L), agar (17 g/L), benomil (100 mg/L), nistatina (200 mg/L) e amoxicilina (15 mg/L). / One of the main limiting factors of the melon production is the occurrence of diseases. The bacteria are among the most important pathogens causing significant losses in the production. Caused by the bacterium Acidovorax avenae subsp. citrulli (Aac), the bacterial-fruit-blotch is a serious disease that affects all types of melon and has worried northeast producers. The main source of inoculum for this bacterium is the infected seed. This work had as objective the development of a semi-selective medium to detect and identify Acidovorax avenae subsp. citrulli in melon seeds, for routine tests in seed pathology laboratories. By means of fungitoxicity tests, qualitative and quantitative antibiograms and biochemical tests the following semi-selective medium to detect Aac in melon seeds was developed: dextrose (5 g/L), NaCl (5 g/L), peptone (5 g/L), KH2PO4 (2 g/L), MgSO4.7H2O (0,2 g/L), phenol red (0,012 g/L), urea (25 g/L), agar (17 g/L), benomil (100 mg/L), nistatina (200 mg/L) and amoxicilin (15 mg/L).

Um dos principais fatores limitantes da produção do melão é a ocorrência de doenças. Entre os patógenos mais importantes estão as bactérias, que causam perdas significativas na produção. Causada pela bactéria Acidovorax avenae subsp. citrulli (Aac), a mancha-aquosa-do-melão, também conhecida como mancha-bacteriana-dofruto, é uma doença grave, que tem preocupado produtores do nordeste, sendo que todos os tipos de melão apresentam suscetibilidade ao patógeno. A principal fonte de inóculo para esta bactéria é a semente infectada. Este trabalho teve como objetivo o desenvolvimento de um meio semi-seletivo para detecção e identificação de Acidovorax avenae subsp. citrulli em sementes de melão, para testes de rotina em laboratórios de patologia de sementes. Por meio de testes de fungitoxicidade, antibiogramas qualitativos e quantitativos e testes bioquímicos foi desenvolvido o seguinte meio semiseletivo para a detecção de Aac em sementes de melão: dextrose (5 g/L), NaCl (5 g/L), peptona (5 g/L), KH2PO4 (2 g/L), MgSO4.7H2O (0,2 g/L), vermelho de fenol (0,012 g/L), uréia (25 g/L), agar (17 g/L), benomil (100 mg/L), nistatina (200 mg/L) e amoxicilina (15 mg/L). / One of the main limiting factors of the melon production is the occurrence of diseases. The bacteria are among the most important pathogens causing significant losses in the production. Caused by the bacterium Acidovorax avenae subsp. citrulli (Aac), the bacterial-fruit-blotch is a serious disease that affects all types of melon and has worried northeast producers. The main source of inoculum for this bacterium is the infected seed. This work had as objective the development of a semi-selective medium to detect and identify Acidovorax avenae subsp. citrulli in melon seeds, for routine tests in seed pathology laboratories. By means of fungitoxicity tests, qualitative and quantitative antibiograms and biochemical tests the following semi-selective medium to detect Aac in melon seeds was developed: dextrose (5 g/L), NaCl (5 g/L), peptone (5 g/L), KH2PO4 (2 g/L), MgSO4.7H2O (0,2 g/L), phenol red (0,012 g/L), urea (25 g/L), agar (17 g/L), benomil (100 mg/L), nistatina (200 mg/L) and amoxicilin (15 mg/L).