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Produção de sintomas e movimento
seguido de agro-inoculação com
ssDNA de “tomato yellow leaf curl
virus” Geminivirus (Tail...
2
INTRODUÇÃO
• Vários gêneros pertencentes a família Geminiviridae
• Geminiviroses: Solanaceae, leguminosas e gramíneas;
• A...
OBJETIVO
• Este estudo procura inicialmente caracterizar o
genoma de um TYLCV isolado da Tailândia,
encontrado infectando ...
MATERIAIS E MÉTODOS
• A purificação do Vírus;
• Clonagem do DNA do genoma do TYLCV;
• Construção do clone por Agroinoculaç...
RESULTADOS E DISCUSSÃO
FIG. 1. As sequências das regiões comuns TYLCV: setas sólidas indicam repetições
comuns, setas trac...
FIG. 2. Southern blot de TYLCV recombinante (Tailândia) clones, pDR9 (putativo componente
A) e pDR7 (putativo componente B...
FIG. 3. TYLCV - Nicotiana benthamiana infectadas 3 semanas após agroinoculação. (a) As
plantas agro-inoculadas com “TYLCV-...
Fig. 4. Análise de Dot blot de TYLCV encontrado infectando plantas de tomate.
Tratamentos 1-4 - somente o componente “A”, ...
FIG. 5. Análise de Southern blot de plantas de Nicotiana benthamiana infectada por TYLCV.
Na (A) linha 1, inoculada com A....
CONCLUSÕES
• Verificou-se que o TYLCV - é um DNA capaz de
infectar através de agroinoculação, mover
sistemicamente, e indu...
LITERATURA CITADA
• ANDERSEN, M. T , RICHARDSON, K A., HARBISON, S. and MORRIS, B. A M (1988).
Nucleotide sequence of the ...
• CZOSNEK. H., BER, R NAV~T, N , ANTIGNUS, Y., COHEN, S., ~~~ZAMIR, D. (1989)
Tomato yellow leaf curl virus DNA forms In t...
• ETESSAMI, P.. WAITS. can cassava mosaic virus coat protein gene deletion mutants dur- Ing Infection
of Nicotiana bentham...
• GARDINER, W. E., SUNTER, G., BRAND, L., ELMER, J. S., ROGERS, S G., and BISARO, D.
M. (1988). Genetic analysis of tomato...
• HAYES, R. J., MACDONALD, J., Courts, R. H. A., and BUCK, K. W (1988) Infection of
Nicoflana benthamana with uncut cloned...
• MACDOW~LL, S W.. MACDONALD, H HAMILTON. W. D 0, COUT~S, R H. A, and BUCK,
K. W. (1985). The nucleotlde sequence of clone...
• Russo, M., COHEN, S., and MARIELLI, G. P. (1980). Virus-like particles in tomato
plants affected by yellow leaf curl dis...
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Produção de sintomas e movimento seguido de agro-inoculação com ssDNA de “tomato yellow leaf curl virus” Geminivirus (Tailândia). - Apresentador : Tony L. Moura - Prof. Milton Luiz da Paz Lima

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ROCHESTER, D.E., KOSITRATANA, W., BEACHY., R.N. Systemic Movement and Symptom Production following Agroinoculation with a Single DNA of Tomato Yellow Leaf Curl Geminivirus (Thailand). VIROLOGY 178,520-526 (1990)

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Produção de sintomas e movimento seguido de agro-inoculação com ssDNA de “tomato yellow leaf curl virus” Geminivirus (Tailândia). - Apresentador : Tony L. Moura - Prof. Milton Luiz da Paz Lima

  1. 1. Produção de sintomas e movimento seguido de agro-inoculação com ssDNA de “tomato yellow leaf curl virus” Geminivirus (Tailândia). Instituto Federal Goiano câmpus Urutaí. Curso de Agronomia Disciplina de Fitopatologia I Apresentador: Tony L. Moura ROCHESTER, D.E., KOSITRATANA, W., BEACHY., R.N. Systemic Movement and Symptom Production following Agroinoculation with a Single DNA of Tomato Yellow Leaf Curl Geminivirus (Thailand). VIROLOGY 178,520-526 (1990) 1
  2. 2. 2
  3. 3. INTRODUÇÃO • Vários gêneros pertencentes a família Geminiviridae • Geminiviroses: Solanaceae, leguminosas e gramíneas; • A espécie - TYLCV em diferentes regiões; • Transmissão: cigarrinha e mosca branca; • Suas sequências usadas para construção de árvores filogenéticas; • Patógeno de grande importância para o tomate no Mediterrâneo. 3
  4. 4. OBJETIVO • Este estudo procura inicialmente caracterizar o genoma de um TYLCV isolado da Tailândia, encontrado infectando tecido de tomate. 4
  5. 5. MATERIAIS E MÉTODOS • A purificação do Vírus; • Clonagem do DNA do genoma do TYLCV; • Construção do clone por Agroinoculação; • Agro-inoculação e análise de plantas infectadas; • Extração de DNA e análise; 5
  6. 6. RESULTADOS E DISCUSSÃO FIG. 1. As sequências das regiões comuns TYLCV: setas sólidas indicam repetições comuns, setas tracejadas indicam a posição estável no caule e uma estrutura de haste e laço estável pode formar. O oligonucleotideo (DR2) usado como um prime secundário abaixo da vertente síntese primordial abaixo da linha. Apenas diferenças de sequência de TYLCV-A TYLCV-B são mostrados. Asteriscos indicam posições onde as lacunas são inseridas para proporcionar o máximo de homologia. 6
  7. 7. FIG. 2. Southern blot de TYLCV recombinante (Tailândia) clones, pDR9 (putativo componente A) e pDR7 (putativo componente B). 1, pDR7 (1 pg), digerido com EcoRI; 2. pDR7 (1 pg), digerido com EcoRI e Sphl; 3. pDR9 (0,5 pg) digerido com EcoRI; 4. pDR9 (1 pg), digerido com EcoRI; 5. pDR9 (1 pg), digerido com EcoRI e BarnHI. 7 RESULTADOS E DISCUSSÃO
  8. 8. FIG. 3. TYLCV - Nicotiana benthamiana infectadas 3 semanas após agroinoculação. (a) As plantas agro-inoculadas com “TYLCV-A" só, “TYLCV-B" sozinho, e “TYLCV A e B. t" (b) Planta que foi agro-inoculada com apenas com o componente A (pDR21: SE). (c) Plantas inoculadas com "A" e "B" componentes (pDR2 1: SE e pDR37: SE, respectivamente). 8 RESULTADOS E DISCUSSÃO
  9. 9. Fig. 4. Análise de Dot blot de TYLCV encontrado infectando plantas de tomate. Tratamentos 1-4 - somente o componente “A”, 5-8 componente B, 9-12 componentes A e B, 13-16 a bactéria responsável pela agro-inoculação (A. tumefaciens 3111 SE), 17-20 não agro-inoculadas com TYLCV. 9 RESULTADOS E DISCUSSÃO 2
  10. 10. FIG. 5. Análise de Southern blot de plantas de Nicotiana benthamiana infectada por TYLCV. Na (A) linha 1, inoculada com A. tumefaciens 3111SE; linha 2, , inoculada com A. tumefaciens do TYLCV-“B”; linha 3, agro-inoculado com TYLCV-“A”; linha 4, agro-inoculado com TYLCV- “A” (enzima “bean nuclease “) ; linhas 5 e 7, agro-inoculado com TYLCV-“A + B”; linhas 6 e 8, agro-inoculado com TYLCV-“A + B” (enzima “bean nuclease “). Linhas l-6 foram hibridizadas com sondas específicas de TYLCV-“A”-e linhas 7, 8, com sondas específicas de TYLCV-“B”. Tipos de DNA : OC, open circle; L, linear; CCC, covalently closed circle; SS, single- stranded. 10 RESULTADOS E DISCUSSÃO
  11. 11. CONCLUSÕES • Verificou-se que o TYLCV - é um DNA capaz de infectar através de agroinoculação, mover sistemicamente, e induzir sintomas como mal- estar, tanto tomate e N. benthamiana. 11
  12. 12. LITERATURA CITADA • ANDERSEN, M. T , RICHARDSON, K A., HARBISON, S. and MORRIS, B. A M (1988). Nucleotide sequence of the geminivIrus Chloris striate mosaic virus. I/iroiogy 164, 443-449. • BOUI.~ON, M. I., STEINKELLNER, J., DONSON, J., MARKHAM, P. G., KING, D. I and DAVIES, J. W. (1989). Mutational analysis of the vinon- sense genes of maize streak virus. /. Gen. Viroi. 70, 2309-2323. • BROUGH. D. L , HAYES, R J., MORGAN, A J., COUTTS, R. H. A., and SUCK, K. W (1988). Effects of mutagenesis in v&o on the ablllty of cloned tomato golden mosaic virus DNA to infect Nicofiana ben- thamiana plants J. Gen. V~rol. 69, 503 -5 14. • COHEN. S., and NITZANY, F. E. (1966). Transmission and host range of the tomato yellow leaf curl virus. Phytopathology 56, 1 127-l 131. • COMMANDEUR, U MATZEIT. V., and GRONENBORN, 6. (1987). The DNA sequence of WDV-C. a wheat dwarf virus Isolate from Czechoslo- vakia. in “Abstracts, VII International Congress of Virology,” p.235. Natlonal Research Council Canada. Ottawa. 12
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