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Registro Completo |
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Biblioteca(s): |
Embrapa Cerrados. |
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Data corrente: |
18/10/2001 |
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Data da última atualização: |
18/10/2001 |
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Autoria: |
LIMA, J. A. A. |
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Título: |
Blackeye cowpea mosaic virus: purification, partial characterization, serology, and immunochemical and cytological techniques for detection of virus-infected legume seeds. |
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Ano de publicação: |
1978 |
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Fonte/Imprenta: |
[S.l]: University of Florida, 1978. |
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Páginas: |
153p. |
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Idioma: |
Inglês |
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Notas: |
PhD. Dissertation. |
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Conteúdo: |
Blackeye cowpea mosaic virus (BICMV) was increased in cowpea, Vigna unguiculata (L.) Walp., 'knuckle purple hull', and infected leaves were used for virus and cytoplasmic inclusion purification. Either n-butanol or a combination of chloroform and carbon tetrachloride was used in the clacification process. Polyacrylamide gel electrophoresis of sodium dodecyl sulfate(SDS) dissociated inclusions and virus revealed that the inclusions were made of a single protein estimated to have e molecular weight (MW) around 70.000 daltons whereas freshly purified BICMV consisted of a main protein component with a MW of 34,000 da itons and two smaller proteins with WMs of 29,000 and 27,000 daltons, Purified BICMV had a 260/280 nm absorption ration of 1.2 and a modal length of 753 nm. Freshly purified BICMV preparations showed a single sedimenting peak with s20 =157-159 s. The purified BICMV cytoplasmic inclusions had absorption spectra characteristic for proteins. Electron microscopy of purified inclusions revealed the presence of tubes showing striations with periodicities of approximately 5nm. Antisera reactive in SDS-immunodiffusion were obtainedagainst untreated virions, pyrrolidine degrasded ceat protein, and untreated BICMV cytoplasmic incluions. Reciprocal double immunodiffusion tests with SDS-treated antigens showed that BICMV is serologically unrelated to seven potyviruses and serologically related to, but distinct from: bean common mosaic virus (BCMV), bean yellow mosaic virus (BYMC), cowpea aphid-borne mosaic virus CAMV), dasheen mosaic (DMV), lettuce mosaic virus (LMV), potato virus y (PVY), soybean mosaic virus (SoyMV), tobacco etch virus (TEV), and watermelon masaic virus-2 (WMV-2). The intregal cross-absorption technique was also used to demonstrate distinction between closely related potyviruses. Ager medium impregnated with a mixture of antisera used for serodiagnosis of BICMV and cowpea mosaic virus in cowpea. Light and electron microscopy of cytoplasmic inclusions induced by BICMV, siratro (Macroptilium atropurpureum (D.C.) Urb.) strain of BCMV (BCMV-S) and CAMV revealed that they are similar to those induced by the potyviruses from Edwardson's subdivision-l. The different reactions induced by BICMV, and CAMV in some cowpea varieties in-dicated that they can also be used as used as differential hosts for these three potyviruses. Sources of resistance for BICMV were found among the cowpea varieties tested. Based on its physical, biological, cytological, and immunochemical properties, BICMV can be differentiated fron any other virus that infects cowpea. Cytoplasmic inclusions induced by BICMV in cowpea and by SoyMV in soybean were detected by serology, light microscopy, and alactron microscopy in hypocotyls of 4-5-day-old seedlings grown from virus-infected seeds. Immunodiffusion tests and serologically specific electrom micro-scopy were used to detect BICMV in hypocotyls of 4-5-day-old cowpea seedlings grown from BICMV-infected seeds. Discs of individual hypocotyls were embedded into the agar medium 4-5 mm away from the antiserum wells. Virus-specific precipitin lines formed between virus-infected hypocotyl discs and antiserum wells, whereas no reactinos were observed with healthy hypocotyls. Precipitin lines were also observed with extracts of mixtures from infected (1 g) and healthy (up to 29 g) tissues. These immunochemical techniques were also used for detecting BCMC in hypocotyls of infdcted 4-5-day-old phaseolus vulgaris L. seedlings and for detecting SoyMV in infected Glycine max (L) Merr. seedlings. Single radial immunodiffusion tests with extracts or discs of cowpea hypocotyls were also useful for detecting BICMV in germinated seeds. The reliability and simplicity of the immunodiffusion tests make them suitable for use in routine seed health testing program in any laboratory. MenosBlackeye cowpea mosaic virus (BICMV) was increased in cowpea, Vigna unguiculata (L.) Walp., 'knuckle purple hull', and infected leaves were used for virus and cytoplasmic inclusion purification. Either n-butanol or a combination of chloroform and carbon tetrachloride was used in the clacification process. Polyacrylamide gel electrophoresis of sodium dodecyl sulfate(SDS) dissociated inclusions and virus revealed that the inclusions were made of a single protein estimated to have e molecular weight (MW) around 70.000 daltons whereas freshly purified BICMV consisted of a main protein component with a MW of 34,000 da itons and two smaller proteins with WMs of 29,000 and 27,000 daltons, Purified BICMV had a 260/280 nm absorption ration of 1.2 and a modal length of 753 nm. Freshly purified BICMV preparations showed a single sedimenting peak with s20 =157-159 s. The purified BICMV cytoplasmic inclusions had absorption spectra characteristic for proteins. Electron microscopy of purified inclusions revealed the presence of tubes showing striations with periodicities of approximately 5nm. Antisera reactive in SDS-immunodiffusion were obtainedagainst untreated virions, pyrrolidine degrasded ceat protein, and untreated BICMV cytoplasmic incluions. Reciprocal double immunodiffusion tests with SDS-treated antigens showed that BICMV is serologically unrelated to seven potyviruses and serologically related to, but distinct from: bean common mosaic virus (BCMV), bean yellow mosaic virus (BYM... Mostrar Tudo |
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Palavras-Chave: |
Cowpea mosaic comovirus; Plant diseases; Seed. |
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Thesagro: |
Doença; Feijão de Corda; Semente; Vigna Sinensis; Vigna Unguiculata; Vírus. |
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Thesaurus Nal: |
cowpeas; viruses. |
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Categoria do assunto: |
-- |
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Marc: |
LEADER 04631nam a2200265 a 4500
001 1560262
005 2001-10-18
008 1978 bl uuuu m 00u1 u #d
100 1 $aLIMA, J. A. A.
245 $aBlackeye cowpea mosaic virus$bpurification, partial characterization, serology, and immunochemical and cytological techniques for detection of virus-infected legume seeds.
260 $a[S.l]: University of Florida$c1978
300 $a153p.
500 $aPhD. Dissertation.
520 $aBlackeye cowpea mosaic virus (BICMV) was increased in cowpea, Vigna unguiculata (L.) Walp., 'knuckle purple hull', and infected leaves were used for virus and cytoplasmic inclusion purification. Either n-butanol or a combination of chloroform and carbon tetrachloride was used in the clacification process. Polyacrylamide gel electrophoresis of sodium dodecyl sulfate(SDS) dissociated inclusions and virus revealed that the inclusions were made of a single protein estimated to have e molecular weight (MW) around 70.000 daltons whereas freshly purified BICMV consisted of a main protein component with a MW of 34,000 da itons and two smaller proteins with WMs of 29,000 and 27,000 daltons, Purified BICMV had a 260/280 nm absorption ration of 1.2 and a modal length of 753 nm. Freshly purified BICMV preparations showed a single sedimenting peak with s20 =157-159 s. The purified BICMV cytoplasmic inclusions had absorption spectra characteristic for proteins. Electron microscopy of purified inclusions revealed the presence of tubes showing striations with periodicities of approximately 5nm. Antisera reactive in SDS-immunodiffusion were obtainedagainst untreated virions, pyrrolidine degrasded ceat protein, and untreated BICMV cytoplasmic incluions. Reciprocal double immunodiffusion tests with SDS-treated antigens showed that BICMV is serologically unrelated to seven potyviruses and serologically related to, but distinct from: bean common mosaic virus (BCMV), bean yellow mosaic virus (BYMC), cowpea aphid-borne mosaic virus CAMV), dasheen mosaic (DMV), lettuce mosaic virus (LMV), potato virus y (PVY), soybean mosaic virus (SoyMV), tobacco etch virus (TEV), and watermelon masaic virus-2 (WMV-2). The intregal cross-absorption technique was also used to demonstrate distinction between closely related potyviruses. Ager medium impregnated with a mixture of antisera used for serodiagnosis of BICMV and cowpea mosaic virus in cowpea. Light and electron microscopy of cytoplasmic inclusions induced by BICMV, siratro (Macroptilium atropurpureum (D.C.) Urb.) strain of BCMV (BCMV-S) and CAMV revealed that they are similar to those induced by the potyviruses from Edwardson's subdivision-l. The different reactions induced by BICMV, and CAMV in some cowpea varieties in-dicated that they can also be used as used as differential hosts for these three potyviruses. Sources of resistance for BICMV were found among the cowpea varieties tested. Based on its physical, biological, cytological, and immunochemical properties, BICMV can be differentiated fron any other virus that infects cowpea. Cytoplasmic inclusions induced by BICMV in cowpea and by SoyMV in soybean were detected by serology, light microscopy, and alactron microscopy in hypocotyls of 4-5-day-old seedlings grown from virus-infected seeds. Immunodiffusion tests and serologically specific electrom micro-scopy were used to detect BICMV in hypocotyls of 4-5-day-old cowpea seedlings grown from BICMV-infected seeds. Discs of individual hypocotyls were embedded into the agar medium 4-5 mm away from the antiserum wells. Virus-specific precipitin lines formed between virus-infected hypocotyl discs and antiserum wells, whereas no reactinos were observed with healthy hypocotyls. Precipitin lines were also observed with extracts of mixtures from infected (1 g) and healthy (up to 29 g) tissues. These immunochemical techniques were also used for detecting BCMC in hypocotyls of infdcted 4-5-day-old phaseolus vulgaris L. seedlings and for detecting SoyMV in infected Glycine max (L) Merr. seedlings. Single radial immunodiffusion tests with extracts or discs of cowpea hypocotyls were also useful for detecting BICMV in germinated seeds. The reliability and simplicity of the immunodiffusion tests make them suitable for use in routine seed health testing program in any laboratory.
650 $acowpeas
650 $aviruses
650 $aDoença
650 $aFeijão de Corda
650 $aSemente
650 $aVigna Sinensis
650 $aVigna Unguiculata
650 $aVírus
653 $aCowpea mosaic comovirus
653 $aPlant diseases
653 $aSeed
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Registro original: |
Embrapa Cerrados (CPAC) |
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 | Acesso ao texto completo restrito à biblioteca da Embrapa Agroenergia. Para informações adicionais entre em contato com cnpae.biblioteca@embrapa.br. |
Registro Completo
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Biblioteca(s): |
Embrapa Agroenergia. |
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Data corrente: |
29/09/2015 |
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Data da última atualização: |
20/09/2017 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Circulação/Nível: |
A - 1 |
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Autoria: |
SANTOS, D. F. K. dos; ISTVAN, P.; NORONHA, E. F.; QUIRINO, B. F.; KRÜGER, R. H. |
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Afiliação: |
Débora Farage Knupp dos Santos, UnB; Paula Istvan, UnB; Eliane Ferreira Noronha, UnB; BETANIA FERRAZ QUIRINO, CNPAE; Ricardo Henrique Krüger, UnB. |
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Título: |
New dioxygenase from metagenomic library from Brazilian soil: insights into antibiotic resistance and bioremediation. |
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Ano de publicação: |
2015 |
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Fonte/Imprenta: |
Biotechnology Letters, v. 37, n. 9, p.1809-1817, 2015. |
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DOI: |
10.1007/s10529-015-1861-x |
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Idioma: |
Inglês |
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Conteúdo: |
Objectives Putative new dioxygenases were identified in a metagenomic b-lactam-resistance screening and, given their key role on aromatic metabolism, we raise the hypothesis that these enzymes maybe concomitantly related to antibiotic resistance and aromatic degradation. Results ORFs of three putative dioxygenases were isolated from resistant metagenomic clones. One of them, CRB2(1), was subcloned into pET24a expression vector and subjected to downstream phenotypic and bioinformatics analyses that demonstrated the ??dual effect?? of our metagenomic dioxygenase, on antibiotic and aromatic resistance. Furthermore, initial characterization assays strongly suggests that CRB2(1) open-reading frame is an extradiol-dioxygenase, most probably a bicupin domain gentisate 1,2-dioxygenase. This observation is, to our knowledge, the first description of a metagenomic dioxygenase and its action on b-lactam resistance. Conclusion Unraveling the diversity of antibiotic resistance elements on the environment could not only identify new genes and mechanisms in which bacteria can resist to antibiotics, but also contribute to biotechnology processes, such as in bioremediation. |
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Palavras-Chave: |
Aromatic metabolism; Aromatic metabolismo; Dioxygenase; Dioxygenase Metagenome; Metagenome; Resistência aos antibióticos. |
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Thesagro: |
Solo. |
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Thesaurus NAL: |
antibiotic resistance; soil. |
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Categoria do assunto: |
-- |
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Marc: |
LEADER 02061naa a2200289 a 4500
001 2025343
005 2017-09-20
008 2015 bl uuuu u00u1 u #d
024 7 $a10.1007/s10529-015-1861-x$2DOI
100 1 $aSANTOS, D. F. K. dos
245 $aNew dioxygenase from metagenomic library from Brazilian soil$binsights into antibiotic resistance and bioremediation.$h[electronic resource]
260 $c2015
520 $aObjectives Putative new dioxygenases were identified in a metagenomic b-lactam-resistance screening and, given their key role on aromatic metabolism, we raise the hypothesis that these enzymes maybe concomitantly related to antibiotic resistance and aromatic degradation. Results ORFs of three putative dioxygenases were isolated from resistant metagenomic clones. One of them, CRB2(1), was subcloned into pET24a expression vector and subjected to downstream phenotypic and bioinformatics analyses that demonstrated the ??dual effect?? of our metagenomic dioxygenase, on antibiotic and aromatic resistance. Furthermore, initial characterization assays strongly suggests that CRB2(1) open-reading frame is an extradiol-dioxygenase, most probably a bicupin domain gentisate 1,2-dioxygenase. This observation is, to our knowledge, the first description of a metagenomic dioxygenase and its action on b-lactam resistance. Conclusion Unraveling the diversity of antibiotic resistance elements on the environment could not only identify new genes and mechanisms in which bacteria can resist to antibiotics, but also contribute to biotechnology processes, such as in bioremediation.
650 $aantibiotic resistance
650 $asoil
650 $aSolo
653 $aAromatic metabolism
653 $aAromatic metabolismo
653 $aDioxygenase
653 $aDioxygenase Metagenome
653 $aMetagenome
653 $aResistência aos antibióticos
700 1 $aISTVAN, P.
700 1 $aNORONHA, E. F.
700 1 $aQUIRINO, B. F.
700 1 $aKRÜGER, R. H.
773 $tBiotechnology Letters$gv. 37, n. 9, p.1809-1817, 2015.
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