Morfogênese in vitro, análise fitoquímica e caracterização anatômica de nim (Azadirachta indica A. Juss)

Detalhes bibliográficos
Autor(a) principal: Rodrigues, Marcelo
Data de Publicação: 2009
Tipo de documento: Dissertação
Idioma: por
Título da fonte: LOCUS Repositório Institucional da UFV
Texto Completo: http://locus.ufv.br/handle/123456789/2516
Resumo: The Azadirachta indica A. Juss species, popularly known as neem, belongs to the Meliaceae family, which has great potential for azadirachtin (AZA) production; however, neem has recalcitrant seeds, cross-pollination and variable content of AZA. Four experiments were conducted in order to understand the in vitro growth limiting factors of these plants and changes in levels of AZA: i) In vitro propagation of neem by culture of collected nodal segments; ii) Evaluation of leaf anatomy of plants grown in three different environments (in vivo, in vitro and acclimatized), using the micromorphometric technique; iii) Evaluation of levels of AZA, which were determined by High Performance Liquid Chromatography (HPLC) in cotyledonary callus, grown in WPM liquid medium with absence and presence of glucose (carbon source), hydrolyzed casein (nitrogen source) and methyl jasmonate (elicitor agent); and, iv) The propagation of somatic embryos from cotyledonary callus. It was found that MS medium has presented the best results for shooting rate, leaflet number and average size of the explants as compared to others culture media (WPM and JADS). It was not induced in vitro rooting of shoots; however, the root system has developed ex vitro during acclimatization, with seedling survival rate of 80%. It was also assayed the general content of lipids, essential oils, resins and oils, tannins and mucilage. It was found that there are significant differences in height between the evaluated leaf tissues, adaxial and abaxial epidermis, as well as in the mesophyll from three environmental conditions. The histochemical assay showed that there is general production of lipids, essential oils and tannins in leaf idioblasts of neem, meanwhile the mucilage test showed negative results for the three evaluated locations. The highest concentrations of AZA (mean 0.2470 mg g-1) were produced in the first and second cultivation week in the presence of these three substances, demonstrating the synergistic effect when the cell mass was cultured in medium supplemented with these. Calli showed high propagation capacity by both regenerative routes organogenesis and embryogenesis starting from the fourth subculture in WPM medium presenting the highest shooting and somatic embryos germination rates. Calli went from a smooth to granular appearance during the subcultures, while color went from dark brown to light brown and white. From the seventh subculture, there was dramatic reduction of in vitro regeneration rate for both types of neem cotyledonary callus. Therefore, this work has contributed for a better understanding of biotechnology techniques aiming the in vitro propagation, cell cultivation for AZA production and anatomical aspects of A. indica.
id UFV_6240c15f806af34c12f8ea27f58c5759
oai_identifier_str oai:locus.ufv.br:123456789/2516
network_acronym_str UFV
network_name_str LOCUS Repositório Institucional da UFV
repository_id_str 2145
spelling Rodrigues, Marcelohttp://lattes.cnpq.br/7327377175498257Buselli, Reginaldo Alves Festuccihttp://lattes.cnpq.br/8711829719442865Silva, Luzimar Campos dahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4799707J8Otoni, Wagner Camposhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4786133Y6Meira, Renata Maria Strozi Alveshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4706996Y7Dias, José Maria Moreirahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783068Z8Xavier, Aloisiohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782565D02015-03-26T13:08:46Z2009-12-212015-03-26T13:08:46Z2009-07-16RODRIGUES, Marcelo. Morphogenesis in vitro, phytochemical analysis and anatomical characterization of neem (Azadirachta indica A. Juss). 2009. 89 f. Dissertação (Mestrado em Botânica estrutural; Ecologia e Sistemática) - Universidade Federal de Viçosa, Viçosa, 2009.http://locus.ufv.br/handle/123456789/2516The Azadirachta indica A. Juss species, popularly known as neem, belongs to the Meliaceae family, which has great potential for azadirachtin (AZA) production; however, neem has recalcitrant seeds, cross-pollination and variable content of AZA. Four experiments were conducted in order to understand the in vitro growth limiting factors of these plants and changes in levels of AZA: i) In vitro propagation of neem by culture of collected nodal segments; ii) Evaluation of leaf anatomy of plants grown in three different environments (in vivo, in vitro and acclimatized), using the micromorphometric technique; iii) Evaluation of levels of AZA, which were determined by High Performance Liquid Chromatography (HPLC) in cotyledonary callus, grown in WPM liquid medium with absence and presence of glucose (carbon source), hydrolyzed casein (nitrogen source) and methyl jasmonate (elicitor agent); and, iv) The propagation of somatic embryos from cotyledonary callus. It was found that MS medium has presented the best results for shooting rate, leaflet number and average size of the explants as compared to others culture media (WPM and JADS). It was not induced in vitro rooting of shoots; however, the root system has developed ex vitro during acclimatization, with seedling survival rate of 80%. It was also assayed the general content of lipids, essential oils, resins and oils, tannins and mucilage. It was found that there are significant differences in height between the evaluated leaf tissues, adaxial and abaxial epidermis, as well as in the mesophyll from three environmental conditions. The histochemical assay showed that there is general production of lipids, essential oils and tannins in leaf idioblasts of neem, meanwhile the mucilage test showed negative results for the three evaluated locations. The highest concentrations of AZA (mean 0.2470 mg g-1) were produced in the first and second cultivation week in the presence of these three substances, demonstrating the synergistic effect when the cell mass was cultured in medium supplemented with these. Calli showed high propagation capacity by both regenerative routes organogenesis and embryogenesis starting from the fourth subculture in WPM medium presenting the highest shooting and somatic embryos germination rates. Calli went from a smooth to granular appearance during the subcultures, while color went from dark brown to light brown and white. From the seventh subculture, there was dramatic reduction of in vitro regeneration rate for both types of neem cotyledonary callus. Therefore, this work has contributed for a better understanding of biotechnology techniques aiming the in vitro propagation, cell cultivation for AZA production and anatomical aspects of A. indica.A espécie Azadirachta indica A. Juss, conhecida popularmente por nim, pertence à família Meliaceae, a qual apresenta grande potencial para a produção de azadirachtina (AZA); contudo, possui sementes recalcitrantes, polinização do tipo cruzada e teor variável de AZA. Com o objetivo de se entender os fatores que limitam o crescimento dessas plantas in vitro e a variação nos teores de AZA, foram conduzidos quatro experimentos: i) A propagação in vitro de nim pela cultura de segmentos nodais coletados; ii) Avaliação das anatômicas foliares de plantas cultivadas em três ambientes distintos (in vivo, in vitro e aclimatizado) por meio da técnica de micromorfometria; iii) Avaliação dos teores de AZA foram determinados por meio de Cromatografia Líquida de Alto Desempenho (HPLC) em calos cotiledonares, cultivados em meio líquido WPM na ausência e presença de glicose (fonte de carbono), caseína hidrolisada (fonte de nitrogênio) e metil jasmonato (agente elicitor); iv) A propagação de embriões somáticos a partir de calos cotiledonares. Constatou-se que o meio MS apresentou os melhores resultados de taxa de brotação, número de folíolos e tamanho médio dos explantes em relação aos demais meios de cultivo (WPM e JADS). Não foi induzido enraizamento in vitro dos brotos, porém, o sistema radicular se desenvolveu ex vitro, durante a aclimatização, apresentando taxa de sobrevivência das mudas de 80%. Foi também analisado o conteúdo de lipídios gerais, óleos essenciais e óleos resinas, tanino e mucilagem. Verificou-se diferenças significativas na altura entre os tecidos foliares avaliados, epiderme adaxial e abaxial, assim como no mesofilo nos três ambientes de cultivo. Os testes histoquímicos evidenciaram que há produção de lipídios gerais, óleos essenciais e taninos em idioblastos foliares de nim enquanto o teste para mucilagem foi negativo nos três ambientes avaliados. As maiores concentrações de AZA (média de 0,2470 μg g-1) foram produzidas na primeira e segunda semana de cultivo na presença dessas três substâncias, demonstrando o efeito sinérgico quando a massa celular foi cultivada em meio suplementado com essas substâncias. Os calos apresentaram alta capacidade de propagação pelas duas vias regenerativas, organogênese e embriogênese a partir do quarto subcultivo, em meio WPM, apresentando as maiores taxas de brotação e germinação de embriões somáticos. Os calos passaram do aspecto liso para o granular ao longo dos subcultivos, enquanto que a coloração passou de marrom escuro para marrom claro e branco. A partir do sétimo subcultivo, houve redução brusca quanto à taxa de regeneração in vitro para ambos os tipos de calos cotiledonares de nim. Portanto, esse trabalho colaborou para maior entendimento das técnicas biotecnológicas visando a propagação in vitro, cultivo de células para produção de AZA e aspectos anatômicos de A. indica.Conselho Nacional de Desenvolvimento Científico e Tecnológicoapplication/pdfporUniversidade Federal de ViçosaMestrado em BotânicaUFVBRBotânica estrutural; Ecologia e SistemáticaAzadirachta indicaNimHPLCAzadirachtinaAnatomiaAzadirachta indicaNeemHPLCAzadirachtinaAnatomyCNPQ::CIENCIAS BIOLOGICAS::BOTANICAMorfogênese in vitro, análise fitoquímica e caracterização anatômica de nim (Azadirachta indica A. Juss)Morphogenesis in vitro, phytochemical analysis and anatomical characterization of neem (Azadirachta indica A. Juss)info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINALtexto completo.pdfapplication/pdf4822847https://locus.ufv.br//bitstream/123456789/2516/1/texto%20completo.pdf4414f0ab2040a7fa5bbcc05b94aceb85MD51TEXTtexto completo.pdf.txttexto completo.pdf.txtExtracted texttext/plain157181https://locus.ufv.br//bitstream/123456789/2516/2/texto%20completo.pdf.txt04497190af906ee6ba126f2e121f020cMD52THUMBNAILtexto completo.pdf.jpgtexto completo.pdf.jpgIM Thumbnailimage/jpeg3741https://locus.ufv.br//bitstream/123456789/2516/3/texto%20completo.pdf.jpgc7c35751984cc479c079de853db2e9c0MD53123456789/25162016-04-08 23:08:19.403oai:locus.ufv.br:123456789/2516Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-09T02:08:19LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false
dc.title.por.fl_str_mv Morfogênese in vitro, análise fitoquímica e caracterização anatômica de nim (Azadirachta indica A. Juss)
dc.title.alternative.eng.fl_str_mv Morphogenesis in vitro, phytochemical analysis and anatomical characterization of neem (Azadirachta indica A. Juss)
title Morfogênese in vitro, análise fitoquímica e caracterização anatômica de nim (Azadirachta indica A. Juss)
spellingShingle Morfogênese in vitro, análise fitoquímica e caracterização anatômica de nim (Azadirachta indica A. Juss)
Rodrigues, Marcelo
Azadirachta indica
Nim
HPLC
Azadirachtina
Anatomia
Azadirachta indica
Neem
HPLC
Azadirachtina
Anatomy
CNPQ::CIENCIAS BIOLOGICAS::BOTANICA
title_short Morfogênese in vitro, análise fitoquímica e caracterização anatômica de nim (Azadirachta indica A. Juss)
title_full Morfogênese in vitro, análise fitoquímica e caracterização anatômica de nim (Azadirachta indica A. Juss)
title_fullStr Morfogênese in vitro, análise fitoquímica e caracterização anatômica de nim (Azadirachta indica A. Juss)
title_full_unstemmed Morfogênese in vitro, análise fitoquímica e caracterização anatômica de nim (Azadirachta indica A. Juss)
title_sort Morfogênese in vitro, análise fitoquímica e caracterização anatômica de nim (Azadirachta indica A. Juss)
author Rodrigues, Marcelo
author_facet Rodrigues, Marcelo
author_role author
dc.contributor.authorLattes.por.fl_str_mv http://lattes.cnpq.br/7327377175498257
dc.contributor.author.fl_str_mv Rodrigues, Marcelo
dc.contributor.advisor-co1.fl_str_mv Buselli, Reginaldo Alves Festucci
dc.contributor.advisor-co1Lattes.fl_str_mv http://lattes.cnpq.br/8711829719442865
dc.contributor.advisor-co2.fl_str_mv Silva, Luzimar Campos da
dc.contributor.advisor-co2Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4799707J8
dc.contributor.advisor1.fl_str_mv Otoni, Wagner Campos
dc.contributor.advisor1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4786133Y6
dc.contributor.referee1.fl_str_mv Meira, Renata Maria Strozi Alves
dc.contributor.referee1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4706996Y7
dc.contributor.referee2.fl_str_mv Dias, José Maria Moreira
dc.contributor.referee2Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783068Z8
dc.contributor.referee3.fl_str_mv Xavier, Aloisio
dc.contributor.referee3Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782565D0
contributor_str_mv Buselli, Reginaldo Alves Festucci
Silva, Luzimar Campos da
Otoni, Wagner Campos
Meira, Renata Maria Strozi Alves
Dias, José Maria Moreira
Xavier, Aloisio
dc.subject.por.fl_str_mv Azadirachta indica
Nim
HPLC
Azadirachtina
Anatomia
topic Azadirachta indica
Nim
HPLC
Azadirachtina
Anatomia
Azadirachta indica
Neem
HPLC
Azadirachtina
Anatomy
CNPQ::CIENCIAS BIOLOGICAS::BOTANICA
dc.subject.eng.fl_str_mv Azadirachta indica
Neem
HPLC
Azadirachtina
Anatomy
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS BIOLOGICAS::BOTANICA
description The Azadirachta indica A. Juss species, popularly known as neem, belongs to the Meliaceae family, which has great potential for azadirachtin (AZA) production; however, neem has recalcitrant seeds, cross-pollination and variable content of AZA. Four experiments were conducted in order to understand the in vitro growth limiting factors of these plants and changes in levels of AZA: i) In vitro propagation of neem by culture of collected nodal segments; ii) Evaluation of leaf anatomy of plants grown in three different environments (in vivo, in vitro and acclimatized), using the micromorphometric technique; iii) Evaluation of levels of AZA, which were determined by High Performance Liquid Chromatography (HPLC) in cotyledonary callus, grown in WPM liquid medium with absence and presence of glucose (carbon source), hydrolyzed casein (nitrogen source) and methyl jasmonate (elicitor agent); and, iv) The propagation of somatic embryos from cotyledonary callus. It was found that MS medium has presented the best results for shooting rate, leaflet number and average size of the explants as compared to others culture media (WPM and JADS). It was not induced in vitro rooting of shoots; however, the root system has developed ex vitro during acclimatization, with seedling survival rate of 80%. It was also assayed the general content of lipids, essential oils, resins and oils, tannins and mucilage. It was found that there are significant differences in height between the evaluated leaf tissues, adaxial and abaxial epidermis, as well as in the mesophyll from three environmental conditions. The histochemical assay showed that there is general production of lipids, essential oils and tannins in leaf idioblasts of neem, meanwhile the mucilage test showed negative results for the three evaluated locations. The highest concentrations of AZA (mean 0.2470 mg g-1) were produced in the first and second cultivation week in the presence of these three substances, demonstrating the synergistic effect when the cell mass was cultured in medium supplemented with these. Calli showed high propagation capacity by both regenerative routes organogenesis and embryogenesis starting from the fourth subculture in WPM medium presenting the highest shooting and somatic embryos germination rates. Calli went from a smooth to granular appearance during the subcultures, while color went from dark brown to light brown and white. From the seventh subculture, there was dramatic reduction of in vitro regeneration rate for both types of neem cotyledonary callus. Therefore, this work has contributed for a better understanding of biotechnology techniques aiming the in vitro propagation, cell cultivation for AZA production and anatomical aspects of A. indica.
publishDate 2009
dc.date.available.fl_str_mv 2009-12-21
2015-03-26T13:08:46Z
dc.date.issued.fl_str_mv 2009-07-16
dc.date.accessioned.fl_str_mv 2015-03-26T13:08:46Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.citation.fl_str_mv RODRIGUES, Marcelo. Morphogenesis in vitro, phytochemical analysis and anatomical characterization of neem (Azadirachta indica A. Juss). 2009. 89 f. Dissertação (Mestrado em Botânica estrutural; Ecologia e Sistemática) - Universidade Federal de Viçosa, Viçosa, 2009.
dc.identifier.uri.fl_str_mv http://locus.ufv.br/handle/123456789/2516
identifier_str_mv RODRIGUES, Marcelo. Morphogenesis in vitro, phytochemical analysis and anatomical characterization of neem (Azadirachta indica A. Juss). 2009. 89 f. Dissertação (Mestrado em Botânica estrutural; Ecologia e Sistemática) - Universidade Federal de Viçosa, Viçosa, 2009.
url http://locus.ufv.br/handle/123456789/2516
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Viçosa
dc.publisher.program.fl_str_mv Mestrado em Botânica
dc.publisher.initials.fl_str_mv UFV
dc.publisher.country.fl_str_mv BR
dc.publisher.department.fl_str_mv Botânica estrutural; Ecologia e Sistemática
publisher.none.fl_str_mv Universidade Federal de Viçosa
dc.source.none.fl_str_mv reponame:LOCUS Repositório Institucional da UFV
instname:Universidade Federal de Viçosa (UFV)
instacron:UFV
instname_str Universidade Federal de Viçosa (UFV)
instacron_str UFV
institution UFV
reponame_str LOCUS Repositório Institucional da UFV
collection LOCUS Repositório Institucional da UFV
bitstream.url.fl_str_mv https://locus.ufv.br//bitstream/123456789/2516/1/texto%20completo.pdf
https://locus.ufv.br//bitstream/123456789/2516/2/texto%20completo.pdf.txt
https://locus.ufv.br//bitstream/123456789/2516/3/texto%20completo.pdf.jpg
bitstream.checksum.fl_str_mv 4414f0ab2040a7fa5bbcc05b94aceb85
04497190af906ee6ba126f2e121f020c
c7c35751984cc479c079de853db2e9c0
bitstream.checksumAlgorithm.fl_str_mv MD5
MD5
MD5
repository.name.fl_str_mv LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)
repository.mail.fl_str_mv fabiojreis@ufv.br
_version_ 1801212893838966784