Expressão do gene ASYMMETRIC LEAVES1 (AS1) em explantes radiculares de Passiflora edulis Sims e organogênese in vitro em Passiflora setacea D.C. (Passifloraceae)
Autor(a) principal: | |
---|---|
Data de Publicação: | 2011 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | LOCUS Repositório Institucional da UFV |
Texto Completo: | http://locus.ufv.br/handle/123456789/2526 |
Resumo: | This study aimed to characterize the expression of ASYMMETRIC LEAVES1 (PeAS1) gene via in situ hybridization, during in vitro regeneration from root segments of Passiflora edulis Sims, a species typically commercial, and establish a protocol which could be repeated, via organogenesis in vitro in Passiflora setacea, a wild species. Root explants from P. edulis were cultured on MS medium for organogenesis induction in branches, supplemented with 2.35 mM BA. For PeAS1 gene detection by in situ hybridization it was produced an antisense probe, using cDNA fragments that had 900 pb. The visualization of the hybridization signal was obtained using a commercial solution of NBT / BCIP as a substrate for alkaline phosphatase. After 12 days on induction medium, meristems were formed via direct organogenesis. In this group of cells, the immunodetection positive reaction on in situ hybridization showed the presence of gene transcripts PeAS1. In buds that has its sources in meristems, PeAS1 gene was expressed in dome and leaf primordia regions, after 25 days of cultivation. For induction of organogenesis in vitro in P. setacea, leaf, hypocotyl and root explants were cultured on MS medium supplemented with 4.44 mM benzylaminopurine (BA), 2.27 mM thidiazuron (TDZ) or in combination with BA (4.44 mM) + TDZ (2. 27 mM) under two distinct light conditions (16h light and dark). For the control treatment, there was no addition of plant growth regulator. Patterns of direct and indirect regeneration were observed in hypocotyls and root. Leaf explants, regeneration only occurred from callus. In hypocotyls, the media supplemented with BA at light condition or BA + TDZ in both incubation conditions provided a higher average of shoots. In root explants, the highest frequency of regeneration was observed when it was cultured in BA + TDZ at light medium. However, under these conditions, the analysis of flow cytometry showed an increase in the average amount of nuclear DNA, in two of five regenerants analyzed, when compared to material derived from seeds. The same medium composition and incubation conditions did not induce polyploidy in regenerating from hypocotyls. It is concluded that the explant type, composition of culture medium and incubation conditions had influence on in vitro morphogenesis of Passiflora setacea. This paper describes, for the first time, information on the molecular aspects of organogenic process from root explants in P. edulis, as well obtaining organogenesis in vitro and characterization of the genetic stability of regenerants from P. setacea. These advances enable the employment of this wild species in breeding programs. |
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Vieira, Lorena Melohttp://lattes.cnpq.br/2835038295258914Silva, Luzimar Campos dahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4799707J8Otoni, Wagner Camposhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4786133Y6Dias, Leonardo Lucas Carnevallihttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4764363H52015-03-26T13:08:48Z2011-12-072015-03-26T13:08:48Z2011-02-16VIEIRA, Lorena Melo. Expression of the gene ASYMMETRIC LEAVES1 (AS1) in root explants from Passiflora edulis Sims and organogenesis in vitro in Passiflora setacea D.C. (Passifloraceae). 2011. 71 f. Dissertação (Mestrado em Botânica estrutural; Ecologia e Sistemática) - Universidade Federal de Viçosa, Viçosa, 2011.http://locus.ufv.br/handle/123456789/2526This study aimed to characterize the expression of ASYMMETRIC LEAVES1 (PeAS1) gene via in situ hybridization, during in vitro regeneration from root segments of Passiflora edulis Sims, a species typically commercial, and establish a protocol which could be repeated, via organogenesis in vitro in Passiflora setacea, a wild species. Root explants from P. edulis were cultured on MS medium for organogenesis induction in branches, supplemented with 2.35 mM BA. For PeAS1 gene detection by in situ hybridization it was produced an antisense probe, using cDNA fragments that had 900 pb. The visualization of the hybridization signal was obtained using a commercial solution of NBT / BCIP as a substrate for alkaline phosphatase. After 12 days on induction medium, meristems were formed via direct organogenesis. In this group of cells, the immunodetection positive reaction on in situ hybridization showed the presence of gene transcripts PeAS1. In buds that has its sources in meristems, PeAS1 gene was expressed in dome and leaf primordia regions, after 25 days of cultivation. For induction of organogenesis in vitro in P. setacea, leaf, hypocotyl and root explants were cultured on MS medium supplemented with 4.44 mM benzylaminopurine (BA), 2.27 mM thidiazuron (TDZ) or in combination with BA (4.44 mM) + TDZ (2. 27 mM) under two distinct light conditions (16h light and dark). For the control treatment, there was no addition of plant growth regulator. Patterns of direct and indirect regeneration were observed in hypocotyls and root. Leaf explants, regeneration only occurred from callus. In hypocotyls, the media supplemented with BA at light condition or BA + TDZ in both incubation conditions provided a higher average of shoots. In root explants, the highest frequency of regeneration was observed when it was cultured in BA + TDZ at light medium. However, under these conditions, the analysis of flow cytometry showed an increase in the average amount of nuclear DNA, in two of five regenerants analyzed, when compared to material derived from seeds. The same medium composition and incubation conditions did not induce polyploidy in regenerating from hypocotyls. It is concluded that the explant type, composition of culture medium and incubation conditions had influence on in vitro morphogenesis of Passiflora setacea. This paper describes, for the first time, information on the molecular aspects of organogenic process from root explants in P. edulis, as well obtaining organogenesis in vitro and characterization of the genetic stability of regenerants from P. setacea. These advances enable the employment of this wild species in breeding programs.O presente estudo teve como objetivos caracterizar a expressão do gene ASYMMETRIC LEAVES1 (AS1), via hibridização in situ, durante a regeneração in vitro de segmentos radiculares de Passiflora edulis Sims, espécie comercialmente cultivada, e estabelecer um protocolo reproduzível via organogênese in vitro em Passiflora setacea, espécie silvestre. Explantes radiculares de P. edulis foram cultivados em meio MS, para indução de organogênese de ramos, suplementado com 2,35 μM de BA. Para detecção do gene PeAS1 pela hibridização in situ foi produzido uma sonda antisense, utilizando-se fragmentos de cDNA com 900 pb. A visualização do sinal de hibridização foi obtida utilizando-se a solução comercial de NBT/BCIP como substrato para fosfatase alcalina. Após 12 dias em meio de indução, meristemóides foram formados, via organogênese direta. Nos meristemóides, a reação positiva a hibridização in situ por imunodetecção evidenciou a presença de transcritos do gene PeAS1. Em gemas formadas a partir dos meristemóides, após 25 dias de cultivo, o gene PeAS1 foi expresso nas regiões do domo e primórdios foliares. Para a indução de organogênese in vitro em P. setacea, explantes foliares, hipocotiledonares e radiculares foram cultivados em meio MS suplementado com 4,44 μM de Benzilaminopurina (BA); 2,27 μM de Tidiazuron (TDZ) ou em combinação com BA (4,44 μM) + TDZ (2,27 μM), sob dois regimes luminosos (16h de luz e escuro). No tratamento controle, não houve adição de fitorregulador. Os padrões de regeneração direta e indireta foram observados em explantes hipocotiledonares e radiculares. Em explantes foliares, a regeneração ocorreu somente a partir de calos. Nos hipocótilos, os meios suplementados por BA na luz ou BA+TDZ em ambas as condições de incubação proporcionaram maior média de brotações. Em explantes radiculares, a maior frequência de regeneração foi observada quando cultivados em meio com BA+TDZ na luz porém, nessas condições, a análise de citometria de fluxo constatou aumento na quantidade média de DNA nuclear, em dois de cinco regenerantes analisados, quando comparado ao material oriundo de sementes. A mesma composição do meio e condição de incubação não induziu poliploidia em regenerantes oriundos de explantes hipocotiledonares. Conclui-se que o tipo de explante, a composição do meio de cultivo e as condições de incubação tiveram influência na morfogênese in vitro de Passiflora setacea. O presente trabalho relata, pela primeira vez, informações de nível molecular do processo organogênico a partir de explantes radiculares em P. edulis, bem como a obtenção de organogênese in vitro e a caracterização da estabilidade genética de regenerantes da espécie silvestre P. setacea, possibilitando a sua utilização em programas de melhoramento genético.Conselho Nacional de Desenvolvimento Científico e Tecnológicoapplication/pdfporUniversidade Federal de ViçosaMestrado em BotânicaUFVBRBotânica estrutural; Ecologia e SistemáticaPassifloraceaeASYMMETRIC LEAVES1Organogênese in vitroCitometria de fluxoPassifloraceaeASYMMETRIC LEAVES1Organogenesis in vitroFlow cytometryCNPQ::CIENCIAS BIOLOGICAS::BOTANICAExpressão do gene ASYMMETRIC LEAVES1 (AS1) em explantes radiculares de Passiflora edulis Sims e organogênese in vitro em Passiflora setacea D.C. (Passifloraceae)Expression of the gene ASYMMETRIC LEAVES1 (AS1) in root explants from Passiflora edulis Sims and organogenesis in vitro in Passiflora setacea D.C. (Passifloraceae)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/pdf2736409https://locus.ufv.br//bitstream/123456789/2526/1/texto%20completo.pdf91157fe9b4d50b90a1ab1f26969eeffaMD51TEXTtexto completo.pdf.txttexto completo.pdf.txtExtracted texttext/plain157150https://locus.ufv.br//bitstream/123456789/2526/2/texto%20completo.pdf.txt1c81aa139b19996472888d521a4d2955MD52THUMBNAILtexto completo.pdf.jpgtexto completo.pdf.jpgIM Thumbnailimage/jpeg3660https://locus.ufv.br//bitstream/123456789/2526/3/texto%20completo.pdf.jpg9f97e664694745baaba2b7132fa63491MD53123456789/25262016-04-08 23:07:44.822oai:locus.ufv.br:123456789/2526Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-09T02:07:44LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false |
dc.title.por.fl_str_mv |
Expressão do gene ASYMMETRIC LEAVES1 (AS1) em explantes radiculares de Passiflora edulis Sims e organogênese in vitro em Passiflora setacea D.C. (Passifloraceae) |
dc.title.alternative.eng.fl_str_mv |
Expression of the gene ASYMMETRIC LEAVES1 (AS1) in root explants from Passiflora edulis Sims and organogenesis in vitro in Passiflora setacea D.C. (Passifloraceae) |
title |
Expressão do gene ASYMMETRIC LEAVES1 (AS1) em explantes radiculares de Passiflora edulis Sims e organogênese in vitro em Passiflora setacea D.C. (Passifloraceae) |
spellingShingle |
Expressão do gene ASYMMETRIC LEAVES1 (AS1) em explantes radiculares de Passiflora edulis Sims e organogênese in vitro em Passiflora setacea D.C. (Passifloraceae) Vieira, Lorena Melo Passifloraceae ASYMMETRIC LEAVES1 Organogênese in vitro Citometria de fluxo Passifloraceae ASYMMETRIC LEAVES1 Organogenesis in vitro Flow cytometry CNPQ::CIENCIAS BIOLOGICAS::BOTANICA |
title_short |
Expressão do gene ASYMMETRIC LEAVES1 (AS1) em explantes radiculares de Passiflora edulis Sims e organogênese in vitro em Passiflora setacea D.C. (Passifloraceae) |
title_full |
Expressão do gene ASYMMETRIC LEAVES1 (AS1) em explantes radiculares de Passiflora edulis Sims e organogênese in vitro em Passiflora setacea D.C. (Passifloraceae) |
title_fullStr |
Expressão do gene ASYMMETRIC LEAVES1 (AS1) em explantes radiculares de Passiflora edulis Sims e organogênese in vitro em Passiflora setacea D.C. (Passifloraceae) |
title_full_unstemmed |
Expressão do gene ASYMMETRIC LEAVES1 (AS1) em explantes radiculares de Passiflora edulis Sims e organogênese in vitro em Passiflora setacea D.C. (Passifloraceae) |
title_sort |
Expressão do gene ASYMMETRIC LEAVES1 (AS1) em explantes radiculares de Passiflora edulis Sims e organogênese in vitro em Passiflora setacea D.C. (Passifloraceae) |
author |
Vieira, Lorena Melo |
author_facet |
Vieira, Lorena Melo |
author_role |
author |
dc.contributor.authorLattes.por.fl_str_mv |
http://lattes.cnpq.br/2835038295258914 |
dc.contributor.author.fl_str_mv |
Vieira, Lorena Melo |
dc.contributor.advisor-co1.fl_str_mv |
Silva, Luzimar Campos da |
dc.contributor.advisor-co1Lattes.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 |
Dias, Leonardo Lucas Carnevalli |
dc.contributor.referee1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4764363H5 |
contributor_str_mv |
Silva, Luzimar Campos da Otoni, Wagner Campos Dias, Leonardo Lucas Carnevalli |
dc.subject.por.fl_str_mv |
Passifloraceae ASYMMETRIC LEAVES1 Organogênese in vitro Citometria de fluxo |
topic |
Passifloraceae ASYMMETRIC LEAVES1 Organogênese in vitro Citometria de fluxo Passifloraceae ASYMMETRIC LEAVES1 Organogenesis in vitro Flow cytometry CNPQ::CIENCIAS BIOLOGICAS::BOTANICA |
dc.subject.eng.fl_str_mv |
Passifloraceae ASYMMETRIC LEAVES1 Organogenesis in vitro Flow cytometry |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS BIOLOGICAS::BOTANICA |
description |
This study aimed to characterize the expression of ASYMMETRIC LEAVES1 (PeAS1) gene via in situ hybridization, during in vitro regeneration from root segments of Passiflora edulis Sims, a species typically commercial, and establish a protocol which could be repeated, via organogenesis in vitro in Passiflora setacea, a wild species. Root explants from P. edulis were cultured on MS medium for organogenesis induction in branches, supplemented with 2.35 mM BA. For PeAS1 gene detection by in situ hybridization it was produced an antisense probe, using cDNA fragments that had 900 pb. The visualization of the hybridization signal was obtained using a commercial solution of NBT / BCIP as a substrate for alkaline phosphatase. After 12 days on induction medium, meristems were formed via direct organogenesis. In this group of cells, the immunodetection positive reaction on in situ hybridization showed the presence of gene transcripts PeAS1. In buds that has its sources in meristems, PeAS1 gene was expressed in dome and leaf primordia regions, after 25 days of cultivation. For induction of organogenesis in vitro in P. setacea, leaf, hypocotyl and root explants were cultured on MS medium supplemented with 4.44 mM benzylaminopurine (BA), 2.27 mM thidiazuron (TDZ) or in combination with BA (4.44 mM) + TDZ (2. 27 mM) under two distinct light conditions (16h light and dark). For the control treatment, there was no addition of plant growth regulator. Patterns of direct and indirect regeneration were observed in hypocotyls and root. Leaf explants, regeneration only occurred from callus. In hypocotyls, the media supplemented with BA at light condition or BA + TDZ in both incubation conditions provided a higher average of shoots. In root explants, the highest frequency of regeneration was observed when it was cultured in BA + TDZ at light medium. However, under these conditions, the analysis of flow cytometry showed an increase in the average amount of nuclear DNA, in two of five regenerants analyzed, when compared to material derived from seeds. The same medium composition and incubation conditions did not induce polyploidy in regenerating from hypocotyls. It is concluded that the explant type, composition of culture medium and incubation conditions had influence on in vitro morphogenesis of Passiflora setacea. This paper describes, for the first time, information on the molecular aspects of organogenic process from root explants in P. edulis, as well obtaining organogenesis in vitro and characterization of the genetic stability of regenerants from P. setacea. These advances enable the employment of this wild species in breeding programs. |
publishDate |
2011 |
dc.date.available.fl_str_mv |
2011-12-07 2015-03-26T13:08:48Z |
dc.date.issued.fl_str_mv |
2011-02-16 |
dc.date.accessioned.fl_str_mv |
2015-03-26T13:08:48Z |
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 |
VIEIRA, Lorena Melo. Expression of the gene ASYMMETRIC LEAVES1 (AS1) in root explants from Passiflora edulis Sims and organogenesis in vitro in Passiflora setacea D.C. (Passifloraceae). 2011. 71 f. Dissertação (Mestrado em Botânica estrutural; Ecologia e Sistemática) - Universidade Federal de Viçosa, Viçosa, 2011. |
dc.identifier.uri.fl_str_mv |
http://locus.ufv.br/handle/123456789/2526 |
identifier_str_mv |
VIEIRA, Lorena Melo. Expression of the gene ASYMMETRIC LEAVES1 (AS1) in root explants from Passiflora edulis Sims and organogenesis in vitro in Passiflora setacea D.C. (Passifloraceae). 2011. 71 f. Dissertação (Mestrado em Botânica estrutural; Ecologia e Sistemática) - Universidade Federal de Viçosa, Viçosa, 2011. |
url |
http://locus.ufv.br/handle/123456789/2526 |
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por |
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por |
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Universidade Federal de Viçosa |
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Mestrado em Botânica |
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UFV |
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BR |
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Botânica estrutural; Ecologia e Sistemática |
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Universidade Federal de Viçosa |
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