Identificação e diversidade genética de rotylenchulus reniformis em áreas de cultivos de meloeiro (cucumis melo l.)
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Digital da Universidade Federal Rural do Semi-Árido (RDU) |
Texto Completo: | https://repositorio.ufersa.edu.br/handle/prefix/847 |
Resumo: | ABSTRACT This research aimed to identify through microscopic and molecular analyzes the presence of the nematode Rotylenchulus reniformis in different melon producing areas and verify the genetic diversity. Soil samples were collected from five farms in the state of Rio Grande do Norte (Dinamarca, Brazil Melon, CY Matsumoto, Itaueira and Agrícola Famosa) and one from Ceará (Mata Fresca). Thirty soil samples were collected in different areas of each farm, which were taken to greenhouse and sown with Vedrantais melon. After 35 days of sowing, the plants were collected, and their roots submitted to the staining process for microscopic identification of R. reniformis. The DNA of the roots of the melon was extracted aiming the molecular analyzes with the universal primers for nematodes and genes specific for R. reniformis. For the genetic diversity study, R. reniformis was isolated from suspensions of soil samples under an optical microscope. Then the DNA from them was extracted through the NucleoSpin Tissue (Macherey-Nagel) ® extraction kit. DNA analyzes were performed from the screening of 20 RAPD primers. The data generated were analyzed by the Genes program. Of the 22 root samples evaluated microscopically, only three showed the presence of R. reniformis females. Regarding the molecular analyzes with the universal primer, 15 areas belonging to five farms were positive for the presence of nematode DNA. The analyzes with specific genes primers revealed that six areas, restricted to two farms (Itaueira and Mata Fresca), were confirmed with the presence of R. reniformis. Only nine of the 20 RAPD primers were selected for diversity analysis, which generated 67 polymorphic bands (95.71%) and three polymorphic bands (4.29%), thus evidencing a high polymorphism, with an average of 7.4 bands polymorphic by primer. The primer with the highest number of polymorphic bands was OPAA-04 with 11 bands whereas OPA-03 was the primer with the lowest number (5). A dendrogram was constructed using the UPGMA hierarchical method. Using a cutoff of 0.55, two groups were found. The first one, was formed by the farms Itaueira and Mata Fresca and the second group by the other farms. The estimates of cophenetic correlation, stress and distortion indicated that the grouping was efficient and with little loss of information. For the dendrogram obtained from 51 R. reniformis samples, the formation of six groups was observed. The largest group formed was V, contemplating 34 samples, approximately 66.7%. The cofenética correlation was estimated at 0.77%. It was found that the study did not indicate any association between the magnitudes of dissimilarities and the origin of the samples. It was verified that the largest (0.92) and the lowest (0.27) value of similarity was found among samples belonging to Fazenda Itaueira. The study indicates that there is variability between samples with a mean dissimilarity coefficient of 0.63 and a coefficient of variation of 14.88%. This is the first genetic variability study with molecular markers performed with samples from the production fields of the main melon exporting region in Brazil |
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Identificação e diversidade genética de rotylenchulus reniformis em áreas de cultivos de meloeiro (cucumis melo l.)NematoideRAPDVariabilidade genéticaNematodeRAPDGenetic variabilityCNPQ::CIENCIAS AMBIENTAIS::MEIO AMBIENTEABSTRACT This research aimed to identify through microscopic and molecular analyzes the presence of the nematode Rotylenchulus reniformis in different melon producing areas and verify the genetic diversity. Soil samples were collected from five farms in the state of Rio Grande do Norte (Dinamarca, Brazil Melon, CY Matsumoto, Itaueira and Agrícola Famosa) and one from Ceará (Mata Fresca). Thirty soil samples were collected in different areas of each farm, which were taken to greenhouse and sown with Vedrantais melon. After 35 days of sowing, the plants were collected, and their roots submitted to the staining process for microscopic identification of R. reniformis. The DNA of the roots of the melon was extracted aiming the molecular analyzes with the universal primers for nematodes and genes specific for R. reniformis. For the genetic diversity study, R. reniformis was isolated from suspensions of soil samples under an optical microscope. Then the DNA from them was extracted through the NucleoSpin Tissue (Macherey-Nagel) ® extraction kit. DNA analyzes were performed from the screening of 20 RAPD primers. The data generated were analyzed by the Genes program. Of the 22 root samples evaluated microscopically, only three showed the presence of R. reniformis females. Regarding the molecular analyzes with the universal primer, 15 areas belonging to five farms were positive for the presence of nematode DNA. The analyzes with specific genes primers revealed that six areas, restricted to two farms (Itaueira and Mata Fresca), were confirmed with the presence of R. reniformis. Only nine of the 20 RAPD primers were selected for diversity analysis, which generated 67 polymorphic bands (95.71%) and three polymorphic bands (4.29%), thus evidencing a high polymorphism, with an average of 7.4 bands polymorphic by primer. The primer with the highest number of polymorphic bands was OPAA-04 with 11 bands whereas OPA-03 was the primer with the lowest number (5). A dendrogram was constructed using the UPGMA hierarchical method. Using a cutoff of 0.55, two groups were found. The first one, was formed by the farms Itaueira and Mata Fresca and the second group by the other farms. The estimates of cophenetic correlation, stress and distortion indicated that the grouping was efficient and with little loss of information. For the dendrogram obtained from 51 R. reniformis samples, the formation of six groups was observed. The largest group formed was V, contemplating 34 samples, approximately 66.7%. The cofenética correlation was estimated at 0.77%. It was found that the study did not indicate any association between the magnitudes of dissimilarities and the origin of the samples. It was verified that the largest (0.92) and the lowest (0.27) value of similarity was found among samples belonging to Fazenda Itaueira. The study indicates that there is variability between samples with a mean dissimilarity coefficient of 0.63 and a coefficient of variation of 14.88%. This is the first genetic variability study with molecular markers performed with samples from the production fields of the main melon exporting region in BrazilEste trabalho objetivou identificar através de análises microscópicas e moleculares a presença do nematoide Rotylenchulus reniformis em diferentes áreas produtoras de melão e verificar a diversidade genética. Foram coletadas 30 amostras de solos em diferentes áreas de cada fazenda, são elas: cinco do estado do Rio Grande do Norte (Dinamarca, Brazil Melon, CY Matsumoto, Itaueira e Agrícola Famosa) e uma do Ceará (Mata Fresca). Onde, as amostras foram levadas para casa de vegetação e semeadas com a cultivar de melão Vedrantais. Após 35 dias da semeadura, as plantas foram coletadas e suas raízes submetidas ao processo de coloração para identificação microscópica de R. reniformis. O DNA das raízes do meloeiro foi extraído visando as análises moleculares com os primers universais para nematoides e genes específicos para R. reniformis. Para o estudo da diversidade genética, R. reniformis foi isolado de suspensões de amostras do solo em microscópio óptico. Em seguida o DNA dos mesmos foi extraído através do kit de extração NucleoSpin Tissue (Macherey-Nagel)®. As análises do DNA foram realizadas a partir da triagem de 20 iniciadores RAPD. Os dados gerados foram analisados pelo programa Genes. Das 22 amostras de raízes avaliadas microscopicamente, apenas três revelaram a presença de fêmeas de R. reniformis. No que tange as análises moleculares com o primer universal, 15 áreas pertencentes a cinco fazendas foram positivas para a presença do DNA de nematoides. Já as análises com primers genes específicos revelaram que seis áreas, restritas a duas fazendas (Itaueira e Mata Fresca), foram confirmadas com a presença de R. reniformis. Apenas nove dos 20 iniciadores RAPD foram selecionados para a análise de diversidade, os quais geraram 67 bandas polimórficas (95,71 %) e três monomórficas (4,29%), evidenciando assim um alto polimorfismo, perfazendo uma média de 7,4 bandas polimórficas por iniciador. O iniciador com maior número de bandas polimórficas foi o OPAA-04 com 11 bandas enquanto o OPA-03 foi o iniciador que apresentou o menor número (5). Um dendrograma foi construído pelo método hierárquico UPGMA. Utilizando um ponto de corte de 0,55, verificou-se dois grupos. O primeiro formado pelas fazendas Iraueira e Mata Fresca e o segundo grupo pelas demais fazendas. As estimativas de correlação cofenética, estresse e distorção indicaram que o agrupamento foi eficiente e com pequena perda de informação. Já para o dendrograma obtido a partir de 51 amostras de R. reniformis, observou-se a formação de seis grupos. O maior grupo formado foi o V, contemplando 34 amostras, aproximadamente 66,7%. A correlação cofenética foi estimada em 0,77%. Verificou-se que o estudo não indicou nenhuma associação entre as magnitudes das dissimilaridades e a origem das amostras. Verificou-se que o maior (0,92) e o menor (0,27) valor de similaridade encontrado foi entre amostras pertencentes à Fazenda Itaueira. O estudo indica que há variabilidade entre as amostras com um coeficiente médio de dissimilaridade de 0,63 e um coeficiente de variação 14,88%. Este é o primeiro estudo da variabilidade genética com marcadores moleculares realizado com amostras oriundas de campos de produção da principal região exportadora de melão no BrasilCoordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESUniversidade Federal Rural do Semi-ÁridoBrasilCentro de Ciências Agrárias - CCAUFERSAPrograma de Pós-Graduação em Ambiente, Tecnologia e SociedadeNunes, Glauber Henrique de Sousa82888884453http://lattes.cnpq.br/6199153570472953Holanda, Ioná Santos Araújo65180062500http://lattes.cnpq.br/2079609001869001Ambrósio, Márcia Michelle de Queiroz96726539487http://lattes.cnpq.br/5986626995999424Bordallo, Patrícia do Nascimento79393977704http://lattes.cnpq.br/4047141046815032Holanda, Ioná Santos Araújo65180062500http://lattes.cnpq.br/2079609001869001Barreto, Alcileide Vieira2018-11-08T21:36:22Z2018-05-042018-11-08T21:36:22Z2018-02-05info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdf(BARRETO, 2018)https://repositorio.ufersa.edu.br/handle/prefix/847porBARRETO, Alcileide Vieira. Identificação e diversidade genética de rotylenchulus reniformis em áreas de cultivos de meloeiro (cucumis melo l.). 2018. 56 f. Dissertação (Mestrado em Ambiente, Tecnologia e Sociedade), Universidade Federal Rural do Semi-Árido, Mossoró, 2018.CC-BY-SAinfo:eu-repo/semantics/openAccessreponame:Repositório Digital da Universidade Federal Rural do Semi-Árido (RDU)instname:Universidade Federal Rural do Semi-Árido (UFERSA)instacron:UFERSA2023-09-14T04:32:28Zoai:repositorio.ufersa.edu.br:prefix/847Repositório Institucionalhttps://repositorio.ufersa.edu.br/PUBhttps://repositorio.ufersa.edu.br/server/oai/requestrepositorio@ufersa.edu.br || admrepositorio@ufersa.edu.bropendoar:2023-09-14T04:32:28Repositório Digital da Universidade Federal Rural do Semi-Árido (RDU) - Universidade Federal Rural do Semi-Árido (UFERSA)false |
dc.title.none.fl_str_mv |
Identificação e diversidade genética de rotylenchulus reniformis em áreas de cultivos de meloeiro (cucumis melo l.) |
title |
Identificação e diversidade genética de rotylenchulus reniformis em áreas de cultivos de meloeiro (cucumis melo l.) |
spellingShingle |
Identificação e diversidade genética de rotylenchulus reniformis em áreas de cultivos de meloeiro (cucumis melo l.) Barreto, Alcileide Vieira Nematoide RAPD Variabilidade genética Nematode RAPD Genetic variability CNPQ::CIENCIAS AMBIENTAIS::MEIO AMBIENTE |
title_short |
Identificação e diversidade genética de rotylenchulus reniformis em áreas de cultivos de meloeiro (cucumis melo l.) |
title_full |
Identificação e diversidade genética de rotylenchulus reniformis em áreas de cultivos de meloeiro (cucumis melo l.) |
title_fullStr |
Identificação e diversidade genética de rotylenchulus reniformis em áreas de cultivos de meloeiro (cucumis melo l.) |
title_full_unstemmed |
Identificação e diversidade genética de rotylenchulus reniformis em áreas de cultivos de meloeiro (cucumis melo l.) |
title_sort |
Identificação e diversidade genética de rotylenchulus reniformis em áreas de cultivos de meloeiro (cucumis melo l.) |
author |
Barreto, Alcileide Vieira |
author_facet |
Barreto, Alcileide Vieira |
author_role |
author |
dc.contributor.none.fl_str_mv |
Nunes, Glauber Henrique de Sousa 82888884453 http://lattes.cnpq.br/6199153570472953 Holanda, Ioná Santos Araújo 65180062500 http://lattes.cnpq.br/2079609001869001 Ambrósio, Márcia Michelle de Queiroz 96726539487 http://lattes.cnpq.br/5986626995999424 Bordallo, Patrícia do Nascimento 79393977704 http://lattes.cnpq.br/4047141046815032 Holanda, Ioná Santos Araújo 65180062500 http://lattes.cnpq.br/2079609001869001 |
dc.contributor.author.fl_str_mv |
Barreto, Alcileide Vieira |
dc.subject.por.fl_str_mv |
Nematoide RAPD Variabilidade genética Nematode RAPD Genetic variability CNPQ::CIENCIAS AMBIENTAIS::MEIO AMBIENTE |
topic |
Nematoide RAPD Variabilidade genética Nematode RAPD Genetic variability CNPQ::CIENCIAS AMBIENTAIS::MEIO AMBIENTE |
description |
ABSTRACT This research aimed to identify through microscopic and molecular analyzes the presence of the nematode Rotylenchulus reniformis in different melon producing areas and verify the genetic diversity. Soil samples were collected from five farms in the state of Rio Grande do Norte (Dinamarca, Brazil Melon, CY Matsumoto, Itaueira and Agrícola Famosa) and one from Ceará (Mata Fresca). Thirty soil samples were collected in different areas of each farm, which were taken to greenhouse and sown with Vedrantais melon. After 35 days of sowing, the plants were collected, and their roots submitted to the staining process for microscopic identification of R. reniformis. The DNA of the roots of the melon was extracted aiming the molecular analyzes with the universal primers for nematodes and genes specific for R. reniformis. For the genetic diversity study, R. reniformis was isolated from suspensions of soil samples under an optical microscope. Then the DNA from them was extracted through the NucleoSpin Tissue (Macherey-Nagel) ® extraction kit. DNA analyzes were performed from the screening of 20 RAPD primers. The data generated were analyzed by the Genes program. Of the 22 root samples evaluated microscopically, only three showed the presence of R. reniformis females. Regarding the molecular analyzes with the universal primer, 15 areas belonging to five farms were positive for the presence of nematode DNA. The analyzes with specific genes primers revealed that six areas, restricted to two farms (Itaueira and Mata Fresca), were confirmed with the presence of R. reniformis. Only nine of the 20 RAPD primers were selected for diversity analysis, which generated 67 polymorphic bands (95.71%) and three polymorphic bands (4.29%), thus evidencing a high polymorphism, with an average of 7.4 bands polymorphic by primer. The primer with the highest number of polymorphic bands was OPAA-04 with 11 bands whereas OPA-03 was the primer with the lowest number (5). A dendrogram was constructed using the UPGMA hierarchical method. Using a cutoff of 0.55, two groups were found. The first one, was formed by the farms Itaueira and Mata Fresca and the second group by the other farms. The estimates of cophenetic correlation, stress and distortion indicated that the grouping was efficient and with little loss of information. For the dendrogram obtained from 51 R. reniformis samples, the formation of six groups was observed. The largest group formed was V, contemplating 34 samples, approximately 66.7%. The cofenética correlation was estimated at 0.77%. It was found that the study did not indicate any association between the magnitudes of dissimilarities and the origin of the samples. It was verified that the largest (0.92) and the lowest (0.27) value of similarity was found among samples belonging to Fazenda Itaueira. The study indicates that there is variability between samples with a mean dissimilarity coefficient of 0.63 and a coefficient of variation of 14.88%. This is the first genetic variability study with molecular markers performed with samples from the production fields of the main melon exporting region in Brazil |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-11-08T21:36:22Z 2018-05-04 2018-11-08T21:36:22Z 2018-02-05 |
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.uri.fl_str_mv |
(BARRETO, 2018) https://repositorio.ufersa.edu.br/handle/prefix/847 |
identifier_str_mv |
(BARRETO, 2018) |
url |
https://repositorio.ufersa.edu.br/handle/prefix/847 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.none.fl_str_mv |
BARRETO, Alcileide Vieira. Identificação e diversidade genética de rotylenchulus reniformis em áreas de cultivos de meloeiro (cucumis melo l.). 2018. 56 f. Dissertação (Mestrado em Ambiente, Tecnologia e Sociedade), Universidade Federal Rural do Semi-Árido, Mossoró, 2018. |
dc.rights.driver.fl_str_mv |
CC-BY-SA info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
CC-BY-SA |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal Rural do Semi-Árido Brasil Centro de Ciências Agrárias - CCA UFERSA Programa de Pós-Graduação em Ambiente, Tecnologia e Sociedade |
publisher.none.fl_str_mv |
Universidade Federal Rural do Semi-Árido Brasil Centro de Ciências Agrárias - CCA UFERSA Programa de Pós-Graduação em Ambiente, Tecnologia e Sociedade |
dc.source.none.fl_str_mv |
reponame:Repositório Digital da Universidade Federal Rural do Semi-Árido (RDU) instname:Universidade Federal Rural do Semi-Árido (UFERSA) instacron:UFERSA |
instname_str |
Universidade Federal Rural do Semi-Árido (UFERSA) |
instacron_str |
UFERSA |
institution |
UFERSA |
reponame_str |
Repositório Digital da Universidade Federal Rural do Semi-Árido (RDU) |
collection |
Repositório Digital da Universidade Federal Rural do Semi-Árido (RDU) |
repository.name.fl_str_mv |
Repositório Digital da Universidade Federal Rural do Semi-Árido (RDU) - Universidade Federal Rural do Semi-Árido (UFERSA) |
repository.mail.fl_str_mv |
repositorio@ufersa.edu.br || admrepositorio@ufersa.edu.br |
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1809747451447345152 |