Proteome analysis of developing seeds of Jatropha curcas L.

Detalhes bibliográficos
Autor(a) principal: Mohibullah Shah
Data de Publicação: 2014
Tipo de documento: Tese
Idioma: eng
Título da fonte: Biblioteca Digital de Teses e Dissertações da UFC
Texto Completo: http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=11638
Resumo: Physic nut (Jatropha curcas L.) is an important crop due to its ability of storing high content of oil in the seeds, which can serve as raw material for biodiesel production. Because of the presence of toxic constituents like phorbol esters (PEs) and curcins, the seed cake produced as a result of oil extraction cannot be utilize for animal feed. Development of the genotypes better suited for the industrial applications and biodiesel production as well as with lower level of toxic constituents is being hampered by a lack of understanding about the a) proteins related to the biosynthesis and degradation of fatty acids (FAs) and triacylglycerides (TAGs), b) role of proteins deposited during seed development and c) proteins related to the synthesis and storage of toxic compounds during seed development. Agreeing with this, we have performed the anatomical analysis of the developing seeds of J. curcas, followed by the proteome analysis of the endosperm isolated from the seeds of J. curcas at five different developmental stages, which resulted into the identification of the 1517, 1256, 1033, 752 and 307 proteins, from Stage 6, 7, 8, 9 and 10, respectively, summing up to a total of 1760 proteins. Proteins with similar expression pattern were grouped into five different clusters and protein quantification based on spectral counts was determined. Besides identification of the proteins involved in the biosynthesis and degradation of the FAs and TAGs, we also identified a large number of proteins involved in the metabolism of the carbohydrates, which are important for supplying energy and carbon source for the synthesis of TAGs in heterotrophic seeds. Among the members of different classes of seed storage proteins (SSPs), we have identified four SSPs named as nutrients reservoir, which in contrast to the other SSPs showed decreasing deposition pattern during seeds development and revealed to have special role during seed development. In addition, peptidases belong to different mechanistic classes were identified, which have a range of functions, highlighting the role in reserve mobilization during germination. Isoforms of curcin were also identified in this proteome analysis which were absent in our previous proteome analysis of the other tissues from these seeds, suggesting that the deposition of these toxic proteins only occur in the endosperm. Similarly, several enzymes involved in the biosynthesis of diterpenoid precursors were identified in this proteome analysis but, like in our previous proteome analysis of the other tissues from J. curcas seeds,we were unable to identify any terpene synthase/cyclase, enzymes responsible for the synthesis of PEs, which collectively suggesting that the synthesis of PEs may not occur in seeds of this plant. In conclusion, the strategy used here enabled us to provide a first in depth proteome analysis of the endosperm from J. curcas developing seeds, which along with providing information regarding important aspects of the seed development, also set the foundation of a proteomic approach to study biotechnologically important plant species.
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spelling info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisProteome analysis of developing seeds of Jatropha curcas L.AnÃlise proteÃmica de desenvolvimento de sementes Jatropha curcas L.2014-02-24Francisco de Assis de Paiva Campos07292082300http://lattes.cnpq.br/7845809799936552FÃbio CÃsar Sousa Nogueira66151899334http://lattes.cnpq.br/9246430154720067Gilberto Barbosa Domont03944204700http://lattes.cnpq.br/1882815704492197Jonas Enrique Aguilar Perales00953260739http://lattes.cnpq.br/5792347469122034Paulo Costa Carvalho04534254709http://lattes.cnpq.br/930073429648257770037320173http://lattes.cnpq.br/3230303572417055Mohibullah ShahUniversidade Federal do CearÃPrograma de PÃs-GraduaÃÃo em BioquÃmicaUFCBRendosperm proteome oilseeds biodiesel mass spectrometry seed development seed proteinsendosperma proteoma oleaginosas biodiesel espectrometria de massa desenvolvimento da semente proteÃnas de sementeBIOQUIMICAPhysic nut (Jatropha curcas L.) is an important crop due to its ability of storing high content of oil in the seeds, which can serve as raw material for biodiesel production. Because of the presence of toxic constituents like phorbol esters (PEs) and curcins, the seed cake produced as a result of oil extraction cannot be utilize for animal feed. Development of the genotypes better suited for the industrial applications and biodiesel production as well as with lower level of toxic constituents is being hampered by a lack of understanding about the a) proteins related to the biosynthesis and degradation of fatty acids (FAs) and triacylglycerides (TAGs), b) role of proteins deposited during seed development and c) proteins related to the synthesis and storage of toxic compounds during seed development. Agreeing with this, we have performed the anatomical analysis of the developing seeds of J. curcas, followed by the proteome analysis of the endosperm isolated from the seeds of J. curcas at five different developmental stages, which resulted into the identification of the 1517, 1256, 1033, 752 and 307 proteins, from Stage 6, 7, 8, 9 and 10, respectively, summing up to a total of 1760 proteins. Proteins with similar expression pattern were grouped into five different clusters and protein quantification based on spectral counts was determined. Besides identification of the proteins involved in the biosynthesis and degradation of the FAs and TAGs, we also identified a large number of proteins involved in the metabolism of the carbohydrates, which are important for supplying energy and carbon source for the synthesis of TAGs in heterotrophic seeds. Among the members of different classes of seed storage proteins (SSPs), we have identified four SSPs named as nutrients reservoir, which in contrast to the other SSPs showed decreasing deposition pattern during seeds development and revealed to have special role during seed development. In addition, peptidases belong to different mechanistic classes were identified, which have a range of functions, highlighting the role in reserve mobilization during germination. Isoforms of curcin were also identified in this proteome analysis which were absent in our previous proteome analysis of the other tissues from these seeds, suggesting that the deposition of these toxic proteins only occur in the endosperm. Similarly, several enzymes involved in the biosynthesis of diterpenoid precursors were identified in this proteome analysis but, like in our previous proteome analysis of the other tissues from J. curcas seeds,we were unable to identify any terpene synthase/cyclase, enzymes responsible for the synthesis of PEs, which collectively suggesting that the synthesis of PEs may not occur in seeds of this plant. In conclusion, the strategy used here enabled us to provide a first in depth proteome analysis of the endosperm from J. curcas developing seeds, which along with providing information regarding important aspects of the seed development, also set the foundation of a proteomic approach to study biotechnologically important plant species.PinhÃo manso (Jatropha curcas L.) à uma cultura importante devido à sua habilidade em armazenar alto conteÃdo de Ãleo nas sementes, as quais podem servir como matÃria-prima para a produÃÃo de biodiesel. Devido à presenÃa de constituintes tÃxicos como Ãsteres de forbol e curcina, a torta da semente produzida como resultado da extraÃÃo do Ãleo nÃo pode ser utilizada na alimentaÃÃo animal. O desenvolvimento de genÃtipos mais adequados a aplicaÃÃes industriais e à produÃÃo de biodiesel assim como apresentando baixos nÃveis de constituintes tÃxicos està sendo prejudicado pela falta de entendimento sobre a) proteÃnas relacionadas a biossÃntese e degradaÃÃo de Ãcidos graxos e triacilglicerÃis, b) o papel de proteÃnas depositadas durante o desenvolvimento da semente e c) proteÃnas relacionadas à sÃntese e reserva de compostos tÃxicos durante o desenvolvimento da semente. Diante disso, nÃs realizamos uma anÃlise anatÃmica de sementes em desenvolvimento de J. curcas, seguido por uma anÃlise proteÃmica do endosperma isolado de sementes dessa espÃcie em cinco diferentes estÃgios de desenvolvimento, o que resultou na identificaÃÃo de 1517, 1256, 1033, 752 e 307 proteÃnas, dos estÃgios 6, 7, 8, 9 e 10, respectivamente, somando um total de 1760 proteÃnas. ProteÃnas com padrÃo de expressÃo similar foram agrupadas em cinco grupos diferentes e a quantificaÃÃo das proteÃnas baseada na contagem dos espectros foi determinada. AlÃm da identificaÃÃo das proteÃnas envolvidas na biossÃntese e degradaÃÃo de FAs e TAGs, nÃs identificamos um grande nÃmero de proteÃnas envolvidas no metabolismo de carboidratos, as quais sÃo importantes para o fornecimento de energia e fontes de carbono para a sÃntese de TAGs em sementes heterotrÃficas. Entre os membros de diferentes classes de proteÃnas de reservas de sementes (SSPs), nÃs identificamos quatro SSPs denominadas reservatÃrios de sementes, que em contraste as outras SSPs mostraram decrÃscimo no padrÃo de deposiÃÃo e revelaram ter um papel especial durante o desenvolvimento da semente. Em adiÃÃo, peptidases pertencentes a diferentes classes mecanÃsticas foram identificadas destacando o papel da mobilizaÃÃo de reservas durante a germinaÃÃo. Isoformas da curcina ausentes em nossas anÃlises proteÃmicas prÃvias de outros tecidos da semente foram identificadas sugerindo que a deposiÃÃo dessas proteÃnas tÃxicas sà ocorre no endosperma. Similarmente, vÃrias enzimas envolvidas na biosÃntese de precursores de diterpenÃides foram identificadas nessa anÃlise proteÃmica, mas como em nossas prÃvias anÃlises proteÃmicas de outros tecidos de sementes de J. curcas, nÃs nÃo fomos capazes de identificar sintases/ciclases de terpenos, enzimas responsÃveis pela sÃntese de PEs, o que coletivamente sugere que a sÃntese desses compostos pode nÃo ocorrer nas sementes dessa planta. Em conclusÃo, a estratÃgia utilizada nos fornece a primeira anÃlise proteÃmica profunda do endosperma de sementes em desenvolvimento de J. curcas, o que alÃm de fornecer informaÃÃes sobre aspectos importantes do desenvolvimento da semente, tambÃm estabelece a base para uma pesquisa proteÃmica com o objetivo de estudar espÃcies vegetais importantes biotecnologicamente.Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgicohttp://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=11638application/pdfinfo:eu-repo/semantics/openAccessengreponame:Biblioteca Digital de Teses e Dissertações da UFCinstname:Universidade Federal do Cearáinstacron:UFC2019-01-21T11:24:58Zmail@mail.com -
dc.title.en.fl_str_mv Proteome analysis of developing seeds of Jatropha curcas L.
dc.title.alternative.pt.fl_str_mv AnÃlise proteÃmica de desenvolvimento de sementes Jatropha curcas L.
title Proteome analysis of developing seeds of Jatropha curcas L.
spellingShingle Proteome analysis of developing seeds of Jatropha curcas L.
Mohibullah Shah
endosperm
proteome
oilseeds
biodiesel
mass spectrometry
seed development
seed proteins
endosperma
proteoma
oleaginosas
biodiesel
espectrometria de massa
desenvolvimento da semente
proteÃnas de semente
BIOQUIMICA
title_short Proteome analysis of developing seeds of Jatropha curcas L.
title_full Proteome analysis of developing seeds of Jatropha curcas L.
title_fullStr Proteome analysis of developing seeds of Jatropha curcas L.
title_full_unstemmed Proteome analysis of developing seeds of Jatropha curcas L.
title_sort Proteome analysis of developing seeds of Jatropha curcas L.
author Mohibullah Shah
author_facet Mohibullah Shah
author_role author
dc.contributor.advisor1.fl_str_mv Francisco de Assis de Paiva Campos
dc.contributor.advisor1ID.fl_str_mv 07292082300
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/7845809799936552
dc.contributor.referee1.fl_str_mv FÃbio CÃsar Sousa Nogueira
dc.contributor.referee1ID.fl_str_mv 66151899334
dc.contributor.referee1Lattes.fl_str_mv http://lattes.cnpq.br/9246430154720067
dc.contributor.referee2.fl_str_mv Gilberto Barbosa Domont
dc.contributor.referee2ID.fl_str_mv 03944204700
dc.contributor.referee2Lattes.fl_str_mv http://lattes.cnpq.br/1882815704492197
dc.contributor.referee3.fl_str_mv Jonas Enrique Aguilar Perales
dc.contributor.referee3ID.fl_str_mv 00953260739
dc.contributor.referee3Lattes.fl_str_mv http://lattes.cnpq.br/5792347469122034
dc.contributor.referee4.fl_str_mv Paulo Costa Carvalho
dc.contributor.referee4ID.fl_str_mv 04534254709
dc.contributor.referee4Lattes.fl_str_mv http://lattes.cnpq.br/9300734296482577
dc.contributor.authorID.fl_str_mv 70037320173
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/3230303572417055
dc.contributor.author.fl_str_mv Mohibullah Shah
contributor_str_mv Francisco de Assis de Paiva Campos
FÃbio CÃsar Sousa Nogueira
Gilberto Barbosa Domont
Jonas Enrique Aguilar Perales
Paulo Costa Carvalho
dc.subject.por.fl_str_mv endosperm
proteome
oilseeds
biodiesel
mass spectrometry
seed development
seed proteins
endosperma
proteoma
oleaginosas
biodiesel
espectrometria de massa
desenvolvimento da semente
proteÃnas de semente
topic endosperm
proteome
oilseeds
biodiesel
mass spectrometry
seed development
seed proteins
endosperma
proteoma
oleaginosas
biodiesel
espectrometria de massa
desenvolvimento da semente
proteÃnas de semente
BIOQUIMICA
dc.subject.cnpq.fl_str_mv BIOQUIMICA
dc.description.sponsorship.fl_txt_mv Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico
dc.description.abstract.por.fl_txt_mv Physic nut (Jatropha curcas L.) is an important crop due to its ability of storing high content of oil in the seeds, which can serve as raw material for biodiesel production. Because of the presence of toxic constituents like phorbol esters (PEs) and curcins, the seed cake produced as a result of oil extraction cannot be utilize for animal feed. Development of the genotypes better suited for the industrial applications and biodiesel production as well as with lower level of toxic constituents is being hampered by a lack of understanding about the a) proteins related to the biosynthesis and degradation of fatty acids (FAs) and triacylglycerides (TAGs), b) role of proteins deposited during seed development and c) proteins related to the synthesis and storage of toxic compounds during seed development. Agreeing with this, we have performed the anatomical analysis of the developing seeds of J. curcas, followed by the proteome analysis of the endosperm isolated from the seeds of J. curcas at five different developmental stages, which resulted into the identification of the 1517, 1256, 1033, 752 and 307 proteins, from Stage 6, 7, 8, 9 and 10, respectively, summing up to a total of 1760 proteins. Proteins with similar expression pattern were grouped into five different clusters and protein quantification based on spectral counts was determined. Besides identification of the proteins involved in the biosynthesis and degradation of the FAs and TAGs, we also identified a large number of proteins involved in the metabolism of the carbohydrates, which are important for supplying energy and carbon source for the synthesis of TAGs in heterotrophic seeds. Among the members of different classes of seed storage proteins (SSPs), we have identified four SSPs named as nutrients reservoir, which in contrast to the other SSPs showed decreasing deposition pattern during seeds development and revealed to have special role during seed development. In addition, peptidases belong to different mechanistic classes were identified, which have a range of functions, highlighting the role in reserve mobilization during germination. Isoforms of curcin were also identified in this proteome analysis which were absent in our previous proteome analysis of the other tissues from these seeds, suggesting that the deposition of these toxic proteins only occur in the endosperm. Similarly, several enzymes involved in the biosynthesis of diterpenoid precursors were identified in this proteome analysis but, like in our previous proteome analysis of the other tissues from J. curcas seeds,we were unable to identify any terpene synthase/cyclase, enzymes responsible for the synthesis of PEs, which collectively suggesting that the synthesis of PEs may not occur in seeds of this plant. In conclusion, the strategy used here enabled us to provide a first in depth proteome analysis of the endosperm from J. curcas developing seeds, which along with providing information regarding important aspects of the seed development, also set the foundation of a proteomic approach to study biotechnologically important plant species.
PinhÃo manso (Jatropha curcas L.) à uma cultura importante devido à sua habilidade em armazenar alto conteÃdo de Ãleo nas sementes, as quais podem servir como matÃria-prima para a produÃÃo de biodiesel. Devido à presenÃa de constituintes tÃxicos como Ãsteres de forbol e curcina, a torta da semente produzida como resultado da extraÃÃo do Ãleo nÃo pode ser utilizada na alimentaÃÃo animal. O desenvolvimento de genÃtipos mais adequados a aplicaÃÃes industriais e à produÃÃo de biodiesel assim como apresentando baixos nÃveis de constituintes tÃxicos està sendo prejudicado pela falta de entendimento sobre a) proteÃnas relacionadas a biossÃntese e degradaÃÃo de Ãcidos graxos e triacilglicerÃis, b) o papel de proteÃnas depositadas durante o desenvolvimento da semente e c) proteÃnas relacionadas à sÃntese e reserva de compostos tÃxicos durante o desenvolvimento da semente. Diante disso, nÃs realizamos uma anÃlise anatÃmica de sementes em desenvolvimento de J. curcas, seguido por uma anÃlise proteÃmica do endosperma isolado de sementes dessa espÃcie em cinco diferentes estÃgios de desenvolvimento, o que resultou na identificaÃÃo de 1517, 1256, 1033, 752 e 307 proteÃnas, dos estÃgios 6, 7, 8, 9 e 10, respectivamente, somando um total de 1760 proteÃnas. ProteÃnas com padrÃo de expressÃo similar foram agrupadas em cinco grupos diferentes e a quantificaÃÃo das proteÃnas baseada na contagem dos espectros foi determinada. AlÃm da identificaÃÃo das proteÃnas envolvidas na biossÃntese e degradaÃÃo de FAs e TAGs, nÃs identificamos um grande nÃmero de proteÃnas envolvidas no metabolismo de carboidratos, as quais sÃo importantes para o fornecimento de energia e fontes de carbono para a sÃntese de TAGs em sementes heterotrÃficas. Entre os membros de diferentes classes de proteÃnas de reservas de sementes (SSPs), nÃs identificamos quatro SSPs denominadas reservatÃrios de sementes, que em contraste as outras SSPs mostraram decrÃscimo no padrÃo de deposiÃÃo e revelaram ter um papel especial durante o desenvolvimento da semente. Em adiÃÃo, peptidases pertencentes a diferentes classes mecanÃsticas foram identificadas destacando o papel da mobilizaÃÃo de reservas durante a germinaÃÃo. Isoformas da curcina ausentes em nossas anÃlises proteÃmicas prÃvias de outros tecidos da semente foram identificadas sugerindo que a deposiÃÃo dessas proteÃnas tÃxicas sà ocorre no endosperma. Similarmente, vÃrias enzimas envolvidas na biosÃntese de precursores de diterpenÃides foram identificadas nessa anÃlise proteÃmica, mas como em nossas prÃvias anÃlises proteÃmicas de outros tecidos de sementes de J. curcas, nÃs nÃo fomos capazes de identificar sintases/ciclases de terpenos, enzimas responsÃveis pela sÃntese de PEs, o que coletivamente sugere que a sÃntese desses compostos pode nÃo ocorrer nas sementes dessa planta. Em conclusÃo, a estratÃgia utilizada nos fornece a primeira anÃlise proteÃmica profunda do endosperma de sementes em desenvolvimento de J. curcas, o que alÃm de fornecer informaÃÃes sobre aspectos importantes do desenvolvimento da semente, tambÃm estabelece a base para uma pesquisa proteÃmica com o objetivo de estudar espÃcies vegetais importantes biotecnologicamente.
description Physic nut (Jatropha curcas L.) is an important crop due to its ability of storing high content of oil in the seeds, which can serve as raw material for biodiesel production. Because of the presence of toxic constituents like phorbol esters (PEs) and curcins, the seed cake produced as a result of oil extraction cannot be utilize for animal feed. Development of the genotypes better suited for the industrial applications and biodiesel production as well as with lower level of toxic constituents is being hampered by a lack of understanding about the a) proteins related to the biosynthesis and degradation of fatty acids (FAs) and triacylglycerides (TAGs), b) role of proteins deposited during seed development and c) proteins related to the synthesis and storage of toxic compounds during seed development. Agreeing with this, we have performed the anatomical analysis of the developing seeds of J. curcas, followed by the proteome analysis of the endosperm isolated from the seeds of J. curcas at five different developmental stages, which resulted into the identification of the 1517, 1256, 1033, 752 and 307 proteins, from Stage 6, 7, 8, 9 and 10, respectively, summing up to a total of 1760 proteins. Proteins with similar expression pattern were grouped into five different clusters and protein quantification based on spectral counts was determined. Besides identification of the proteins involved in the biosynthesis and degradation of the FAs and TAGs, we also identified a large number of proteins involved in the metabolism of the carbohydrates, which are important for supplying energy and carbon source for the synthesis of TAGs in heterotrophic seeds. Among the members of different classes of seed storage proteins (SSPs), we have identified four SSPs named as nutrients reservoir, which in contrast to the other SSPs showed decreasing deposition pattern during seeds development and revealed to have special role during seed development. In addition, peptidases belong to different mechanistic classes were identified, which have a range of functions, highlighting the role in reserve mobilization during germination. Isoforms of curcin were also identified in this proteome analysis which were absent in our previous proteome analysis of the other tissues from these seeds, suggesting that the deposition of these toxic proteins only occur in the endosperm. Similarly, several enzymes involved in the biosynthesis of diterpenoid precursors were identified in this proteome analysis but, like in our previous proteome analysis of the other tissues from J. curcas seeds,we were unable to identify any terpene synthase/cyclase, enzymes responsible for the synthesis of PEs, which collectively suggesting that the synthesis of PEs may not occur in seeds of this plant. In conclusion, the strategy used here enabled us to provide a first in depth proteome analysis of the endosperm from J. curcas developing seeds, which along with providing information regarding important aspects of the seed development, also set the foundation of a proteomic approach to study biotechnologically important plant species.
publishDate 2014
dc.date.issued.fl_str_mv 2014-02-24
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
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status_str publishedVersion
format doctoralThesis
dc.identifier.uri.fl_str_mv http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=11638
url http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=11638
dc.language.iso.fl_str_mv eng
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dc.publisher.none.fl_str_mv Universidade Federal do CearÃ
dc.publisher.program.fl_str_mv Programa de PÃs-GraduaÃÃo em BioquÃmica
dc.publisher.initials.fl_str_mv UFC
dc.publisher.country.fl_str_mv BR
publisher.none.fl_str_mv Universidade Federal do CearÃ
dc.source.none.fl_str_mv reponame:Biblioteca Digital de Teses e Dissertações da UFC
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instname_str Universidade Federal do Ceará
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