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Conserva??o in vitro DE Hyptis ramosa Pohl ex Benth. (LAMIACEAE)Santos, Suzivany Almeida dos 29 September 2017 (has links)
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Previous issue date: 2017-09-29 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / The genus Hyptis is composed of about 400 species distributed throughout several countries, the majority being in a native and non-domesticated state. The genus has great importance for the pharmaceutical industry because of the high diversity found in its species. It has excellent pharmacological potential and is of great economic interest. Biological properties like it?s antimicrobial, anti - inflammatory, anesthetic, and insecticidal potential have already been demonstrated. However, in addition to the endemism present in many species, there are few works related to conservation of the genus and more research is needed to elucidate potential methods for its preservation. The objective of this research was the in vitro conservation of the species Hyptis ramosa. In addition to adjusting the protocols of cryopreservation of axillary guides and shoots, we aimed to contribute to the conservation of germplasm of this species. The microplants used came from the germination of seeds in MS1/2 medium. For the in vitro conservation, 10 treatments were evaluated, with different combinations between sucrose, mannitol, and sorbitol. After inoculation, the plants were maintained in a growth room for 240 days and evaluated at 60, 120, 180, and 240 days. Each evaluation quantified the number of shoots, shoot length, root number, and root length. For cryopreservation, the techniques of vitrification for axillary calluses and buds and for encapsulation of axillary shoots were evaluated. The tests were carried out in the Laboratory of Germination (LAGER) and Vegetable Tissue Culture (LCTV) of the Horto Florestal Experimental Unit da Universidade Estadual de Feira de Santana. With the results obtained, we concluded that in vitro conservation of microplants of this species is possible using the combination of 87.64 mM sucrose combined with 87.64 mM mannitol in MS medium (MURASHIGE; SKOOG, 1969). With the methodology used, cryopreservation of calluses and shoots of the species was not possible. Cell death occurred in the first stages of the callus vitrification process and in the cryogenic stage for the shoots. / O g?nero Hyptis abrange cerca de 400 esp?cies distribu?das em v?rios pa?ses, sendo a grande maioria ainda encontrada em estado nativo e n?o domesticado. O g?nero apresenta grande import?ncia para a ind?stria farmac?utica por apresentar uma grande diversidade de esp?cies com grande potencial farmacol?gico e de interesse econ?mico, com diversas atividades biol?gicas j? comprovadas, como antimicrobiana, anti-inflamat?ria, anest?sica e inseticida. Contudo, al?m do endemismo presente em muitas esp?cies existem poucos trabalhos relacionados com a sua conserva??o, havendo a necessidade de mais pesquisas. No presente trabalho objetivou-se a conserva??o in vitro de plantas de Hyptis ramosa, al?m de ajustar protocolos para crioconserva??o de calos e gemas axilares, visando contribuir para a conserva??o de germoplasma desta esp?cie. As microplantas utilizadas foram provenientes da germina??o de sementes em meio MS1/2. Para conserva??o in vitro foram avaliados 10 tratamentos, com diferentes combina??es entre sacarose, manitol e sorbitol. Ap?s a inocula??o as plantas foram mantidas em sala de crescimento por 240 dias, com avalia??es aos 60, 120, 180 e 240 dias, quantificando-se o n?mero de brotos, comprimento da parte a?rea, n?mero de raiz e comprimento da raiz. Para a crioconserva??o foram avaliadas as t?cnicas de vitrifica??o para calos e gemas axilares e de encapsulamento de gemas axilares. Os ensaios foram realizados nos Laborat?rios de Germina??o (LAGER) e de Cultura de Tecidos Vegetais (LCTV) da Unidade Experimental Horto Florestal da Universidade Estadual de Feira de Santana. Com os resultados obtidos p?de-se concluir que ? poss?vel a conserva??o in vitro de microplantas dessa esp?cie, utilizando-se a combina??o de 87,64mM de sacarose combinado com 87,64mM de manitol, em meio MS (MURASHIGE; SKOOG, 1969). Com a metodologia utilizada n?o foi poss?vel a crioconserva??o de calos e gemas da esp?cie, havendo a morte celular nas primeiras etapas do processo de vitrifica??o dos calos e na etapa criog?nica para as gemas.
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Sistemas modelos de forma??o de lignina utilizando recursos sint?ticos e celulares de Eucalyptus grandis (Hill ex Maiden). / Systems models of lignin formation using synthetic and cellular resources of Eucalyptus grandis (Hill ex Maiden).Pereira, Regina Paula Willemen 28 May 2009 (has links)
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Previous issue date: 2009-05-28 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The main objective of this research was to work out models for the study of the
lignification. Eucalyptus grandis seedlings were produced in vitro, and the stem
segments explants were used to obtain friable callus. For such, the auxins tested were
TDZ, 2,4-D, NAA and IAA. The treatments with 50,0 ?M 2,4-D; 3,0 ?M TDZ and in
the absence of growth regulator didn't present callus formation. After 210 days, it was
chosen a callus formed in the culture medium added with 2,5 ?M of TDZ. The selected
callus was maintained in the same treatment, however, in liquid medium and under
agitation for 60 days at 25 ?C in the darkness, to induce cellular wall and extracellular
lignin formation. After this period, the cells suspensions were treated in the following
order: MS medium supplemented with sucrose, 2,4-D + kinetin, coumaric acid and
without extra supplement. The cells were maintained in these treatments for 30 days. It
was used a completely randomized design with four replications. Each plot consisted of
an Erlenmeyer with 125 ml of cells suspension culture. For cellular wall lignin
extraction and determination, it was used the method of lignin thyoglicolate. The
concentration of cellular wall and extracellular lignins were evaluated by UR. For data
normality verification it was used the Lilliefors test. The results were statistically
analyzed by variance analysis and by Tukey test at 5% level of significance. The highest
lignin concentration in the cellular wall (371.4200 ppm) was verified in the treatment
EG1pc 0,0584 ?M of sucrose. Extracellular lignins were also analyzed with Wiesner
reagent, H NMR and 13C NMR. The highest concentration (134.3167 ppm) of
extracellular lignins was verified in the treatment EG3e of 100?M coumaric acid).
However, with the results obtained in H NMR and ??CNMR analysis, it was not possible
to characterize the polymerization of pure extracellular lignin. A very useful tool for
lignification study is the dehydrogenized polymerization (DHP) in vitro. DHPs were
produce in cells suspension filtered medium (substrate) at different treatments. For each
treatment, it was added H2O2, peroxidase or H2O2 + peroxidase. The DHPs production
was done in MS culture medium (without the prior cultivation of cells) as a substrate. It
was added to MS medium three different solutions (coniferyl or sinapyl alcohol,
peroxidase and the H2O2) by dripping. The products of DHP and the filtered were
analyzed by IR and H NMR. The DHPs produced in medium MS without the previous
growth of the cells, were analyzed by H NMR and ??CNMR. The IR spectra didn t
present any sign in the zone of 1500 cm-1. H NMR also showed no formation of DHPs.
Nevertheless, in the, cells suspensions both in H NMR and in ??CNMR analysis
presented signs of DHPs, except the DHP1c (MS + 0,0584 ?M sucrose). H NMR
presented: 3.86; 6.92; and 4.71 ppm and the ??CNMR: 134,26; 88,21; 65,17; 55,90; and
20,75 ppm. / O objetivo principal desse trabalho foi a elabora??o de modelos para o estudo da
lignifica??o. Mudas de Eucalyptus grandis, obtidas a partir da germina??o de sementes
in vitro, foram desenvolvidas visando ? obten??o de explantes de segmentos caulinares.
Para tal, foram testadas as auxinas TDZ, 2,4-D, ANA e AIA. N?o foi constatada a
forma??o de calos nos tratamentos contendo 50,0 ?M de 2,4-D; 3,0 ?M de TDZ e na
aus?ncia de regulador de crescimento. Ap?s 210 dias, foi selecionado um calo formado
no tratamento contendo 2,5 ?M de TDZ. Este calo foi cultivado no mesmo tratamento,
por?m, em meio l?quido e sob agita??o por 60 dias a 25 ?C no escuro, para indu??o da
forma??o de lignina da parede celular e ligninas extracelulares. Ap?s este per?odo, as
c?lulas em suspens?o foram submetidas aos seguintes tratamentos: meio MS
suplementado com sacarose, 2,4-D + cinetina, ?cido p-cum?rico e sem suplemento
extra. As c?lulas permaneceram nestes tratamentos por 30 dias. O delineamento
experimental utilizado foi o inteiramente casualizado, com 4 repeti??es, sendo a parcela
experimental constitu?da por um Erlenmeyer contendo 125 ml de cultura de c?lulas em
suspens?o. Para extra??o e determina??o da lignina da parede celular, foi utilizada a
t?cnica baseada na lignina tioglicolato. A concentra??o de lignina da parede das c?lulas
e das ligninas extracelulares foi avaliada em UV. Para verificar a normalidade dos dados
foi usado o teste Lilliefors. Os dados foram submetidos ? an?lise de vari?ncia e as
m?dias comparadas pelo teste de Tukey a 5% de probabilidade. A maior concentra??o
de lignina na parede celular (371.4200 ppm) foi verificada no tratamento EG1pc 0,0584
?M de sacarose. Ligninas extracelulares tamb?m foram analisadas com o reagente
Wiesner, RMN H e RMN13C. A maior concentra??o (134.3167 ppm) de ligninas
extracelulares foi constatada no tratamento EG3e (100?M ?cido p-cum?rico). Por?m, os
resultados de RMN H e RMN13C, n?o permitiram caracterizar a polimeriza??o de
lignina extracelular pura. No estudo da lignifica??o utilizou-se a t?cnica de
polimeriza??o desidrogenativa (DHP) in vitro. DHPs foram elaborados em meios
obtidos a partir de filtrados das culturas de c?lulas em suspens?o em diferentes
tratamentos. Para cada tratamento, foi adicionado H2O2, peroxidase ou H2O2 +
peroxidase. A produ??o de DHPs foi efetuada em meio MS (sem o pr?vio cultivo de
c?lulas) como substrato no qual tr?s solu??es (?lcool conifer?lico ou sinap?lico,
peroxidase e o H2O2) foram adicionadas por gotejamento. Os produtos das DHPs,
juntamente com os filtrados, foram analisados por IV e RMN H. Nos tratamentos
realizados em meio MS sem o pr?vio cultivo de c?lulas em suspens?o, os DHPs
produzidos foram analisados por RMN H e RMN13C. Os espectros de IV n?o
apresentaram sinal na regi?o de 1500 cm-1, bem como as an?lises em RMN H
mostraram que n?o houve forma??o de DHPs. Por?m, nas suspens?es celulares tanto em
RMN H quanto RMN13C apresentaram sinais de DHP, exceto no tratamento com
DHP1c (MS + 0,0584 ?M sacarose). As an?lises em RMN H apresentaram: 3.86; 6.92 e
4.71 ppm e de RMN13C: 134,26; 88,21; 65,17; 55,90 e 20,75 ppm.
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Lignifica??o comparativa de Eucalyptus urophylla S. T. Blake por ferramentas biotecnol?gicas e polimeriza??o in vitro. / Comparative lignification of Eucalyptus urophylla S.T.Blake by biotechnological tools and polymerization in vitro.Monteiro, Maria Beatriz de Oliveira 29 May 2009 (has links)
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Previous issue date: 2009-05-29 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior-CAPES / In spite of the technological progresses, the understanding of the lignin structural
formation is still matter of scientific investigations. This research aimed to utilize
Eucalyptus urophylla S.T. Blake callus for lignin production. This callus were obtained
from stems segments explants grown in culture medium added with a combination of
the cytokine TDZ and the auxins acid indole-3-acetic (IAA), acid ?-naphthaleneacetic
(NAA) and acid 2,4-dichlorophenoxyacetic (2,4-D). This growth regulators were
utilized either alone or mixtured. For cell suspension production it was utilized callus
obtained in medium culture added with 20?M IAA + 3?M TDZ, after 30 days of
growth in vitro. Once the cells suspension was obtained, the lignin production was
induced by four elicitors: jasmonic acid (JA), NAA, sucrose and control (without
elicitor). It was used a completely randomized design with four replications. Each plot
consisted of an Erlenmeyer with 125 ml of cells suspension culture. The Wiesner test
confirmed the lignin presence in all treatments. In 3 of the 4 replicatons it was
performed another evaluation to the production of DHPs (polymers by oxidative
dehydrogenation) utilizing suspension filtrate added with H2O2, H2O2 + peroxidase and
peroxidase. These new treatments were analyzed through polilignols production
utilizing infrared (IR) and nuclear magnetic resonance of the hydrogen (NMR H). The
suspension filtrate analysis of the 4th replication through ultraviolet (UV), IR, NMR 13C
and NMR H evidenced the production of extra cellular lignin. Of this, the largest
content was obtained in presence of sucrose as elicitor, followed by ANA and AJ. The
cells in suspension increased the cellular wall lignin content in all treatments in relation
to the control and the largest values were with the medium containing sucrose. DHPs
were also analyzed utilizing as mattress the MS medium added of the same elicitors
tested in the cellular suspension phase. For this, it was utilized as precursors the
following alcohols: coniferyl or sinapyl, H2O2 and peroxidase; amd the analyses were
done in RMN H and RMN 13C. The results showed DHPs synthesis of both coniferyl
alcohol (DHP1c, DHP2c and DHP4c) and sinapyl alcohol (DHP2s). Nevertheless, It
was not synthesized DHP in the treatment containing sucrose when the precursor was
the coniferyl alcohol. On the other hand, when the sinapyl alcohol was the precursor,
DHPs were only synthesized in the presence of ANA as elicitor. It was concluded that
sucrose is an appropriate elicitor for the lignin production in cells suspension both at
cellular and extra cellular level. However, this result was not observed in relation to
DHP production. NAA auxin had a better functionality in the DHP2c and DHP2s
formation. These results may be considered a progress in the lignification studies with
the use of E. urophylla cell suspension. Once all these questions were answered and
solved, it will be possible to develop E.urophylla suitable plants with better quality
forest products and able to cause smaller environmental impact in the industrial
processes. / Apesar dos avan?os tecnol?gicos, a compreens?o da forma??o estrutural da lignina
ainda ? alvo de in?meras investiga??es cient?ficas. Esta pesquisa foi realizada com calos
de Eucalyptus urophylla S.T. Blake para a produ??o de lignina. Os calos foram obtidos
a partir de explantes de segmentos caulinares desenvolvidos em meios de cultura
acrescidos de uma combina??o da citocinina TDZ e das auxinas ?cidos: 3-indolac?tico
(AIA), ?-naftalenoac?tico (ANA) e diclorofenoxiac?tico (2,4-D), nas formas isoladas e
conjugadas. Para a produ??o de c?lulas em suspens?o foram utilizados os calos
formados no tratamento contendo 20?M de AIA + 3?M de TDZ, ap?s 30 dias de cultivo
in vitro. Obtidas as c?lulas em suspens?o, a produ??o de lignina foi induzida
empregando-se quatro elicitores: ?cido jasm?nico (AJ), ANA, sacarose e testemunha
(sem o emprego de elicitores). O delineamento experimental utilizado foi o inteiramente
casualizado, 4 repeti??es e um Erlenmeyer contendo 125 ml de cultura de c?lulas em
suspens?o. O teste de Wiesner confirmou a presen?a de lignina, em todos os tratamentos
testados. Em 3 das 4 repeti??es foram realizados subtratamentos para a produ??o de
DHPs (pol?meros por desidrogena??o oxidativa) a partir do filtrado da suspens?o com
H2O2, H2O2 + peroxidase e peroxidase. A an?lise desses subtratamentos foi realizada
pela detec??o da produ??o de polilign?is atrav?s de raios infravermelho (IV) e
resson?ncia magn?tica nuclear do hidrog?nio (RMN H). O filtrado da suspens?o da
repeti??o 4 foi analisado por ultravioleta (UV), IV, RMN 13C e RMN H, sendo
constatada a presen?a de lignina extracelular, com o maior teor sendo observado na
presen?a do elicitor sacarose, seguido do ANA e AJ. As c?lulas em suspens?o
apresentaram aumento no teor de lignina na parede celular em todos os tratamentos em
rela??o ? testemunha e os maiores valores foram com o meio contendo sacarose. Foram
analisadas tamb?m as DHPs tendo como colch?o o meio de cultura MS acrescido dos
mesmos elicitores testados na fase de suspens?o celular. Para isto foram utilizados como
precursores os ?lcoois conifer?lico ou sinap?lico, H2O2 e peroxidase, com a lignina
analisada em RMN H e RMN 13C. Os resultados mostraram que houve s?ntese de DHPs
do ?lcool conifer?lico (DHP1c, DHP2c e DHP4c) e do ?lcool sinap?lico (DHP2s).
Porem quando se utilizou como precursor o ?lcool conifer?lico n?o foram sintetizadas
DHPs no tratamento contendo sacarose como elicitor. E, quando foi utilizado como
precursor o ?lcool sinap?lico, somente foram formadas DHPs na presen?a do elicitor
ANA. Concluiu-se, ent?o, que a sacarose apresentou-se como um elicitor adequado para
a produ??o de lignina nas c?lulas em suspens?o, tanto em n?vel celular quanto
extracelular. Entretanto, isto n?o foi observado em rela??o ? produ??o de DHP. A
auxina ANA teve funcionalidade maior na forma??o de DHP2c e DHP2s. Estes
resultados devem ser considerados como um avan?o nos estudos de lignifica??o com a
utiliza??o de c?lulas em suspens?o de E. urophylla. Quando esses questionamentos
forem identificados e solucionados ser? poss?vel o desenvolvimento de novos
indiv?duos desta esp?cie que conduzam ? produ??o de produtos florestais de melhor
qualidade, com menor impacto ambiental nos processos industriais.
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Lignifica??o comparativa de Eucalyptus urophylla S. T. Blake por ferramentas biotecnol?gicas e polimeriza??o in vitro / Comparative lignification of Eucalyptus urophylla S.T.Blake by biotechnological tools and polymerization in vitroMONTEIRO, Maria Beatriz de Oliveira 29 May 2009 (has links)
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2009 - Maria Beatriz de Oliveira Monteiro.pdf: 6859044 bytes, checksum: 68c2b98ad22b807fdd1e5779b401a4aa (MD5)
Previous issue date: 2009-05-29 / CAPES / In spite of the technological progresses, the understanding of the lignin structural formation is still matter of scientific investigations. This research aimed to utilize Eucalyptus urophylla S.T. Blake callus for lignin production. This callus were obtained from stems segments explants grown in culture medium added with a combination of the cytokine TDZ and the auxins acid indole-3-acetic (IAA), acid ?-naphthaleneacetic (NAA) and acid 2,4-dichlorophenoxyacetic (2,4-D). This growth regulators were utilized either alone or mixtured. For cell suspension production it was utilized callus obtained in medium culture added with 20?M IAA + 3?M TDZ, after 30 days of growth in vitro. Once the cells suspension was obtained, the lignin production was induced by four elicitors: jasmonic acid (JA), NAA, sucrose and control (without elicitor). It was used a completely randomized design with four replications. Each plot consisted of an Erlenmeyer with 125 ml of cells suspension culture. The Wiesner test confirmed the lignin presence in all treatments. In 3 of the 4 replicatons it was performed another evaluation to the production of DHPs (polymers by oxidative dehydrogenation) utilizing suspension filtrate added with H2O2, H2O2 + peroxidase and peroxidase. These new treatments were analyzed through polilignols production utilizing infrared (IR) and nuclear magnetic resonance of the hydrogen (NMR H). The suspension filtrate analysis of the 4th replication through ultraviolet (UV), IR, NMR 13C and NMR H evidenced the production of extra cellular lignin. Of this, the largest content was obtained in presence of sucrose as elicitor, followed by ANA and AJ. The cells in suspension increased the cellular wall lignin content in all treatments in relation to the control and the largest values were with the medium containing sucrose. DHPs were also analyzed utilizing as mattress the MS medium added of the same elicitors tested in the cellular suspension phase. For this, it was utilized as precursors the following alcohols: coniferyl or sinapyl, H2O2 and peroxidase; amd the analyses were done in RMN H and RMN 13C. The results showed DHPs synthesis of both coniferyl alcohol (DHP1c, DHP2c and DHP4c) and sinapyl alcohol (DHP2s). Nevertheless, It was not synthesized DHP in the treatment containing sucrose when the precursor was the coniferyl alcohol. On the other hand, when the sinapyl alcohol was the precursor, DHPs were only synthesized in the presence of ANA as elicitor. It was concluded that sucrose is an appropriate elicitor for the lignin production in cells suspension both at cellular and extra cellular level. However, this result was not observed in relation to DHP production. NAA auxin had a better functionality in the DHP2c and DHP2s formation. These results may be considered a progress in the lignification studies with the use of E. urophylla cell suspension. Once all these questions were answered and solved, it will be possible to develop E.urophylla suitable plants with better quality forest products and able to cause smaller environmental impact in the industrial processes. / Apesar dos avan?os tecnol?gicos, a compreens?o da forma??o estrutural da lignina ainda ? alvo de in?meras investiga??es cient?ficas. Esta pesquisa foi realizada com calos de Eucalyptus urophylla S.T. Blake para a produ??o de lignina. Os calos foram obtidos a partir de explantes de segmentos caulinares desenvolvidos em meios de cultura acrescidos de uma combina??o da citocinina TDZ e das auxinas ?cidos: 3-indolac?tico (AIA), ?-naftalenoac?tico (ANA) e diclorofenoxiac?tico (2,4-D), nas formas isoladas e conjugadas. Para a produ??o de c?lulas em suspens?o foram utilizados os calos formados no tratamento contendo 20?M de AIA + 3?M de TDZ, ap?s 30 dias de cultivo in vitro. Obtidas as c?lulas em suspens?o, a produ??o de lignina foi induzida empregando-se quatro elicitores: ?cido jasm?nico (AJ), ANA, sacarose e testemunha (sem o emprego de elicitores). O delineamento experimental utilizado foi o inteiramente casualizado, 4 repeti??es e um Erlenmeyer contendo 125 ml de cultura de c?lulas em suspens?o. O teste de Wiesner confirmou a presen?a de lignina, em todos os tratamentos testados. Em 3 das 4 repeti??es foram realizados subtratamentos para a produ??o de DHPs (pol?meros por desidrogena??o oxidativa) a partir do filtrado da suspens?o com H2O2, H2O2 + peroxidase e peroxidase. A an?lise desses subtratamentos foi realizada pela detec??o da produ??o de polilign?is atrav?s de raios infravermelho (IV) e resson?ncia magn?tica nuclear do hidrog?nio (RMN H). O filtrado da suspens?o da repeti??o 4 foi analisado por ultravioleta (UV), IV, RMN 13C e RMN H, sendo constatada a presen?a de lignina extracelular, com o maior teor sendo observado na presen?a do elicitor sacarose, seguido do ANA e AJ. As c?lulas em suspens?o apresentaram aumento no teor de lignina na parede celular em todos os tratamentos em rela??o ? testemunha e os maiores valores foram com o meio contendo sacarose. Foram analisadas tamb?m as DHPs tendo como colch?o o meio de cultura MS acrescido dos mesmos elicitores testados na fase de suspens?o celular. Para isto foram utilizados como precursores os ?lcoois conifer?lico ou sinap?lico, H2O2 e peroxidase, com a lignina analisada em RMN H e RMN 13C. Os resultados mostraram que houve s?ntese de DHPs do ?lcool conifer?lico (DHP1c, DHP2c e DHP4c) e do ?lcool sinap?lico (DHP2s). Porem quando se utilizou como precursor o ?lcool conifer?lico n?o foram sintetizadas DHPs no tratamento contendo sacarose como elicitor. E, quando foi utilizado como precursor o ?lcool sinap?lico, somente foram formadas DHPs na presen?a do elicitor ANA. Concluiu-se, ent?o, que a sacarose apresentou-se como um elicitor adequado para a produ??o de lignina nas c?lulas em suspens?o, tanto em n?vel celular quanto extracelular. Entretanto, isto n?o foi observado em rela??o ? produ??o de DHP. A auxina ANA teve funcionalidade maior na forma??o de DHP2c e DHP2s. Estes resultados devem ser considerados como um avan?o nos estudos de lignifica??o com a utiliza??o de c?lulas em suspens?o de E. urophylla. Quando esses questionamentos forem identificados e solucionados ser? poss?vel o desenvolvimento de novos indiv?duos desta esp?cie que conduzam ? produ??o de produtos florestais de melhor qualidade, com menor impacto ambiental nos processos industriais.
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Micropropaga??o e conserva??o in vitro de Orthophytum mucugense Wand. e Concei??oLima, Andressa Priscila Pianc? Santos 23 March 2016 (has links)
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DISSERTA??O ANDRESSA PRISCILA PIANC? S. LIMA.pdf: 1903052 bytes, checksum: 3d8a27256e27d81bbdd1cf3ad9eee48f (MD5) / Made available in DSpace on 2016-07-19T23:28:56Z (GMT). No. of bitstreams: 1
DISSERTA??O ANDRESSA PRISCILA PIANC? S. LIMA.pdf: 1903052 bytes, checksum: 3d8a27256e27d81bbdd1cf3ad9eee48f (MD5)
Previous issue date: 2016-03-23 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Chapada Diamantina ? BA presents endemic especies with ornamental potential among which is Orthophytum mucugense, which occurs in the municipality Mucug?. This species is a target of extractivism and it is considered vulnerable, rendering studies of propagation and conservation of the species necessary. Therefore, the objective of this study was to establish protocols of micropropagation and in vitro conservation of O. mucugense. To that end, morphogenetic responses were evaluated in three types of explants, stem, root and leaf under the effect of different concentrations of the plant growth regulators NAA, 2,4-D and BAP. Sprouts formation in stem explants by direct organogenesis, in basal leaf explants by indirect organogenesis and callus formation in basal root and leaf explants were obtained. The rooting of the shoots was carried out with 1 coal g.L-1 activated for 30 days, and microplants acclimatized in a greenhouse with direct exposure to the environment. These results evince that tissue culture is a viable tool for in vitro propagation of O. mucugense. In in vitro conservation the effect of the medium salts reduction, of osmotic agents and of the retardant ancymidol in the minimum growth of the plants were tested; after 300 days of cultivation, analysis of the plant growth, of the chlorophyll content and of the regenerative capacity were carried out. On the basis of these assessments, the ancymidol, in the concentrations used, is not efficient in minimal growth induction, and the reduction of MS salts to ?3, with the combination of 45 g.L-1 of sucrose with 7.8 g.L-1 of mannitol is indicated for in vitro conservation of O. mucugense. / A Chapada Diamantina ? BA apresenta esp?cies end?micas com potencial ornamental dentre as quais est? a Orthophytum mucugense, que ocorre no munic?pio de Mucug?. Esta esp?cie ? alvo de extrativismo e considerada como vulner?vel, sendo necess?rios estudos de propaga??o e conserva??o da mesma. Portanto, objetivou-se neste trabalho estabelecer protocolos de micropropaga??o e conserva??o in vitro de O. mucugense. Para isto foram avaliadas as respostas morfog?nicas em tr?s tipos de explantes, caulinar, radicular e foliar, sob efeito de diferentes concentra??es dos reguladores vegetais ANA, 2,4-D e BAP. Foram obtidas forma??o de brotos em explante caulinar por organog?nese direta, em explante foliar basal por organog?nese indireta e forma??o de calo em explante foliar basal e radicular. O enraizamento dos brotos foi realizado com 1 g.L-1 de carv?o ativado por 30 dias, e as microplantas aclimatizadas em casa de vegeta??o com exposi??o direta ao ambiente. Estes dados demonstram que a cultura de tecidos ? uma ferramenta vi?vel para a propaga??o in vitro de O. mucugense. Na conserva??o in vitro foram testados o efeito da redu??o de sais do meio, de agentes osm?ticos e do retardante ancymidol no crescimento m?nimo das plantas; ap?s 300 dias de cultivo foram realizadas an?lises de crescimento das mesmas, do teor de clorofila e da capacidade regenerativa. Com base nestas avalia??es o ancymidol, nas concentra??es utilizadas, n?o ? eficiente na indu??o do crescimento m?nimo, e a redu??o de sais MS para ?3 com a combina??o de 45 g.L-1 de sacarose com 7,8 g.L-1 de manitol ? indicada para conserva??o in vitro de O. mucugense.
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Micropropaga??o de Hyptis ramosa Pohl ex Benth. (Lamiaceae)Sousa, Fl?via Pereira de 20 March 2015 (has links)
Submitted by Ricardo Cedraz Duque Moliterno (ricardo.moliterno@uefs.br) on 2016-08-24T20:52:09Z
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Disserta??o Final_Fl?via_PPGRGV (2).pdf: 1857571 bytes, checksum: 967f2b7d728622a11a98a803f13ff0de (MD5) / Made available in DSpace on 2016-08-24T20:52:09Z (GMT). No. of bitstreams: 1
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Previous issue date: 2015-03-20 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Hyptis ramosa Pohl ex Benth (Lamiaceae) is native and endemic to the semi-arid northeast, with its unknown phytochemical constitution so far. Considering the pharmacological importance of the species of this family, the development of forms of propagation and in vitro culture can contribute to the inclusion of these species in sustainable production systems and the conservation of the same. Thus, the objective of this work was to study the in vitro propagation of the species H. ramosa, through direct organogenesis and callus formation, and the biochemical characterization of the obtained calluses, thus allowing the establishment of strategies for their conservation and sustainable use. To this, H. ramosa seeds were disinfected and established in medium MS / 2 culture. In the multiplication phase was tested the influence of cytokinins BAP, CIN and TDZ on different explants. The obtained shoots were individualized and transferred to MS / 2 medium containing different concentrations of auxin IBA and activated carbon for rooting them. Regenerated and rooted in vitro microplants were subjected to pre-acclimatization in different cultivation container closure and then were transferred to ex vitro conditions in commercial substrate Plantmax? being quantified plant survival rate at 30 days after transfer. For callus induction, we used explants and different concentrations of 2,4-D and BAP, determining the growth curve from the fresh weight of callus until the 28th day of cultivation, at intervals of seven days. Concurrent with obtaining the growth curve it is quantified in calluses obtained the total soluble sugar content, reducing sugar and crude protein. The in vitro propagation of H. ramosa is possible using the nodal segment as a source of explants in MS medium supplemented with BAP. In vitro rooting occurs, even in free auxin. The species showed survival of 100%, regardless of the day of pre-acclimatization phase. For callus induction the best explant is the nodal segment, and the combination of 2.4-D and BAP favor the formation of the same. The callus growth curve showed quadratic behavior with two different phases and biochemical analysis showed the maximum level of total soluble sugars, reducing sugars and crude protein at 14 ?, 21 ? and 14 ? days, respectively. / Hyptis ramosa Pohl ex Benth (Lamiaceae) ? uma esp?cie nativa e end?mica do semi?rido nordestino, sendo sua constitui??o fitoqu?mica desconhecida at? o momento. Considerando a import?ncia farmacol?gica das esp?cies dessa fam?lia, o desenvolvimento de formas de propaga??o e cultivo in vitro poder? contribuir para a inser??o dessas esp?cies em sistemas de produ??o sustent?veis e a conserva??o das mesmas. Diante disso, o objetivo deste trabalho foi estudar a propaga??o in vitro da esp?cie H. ramosa, atrav?s de organog?nese direta e calog?nese, bem como a caracteriza??o bioqu?mica dos calos obtidos, permitindo assim o estabelecimento de estrat?gias para a sua conserva??o e explora??o sustent?vel. Para isso, sementes de H. ramosa foram desinfestadas e estabelecidas em meio de cultura MS/2. Na fase de multiplica??o foi testada a influ?ncia das citocininas BAP, CIN e TDZ sobre diferentes explantes. As brota??es obtidas foram individualizadas e transferidas para meio MS/2 contendo diferentes concentra??es da auxina AIB e de carv?o ativo para o enraizamento das mesmas. As microplantas regeneradas e enraizadas in vitro foram submetidas ? pr?-aclimatiza??o em diferentes tipos de fechamento do recipiente de cultivo e, posteriormente, foram transferidas para a condi??o ex vitro em substrato comercial Plantmax?, sendo quantificada a taxa de sobreviv?ncia das plantas aos 30 dias ap?s a transfer?ncia. Para a indu??o de calos utilizou-se diferentes explantes e concentra??es de 2,4-D e BAP, determinando-se a curva de crescimento a partir da mat?ria fresca dos calos at? o 28o dia de cultivo, em intervalos de sete dias. Concomitante com a obten??o da curva de crescimento quantificou-se nos calos obtidos o teor de a??cares sol?veis totais, a??cares redutores e prote?na bruta. A propaga??o in vitro de H. ramosa ? poss?vel utilizando-se o segmento nodal como fonte de explante, em meio de cultura MS suplementado com BAP. O enraizamento in vitro ocorre, mesmo em meio isento de auxina. A esp?cie apresentou sobreviv?ncia de 100%, independentemente da realiza??o da fase de pr?-aclimatiza??o. Para indu??o de calos o melhor explante ? o segmento nodal, sendo a combina??o de 2.4-D e BAP favor?vel a forma??o dos mesmos. A curva de crescimento de calos mostrou comportamento quadr?tico com duas fases distintas e a an?lise bioqu?mica evidenciou o teor m?ximo de a??cares sol?veis totais, a??cares redutores e prote?na bruta aos 14?, 21? e 14? dias, respectivamente.
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Indu??o de calos em anteras e poliploidia em gen?tipos de melanciaSilva, Carla Maria de Jesus 29 March 2018 (has links)
Submitted by Verena Pereira (verenagoncalves@uefs.br) on 2018-07-03T23:23:34Z
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Previous issue date: 2018-03-29 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / The watermelon (Citrullus lanatus) is a very important vegetable for a region of Northeast Brazil due to its adaptation as the natural conditions and the good characteristics of the fruit that are obtained. The aims of the current study are to assess watermelon genotype responses to calluses in anthers by using growth and temperature regulators, and to induce polyploidy through colchicine use (at different concentrations), exposure time, mechanical scarification and application methods. Anthers of Smile and Sugar Baby lines were inoculated in MS medium at different concentrations of 2.4-D or of BAP with 2.4-D, in combination with the pre-treatment (low temperature). Crimson Sweet cultivar seedlings were treated with different colchicine concentrations at two different times, with and without scarification, in order to induce polyploidy. Line LDRO was subjected to different colchicine concentrations at two different times and application methods: a) direct in the seed method (with, and without, scarification) (DSM, WE and WOE), b) Radicle emission method (ERM), c) Hypocotyl and root insertion point method (HRIM), d) At the apex of the seedling method (ASM) and e) Inverted hypocotyl method (IHM). Crimson Sweet flower buds were subjected to colchicine, at two different times, in order to induce polyploidy. Results have shown that 2.4-D often induced callus formation in both lines, but BAP/2.4-D interaction and pre-treatment did not increase the induction frequency. Crimson Sweet showed higher induction rate at 0.2% colchicine for 48h, WE. Line LDRO presented plants with tetraploid cells at 0.2% colchicine for 24 and 48h. The method 0.2% DSM, WE and WOE also generated plants with tetraploid cells. The diameter of treated pollen grains in flower buds have increased; the higher rate of non-reduced gametes induction was 16.07% and flower bud diameter (1.5mm) was estimated as adequate for induction / A melancia (Citrullus lanatus) ? uma hortali?a muito importante para a regi?o Nordeste do Brasil por sua adapta??o as condi??es clim?ticas e pelas boas caracter?sticas de fruto que se obt?m. Os objetivos desse trabalho foram avaliar as respostas de gen?tipos de melancia quanto ? indu??o de calos em anteras utilizando reguladores de crescimento e temperatura; e induzir a poliploidia mediante o uso da colchicina em diferentes concentra??es, tempos de exposi??o, escarifica??o mec?nica e m?todos de aplica??o. Anteras das linhagens de Smile e Sugar Baby foram inoculadas em meio MS sob diferentes concentra??es de 2,4-D ou BAP com 2,4-D, associado ao pr?-tratamento (baixa temperatura). Para indu??o de poliploidia, sementes da cultivar Crimson Sweet foram tratadas com diferentes concentra??es de colchicina, em dois tempos, com e sem escarifica??o. Para a linhagem LDRO, utilizaram-se diferentes concentra??es de colchicina em dois tempos e m?todos de aplica??o: a) M?todo direto na semente (com escarifica??o e sem escarifica??o) (MDS, CE e SE), b) M?todo da semente com emiss?o da rad?cula (MER), c) M?todo no ponto de inser??o do hipoc?tilo e raiz (MIHR), d) M?todo no ?pice da pl?ntula (MAP) e e) M?todo do hipoc?tilo invertido (MHI). Na indu??o de poliploidia em bot?es florais de Crimson Sweet utilizou-se colchicina em dois tempos. Os resultados mostraram que o 2,4-D induziu a maior frequ?ncia de calos nas duas linhagens, a intera??o BAP com 2,4-D e o pr?-tratamento, n?o aumentaram a frequ?ncia de indu??o. Para Crimson Sweet, o maior percentual de indu??o foi obtido com colchicina a 0,2% por 48 h, SE. Para LDRO, observou-se uma frequ?ncia de plantas com c?lulas tetraploides com colchicina a 0,2% por 24 h e 48 h. Nos m?todos 0,2% MDS CE e MHI observou-se tamb?m plantas com c?lulas tetraploides. Em bot?es florais, o di?metro dos gr?os de p?len tratados aumentou; o maior percentual de indu??o de gametas n?o reduzidos foi de 16,07% e; o di?metro em torno de 1,5 mm do bot?o floral foi estimado como adequado para indu??o
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