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Sitka spruce (Picea sitchensis [Bong.] Carr.) seed germination in relation to seed development, dormancy and storageJones, Stephen Keith January 1995 (has links)
No description available.
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Mecanismos de controle de tolerância à dissecação em sementes de Caesalpinia echinata LAM. (Pau-Brasil) e Caesalpinia peltophoroides BENTH. (Sibipiruna)Silva, João Paulo Naldi [UNESP] 22 February 2010 (has links) (PDF)
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silva_jpn_me_botib.pdf: 1461698 bytes, checksum: f3d2da00e4477125c12042e4a44960b3 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / A aquisição da tolerância à dessecação ocorre durante a maturação da semente, após acúmulo de reservas e antes do início do processo natural de secagem, porém, com o avanço da germinação a sensibilidade a dessecação aumenta. Estresses hídricos podem induzir ou restabelecer a tolerância à dessecação em sementes, porém, não se sabe em quais processos ele atua, como na ativação de genes. O objetivo deste trabalho foi avaliar a aquisição e perda de tolerância à dessecação em sementes de Caesalpinia echinata Lam. (pau-brasil), e a possibilidade de indução deste processo avaliando as modificações nos carboidratos solúveis e de ciclitóis, nos níveis endógenos de ABA e na expressão de aquaporinas. Foram observadas diferenças na aquisição de tolerância em sementes imaturas de mesma idade, produzidas nas mesmas matrizes em 2007 e 2008, com diferentes composições de carboidratos solúveis. Sementes maduras perdem a tolerância à dessecação logo após protrusão da raiz primária, em comprimentos dependentes do grau de deterioração da semente. Sementes imaturas e maduras tolerantes de C. echinata suportaram a desidratação de formas diferentes, utilizando reservas de amido e ciclitóis, com papel da sacarose quando secas. Esses resultados foram comparados com sementes de Caesalpinia peltophoroides Benth (sibipiruna), demonstrando comportamentos fisiológicos distintos, provavelmente relacionados com o tipo de reserva que acumulam / The acquisition of desiccation tolerance occurs during seed maturation, after reserves accumulation and before the natural process of drying, however, with seed germination the desiccation sensitivity increases. Water stress can induce or restore the desiccation tolerance in seeds, but it is unclear what processes it acts, like activation of genes. The objective of this work was to evaluate the acquisition and loss of desiccation tolerance in Caesalpinia echinata Lam (brazil-wood) seeds, and the possibility to induction this process, evaluating the soluble carbohydrates and cyclitols changes, the ABA endogenous levels and expression of aquaporins. Differences were observed in the tolerance acquisition in immature seeds of the same age, produced by the same trees in 2007 and 2008, showing different compositions of soluble carbohydrates. Mature seeds lose desiccation tolerance soon after radicle protrusion in length depending on the degree of seed deterioration lot. Immature and mature tolerant seeds of C. echinata support the dehydration by different ways, using starch reserves and cyclitols, seen sucrose when the seed dry. These results were compared with seeds of Caesalpinia peltophoroides (sibipiruna), showing different physiological parameters, probably related to the type of reserves that they accumulate
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A proteomic investigation of the rhizomes of the resurrection fern Mohria caffrorum L. (Desv.) in response to desiccationShoko, Ryman January 2015 (has links)
Includes bibliographical references / As there is limited information on the mechanisms of vegetative desiccation-tolerance in resurrection plant rhizomes, this work was undertaken to study the mechanisms of desiccation-tolerance in Mohria caffrorum rhizomes. Fronds of this plant have been previously characterized as being desiccation-tolerant in summer and desiccation-sensitive in winter. Since fern rhizomes are perennial organs, it was of interest to establish whether these organs are also perennially desiccation-tolerant and, whether or not the rhizomes regulate desiccation-tolerance in the fronds. Ultra-structural evidence using transmission electron microscopy and viability studies using electrolyte leakage analysis showed that the rhizomes were desiccation-tolerant throughout the seasons. Quantitative proteomics analysis using isobaric tags for relative and absolute quantification was employed to investigate molecular mechanisms of desiccation-tolerance in the rhizomes of this plant. Using a custom fern rhizome specific peptide sequence database, 236 proteins were identified. Of these, 16 proteins increased in abundance while 14 declined, in the summer collected rhizomes. On the other hand, 16 proteins increased in abundance and 20 declined in the winter form. Western blot analysis confirmed the expression trends of heat shock protein 70-2 and superoxide dismutase-[Cu-Zn], which were among the differentially expressed proteins. Bioinformatics analysis of the differentially expressed proteins was carried out using network enrichment tools, to identify key molecular processes and pathways involved in the rhizome response to desiccation stress. Results indicate that the rhizomes use different molecular mechanisms to achieve desiccation-tolerance in winter and summer. Potential cross-talks and cross-tolerances were identified in which mechanisms protecting the rhizomes against desiccation-tolerance appeared to also protect them against heat stress, and in winter an apparent cross-talk against desiccation and pathogen stresses was also identified. This study is the first report of evidence that M.caffrorum rhizomes are the 'master-regulator organs' responsible for regulating desiccation-tolerance in the fronds. This role was inferred from the rhizome's predicted up-/down-regulation of biological processes and pathways that relate to leaf senescence, shoot system morphogenesis and gametophyte development, among others.
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Molecular Biology of Desiccation Tolerance in the Cyanobacterium Nostoc communeWright, Deborah J. 13 February 2004 (has links)
The molecular biology of desiccation tolerance was investigated in the cyanobacteria with emphasis on Nostoc commune. Analysis of DNA from 41 samples of desiccated Nostoc spp. of varied age and global distribution led to the amplification of 43 independent tRNALEU(UAA) group 1 intron sequences. Phylogenetic analysis of the entire data set made it possible to define the form species Nostoc commune.
The synthase (spsA) and phosphatase (sppA) genes required for the synthesis of sucrose were isolated from cyanobacterium Synechocystis sp. strain PCC 6803 and overexpressed in E. coli in two different vector constructions. Transformants had a marked increased capacity for desiccation tolerance. Sucrose synthesis was confirmed through thin layer chromatography (TLC) analysis of cell extracts from transformants.
Long-term stability of DNA in desiccated Nostoc samples was demonstrated by the ability to amplify selected gene loci from samples stored dry for decades. Successful amplification in some samples was possible only after treatment with phenacylthiazolium bromide, a reagent that disrupts covalent cross-links; indicating that the DNA was modified by cross-links that occurred between reducing sugars and the primary amines on the DNA.
Abundant superoxide dismutase was released following rehydration of desiccated field material N. commune CHEN after 13 years in the dry state. sodF mRNA was present in the dry material but was turned over within 15 min of rehydration. mRNA levels then rose and appeared to reach steady state levels after 3 hours and remained abundant after 24 hours of rehydration. / Master of Science
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Mecanismos de controle de tolerância à dissecação em sementes de Caesalpinia echinata LAM. (Pau-Brasil) e Caesalpinia peltophoroides BENTH. (Sibipiruna) /Silva, Joao Paulo Naldi. January 2010 (has links)
Orientador: Claudio José Barbedo / Banca: Jose Maria Rocha Faria / Banca: João Rodrigues Domingos / Resumo: A aquisição da tolerância à dessecação ocorre durante a maturação da semente, após acúmulo de reservas e antes do início do processo natural de secagem, porém, com o avanço da germinação a sensibilidade a dessecação aumenta. Estresses hídricos podem induzir ou restabelecer a tolerância à dessecação em sementes, porém, não se sabe em quais processos ele atua, como na ativação de genes. O objetivo deste trabalho foi avaliar a aquisição e perda de tolerância à dessecação em sementes de Caesalpinia echinata Lam. (pau-brasil), e a possibilidade de indução deste processo avaliando as modificações nos carboidratos solúveis e de ciclitóis, nos níveis endógenos de ABA e na expressão de aquaporinas. Foram observadas diferenças na aquisição de tolerância em sementes imaturas de mesma idade, produzidas nas mesmas matrizes em 2007 e 2008, com diferentes composições de carboidratos solúveis. Sementes maduras perdem a tolerância à dessecação logo após protrusão da raiz primária, em comprimentos dependentes do grau de deterioração da semente. Sementes imaturas e maduras tolerantes de C. echinata suportaram a desidratação de formas diferentes, utilizando reservas de amido e ciclitóis, com papel da sacarose quando secas. Esses resultados foram comparados com sementes de Caesalpinia peltophoroides Benth (sibipiruna), demonstrando comportamentos fisiológicos distintos, provavelmente relacionados com o tipo de reserva que acumulam / Abstract: The acquisition of desiccation tolerance occurs during seed maturation, after reserves accumulation and before the natural process of drying, however, with seed germination the desiccation sensitivity increases. Water stress can induce or restore the desiccation tolerance in seeds, but it is unclear what processes it acts, like activation of genes. The objective of this work was to evaluate the acquisition and loss of desiccation tolerance in Caesalpinia echinata Lam (brazil-wood) seeds, and the possibility to induction this process, evaluating the soluble carbohydrates and cyclitols changes, the ABA endogenous levels and expression of aquaporins. Differences were observed in the tolerance acquisition in immature seeds of the same age, produced by the same trees in 2007 and 2008, showing different compositions of soluble carbohydrates. Mature seeds lose desiccation tolerance soon after radicle protrusion in length depending on the degree of seed deterioration lot. Immature and mature tolerant seeds of C. echinata support the dehydration by different ways, using starch reserves and cyclitols, seen sucrose when the seed dry. These results were compared with seeds of Caesalpinia peltophoroides (sibipiruna), showing different physiological parameters, probably related to the type of reserves that they accumulate / Mestre
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Isolation and partial characterization of a water stress protein of the desiccation-tolerant cyanobacterium Nostoc commune UTEX 584 expressed in Escherichia coliSines, Brian James 30 December 2008 (has links)
A desiccation-tolerant cyanobacterium <i>Nostoc commune</i> accumulates a novel group of water stress proteins (Wsp) in response to cycles of repeated drying and rehydration. Antibodies, specific for Wsp, were used to screen a lambdafix II library of <i>N. commune</i> UTEX 584 Bam H1 DNA fragments and an 8.5-kb fragment, containing a gene cluster that synthesized a 59-kDa cross-reactive protein. The cloned fragment comprised five ORF’s. The ORF’s 59, 24, 22, 36, and 70, each potentially encode products of molecular weights of 59, 24, 22, 36, and 70-kDa, respectively. The 59 and 24 ORF products were found to be expressed in <i>E. coli</i>. The 59-kDa product of this fragment gives the strongest cross-reaction with the Wsp antiserum. The 59-kDa protein was partially purified. The 24-kDa product was successfully purified to homogeneity and partially characterized.
This study used <i>E. coli</i> strain DH10B transformed with the pTrc 99A plasmid. The pTre 99A contains the 8.5-kb gene cluster fragment of interest. The products of ORF 24 and 59 were isolated using an initial 40-60 % ammonium sulfate precipitation of a clarified <i>E. coli</i> cell lysate. The clarified cell lysate was then subjected to streptomycin sulfate precipitation. The cell lysate was then dialyzed extensively. The cell lysate was then applied to a Mono Q HR 5/5 anion exchange column using a 2 M KCl gradient elution procedure. The Mono Q column yielded a fraction containing both ORF products which eluted with approximately 400 mM KCl. This fraction was then applied to a Superose 12 HR 10/30 gel filtration column. The eluent fraction containing the ORF 24 product was then reapplied to the Superose 12 to yield the final fraction containing only the ORF 24 product. The final fraction of ORF 24 was purified to homogeneity as determined by SDS-PAGE analysis. Approximately 750 μg of ORF 24 was isolated. This preparation was used for characterization studies.
Characterization studies of ORF 24 consisted of an amino-terminal sequence analysis, an estimation of the molecular weight using gel filtration chromatography and SDS-PAGE analysis, and an analysis of enzymatic activity as suggested by amino acid sequence homologies. The amino-terminal sequence of ORF 24 is P V E Q R S H D. The molecular weight of ORF 24 using gel-filtration chromatography and SDS-PAGE analysis is 26-kDa and 23-kDa, respectively. From gene sequence analysis, the molecular weight of ORF 24 is known to be 24,340-Da. These data indicate that ORF 24 is a monomer. ORF 24 was found to have amino acid sequence homologies with a pectate lyase (E 4.2.2.2) periplasmic precursor from <i>Erwinia caratovora</i> subspecies and a dextransucrase (EC 2.4.1.5) precursor from <i>Streptoccocus mutans</i> GS-5. However, pectate lyase activity was not detected in cellular extracts over a 24 hour period. In addition, ORF 24 was not found to interact with 10 % substrate solutions of N-acetylglucosamine, pectin, UTEX 584 sheath material, DRH1 sheath material, sucrose, or glucose using thin layer chromatography. These studies indicate that the enzymatic activities proposed from amino acid sequence homologies have not been detected. The suggestion that ORF 24 is a water stress protein with a protective function on a structural level with regards to desiccation-tolerance requires further study. / Master of Science
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Microgramma squamulosa (Kaulf.) de la Sota e Pleopeltis hirsutissima (Raddi) de la Sota: estratégias contrastantes de uso da água em um ambiente epifítico / Microgramma squamulosa (Kaulf.) de la Sota and Pleopeltis hirsutissima (Raddi) de la Sota: two epiphytes with contrasting water use strategiesBarbosa, Joyce Marques 15 October 2012 (has links)
Entre os vegetais existem duas estratégias distintas para superar as condições de estresse hídrico que condicionam a sensibilidade a dessecação (SD) que está associada a mecanismos de evitação e a tolerância a dessecação (TD) que envolve suportar o estresse hídrico diretamente no nível dos tecidos e no nível subcelular. Sabe-se que os custos acoplados aos mecanismos de dessecação e reidratação em espécies TD são elevados, e provavelmente superiores aos de espécies SD. Desta forma, a coexistência de espécies com estas estratégias, como ocorre no ambiente epifítico da Cidade Universitária Armando Sales de Oliveira (CUASO), induzem questões quanto a as vantagens relativas de espécies TD sobre SD. Devido aos altos custos da estratégia TD é possível propor que estas deveriam apresentar um ganho médio de carbono superior a de espécies SD durante o período favorável (com disponibilidade hídrica). Além disso, também é razoável esperar que as distintas formas de uso da água, vinculadas às estratégias, poderiam facilitar o convívio de espécies TD e SD através da segregação de nichos. Com intuito de verificar essas hipóteses, foram realizadas análises relacionadas a estrutura de comunidade de epífitas da CUASO e, a partir destas, a detecção da associação espacial mais freqüente, que resultou composta por Pleopeltis hirsutissima (TD) e Microgramma squamulosa (SD). Estas espécies foram então selecionadas para avaliar parâmetros associados à dinâmica do uso da água, assimilação líquida de carbono, dinâmica da capacidade de uso da luz, estado de hidratação foliar e pigmentos fotossintéticos. Os resultados apontaram que espécies de ambas as estratégias não apresentam sinais de estresse crônico no ambiente estudado. Os dados reforçaram as respostas contrastantes de TD e SD, refletindo as formas distintas de lidar com os recursos e condições do ambiente. Ao contrário do esperado, P. hirsutissima não apresentou uma assimilação de carbono superior a M. squamulosa, indicando que o ganho de carbono não é um dos fatores possíveis na compensação dos custos acoplados a sua estratégia (TD). Porém, os resultados apresentados quanto a dinâmica da capacidade de uso da luz sugerem que a coexistência é facilitada pela segregação de nichos de luz, sendo a espécie TD mais heliófila do que a espécie SD / In land plants the resistance to drought can be accomplished by two main strategies. One involves mainly dehydration avoidance mechanisms and the other is based on the ability to withstand drought at the tissues and down to subcellular levels. The former is found on the desiccation sensitive plants (DS) and the last describes the desiccation tolerant plants (DT). The metabolic costs of the desiccation tolerance mechanisms are probably high, at least when considering the cessation of photosynthesis during anabiosis - a condition that is absent in drought avoidance strategies. The coexistence of drought avoiders and desiccation tolerant plants in the same substrate and condition can rise some questions about the relative advantages of each strategy when submitted to similar water regime. We ask whether desiccation tolerant plants have a compensatory carbon gain under a watering regime that induces frequent anabiosis or cumulative damage in drought avoidance have a detrimental effect comparable to the costs of the desiccation tolerance. An analysis of the epiphytic plant community associated to the trees of the CUASO urban landscape showed a very frequent co-occurring species pair including the desiccation sensitive Microgramma squamulosa and a desiccation tolerant Pleopeltis hirsutissima, both cryptogams of the same plant family. These species were thoroughly examined regarding the patterns of light, water and carbon dynamics together with pigments concentration during cycles of natural drought and wet climate field conditions. These observations were complemented with experimental induction of different water and light regimes. The results showed no sign of chronic stress in both plants living at the same condition but allowed us to identify large distinctions in resource use patterns. A compensatory carbon assimilation was not found in P.hirsutissima. Both species have similar CO2 assimilation values. Experimental induction of drought in different light regimes allowed to verify a relative advantage in desiccation tolerant P.hirsutissima in comparison to the desiccation sensitive M.squamulosa under high irradiance values. Furthermore, light curve analysis suggest that they share the same tree but their niches differ in light intensity regime allowing the high frequency of co-occurrence observed in the field
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Aquisição da qualidade fisiológica de sementes de feijão-caupi (Vigna unguiculata (L) Walper) / Acquisition of the physiological quality of seeds of feijão-caupi (Vigna unguiculata (L) Walper)Moreno, Leticia de Aguila [UNESP] 27 January 2017 (has links)
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Previous issue date: 2017-01-27 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O feijão-caupi (Vigna unguiculata) é uma espécie relativamente bem estudada, contudo faltam informações sobre a aquisição da qualidade fisiológica ao longo da maturação. Determinar o período em que cada componente da qualidade fisiológica é adquirido permite um ajuste no momento ideal da colheita e consequentemente uma colheita no período em que a semente se encontra com o máximo de qualidade fisiológica. Este trabalho teve como objetivo, determinar quando sementes de Vigna unguiculata adquirem qualidade fisiológica, incluindo a aquisição de tolerância à dessecação e a longevidade. O estudo foi realizado na Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP), Faculdade de Ciências Agronômicas - Laboratório de Análise de Sementes - Departamento de Produção e Melhoramento Vegetal (DPMV), Campus de Botucatu-SP. A produção de sementes foi realizada em abril de 2015, seguida de coleta e caracterização morfológica das sementes nos estádios reprodutivos 17, 20, 23, 25, 28, 31, 34, 37 e 40 dias após a antese (DAA). Para cada estádio foi determinado o teor de água, massa fresca, massa seca, germinação e vigor nas sementes frescas. Em seguida sementes foram submetidas a secagem e foi realizado o teste de germinação para se determinar a aquisição de tolerância à dessecação. As sementes secas foram então armazenadas à 35°C e 75% umidade relativa, para caracterizar a aquisição de longevidade. A qualidade fisiológica (germinação, tolerância à dessecação, vigor e longevidade) em feijão-caupi é adquirida ao longo da maturação da semente. O acúmulo de massa seca se iniciou próximo aos 17 DAA e atingiu o máximo aos 31 DAA. A germinação foi iniciada aos 28 DAA e atingiu seu máximo aos 37 DAA. A tolerância à dessecação foi iniciada a partir dos 28 DAA atingindo seu máximo aos 31 DAA. O vigor e a longevidade foram iniciados a partir dos 31 DAA apresentando seus máximos aos 37 DAA. / Cowpea (Vigna unguiculata) is a relatively well-studied specie, but lack information on the acquisition of physiological quality throughout maturation. Determine the period in which each physiologic component is obtained allows an adjustment in the optimal time of harvesting a crop and consequently the period in which seed acquire the maximum physiological quality. This study aimed to determine when Vigna unguiculata seeds acquire physiological quality, including acquisition of desiccation tolerance and longevity. The research was conducted at the Universidade Estadual Paulista Julio de Mesquita Filho (UNESP), Faculty of Agricultural Science - Laboratory of Seed Analysis - Department of Plant Breeding and Production, Botucatu-SP. Seed production was performed at the beginning of the year 2015, followed by collection and morphological characterization of the seeds in the reproductive stages 17, 20, 23, 25, 28, 31, 34, 37 and 40 days after anthesis (DAA). For each stage was determined the water content, fresh weight, dry weight, germination and vigor on fresh seeds. Then seeds were submitted to drying and carried out the germination test to determine the acquisition of desiccation tolerance and longevity. The dried seeds were then stored at 35 ° C and 75% relative humidity, to characterize the acquisition of longevity. The physiological quality (germination, desiccation tolerance, vigor and longevity) of Vigna unguiculata is acquired during seed maturation, after the maximum dry matter accumulation. The dry matter accumulation began close to 17 DAA and reached the maximum at 31 DAA. Germination was acquired at 28 DAA and reached its maximum at 37 DAA. The desiccation tolerance was acquired from 28 DAA reaching its maximum at 31 DAA. The vigor and longevity were acquired from 31 DAA showing their maximum to 37 DAA.
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Microgramma squamulosa (Kaulf.) de la Sota e Pleopeltis hirsutissima (Raddi) de la Sota: estratégias contrastantes de uso da água em um ambiente epifítico / Microgramma squamulosa (Kaulf.) de la Sota and Pleopeltis hirsutissima (Raddi) de la Sota: two epiphytes with contrasting water use strategiesJoyce Marques Barbosa 15 October 2012 (has links)
Entre os vegetais existem duas estratégias distintas para superar as condições de estresse hídrico que condicionam a sensibilidade a dessecação (SD) que está associada a mecanismos de evitação e a tolerância a dessecação (TD) que envolve suportar o estresse hídrico diretamente no nível dos tecidos e no nível subcelular. Sabe-se que os custos acoplados aos mecanismos de dessecação e reidratação em espécies TD são elevados, e provavelmente superiores aos de espécies SD. Desta forma, a coexistência de espécies com estas estratégias, como ocorre no ambiente epifítico da Cidade Universitária Armando Sales de Oliveira (CUASO), induzem questões quanto a as vantagens relativas de espécies TD sobre SD. Devido aos altos custos da estratégia TD é possível propor que estas deveriam apresentar um ganho médio de carbono superior a de espécies SD durante o período favorável (com disponibilidade hídrica). Além disso, também é razoável esperar que as distintas formas de uso da água, vinculadas às estratégias, poderiam facilitar o convívio de espécies TD e SD através da segregação de nichos. Com intuito de verificar essas hipóteses, foram realizadas análises relacionadas a estrutura de comunidade de epífitas da CUASO e, a partir destas, a detecção da associação espacial mais freqüente, que resultou composta por Pleopeltis hirsutissima (TD) e Microgramma squamulosa (SD). Estas espécies foram então selecionadas para avaliar parâmetros associados à dinâmica do uso da água, assimilação líquida de carbono, dinâmica da capacidade de uso da luz, estado de hidratação foliar e pigmentos fotossintéticos. Os resultados apontaram que espécies de ambas as estratégias não apresentam sinais de estresse crônico no ambiente estudado. Os dados reforçaram as respostas contrastantes de TD e SD, refletindo as formas distintas de lidar com os recursos e condições do ambiente. Ao contrário do esperado, P. hirsutissima não apresentou uma assimilação de carbono superior a M. squamulosa, indicando que o ganho de carbono não é um dos fatores possíveis na compensação dos custos acoplados a sua estratégia (TD). Porém, os resultados apresentados quanto a dinâmica da capacidade de uso da luz sugerem que a coexistência é facilitada pela segregação de nichos de luz, sendo a espécie TD mais heliófila do que a espécie SD / In land plants the resistance to drought can be accomplished by two main strategies. One involves mainly dehydration avoidance mechanisms and the other is based on the ability to withstand drought at the tissues and down to subcellular levels. The former is found on the desiccation sensitive plants (DS) and the last describes the desiccation tolerant plants (DT). The metabolic costs of the desiccation tolerance mechanisms are probably high, at least when considering the cessation of photosynthesis during anabiosis - a condition that is absent in drought avoidance strategies. The coexistence of drought avoiders and desiccation tolerant plants in the same substrate and condition can rise some questions about the relative advantages of each strategy when submitted to similar water regime. We ask whether desiccation tolerant plants have a compensatory carbon gain under a watering regime that induces frequent anabiosis or cumulative damage in drought avoidance have a detrimental effect comparable to the costs of the desiccation tolerance. An analysis of the epiphytic plant community associated to the trees of the CUASO urban landscape showed a very frequent co-occurring species pair including the desiccation sensitive Microgramma squamulosa and a desiccation tolerant Pleopeltis hirsutissima, both cryptogams of the same plant family. These species were thoroughly examined regarding the patterns of light, water and carbon dynamics together with pigments concentration during cycles of natural drought and wet climate field conditions. These observations were complemented with experimental induction of different water and light regimes. The results showed no sign of chronic stress in both plants living at the same condition but allowed us to identify large distinctions in resource use patterns. A compensatory carbon assimilation was not found in P.hirsutissima. Both species have similar CO2 assimilation values. Experimental induction of drought in different light regimes allowed to verify a relative advantage in desiccation tolerant P.hirsutissima in comparison to the desiccation sensitive M.squamulosa under high irradiance values. Furthermore, light curve analysis suggest that they share the same tree but their niches differ in light intensity regime allowing the high frequency of co-occurrence observed in the field
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Spectroscopic & thermodynamic investigations of the physical basis of anhydrobiosis in caenorhabditis elegans dauer larvaeAbu Sharkh, Sawsan E. 17 April 2015 (has links) (PDF)
Anhydrobiotic organisms have the remarkable ability to lose extensive amounts of body water and survive in an ametabolic, suspended animation state. Distributed to various taxa of life, these organisms have evolved strategies to efficiently protect their cell membranes and proteins against extreme water loss. At the molecular level, a variety of mutually non-exclusive mechanisms have been proposed to account particularly for preserving the integrity of the cell membranes in the desiccated state. Recently, it has been shown that the dauer larva of the nematode Caenorhabditis elegans is anhydrobiotic and accumulates high amounts of trehalose during preparation for harsh desiccation (preconditioning), thereby allowing for a reversible desiccation / rehydration cycle.
Here, we have used this genetic model to study the biophysical manifestations of anhydrobiosis and show that, in addition to trehalose accumulation, the dauer larvae exhibit a systemic chemical response upon preconditioning by dramatically reducing their phosphatidylcholine (PC) content. The C. elegans strain daf-2 was chosen for these studies, because it forms a constitutive dauer state under appropriate growth conditions. Using complementary approaches such as chemical analysis, time-resolved FTIR-spectroscopy, Langmuir-Blodgett monolayers, and fluorescence spectroscopy, it is shown that this chemical adaptation of the phospholipid (PL) composition has key consequences for their interaction with trehalose. Infrared-spectroscopic experiments were designed and automated to particularly address structural changes during fast hydration transients.
Importantly, the coupling of headgroup hydration to acyl chain order at low humidity was found to be altered on the environmentally relevant time scale of seconds. PLs from preconditioned larvae with reduced PC content exhibit a higher trehalose affinity, a stronger hydration-induced gain in acyl chain free volume, and a wider spread of structural relaxation rates during lyotropic transitions and sub- headgroup H-bond interactions as compared to PLs from non-preconditioned larvae. The effects are related to the intrinsically different hydration properties of PC and phosphatidylethanolamine (PE) headgroups, and lead to a larger hydration-dependent rearrangement of trehalose-mediated H-bond network in PLs from preconditioned larvae. This results in a lipid compressibility modulus of ∼0.5 mN/m and 1.2 mN/m for PLs derived from preconditioned and non-preconditioned larvae, respectively.
The ensemble of these changes evidences a genetically controlled chemical tuning of the native lipid composition of a true anhydrobiote to functionally interact with a ubiquitous protective disaccharide. The biological relevance of this adaptation is the preservation of plasma membrane integrity by relieving mechanical strain from desiccated trehalose- containing cells during fast rehydration. Finally, the thermo-tropic lipid phase behavior was studied by temperature-dependent ATR-FTIR and fluorescence spectroscopy of LAURDAN-labeled PLs. The results show that the adaptation to drought, which is accomplished to a significant part by the reduction of the PC content, relies on reducing thermo-tropic and enhancing lyotropic phase transitions. The data are interpreted on a molecular level emphasizing the influence of trehalose on the lipid phase transition under biologically relevant conditions by a detailed analysis of the lipid C=O H-bond environment.
The salient feature of the deduced model is a dynamic interaction of trehalose at the PL headgroup region. It is proposed here that the location of trehalose is changed from a more peripheral to a more sub-headgroup-associated position. This appears to be particularly pronounced in PLs from preconditioned worms. The sugar slides deeper into the inter-headgroup space during hydration and thereby supports a quick lateral expansion such that membranes can more readily adapt to the volume changes in the swelling biological material at reduced humidity. The data show that the nature of the headgroup is crucial for its interaction with trehalose and there is no general mechanism by which the sugar affects lipidic phase transitions. The intercalation into a phosphatidylethanolamine-rich membrane appears to be unique.
In this case, neither the phase transition temperature nor its width is affected by the protective sugar, whereas strong effects on these parameters were observed with other model lipids. With respect to membrane preservation, desiccation tolerance may be largely dependent on reducing phosphatidylcholine and increasing the phsophatidylethanolamine content in order to optimize trehalose headgroup interactions. As a consequence, fast mechanical adaptation of cell membranes to hydration-induced strain can be realized.
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