La dynamique mitochondriale est impliquée dans la maintenance et la fonction des mitochondries. Dans les graines sèches tout les processus cellulaires sont arrêtés du fait de la faible teneur en eau des tissues, et la transition développementale que représente la germination requiert la réactivation de la dynamique cellulaire. Une approche de bio-imagerie sur la plante modèle Arabidopsis a été utilisée afin d’étudier la réactivation des mitochondries nécessaire à la germination. La réactivation bioénergétique des mitochondries, mesurée par la présence du potentiel membranaire, intervient dès le début de l’hydratation des tissus. Cependant les mitochondries restent statiques et la dynamique mitochondriale ne reprend que plus tardivement. La réactivation des mitochondries provoque une réorganisation du chondriome impliquant la biogenèse de membranes et une fusion massive menant à la formation de structures réticulaires et périnucléaires, qui permet le mélange des nucléoïdes d’ADNmt précédemment isolés en unités discrètes. La mitophagie, un indicateur de la qualité mitochondriale, est réactivée de manière concomitante à la dynamique, alors qu’elle est réprimée durant la biogenèse des mitochondries. La fin de la germination coïncide avec la fragmentation du chondriome tubulaire, menant au doublement du nombre de mitochondrie et à une redistribution hétérogène des nucléoïdes dans le chondriome, générant une population de mitochondrie adaptée à la croissance des plantules. Cette thèse met en évidence l’imbrication des processus de dynamique mitochondriale, de biogenèse et de contrôle qualité des mitochondries requis pour la germination et pour la transition vers l’autotrophie. / Mitochondrial dynamics underpin their function and maintenance. In dry seeds, all cellular processes are in stasis due to a low water content. Thus, the developmental switch leading to germination necessarily involves a reactivation of cellular dynamics. In order tobetter understand the role played by mitochondrial dynamics during germination we used Arabidopsis as a model for a bioimaging approach to investigate the rapid reactivation of mitochondria that is required in order to provide ATP to support germination. Bioenergetic reactivation, visualised as the presence of a mitochondrial membrane potential, is almost immediate upon rehydration. However, the reactivation of mitochondrial dynamics only occurs after several hours of rehydration. The reactivation of mitochondrialbioenergetics and dynamics lead to a dramatic reorganisation of the chondriome involving massive fusion and membrane biogenesis to form a perinuclear tubuloreticular structure enabling mixing of previously discrete mtDNA nucleoids. Mitophagy, an indicator of mitochondrial quality, is reactivated concomitant with a reactivation of mitochondrial dynamics, but is repressed at time of mitochondrial biogenesis. The end of germination coincides with fragmentation of the tubular chondriome leading to a doubling of mitochondrialnumber and heterogeneous redistribution of the nucleoids amongst the mitochondria, generating a population of mitochondria tailored to seedling growth. This thesis provides strong evidence for the tight interweaving of mitochondrial dynamics, mitochondrialbiogenesis and mitochondrial quality control that is required to ensure effective germination and the transition to autotrophy.

Orchardgrass (<i>Dactylis glomerata</i> L.) and western wheatgrass (<i>Pascopyrum smithii </i>(Rydb.) L.) seeds have different degrees of dormancy that result in non-uniform seedling emergence in the field. Seed dormancy of the two species, in part, causes disagreement between germination tests in the laboratory and seedling emergence in the field. Experiments were conducted over two years in the laboratory and in the field to determine the effects of alternating temperatures on changes in seed dormancy and germination of orchardgrass and western wheatgrass. The two western wheatgrass cultivars (Walsh and LC9078a) had deeper dormancy than the two orchardgrass cultivars (Arctic and Lineta). Dormancy of both species was broken by temperatures with 10oC amplitude; this temperature variation was similar to that which occurred at a 1 cm depth in the soil. Optimal temperatures for germination of orchardgrass (10-25oC) were broader than those for western wheatgrass (15-20oC). Seedling emergence of orchardgrass was less sensitive to seeding date in the spring than western wheatgrass; seedling emergence of western wheatgrass increased as seeding date was delayed from early to late May if soil water was not limiting. The rate of seedling emergence increased with increasing temperature in both species, therefore, faster and more uniform seedling emergence can be expected from late spring seeding dates. Seeds were often exposed to light during germination tests in the laboratory while planting seeds in the soil usually prevented exposure of seeds to light. Seedling emergence of orchardgrass in the field was usually less than the germination percentage obtained in the laboratory because of light exposure during germination tests could break dormancy in orchardgrass seeds and the small seeds of orchardgrass had limited energy reserves for pre-emergence seedling growth. On the other hand, germination of western wheatgrass seeds was reduced by exposure to light during germination and seeds were larger than those of orchardgrass. Therefore, seedling emergence of western wheatgrass in the field was usually greater than germination tests would predict. The use of thermal time models to study seed dormancy changes and germination revealed the dual effects of temperature on these processes. The modified thermal time model takes the difference between germination and seedling emergence into account and can accurately predict seedling emergence in the field (R2=0.88 to 0.99). Thermal time models for predicting seedling emergence in the field can also be developed for other forages, however, cultivar- and species-specific parameters must be developed for the models.

Orchardgrass (<i>Dactylis glomerata</i> L.) and western wheatgrass (<i>Pascopyrum smithii </i>(Rydb.) L.) seeds have different degrees of dormancy that result in non-uniform seedling emergence in the field. Seed dormancy of the two species, in part, causes disagreement between germination tests in the laboratory and seedling emergence in the field. Experiments were conducted over two years in the laboratory and in the field to determine the effects of alternating temperatures on changes in seed dormancy and germination of orchardgrass and western wheatgrass. The two western wheatgrass cultivars (Walsh and LC9078a) had deeper dormancy than the two orchardgrass cultivars (Arctic and Lineta). Dormancy of both species was broken by temperatures with 10oC amplitude; this temperature variation was similar to that which occurred at a 1 cm depth in the soil. Optimal temperatures for germination of orchardgrass (10-25oC) were broader than those for western wheatgrass (15-20oC). Seedling emergence of orchardgrass was less sensitive to seeding date in the spring than western wheatgrass; seedling emergence of western wheatgrass increased as seeding date was delayed from early to late May if soil water was not limiting. The rate of seedling emergence increased with increasing temperature in both species, therefore, faster and more uniform seedling emergence can be expected from late spring seeding dates. Seeds were often exposed to light during germination tests in the laboratory while planting seeds in the soil usually prevented exposure of seeds to light. Seedling emergence of orchardgrass in the field was usually less than the germination percentage obtained in the laboratory because of light exposure during germination tests could break dormancy in orchardgrass seeds and the small seeds of orchardgrass had limited energy reserves for pre-emergence seedling growth. On the other hand, germination of western wheatgrass seeds was reduced by exposure to light during germination and seeds were larger than those of orchardgrass. Therefore, seedling emergence of western wheatgrass in the field was usually greater than germination tests would predict. The use of thermal time models to study seed dormancy changes and germination revealed the dual effects of temperature on these processes. The modified thermal time model takes the difference between germination and seedling emergence into account and can accurately predict seedling emergence in the field (R2=0.88 to 0.99). Thermal time models for predicting seedling emergence in the field can also be developed for other forages, however, cultivar- and species-specific parameters must be developed for the models.

Seed maturation is an important process that is evolutionarily advantageous, allowing for seed dispersal and germination under favourable growth conditions. The B3-domain transcription factor FUSCA3 (FUS3) is a master regulator of seed maturation and controls developmental phase transitions through hormonal regulation in Arabidopsis thaliana. The aim of this study was to determine the post-translational regulation of FUS3 during embryonic and vegetative development. Here, FUS3 was found to interact with the E3 ubiquitin ligase ABI3-INTERACTING PROTEIN2 (AIP2) in yeast two-hybrid, in vitro, and in planta assays. Analysis of transcriptional and translational reporters also showed overlapping spatial and temporal expression patterns of AIP2 and FUS3. Furthermore, in vitro FUS3 degradation was delayed in aip2-1 mutant and increased FUS3-GFP levels were observed during mid-embryogenesis in aip2-1. Finally, double transgenic plants overexpressing AIP2 and FUS3 showed reduced FUS3 levels and reversion of the gain-of-function FUS3 phenotypes back to WT. Together, these results indicate that AIP2 is a negative regulator of FUS3.

Seed germination has many impacts on the uses of seeds, and is an important subject for study. Seed germination is regulated at both transcriptional and post-transcriptional levels. Therefore, it is important to study how polyadenylation regulates gene expression during seed germination. To this end, a modified Illumina GAIIx sequencing protocol (described in Chapter Two) was developed that allows deep coverage of poly(A) site position and distribution. Alternative polyadenylation (APA) regulates gene expression by choosing one potential poly(A) site on a precursor RNA consequentially shortening/lengthening the mRNA relative to other possible sites. To further explore this phenomenon, genes affected by APA during seed germination and other developmental stages were identified (Chapter Three). These genes were categorized based on the location of poly(A) sites. Several genes were chosen to demonstrate how APA, especially that occurring in the coding regions and 5’ untranslated regions, might down regulate gene expression by generating truncated transcripts. In animal oocytes, maternally-derived mRNAs are stored with short poly(A) tails and reactivated by the cytoplasmic polyadenylation complex. It has been reported that seeds also contain stored mRNAs. Moreover, germination and its completion are less sensitive to de novo transcription inhibitors than to poly(A) polymerase inhibitors. Together, these considerations suggest that stored RNA without or with a short poly(A) tail (stored, unadenylated RNA) may be present in dry seed and function in seed germination upon reactivation by cytoplasmic polyadenylation. To further explore this, in Chapter Four, mRNA polyadenylation was studied through the course of germination using a combination of transcriptional inhibitors and the modified sequencing protocol described in Chapter Two. 273 putative stored, unadenylated RNAs were identified. Gene ontology analysis revealed that genes whose products are involved in translation are overrepresented; these genes encode 21 60S- and 10 40S-ribosomal proteins. These results indicate that transcripts whose products are involved in translation might be a major component of the stored, unadenylated RNA pool and, more importantly, translation might be the first cellular process to be activated during seed germination.

Fire, a natural disturbance, regulates species composition in Fescue Prairie. However, little is known about the contribution of different regeneration strategies in altering species composition after burning in Fescue Prairie. The present study was conducted to determine if and how fire and associated fire cues regulate species composition in Fescue Prairie through their effects on seedling recruitment. The effects of fire and various fire cues, including smoke, ash, and smoke plus ash on seedlings emerging in the field and/or from litter, 0-1 cm, and 1-5 cm layers of the soil seed banks were therefore examined. These studies were complimented by a study of testing effects of smoke originated from different plant materials on seed germination and seedling growth of species from Fescue Prairie. Chemical analyses were also conducted to determine whether different active compounds existed in smoke made from different materials, which in turn affect germination and seedling growth differently. Burning increased densities, richness, and diversity of seedlings emerging in the field. This was possibly attributed to direct fire cues of burning. Seedling densities of native forbs and non-native graminoids emerging from the soil seed bank were increased and decreased by burning, respectively. Ash and smoke plus ash increased density of forbs emerging from the soil seed bank. Species composition of seedlings emerging in the field and from the soil seed bank was altered by burning. Complex responses were observed for the effects of smoke on seedling establishment, which depended on the type and dilution of smoke solutions, as well as germination conditions. Smoke solutions partly substituted light requirement for germination of Artemisia ludoviciana. Germination of Cirsium arvense and Conyza canadensis only responded to smoke solutions at 25/15°C, but not at 10/0 °C. Diluted smoke solutions increased radical length of Artemisia ludoviciana. Karrikinolide (KAR1) was in the smoke made from prairie hay and wheat straw, but not in that made from alfalfa. This is the first report that different active compounds existed in smoke made from different materials. Highly concentrated smoke solutions made from alfalfa increased germination and radical length of Conyza canadensis, while the same concentrated smoke solutions made from prairie hay and wheat straw reduced germination of Conyza canadensis at 25/15 °C in darkness. Priming in KAR1 solutions and active fractions obtained from prairie hay and/or wheat straw increased germination of Artemisia frigida, Artemisia ludoviciana, and Conyza canadensis at certain germination conditions. In summary, fire and direct fire cues, smoke and ash specifically, stimulated recruitment of some species, especially early seral species and native forbs, contributing to potential changes in species composition of the Fescue Prairie. Different compounds existed in smoke solutions made from alfalfa as compared with those from prairie hay and wheat straw, showing different effects on seed germination and seedling growth. KAR1, the most important active compound discovered in smoke, was present in the smoke made from prairie hay and wheat straw, but was not in that made from alfalfa.

Germination course and mycorrhizal partners during germination and adulthood were studied in Neottia ovata and N. cordata. Neottia ovata was studied in two different biotops: meadows and forests. Research was conducted in in situ conditions in the Šumava and the Smrčiny mountains.

Made available in DSpace on 2014-06-11T19:23:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-03-05Bitstream added on 2014-06-13T19:49:38Z : No. of bitstreams: 1 guaraldo_ac_me_rcla.pdf: 708575 bytes, checksum: eb1ceaebc23d768a26ad3c2bdff138a0 (MD5) / As epífitas são responsáveis por grande parte da diversidade vegetal das florestas neotropicais. São também importantes para a diversidade faunística dessas regiões, pois fornecem recursos alimentares (e.g., flores, frutos e sementes) e micro-ambientes propícios à sobrevivência de vários animais. Rhipsalis (Cactaceae) é um gênero de plantas holoepífitas cujos frutos são consumidos principalmente por um único gênero de aves, as Euphonia. Para verificar a hipótese de especialização na dispersão de sementes de Rhipsalis pelas Euphonia, inicialmente foi estudada a fenologia reprodutiva dessa epífita. Uma vez conhecido o período de disponibilidade de frutos maduros aos frugívoros, foram realizados registros eventuais de frugivoria e observações focais em indivíduos de Rhipsalis para definir a assembléia de frugívoros consumidores de seus frutos. Posteriormente, frutos de cinco espécies de Rhipsalis foram fornecidos a aves frugívoras mantidas em cativeiro. Testes de germinação foram realizados com as sementes que passaram pelo trato digestório dessas aves com o objetivo de verificar seu potencial como dispersoras efetivas das sementes de Rhipsalis. Os resultados obtidos demonstraram que o gênero Rhipsalis apresenta sincronia na produção das fenofases reprodutivas entre os indivíduos e atividade reprodutiva ao longo de todo o ano. A produção de frutos maduros teve duração longa entre os meses de junho e fevereiro. Todas as espécies apresentaram fenofases reprodutivas sazonais. As observações de frugivoria e os testes de germinação realizados confirmaram que as sementes de Rhipsalis podem ser dispersas por endozoocoria. O processamento dos frutos pelas aves frugívoras, especificamente a remoção da polpa que recobre as sementes, não apenas incrementou as taxas de germinação e germinabilidade das sementes, mas também se mostrou importante para... / Epiphytes are responsible for a great part of the plant diversity in Neotropical Forests. They are also important for animal diversity due to the production of food resources (e.g. flowers, fruits and leaves) and microhabitats used by a myriad of animal species. Rhipsalis (Cactaceae) is a holoepiphytic plant genus whose fruits are eaten mostly by a single bird genus, the Euphonia. To test the hypothesis that Rhipsalis presents a specialized seed dispersal system, firstly it was studied the reproductive phenology of this epiphyte. Once the availability of ripe fruits to birds was known, occasional frugivory records and focal observation on fruiting individuals of Rhipsalis were made to record the assemblage of animals that eat its fruits. Fruits of five species of Rhipsalis were offered to captive frugivorous birds to access their role as effective dispersers of Rhipsalis seeds. For this, germination tests were made with the seeds after they had passed through the birds’ digestive system. Results showed that Rhipsalis individuals have synchronized reproductive phenophases and the genus has reproductive activity along the year. Ripe fruit production lasted for a long period, between June and February. All species showed significant seasonality in the reproductive phenophases. Frugivory records and germination tests confirmed that the seeds of Rhipsalis may be dispersed by endozoochory. Additionally, the manipulation of the fruits by birds – specifically the removal of the pulp that recovers the seeds – provided higher germination rates and germinability to the seeds, suggesting that the passage though a bird’ gut is important to reactivate the metabolism of the seeds’ embryo. Although 31 animal species were recorded eating Rhipsalis fruits, Euphonia birds may be considered as the main consumer, being responsible for around 60% of feeding visits. Seed dispersal systems involving... (Complete abstract click electronic access below)

Orientador: Marco Aurélio Pizo / Banca: Wesley Rodrigues Silva / Banca: Leonor Patricia Cerdeira Morellato / Resumo: As epífitas são responsáveis por grande parte da diversidade vegetal das florestas neotropicais. São também importantes para a diversidade faunística dessas regiões, pois fornecem recursos alimentares (e.g., flores, frutos e sementes) e micro-ambientes propícios à sobrevivência de vários animais. Rhipsalis (Cactaceae) é um gênero de plantas holoepífitas cujos frutos são consumidos principalmente por um único gênero de aves, as Euphonia. Para verificar a hipótese de especialização na dispersão de sementes de Rhipsalis pelas Euphonia, inicialmente foi estudada a fenologia reprodutiva dessa epífita. Uma vez conhecido o período de disponibilidade de frutos maduros aos frugívoros, foram realizados registros eventuais de frugivoria e observações focais em indivíduos de Rhipsalis para definir a assembléia de frugívoros consumidores de seus frutos. Posteriormente, frutos de cinco espécies de Rhipsalis foram fornecidos a aves frugívoras mantidas em cativeiro. Testes de germinação foram realizados com as sementes que passaram pelo trato digestório dessas aves com o objetivo de verificar seu potencial como dispersoras efetivas das sementes de Rhipsalis. Os resultados obtidos demonstraram que o gênero Rhipsalis apresenta sincronia na produção das fenofases reprodutivas entre os indivíduos e atividade reprodutiva ao longo de todo o ano. A produção de frutos maduros teve duração longa entre os meses de junho e fevereiro. Todas as espécies apresentaram fenofases reprodutivas sazonais. As observações de frugivoria e os testes de germinação realizados confirmaram que as sementes de Rhipsalis podem ser dispersas por endozoocoria. O processamento dos frutos pelas aves frugívoras, especificamente a remoção da polpa que recobre as sementes, não apenas incrementou as taxas de germinação e germinabilidade das sementes, mas também se mostrou importante para... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Epiphytes are responsible for a great part of the plant diversity in Neotropical Forests. They are also important for animal diversity due to the production of food resources (e.g. flowers, fruits and leaves) and microhabitats used by a myriad of animal species. Rhipsalis (Cactaceae) is a holoepiphytic plant genus whose fruits are eaten mostly by a single bird genus, the Euphonia. To test the hypothesis that Rhipsalis presents a specialized seed dispersal system, firstly it was studied the reproductive phenology of this epiphyte. Once the availability of ripe fruits to birds was known, occasional frugivory records and focal observation on fruiting individuals of Rhipsalis were made to record the assemblage of animals that eat its fruits. Fruits of five species of Rhipsalis were offered to captive frugivorous birds to access their role as effective dispersers of Rhipsalis seeds. For this, germination tests were made with the seeds after they had passed through the birds' digestive system. Results showed that Rhipsalis individuals have synchronized reproductive phenophases and the genus has reproductive activity along the year. Ripe fruit production lasted for a long period, between June and February. All species showed significant seasonality in the reproductive phenophases. Frugivory records and germination tests confirmed that the seeds of Rhipsalis may be dispersed by endozoochory. Additionally, the manipulation of the fruits by birds - specifically the removal of the pulp that recovers the seeds - provided higher germination rates and germinability to the seeds, suggesting that the passage though a bird' gut is important to reactivate the metabolism of the seeds' embryo. Although 31 animal species were recorded eating Rhipsalis fruits, Euphonia birds may be considered as the main consumer, being responsible for around 60% of feeding visits. Seed dispersal systems involving... (Complete abstract click electronic access below) / Mestre

Submitted by (edna.saturno@ufrpe.br) on 2016-07-05T13:44:57Z No. of bitstreams: 1 Giseli Maria de Araujo.pdf: 2688552 bytes, checksum: 01677236b208b452a93a129565eea111 (MD5) / Made available in DSpace on 2016-07-05T13:44:57Z (GMT). No. of bitstreams: 1 Giseli Maria de Araujo.pdf: 2688552 bytes, checksum: 01677236b208b452a93a129565eea111 (MD5) Previous issue date: 2009-02-20 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Caatinga has suffered strong degradation due to the people growth and the need for expansion of lands for agriculture, cattle, housing and leisure. Among several environments in the caatinga, the riparian areas are target of devastation due to the proximity of the water, soil fertility and warm weather. The aim of this work was realize a floristic survey of two areas of riparian vegetation in the caatinga and to study the germination of four species that occur in that vegetation. The floristic study was carried out in the municipality of Glória (0253155/9006452 UTM), in Bahia, and Petrolândia (0559297/9005180 UTM), in Pernambuco. The germination study was realized for Albizia inundata (Mart.) Barneby & J.W. Grimes, Piptadenia moniliformis Benth., Triplaris pachau Mart. e Trischidium molle (Benth) H.E. Ireland. The seeds were collected directly from the adult plant, manipulated, processed and submitted to the control treatment, mechanical scarification, 12 hours photoperiod and light absence. 33 families and 59 species were registered. The families with higher species richness in both areas were Cactaceae (6 species), Euphorbiaceae (6species) and Convolvulaceae (5 species). The families with higher richness for the studied areas are the same for non riparian caatinga areas of sandy and rocky soils. The floristic similarity between the two areas was 74%, indicating that those areas are very similar. In relation to the germination, Albizia inundata shows the break of dormancy mechanism after the 12 hours photoperiod treatment. The seeds of Trischidium molle have their percentage of germination enhanced when submitted to the mechanical scarification. The seeds of Triplaris pachau and Piptadenia moniliformis showed similar percentage of germination with the control and scarification treatments, indicating that there is no need for the application of the treatment of dormancy break for the seedling production. / A caatinga tem sofrido forte degradação em decorrência do crescimento populacional e da necessidade de expansão de áreas agricultáveis, pecuarísticas, de moradia e lazer. Dentre os diversos ambientes presentes na caatinga, as áreas ciliares são alvo de grande devastação devido à proximidade do recurso hídrico, fertilidade do solo e condição de clima mais amena. Diante desta realidade objetivou-se realizar o levantamento florístico de duas áreas ciliares da vegetação da caatinga e realizar um estudo de germinação de quatro espécies de ocorrência na vegetação ciliar da caatinga. O estudo florístico foi realizado nos municípios de Glória (0253155/9006452 UTM), na Bahia, e de Petrolândia(0559297/9005180 UTM), em Pernambuco. O estudo de germinação foi realizado para as espécies Albizia inundata (Mart.) Barneby & J.W. Grimes, Piptadenia moniliformis Benth., Triplaris pachau Mart. e Trischidium molle (Benth) H.E. Ireland. As sementes foram coletadas diretamente na planta-mãe, beneficiadas, triadas e submetidas aos tratamentos controle, escarificação mecânica, fotoperíodo de 12 horas e ausência de luz. Foram registradas 33 famílias botânicas e 59 espécies. As famílias queapresentaram maior riqueza de espécies em ambas as áreas foram Cactaceae (6 espécies), Euphorbiaceae (6 espécies) e Convolvulaceae (5 espécies). As famílias de maior riqueza constatada para as áreas ciliares estudadas são as mesmas encontradas nas áreas de caatinga não ciliares de solos arenosos e também nos pedregosos. A similaridade florística entre as duas áreas analisadas foi de 74%, indicando que as áreas são bastante semelhantes. Em relação à germinação, constatou-se que Albizia inundata exibe mecanismo de dormência superada pelo tratamento de fotoperíodo de 12 horas. As sementes de Trischidium molle têm seu percentual de germinação aumentado se submetidas à escarificação mecânica. As sementes de Triplaris pachau e Piptadenia moniliformis apresentaram similares percentuais de germinação no tratamento controle e escarificação, indicando não existir necessidade de aplicação de tratamento de quebra de dormência para a produção de mudas.