Title page, contents and abstract only. The complete thesis in print form is available from the University of Adelaide Library. / Pre-harvest sprouting (PHS) is an important economic problem which affects a significant proportion of the Australian wheat crop through quality downgrading. Grain dormancy is the most effective means of overcoming germination in the wheat spikelet at harvest maturity. It has been a consistent observation over a long period of time that dormant red-grained wheat genotypes are almost more dormant than dormant white-grained genotypes. In white-grained wheat, there are two factors which contribute to dormancy, embryo sensitivity to abscisic acid (ABA) and an interacting and unknown seed coat factor. The proposed dormancy model is that complete dormancy can only be achieved with the coordinate expression of these two factors. This primary objective of this project was to determine the role of this putative seed coat factor in grain dormancy of white-grained wheat."--Abstract. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1259900 / Thesis (Ph.D.) -- University of Adelaide, School of Agriculture, Food and Wine, 2006

Title page, contents and abstract only. The complete thesis in print form is available from the University of Adelaide Library. / Pre-harvest sprouting (PHS) is an important economic problem which affects a significant proportion of the Australian wheat crop through quality downgrading. Grain dormancy is the most effective means of overcoming germination in the wheat spikelet at harvest maturity. It has been a consistent observation over a long period of time that dormant red-grained wheat genotypes are almost more dormant than dormant white-grained genotypes. In white-grained wheat, there are two factors which contribute to dormancy, embryo sensitivity to abscisic acid (ABA) and an interacting and unknown seed coat factor. The proposed dormancy model is that complete dormancy can only be achieved with the coordinate expression of these two factors. This primary objective of this project was to determine the role of this putative seed coat factor in grain dormancy of white-grained wheat."--Abstract. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1259900 / Thesis (Ph.D.) -- University of Adelaide, School of Agriculture, Food and Wine, 2006

Phytophthora cinnamomi has been recognised as a key threatening process to Australia’s biodiversity by the Commonwealth’s Environment Protection and Biodiversity Conservation Act 1999. Despite over 80 years of extensive research, its exact mode of survival is still poorly understood. It is widely accepted that thin- and thick-walled chlamydospores are the main survival propagules while oospores are assumed to play no role in the survival of the pathogen in the Australian environment, yet evidence is limited. The saprophytic ability of the pathogen is still unresolved despite the important role this could play in the ability of the pathogen to survive in the absence of susceptible hosts. This thesis aimed to investigate chlamydospores, oospores and the saprophytic ability of P. cinnamomi to determine their contribution to survival. Phytophthora cinnamomi did not show saprophytic ability in non-sterile soils. The production of thick-walled chlamydospores and selfed oospores of P. cinnamomi in vitro was documented. Thick-walled chlamydospores were sporadically formed under sterile and non-sterile conditions in vitro but exact conditions for stimulating their formation could not be determined. The formation of thick-walled chlamydospores emerging from mycelium of similar wall thickness was observed, challenging the current knowledge of chlamydospore formation. Selfed oospores were abundant in vitro on modified Ribeiro’s minimal medium in one isolate. Three other isolates tested also produced oospores but not in large numbers. Although the selfed oospores did not germinate on a range of media, at least 16 % were found to be viable using Thiozolyl Blue Tetrazolium Bromide staining and staining of the nuclei with 4´, 6-diamidino-2-phenylindole.2HCl (DAPI). This indicated the potential of selfed oospores as survival structures and their ability to exist dormantly. The ability of phosphite to kill chlamydospores and selfed oospores was studied in vitro. Results challenged the efficacy of this chemical and revealed the necessity for further study of its effect on survival propagules of P. cinnamomi in the natural environment. Phosphite was shown to induce dormancy in thin-walled chlamydospores if present during their formation in vitro. Interestingly, dormancy was only induced by phosphite in isolates previously reported as sensitive to phosphite and not those reported as tolerant. Chlamydospores were produced uniformly across the radius of the colony on control modified Ribeiro’s minimal medium but on medium containing phosphite (40 or 100 µg ml-1), chlamydospore production was initially inhibited before being stimulated during the log phase of growth. This corresponded to a point in the colony morphology where mycelial density changed from tightly packed mycelium to sparse on medium containing phosphite. This change in morphology did not occur when the pathogen was grown on liquid media refreshed every four days, and chlamydospores were evenly distributed across the radius of these colonies. This trend was not observed in selfed oospores produced in the presence of phosphite. Selfed oospore production was found to be inhibited by phosphite at the same concentrations that stimulated chlamydospore production. Isolates of P. cinnamomi were transformed using a protoplast/ polyethylene glycol method to contain the Green Fluorescent Protein and geneticin resistance genes to aid in future studies on survival properties of the organism. Although time constraints meant the stability of the transgene could not be determined, it was effective in differentiating propagules of the transformed P. cinnamomi from spores of other microrganisms in a non-sterile environment. Two different sized chlamydospores (approximately 30 µm diameter and < 20 µm diameter) were observed in preliminary trials of transformed P. cinnamomi inoculated lupin roots floated in non-sterile soil extracts and these were easily distinguished from microbial propagules of other species. The growth and pathogenicity was reduced in two putative transformants and their ability to fluoresce declined over ten subcultures but they still remained resistant to geneticin. This study has improved our knowledge on the survival abilities of P. cinnamomi in vitro and has provided a useful tool for studying these abilities under more natural glasshouse conditions. Important implications of phosphite as a control have been raised.

Thesis (MScAgric)--University of Stellenbosch, 2002. / ENGLISH ABSTRACT: In order to study the dormancy of apple buds in conditions of inadequate chilling a number of trails were done during 2000 and 2001. Year-old, unbranched shoots of 'Royal Gala', 'Braeburn', 'Cripps' Pink' and 'Granny Smith' apple were harvested randomly from bearing commercial orchards in the Koue Bokkeveld [33°S, 945m, ca.1300 Utah model chili units (CU)) and Elgin (34 "S, 305m, ca.750CU) regions of the Western Cape, South Africa, respectively. Shoots were chilled at 5-rC. Two replicate bundles were removed from the cold room, prepared and forced at 25°C with continuous illumination until budburst had occurred on at least 25% of the shoots per bundle. The change in the rate of budburst over time was calculated for each orchard and to this response; either a linear or a quadratic function was fitted. Poorly correlated variables were selected that best describe these functions. Using these variables, the orchards were separated into cluster groupings that represented a dormancy pattern. The first split separated the lower chilling requirement cultivars from the higher cultivars. The second and third split separated the orchards according to area differences. The clusters representing the warmer area orchards initially entered deeper into dormancy before exiting. The clusters for the colder area immediately had an increased budburst rate. This data confirm that the chilling requirement includes a period of dormancy induction. An important genotype and environment interaction, other than cold unit accumulation, was observed that could be responsible for terminating bud dormancy. Terminal apple buds from 'Royal Gala' Braeburn', 'Cripps' Pink' and 'Granny Smith' apples were cut from orchards in the Koue Bokkeveld and Elgin regions of the Western Cape, South Africa. Buds were harvested every two weeks during the dormant period. The buds were cut in half and leaf scales removed before the water potential were measured. Fresh and dry weights of the buds were also determined. The data presented confirms the changes in availability of free water in dormant buds and that it could be measured in this way. A definite influence of temperature was illustrated. The water potential from buds in a cold production area (Koue Bokkeveld) behaved more "normally" - water is in a bound form during most of the winter and change to an available form later in winter - whereas buds from a warmer production area did not change much in water potential or content. In the trial, two-year-old proleptic-branched shoots, ca. 500mm long, were selected from a 'Royal Gala' orchard in the Koue Bokkeveld region in the Western Cape, South Africa. During the dormancy period of 2000, shoots received two cold treatments; chilling in a cold room at 5-7°C and the natural chilling received in the field. In 2001, the trail was repeated, but only with the field chilling. The shoots received five dormant pruning treatments: control (not pruned), pruning back to the fourth lateral (heading) before or after chilling; and removal of the 2nd and 3rd laterals (thinning) before or after chilling. After pruning and chilling treatments, the shoots were removed from the orchard or cold room every two weeks and forced in a growth chamber. The rate of bud burst (1/days to 50% bud burst) was calculated for the terminal buds of the lateral shoots. Laterals were categorisation according to position: the terminal extension shoot, the 4th lateral, and all other laterals were pooled. Removing distal tissue by pruning (heading more than thinning) promoted bud burst on laterals. Pruning before chilling was more effective than after chilling. The correlative phenomena that inhibit bud burst on proximal shoots within two-yearold branches were manipulated by pruning. The branching response of one-year-old unbranched shoots, 0.5m long, from 'Royal Gala' and 'Cripps' Pink' apple and 'Rosemary' pear were studied after physical manipulation treatments. Shoots for treatment a to d were re-orientated from either a horizontal or vertical position or left in the original position as control, treatment e to h involved the same re-orientation of shoots and were headed. The amount of growth (in mm) from each node was recorded as well as the position from the terminal bud. The 'Cripps' Pink' had a definite shift in the acrotonic branching pattern (for headed and unheaded), towards a more basitonic response. The reduced effect on 'Royal Gala' and 'Rosemary' suggest a difference in genotype response to the treatments as well as time of treatment. / AFRIKAANSE OPSOMMING: Ter wille van die navorsing oor die invloed van gebrekige koue op dormansie van apple knoppe en die gepaardgaande probleme is 'n reeks proewe gedoen gedurende 2000 en 2001. Jaar oue onvertakte lote van 'Royal Gala', 'Braeburn' , 'Cripps' Pink' en 'Granny Smith' appels is ewekansig geoes vanaf komersieële boorde in die Koue Bokkeveld [33°S, 945m, ca.1300 Utah koue eenhede (CU)] en Elgin (34°S, 305m, ca.750CU) omgewings van die Wes Kaap , Suid Afrika. Die lote is daarna verkoel gehou by 5- 7°C. Lote is elke twee weke vanuit die koue kamer geneem en geforseer met 25°C en deurlopend belig. Die aantal knoppe wat groenpunt bereik het is genoteer totdat 25% van die lote begin bot. Die verandering oor tyd vir elke boord is bereken en 'n liniëre of kwadratiese funksie is daarop gepas. Swak gekorreleerde waardes is gekies wat die funksies die beste beskryf. D.m.v hierdie die waardes is die boorde in groepe ingedeel wat 'n dormansie patroon verteenwoordig. Na die eerste vedeeling is die hoë en lae koue behoefte kultivars geskei. In opvolgende verdelings is die boorde verder in die twee areas geskei met elk 'n spesifieke dormansie patroon. Die groepe wat die warmer area se boorde bevat het aanvanklik dieper in dormansie in beweeg voor dit 'n styging in groei potensiaal getoon het. Die groepe vanaf die kouer produksie area het onmiddelik 'n verhooging in bot tempo getoon. Die data bevestig dat 'n koue behoefte 'n periode van dormansie induksie insluit. 'n Belangrike kultivaromgewing- interaksie, ten spyte van koue eenhede akkumulasie, is waargeneem wat verantwoordlik kan wees vir beëindiging van dormansie. Om die verandering van water status in dormante appel knoppe te bestudeer, was die volgende proef uitgevoer. Terminale apple knoppe van dieselfde vier kultivars en vanaf die selfde twee areas in die Wes Kaap as bogenoemde, is elke tweede week gedurende die winter 2001 gesny vanaf komersieële boorde. Daarna is die knoppe middel deur gesny en die skudblare verwyder voor die vars massa gemeet en daarna die waterpotensiaal bepaal is. Die data het bevestig dat daar veranderings in beskikbaarheid van vry water in dormante knoppe gedurende dormansie plaasvind. 'n Definitiewe invloed van temperatuur op waterpotensiaal is geïllustreer. Die waterpotensiaal van knoppe in die kouer produksie area toon 'n meer normale respons (gedurende die winter is die water in 'n gebonde vorm, wat later in die winter na vry water verander). Daar teenoor is daar in die knoppe van die warm produksie area nie veel verandering in waterpotensiaal of water massa getoon nie. Gedurende die winter van 2000 is twee jaar oue proleptiese vertakte 'Royal Gala' lote, ongeveer 500mm lank, gekies vanaf 'n boord in die Koue Bokkeveld. Die lote is verdeel en het twee koue behandelings ontvang. Koue kamer by 5-T'C en natuurlike koue in die boord. In 2001 is net die natuurlike koue behandeling herhaal. Daar is vyf dormante snoei behandeling op die lote gedoen; kontrole (geen), snoei terug tot die vierde lateraal voor en na die koue; verwydering van die 2de en 3de laterale voor en na koue. Na koue en snoei is die lote verwyder en in die groeikamer geforseer by 25°C en met konstante illuminasie. Die tempo waarteen die laterale gebot het is bereken (1\dae to 50% groen punt) waarna die laterale in klasse gedeel is; terminale knop, die vierde lateraal en die res van die laterale saam. Verwydering van distale weefsel d.m.V. snoei verhoog die tempo waarteen laterale knoppe groenpunt bereik. Snoei voor die koue behandeling was meer effektief as daarna. Die korrelatiewe fenomeen wat bot inhibeer van proksimale lote kan gemanipuleer word in die twee jaar apple loot. Die vertakkings gedrag van een jaar oue onvertakte 'Royal Gala', 'Cripps' Pink' apple en 'Rosemary' peer lote, 0.5m lank, is ondersoek na fisiese manipulasies gedoen is gedurende mid winter 2000. Vir die eerste vier behandelings (a,b,c,d) is die lote in 'n horisontale of vertikale possisie gelos as kontrole, of horisontale of vertikale gebuig en daar gehou d.m.v. binddraad. Behandelings e tot h het dieselfde behels maar die lote is ook nog in die helfte deur gesny (getop). Die hoeveelheid groei in mm vir elke node is aangeteken asook die posisie vanaf die terminale knop. Die 'Cripps' Pink' het 'n definitiewe veskuiwing vanaf 'n akrotoniese na 'n basitoniese vertakkingspatroon getoon. Die reaksies op die manipulasies of die gebrek daaraan ('Royal Gala' en 'Rosemary' ) kan 'n aanduiding wees dat genotipes verskillend reageer op die behandelings asook op die tyd wat dit gedoen was.

Made available in DSpace on 2014-06-11T19:30:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-02-25Bitstream added on 2014-06-13T21:01:06Z : No. of bitstreams: 1 000755167.pdf: 2367968 bytes, checksum: 406c0b563423541bacf6a7c59f70fbe2 (MD5) / A ocorrência de dormência se opõe ao pleno sucesso da propagação futura das espécies nativas de interesse para fins comerciais. Assim, o principal objetivo é o estudo fisiológico e molecular de sementes (embrião) de Annona crassiflora durante a superação da dormência morfofisiológica natural no campo. As sementes foram coletadas de áreas do cerrado do Estado de Minas Gerais, secas em temperatura entre 20-25ºC, em condições de laboratório, até atingirem umidade de 10% com base úmida. Foram feitas no mesmo lote de sementes a determinação da curva de embebição em água, germinação em campo, teor de água e crescimento do embrião mensal. As sementes foram colocadas em sacos de poliester com orifícios de 0,5 cm de diâmetro e, enterradas no mês de abril. Em paralelo, sementes foram colocadas para embeber em solução de giberelina (GA4) na concentração de 100μM, por oito dias e tiveram o comprimento do embrião e teor de água avaliados. Em novembro, as sementes enterradas a campo foram desenterradas e as que apresentaram rupturas no tegumento (sem protrusão radicular) tiveram o embrião isolados para a extração de RNA. Em seguida a integridade do RNA foi verificada em Bioanalyzer (Agilent 2100) e as bibliotecas foram preparadas utilizando o protocolo TruSeq RNA sample prep v2. O sequenciamento foi realizado no HiScanSQ (Illumina) e os dados gerados analisados pelo plataforma CLC Genomics Workbench versão 6.0.2 (CLC bio, Denmark) para montagem do transcriptoma referência e quantificação da expressão por valores de RPKM (Reads per Kilobase per Million of mapped reads). Os contigs foram anotados usando o programa BLAST2Go e manualmente. A semente de A. crassiflora estava totalmente embebida em torno de sete dias de embebição e o teor de água variou de acordo com as condições de umidade do solo, sendo maior no período de germinação. O crescimento do embrião iniciou-se... / The occurrence of dormancy opposes to the success of the future propagation of species. Thus, the objective of this study was to performe physiological and molecular studies em seed of Annona crassiflora during overcoming morphological dormancy under field onditions. The seeds were collected from areas of the Cerrado biome from the Minas Gerais State, dried at room temperature until the moisture content of 10% on wet basis. The imbibtion curve, germination in the field, water content and growth of the embryo were made in the same batch of seeds. The seeds were placed in polyester bags with holes of 0.5 cm diameter, buried in April. In parallel, seeds were also imbibed in gibberellin (GA4) at a concentration of 100μM during seven days and had the embryo length measured. In October, embryos were isolated from seeds showing growth of the embryo but without radicle protrusion for RNA extraction. The integrity of the RNAs was verified in Bioanalyzer (Agilent 2100). Following, cDNA libraries were prepared using the TruSeq RNA sample prep protocol v2. Sequencing was performed in HiScanSQ (Illumina) and the analyses of the data was performed by using the CLC Genomics Workbench platform version 6.0.2 (CLC bio, Denmark) for transcriptome assembly and quantification of the expression values for RPKM (Reads per kilobase per Million of mapped reads). The contigs were anotados using the program BLAST2Go and manually. The seed is fully imbibed at seven days imbibitions, and the water content varies with soil conditions being higher during germination. The growth of the embryo starts in August and maintained until January, with peak of germination observed in November. After this period, the seeds that did not germinate formed a soil seed bank and there was no growth of the embryo. After 28 days from the start of imbibitions in GA4 the first seeds showed testa rupture indicating growth of the embryo. From the ...

Made available in DSpace on 2014-12-02T11:16:45Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-07-30Bitstream added on 2014-12-02T11:21:41Z : No. of bitstreams: 1 000800296.pdf: 1283716 bytes, checksum: 1d4a030d4a8f9b3af5d83c8002c481d7 (MD5) / A Brachiaria humidicola cv. BRS Tupi é a primeira cultivar de B. humidicola indicada para uso no Brasil e surge como mais uma opção para áreas úmidas de baixa e média fertilidades. C omo a maioria das gramíneas forrageiras tropicais, sementes recém - colhidas dessa cultivar são afetadas pelo fenômeno da dormência . Apesar da evidente importância da dormência em sementes recém - colhidas de Brachiaria , os mecanismos que estão envolvidos na superação da dormência que ocorre durante o armazenamento ainda são pouco elucidados. Assim, este estudo foi conduzido com o objetivo de avaliar a superação da dormência em sementes de B. humidicola cv. BRS Tupi durante diferentes períodos de armazenament o (0, 3, 6, 9, e 12 meses). Sementes de B. humidicola cv. BRS Tupi foram multiplicadas em campo e, após o processo de secagem em ambiente natural e beneficiamento , foram armazenadas. Após os períodos de armazenamento , as sementes foram avaliadas quanto as suas alterações fisiológicas através de curvas de embebição, do teor de água das partes da semente (embrião e endosperma) durante a embebição, germinação de sementes inteiras , germinação de sementes inteiras e cariops es nuas em diferentes concentrações de ácido giberélico, potencial de crescimento do embrião de cariopses nuas em restrição hídric a e germinação de sementes escarificadas. O armazenamento favorece o aumento do teor de água das partes da semente durante a e mbebição. A superação da dormência ocorre com a retirada das coberturas da semente indicando possíveis impedimentos mecânicos que são superados durante o armazenamento devido ao aumento no potencial de crescimento do embrião, provavelmente relacionado à aç ão de GA nas sementes . A escarificação química não foi eficiente na superação da dormência durante o armazenamento / The Brachiaria humidicola cv. BRS Tupi is the first cultivar of B. humidicola indicated for use in Brazil and comes up as one more option for the humid areas of low and average fertility. As the majority of the tropical forage grass, freshly harvested se eds of this cultivar are affected by the phenomenon of dormancy. Despite the obvious importance of dormancy in freshly harvested seeds of Brachiaria , the mechanisms that are involved in overcoming dormancy that occurs during storage are still poorly unders tood. Thus, the present study was carried out with the aim to evaluate the overcoming dormancy in seeds of B. humidicola cv. BRS Tupi during different storage periods (0, 3, 6, 9, and 12 months). Seeds of B. humidicola cv. BRS Tupi were multiplied in the f ield, and after the drying process in the natural environment and processing, were stored. After the storage periods the seeds were evaluated according to its physiological changes through imbibition curves, evaluation of the water content of the seed part s (embryo and endosperm) during imbibition, germination of whole seeds , germination in different concentrations of gibberellic acid of whole seeds and naked caryopses, potential of growth of the embryo of naked caryopses in water restriction and germination of scarified seeds. The storage favors the increase of the water conten t of the parts of the seed during the imbibitions. The overcoming of dormancy happens with the removal of the seed coat showing possible mechanical barrier which is overcome during the storage due to the raise in the potential of growth of the embryo, possibly related to the action of GA on the seeds. The chemical scarific ation was not efficient in overcoming the dormancy during the storage

Worldwide production of potatoes is in excess of 350 million metric tonnes per annum, of which 60% is intended for human consumption. Since the period of tuber dormancy before tuber buds sprout is usually shorter than the optimal market storage period, control of sprouting is essential. To prolong dormancy, tubers are either stored at low temperatures and/or are treated with chemical sprout inhibitors, the use of which subject to increasing scrutiny in the European Union due to their impact upon the environment. Cytokinins, a group of plant growth regulators, are known to play a central role in tuber bud sprouting and tuber initiation from stolon tips in Solanum tuberosum L. although it is unclear when and how cytokinins act to regulate dormancy. The interconversion of cytokinins is incompletely understood. Enzymes identified to date have higher affinities for aminopurines than their cytokinin equivalents. A novel cytokinin binding protein Solanum tuberosum Cytokinin Phosphorylase 1 (StCKP1), has been identified in tuberising stolon tips which shares regions of homology with members of the nucleosidase and phosphoribosyl transferase family. The composition of cytokinin N9 conjugates in tuber bud and surrounding tissue is known to change on transition from a dormant state, with an increase in base and riboside types observed. Analysis of transcripts indicates an increase in abundance of StCKP1 on tuberisation of stolon tips, and high abundance in the periderm of dormant tubers. Analysis of protein abundance by immunoblotting echoes this finding and indicates StCKP1 begins to accumulate in stolon tips shortly before tuberisation, matching binding activity. Transgenic analysis of the cytological reporter gene uidA under the control of two identified promoter regions indicates StCKP1 is expressed predominantly in tuber tissue. Analysis of StCKP1 activity by HPLC and LC-MS-MS shows that StCKP1 catalyses the interconversion of free base and riboside. KMs determined for cytokinin and aminopurine substrates indicate that StCKP1 has a higher affinity for cytokinin substrates and, of these cytokinins, displays a higher affinity for the free base catalysing ribosylation of the N9 to form the corresponding riboside. Desiree cultivars over-expressing StCKP1 under the CaMV 35S promoter exhibited an increased rate of tuberisation of stolon tips and an increase in the length of the dormant period following lifting. Over-expression of StCKP1 was found in particular to increase the chill sensitive period of dormancy, confirming results of StCKP1 knock-down by RNAi. Transcript abundance of StCKP1 at tuberisation in other cultivars including King Edward and Maris Peer was found to correlate with the dormancy characteristics prescribed by the European Cultivated Potato Database and the British potato variety database.

Orientador: Edvaldo Aparecido Amaral da Silva / Banca: Juliana Pereira Bravo / Banca: Adriana Rodrigues Passos / Banca: João Nakagawa / Banca: Alessandro de Mello Varani / Resumo: A ocorrência de dormência se opõe ao pleno sucesso da propagação futura das espécies nativas de interesse para fins comerciais. Assim, o principal objetivo é o estudo fisiológico e molecular de sementes (embrião) de Annona crassiflora durante a superação da dormência morfofisiológica natural no campo. As sementes foram coletadas de áreas do cerrado do Estado de Minas Gerais, secas em temperatura entre 20-25ºC, em condições de laboratório, até atingirem umidade de 10% com base úmida. Foram feitas no mesmo lote de sementes a determinação da curva de embebição em água, germinação em campo, teor de água e crescimento do embrião mensal. As sementes foram colocadas em sacos de poliester com orifícios de 0,5 cm de diâmetro e, enterradas no mês de abril. Em paralelo, sementes foram colocadas para embeber em solução de giberelina (GA4) na concentração de 100μM, por oito dias e tiveram o comprimento do embrião e teor de água avaliados. Em novembro, as sementes enterradas a campo foram desenterradas e as que apresentaram rupturas no tegumento (sem protrusão radicular) tiveram o embrião isolados para a extração de RNA. Em seguida a integridade do RNA foi verificada em Bioanalyzer (Agilent 2100) e as bibliotecas foram preparadas utilizando o protocolo TruSeq RNA sample prep v2. O sequenciamento foi realizado no HiScanSQ (Illumina) e os dados gerados analisados pelo plataforma CLC Genomics Workbench versão 6.0.2 (CLC bio, Denmark) para montagem do transcriptoma referência e quantificação da expressão por valores de RPKM (Reads per Kilobase per Million of mapped reads). Os contigs foram anotados usando o programa BLAST2Go e manualmente. A semente de A. crassiflora estava totalmente embebida em torno de sete dias de embebição e o teor de água variou de acordo com as condições de umidade do solo, sendo maior no período de germinação. O crescimento do embrião iniciou-se... / Abstract: The occurrence of dormancy opposes to the success of the future propagation of species. Thus, the objective of this study was to performe physiological and molecular studies em seed of Annona crassiflora during overcoming morphological dormancy under field onditions. The seeds were collected from areas of the Cerrado biome from the Minas Gerais State, dried at room temperature until the moisture content of 10% on wet basis. The imbibtion curve, germination in the field, water content and growth of the embryo were made in the same batch of seeds. The seeds were placed in polyester bags with holes of 0.5 cm diameter, buried in April. In parallel, seeds were also imbibed in gibberellin (GA4) at a concentration of 100μM during seven days and had the embryo length measured. In October, embryos were isolated from seeds showing growth of the embryo but without radicle protrusion for RNA extraction. The integrity of the RNAs was verified in Bioanalyzer (Agilent 2100). Following, cDNA libraries were prepared using the TruSeq RNA sample prep protocol v2. Sequencing was performed in HiScanSQ (Illumina) and the analyses of the data was performed by using the CLC Genomics Workbench platform version 6.0.2 (CLC bio, Denmark) for transcriptome assembly and quantification of the expression values for RPKM (Reads per kilobase per Million of mapped reads). The contigs were anotados using the program BLAST2Go and manually. The seed is fully imbibed at seven days imbibitions, and the water content varies with soil conditions being higher during germination. The growth of the embryo starts in August and maintained until January, with peak of germination observed in November. After this period, the seeds that did not germinate formed a soil seed bank and there was no growth of the embryo. After 28 days from the start of imbibitions in GA4 the first seeds showed testa rupture indicating growth of the embryo. From the ... / Doutor

La graine est le produit de la reproduction sexuée chez les Angiospermes. Elle assure la survie et la dispersion de l'espèce. La germination des graines est la première étape de la croissance des plantes. La transition entre l'état de dormance des graines et leur germination est une étape clef dans le cycle de vie des plantes qui a des conséquences écologique et commerciale. Depuis plusieurs décennies, de nombreux facteurs de l'environnement ont été étudiés pour leurs implications et leurs conséquences dans le processus de dormance et de germination des graines. Les études sur la réponse des semences aux changements de température en liens avec le réchauffement climatique ont un intérêt primordial. Le but de ce travail a été d'étudier la régulation de la dormance et de la germination des graines de tournesol par la température. Une analyse protéomique et un profilage enzymatique ont été réalisés afin de mieux comprendre la régulation du métabolisme pendant la levée de dormance par la température. L'utilisation d'approches moléculaires et cytologiques, nous ont permis d'appréhender l'interaction entre la température et les phytohormones impliquées dans ce processus. Nos résultats ont révélé le rôle joué par la température comme facteur externe affectant la dormance et la germination des graines en agissant d'une part sur le métabolisme et d'autre part sur la quantité et la localisation cellulaire des principales hormones endogènes. / A seed is the product of sexual reproduction and the means by which the new individual is dispersed by angiosperms. Seed germination being the first step of plant establishment, the ultimate role of the transition between seed dormancy and germination during plant lifecycle is an important ecological and commercial trait. Last several decades, several environment factors have been reviewed to strongly effect the process of seed dormancy and germination. However, studies about seed response to temperature change are acute with the global warming. The aim of this work was to investigate temperature regulation of dormancy and germination in sunflower seeds. Proteomic analysis and enzyme profiling have been used to study metabolism regulation during seed dormancy release by temperature. Moreover, using molecular and cytological approaches, we investigate the interaction between temperature and phytohormones involved in this process. Our results revealed that temperature as an external factor to effect seed dormancy and germination by affecting, in one hand, the metabolism, and in the other hand the level and localization of major endogenous hormones.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / Este estudo apresenta a hipÃtese de que P. stipulacea à uma espÃcie caracterÃstica de estÃgio inicial de sucessÃo, enquanto A. colubrina de estÃgios mais avanÃados. Para caracterizar as duas espÃcies, estudaram-se alguns atributos utilizados na classificaÃÃo de grupos ecolÃgicos de florestas tropicais Ãmidas, respondendo as seguintes questÃes: a) as espÃcies apresentam dormÃncia de sementes? b) quais as condiÃÃes de temperatura e luz para germinaÃÃo? c) as sementes apresentam tolerÃncia à dessecaÃÃo? d) quais as caracterÃsticas de crescimento das plÃntulas sob diferentes graus de irradiÃncia? Dessa forma, o trabalho apresentou as seguintes previsÃes: a) as sementes de P. stipulacea apresentam dormÃncia, enquanto as de A. colubrina nÃo apresentam; b) as sementes de P. stipulacea germinam em uma ampla faixa de temperatura, enquanto as de A. colubrina germinam em uma faixa de temperatura mais estreita; c) as sementes de P. stipulacea toleram baixos nÃveis de dessecaÃÃo, enquanto as de A. colubrina nÃo toleram; d) as plÃntulas de P. stipulacea crescem melhor em maiores nÃveis de irradiÃncia, enquanto as de A. colubrina crescem melhor em menores nÃveis de irradiÃncia. Os frutos foram coletados em Ãrea de caatinga situada na fazenda âNÃo Me Deixesâ, no municÃpio de QuixadÃ-CE, e os experimentos foram realizados no LaboratÃrio de AnÃlise de Sementes e no NÃcleo de Ensino e Pesquisa em Agricultura Urbana. Resultados do teste preliminar de germinaÃÃo mostraram que sementes de P. stipulacea apresentam dormÃncia, sendo melhor superada com escarificaÃÃo quÃmica por 10 minutos. Jà as sementes de A. colubrina nÃo possuem dormÃncia. O experimento de temperatura e luz revelou que as duas espÃcies apresentam a mesma amplitude de germinaÃÃo (10ÂC a 40ÂC), com melhor germinaÃÃo a 30ÂC, porÃm, nas menores temperaturas P. stipulacea apresentou germinaÃÃo muito baixa. Quanto à tolerÃncia a dessecaÃÃo, as sementes de P. stipulacea suportaram secagem atà 5,16% de umidade sem influenciar a germinaÃÃo, representando comportamento ortodoxo. Jà com A. colubrina a germinaÃÃo diminuiu à medida que as sementes perderam Ãgua, porÃm, mesmo com teor de Ãgua de 5,81%, a germinaÃÃo permaneceu acima de 80%, indicando que esta espÃcie tambÃm apresenta sementes ortodoxas. No experimento de irradiÃncia, plÃntulas de P. stipulacea apresentaram maior diÃmetro a pleno sol e, em todos os tratamentos, acumularam maior biomassa para o caule. A. colubrina apresentou maior altura e diÃmetro em 50% de sombra e maior nÃmero de folhas e biomassa da parte aÃrea em 70% de sombra, indicando que esta espÃcie tolera nÃveis intermediÃrios de sombra. Sugere-se que caracterÃsticas utilizadas na classificaÃÃo de grupos ecolÃgicos de florestas tropicais Ãmidas tambÃm possam ser utilizadas para subsidiar a identificaÃÃo de qual estÃgio sucessional pertencem determinadas espÃcies da caatinga, com o objetivo de entender sua ocorrÃncia, distribuiÃÃo e desenvolvimento, para melhor conservaÃÃo e uso adequado no reflorestamento de Ãreas degradadas. / This study presents the hypothesis that P. stipulacea is a species characteristic of early stages of succession, while A. colubrina in later stages. To characterize the two species studied are some attributes used in classification of ecological groups of tropical rainforests, answering the following questions: a) species have seed dormancy? b) what conditions of temperature and light for germination? c) the seeds are desiccation tolerance? d) the characteristics of seedling growth under different levels of irradiance? Thus, the work made the following predictions: a) seeds of P. stipulacea present dormancy, while those of A. colubrina not present; b) seeds of P. stipulacea germinate over a wide temperature range, while those of A. colubrina germinate in a narrow temperature range; c) the seeds of P. stipulacea tolerate low levels of desiccation, while those of A. colubrina not tolerate; d) seedlings of P. stipulacea grow best at higher irradiance levels, while those of A. colubrina grow best at lower levels of irradiance. Fruits were collected in the caatinga area located on the farm "NÃo Me Deixesâ city of Quixada-CE, and the experiments were performed at the Seed Analysis Laboratory and the Core Teaching and Research in Urban Agriculture. Results of preliminary testing of germination showed that seeds of P. stipulacea present dormancy and are better solved with chemical scarification for 10 minutes. The seeds of A. colubrina have no dormancy. The experiment temperature and light was revealed that the two species have the same range of germination (10ÂC to 40ÂC), with best germination at 30ÂC, but at lower temperatures P. stipulacea showed germination very low. As for tolerance to desiccation, seeds of P. stipulacea bore drying up 5.16% moisture without influencing germination, representing orthodox behavior. Already A. colubrina germination decreased as the seeds lost water, but even with a water content of 5.81%, germination remained above 80%, indicating that this species also has orthodox seeds. In the irradiance experiment, seedlings of P. stipulacea showed larger diameter in full sun, and all treatments accumulated more biomass to the stem. A. colubrina showed greater height and diameter in 50% shade and a larger number of leaves and shoot biomass by 70% shade, indicating that this species tolerates intermediate levels of shade. It is suggested that features used for classification of ecological groups of tropical forests could also be used to support the identification of which stage of succession belongs to certain species of caatinga, with the aim of understanding their occurrence, distribution and development, for better conservation and use suitable for the reforestation of degraded areas.