The primary aims of this dissertation were to (1) identify and characterize the water-gap complex in seeds of Geraniaceae, (2) investigate its role in physical dormancy (PY) break and (3) develop a new classification system for water-gap complexes in seeds of angiosperms. The winter annuals Geranium carolinianum and G. dissectum were selected as the main representative species for the study, and seeds of an additional 29 species from the Geraniaceae were used to compare the water-gap complex within the family. A new classification system for water-gap complexes in species with PY was developed by comparing the morpho-anatomical features of PY seeds and fruits of 16 families. The water-gap complex of G. carolinianum was identified as a micropyle-hinged valve gap complex, and only a slight morpho-anatomical variation was observed within the family. Ontogenetic studies of the seed coat of G. carolinianum revealed that the water-gap region of Geraniaceae develops as an entity of the micropyle. The timing of seed germination with the onset of autumn can be explained by PY-breaking processes involving (a) two-temperature-dependent steps in G. carolinianum, and (b) one or two moisture-dependent step(s) along with the inability to germinate under high temperatures in G. dissectum. Step-I and step-II in PY-breaking of G. carolinianum are controlled by chemical and physical processes, respectively. This study indicates the feasibility of applying the developed thermal time model to predict or manipulate sensitivity induction in seeds with two-step PY-breaking processes. The model is the first and the most detailed one yet developed for sensitivity induction in PY-break. Based on the morpho-anatomical features, three basic water-gap complexes (types I, II and III) were identified in species with PY in 16 families. Depending on the number of openings involved in initial imbibition, the water-gap complexes were subdivided into simple and compound. The new classification system enables the understanding of relationships between water-gap complexes of taxonomically unrelated species with PY.

The biology of seed dormancy and germination of 46 species representing 11 of the 12 tribes in Convolvulaceae were compared in laboratory (mostly), field and greenhouse experiments. Seeds were tested for kind of dormancy and storage behavior; artificial or simulated natural treatments were applied to break physical dormancy (PY); the initial route of water entry (“water gap”) into seeds was identified; the morphoanatomy of the water gap was compared in seeds of 17 species; ontogenetical differences between water gap and seed coat away from the hilum were described in Ipomoea lacunosa seeds; cycling of sensitivity to dormancy break was elucidated in seeds of I. lacunosa, I. hederacea, Cuscuta australis and Jaquemontia ovalifolia; and mechanism of opening of the water gap was determined for seeds of I. lacunosa and of I hederacea. Seeds of only three of the 46 species were nondormant. Two of these were recalcitrant (Maripa panamensis and Erycibe henryi), and the other one was orthodox (Bonamia menziesii). Seeds of the other 43 species were orthodox and had PY except those of Cuscuta europea, which also had physiological dormancy (PD) i.e. combinational dormancy (PY + PD). Two bulges adjacent to the micropyle were identified as the water gap in all seeds with PY except those of Cuscuta, in which the hilar fissure is the water gap. Anatomy of the bulges (water gap) adjacent to the micropyle differs from that of seed coat away from the bulges. A different sequence and phase of anticlinal and periclinal cell divisions during development created weak transitional zones between bulge - hilum and bulge - seed coat away from hilum. Water vapor pressure changes below the bulges caused formation of the opening(s) in water gap. Seeds of I. lacunosa I. hederacea, C. australis and J. ovalifolia cycle between sensitive and insensitive states to dormancy break, but not between PY and nondormancy. Seed dormancy and storage characteristics and anatomy and morphology of dormancy of seeds of Convolvulaceae closely follow the molecular phylogeny of the family. I suggest that PY in seeds of subfamily Convolvuloideae evolved from nondormant recalcitrant seeds of an ancestor closely related to Erycibeae.

Submitted by LUÍS FELIPE DAIBES DE ANDRADE null (luipedaibes@hotmail.com) on 2017-12-09T19:13:02Z No. of bitstreams: 1 TeseVersaoFinal_LFD.pdf: 4542971 bytes, checksum: dc820c668f873e34980831a5f5f98979 (MD5) / Approved for entry into archive by Adriana Aparecida Puerta null (dripuerta@rc.unesp.br) on 2017-12-11T16:56:37Z (GMT) No. of bitstreams: 1 andrade_lfd_dr_rcla.pdf: 4542971 bytes, checksum: dc820c668f873e34980831a5f5f98979 (MD5) / Made available in DSpace on 2017-12-11T16:56:37Z (GMT). No. of bitstreams: 1 andrade_lfd_dr_rcla.pdf: 4542971 bytes, checksum: dc820c668f873e34980831a5f5f98979 (MD5) Previous issue date: 2017-12-01 / Outra / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O fogo é o principal distúrbio em diversas vegetações ao redor do mundo, denominadas ecossistemas inflamáveis. Neste contexto, muitas espécies possuem estratégias de regeneração e colonização do ambiente pós-fogo, o que tipicamente envolve a sobrevivência (tolerância) das sementes e/ou quebra da dormência. Em especial, muitas sementes de leguminosas possuem o tegumento impermeável (dormência física), que pode ser rompido por meio de choques térmicos relacionados ao fogo. Outro fator que pode auxiliar no processo de quebra da dormência física é a flutuação térmica no solo, cuja amplitude é aumentada nas clareiras formadas pela remoção da vegetação durante a queima. Ambos os fatores, fogo e flutuação térmica, são relativamente bem estudados nos ecossistemas inflamáveis da Austrália e em vegetações Mediterrâneas. Por outro lado, os padrões relacionados à quebra da dormência e germinação das sementes ainda são controversos e menos conhecidos nas savanas tropicais da África e da América do Sul. Assim, esta Tese de doutorado teve como objetivo avaliar o papel do fogo na germinação e sobrevivência das sementes de leguminosas do Cerrado. Para tanto, realizamos tratamentos em campo, incluindo queimas experimentais, e também aplicamos tratamentos no laboratório, simulando a flutuação térmica nas clareiras e a passagem do fogo (choques térmicos). Após os tratamentos, observamos a germinação em condições ótimas, fazendo contagens três vezes por semana, e realizamos testes de viabilidade ao final de 30 dias dos experimentos. As análises estatísticas consistiram basicamente em GLMMs com distribuição binomial, considerando as réplicas como efeitos aleatórios. Os resultados apontam que as sementes morrem quando diretamente expostas ao fogo na superfície do solo. Por outro lado, há uma maior probabilidade de sobrevivência quando as sementes estão localizadas em clareiras (gaps) da vegetação. Nos gaps, a maior porcentagem de solo nu proporciona temperaturas do fogo mais amenas, queimando por menos tempo. Quando enterradas 1-cm sob o solo, as sementes sempre sobrevivem e pode haver quebra de dormência. A flutuação térmica também pode quebrar uma proporção significativa da dormência em condições de campo, especialmente em Mimosa leiocephala. Em laboratório, ao contrário do esperado, não há quebra da dormência, indicando que a flutuação térmica em si não consiste um mecanismo para quebra da dormência física em espécies do Cerrado. Nos choques térmicos, poucas espécies (seis de 46) apresentaram quebra da dormência. Observamos também que a mortalidade das sementes está relacionada a um trade-off entre forma de crescimento, tamanho das sementes e presença de dormência. Isso se deve ao tamanho pequeno das sementes de arbustos, que a despeito de tipicamente ocorrerem em savana aberta, podem morrer sob condições severas dos choques térmicos, mais do que as espécies arbóreas. Dentre as espécies arbóreas, a filogenia influencia no tamanho e, consequentemente, na mortalidade das sementes frente ao fogo. Concluímos que a germinação das sementes não está diretamente ligada à presença do fogo no mosaico Cerrado-floresta, contrastando com os padrões reconhecidos para outros ecossistemas inflamáveis. / Fire is the principal disturbance in several vegetation types around the world, being called flammable ecosystems. In this context, many species have strategies to regenerate and colonize the post-fire environment, which typically involve seed survival (tolerance) and/or breaking of dormancy. In special, legume seeds usually have an impermeable seed coat (physical dormancy), which may be disrupted by fire-related heat shocks. Another factor that can help in the process of dormancy-breaking is the temperature fluctuation in the soil, which amplitude is increased in gaps formed by the removal of vegetation as a result of the passage of fire. Both factors, fire and temperature fluctuation, are relatively well studied in flammable ecosystems from Australia and Mediterranean vegetation. On the other hand, the patterns related to dormancy-breaking and seed germination are still controversial and less known in tropical savannas from Africa and South America. Therefore, this doctoral Thesis has aimed to evaluate the role of fire on germination and survival of legume seeds from Cerrado. Therefore, we conducted experiments in the field, including experimental burning, and also applied treatments in the laboratory, simulating temperature fluctuation in the gaps and fire passage (heat shocks). After the treatments, we observed germination under optimal conditions, making counting three times a week, and accomplishing viability tests by the end of 30 days of the experiment. The statistical analyses basically consisted on GLMMS with binomial distribution, considering replicates as random effects. Results pointed that seeds die when directly exposed to fire in the soil surface. On the other hand, there is a higher probability of survival when seeds are placed in vegetation gaps. In the gaps, the higher percentage of bare soil provides milder fire temperatures, lasting for less time. When buried at 1-cm belowground, seeds always survive and dormancy-breaking may occur. Temperature fluctuation can also break a significant proportion of dormancy under field conditions, especially for Mimosa leiocephala. In the laboratory, there is no dormancy-breaking, showing that temperature fluctuation itself might not be a mechanism for physical dormancy-breaking in Cerrado species. When heat shocks were applied, six out of 46 species showed dormancy-breaking. We also observed that seed mortality is related to a trade-off between growth-form, seed size and the presence of dormancy. This is due to the small size of seeds from shrubs, that although typically occurring in the open savannas, they might die under severe conditions of temperatures. Among the tree species, phylogeny influences on size and, consequently, mortality of seeds in face to fire. We concluded that seed germination is not directly linked to the presence of fire in the Cerrado-forest mosaic, contrasting to the patterns recognized for other flammable ecosystems. / Fundação Grupo Boticário 0153_2011_PR / FAPESP 2015/06743-0 / CAPES/PDSE 88881.131702/2016-01 / CNPq 455183/2014-7