Preharvest sprouting (PHS), the germination of seeds on the maternal plant before harvest, is a big challenge for barley producers worldwide. It is attributed mainly to low seed dormancy. The balance between two classical plant hormones, gibberellin (GA) and abscisic acid (ABA) regulates seed dormancy and germination, and the endogenous level of these two hormones in plants is determined by their biosynthesis and catabolism. This thesis characterized the expression patterns of the major GA and ABA metabolism genes in barley cv. Betzes during seed development, and germination in both dormant and non-dormant seeds. The results indicate that specific gene family members of the two hormones play distinct temporal roles in regulating seed development, dormancy onset and release, and germination. Since only two genes encoding the GA deactivating GA 2-oxidase enzyme have been known so far in barley, this study also identified two new GA2ox genes designated as HvGA2ox1 and HvGA2ox3.

Preharvest sprouting (PHS), the germination of seeds on the maternal plant before harvest, is a big challenge for barley producers worldwide. It is attributed mainly to low seed dormancy. The balance between two classical plant hormones, gibberellin (GA) and abscisic acid (ABA) regulates seed dormancy and germination, and the endogenous level of these two hormones in plants is determined by their biosynthesis and catabolism. This thesis characterized the expression patterns of the major GA and ABA metabolism genes in barley cv. Betzes during seed development, and germination in both dormant and non-dormant seeds. The results indicate that specific gene family members of the two hormones play distinct temporal roles in regulating seed development, dormancy onset and release, and germination. Since only two genes encoding the GA deactivating GA 2-oxidase enzyme have been known so far in barley, this study also identified two new GA2ox genes designated as HvGA2ox1 and HvGA2ox3.

After-ripening, the process of seed dormancy loss in dry storage is associated with a decrease in the mean base water potential, one of the parameters of hydrothermal time. The rate of change of the mean base water potential is assumed to be a linear function of temperature above a specific base temperature and as a result can be described by a thermal after-ripening (TAR) time model, an extension of hydrothermal modelling. The thermal requirement for after-ripening is the thermal time necessary for the modelling base water potential of the seed to shift from its original value to its final value. In order to include the effects of water potential on the rate of dormancy loss, a hydrothermal after-ripening (HTAR) time model was developed. Laboratory and field studies were conducted using seeds of Bromus tectorum. These studies identified four important ranges of water potential that influence the rate of dormancy loss. The ranges are identified as follows: seeds experiencing soil water potentials seeds experiencing soil water potentials <-400 MPa do not after-ripen, between -400 MPa and -150 MPa seeds after-ripen as a function of temperature (T) and water potential (Ψ), seeds experiencing water potentials >-150 MPa after-ripen as a linear function of temperature, and somewhere above -40 MPa seeds are too wet to after-ripen. These ranges suggest that specific reaction thresholds associated with non-fully imbibed seeds also apply to the process of after-ripening. The HTAR model for B. tectorum seeds generally improved predictions of dormancy loss in the field under soil conditions that were too dry for TAR alone. Reduced after-ripening rate under extremely dry conditions is ecologically relevant in explaining how seeds may prolong dormancy under high soil temperature conditions.

Rôle du métabolisme post-transcriptionnel des ARNm dans la régulation de la dormance des semences d’Arabidopsis thaliana.Une étude physiologique nous a permis d'identifier l'influence de la température et de l'humidité relative (HR) lors du stockage des graines dormantes d’Arabidopsis. Après une levée de dormance atteinte en 7 semaines avec des cinétiques variables selon les conditions, on observe une induction de la dormance secondaire à faible HR et une perte progressive de la viabilité à forte HR. La levée et l’induction de la dormance sont associées à la régulation de gènes liés aux voies de l'acide abscissique et des gibbérellines. Nous avons étudié la dynamique d’association des ARNm aux polysomes et comparé la transcription et la traduction des graines dormantes et non dormantes au cours de l’imbibition. Nous montrons qu'il n'y a pas de corrélation entre transcriptome et traductome et que la régulation de la germination est principalement liée à la traduction. Ceci suppose un recrutement sélectif et dynamique des ARNm liés aux polysomes dans les graines dormantes et non dormantes. Certaines caractéristiques de la région 5'UTR des transcrits semble impliquées dans la sélection des ARNm traduits pendant la germination. Les phénotypes de mutants d’éléments du catabolisme des ARN montrent que la dormance est également associée à une dégradation sélective des ARNm. Les P-bodies (localisés dans des lignées YFP-DCP1) sont d’ailleurs en quantité plus importante dans les graines non-dormantes. La comparaison des transcriptomes des mutants vcs-8 et xrn4-5 a permis l'identification de plusieurs transcrits dégradés via VCS ou XRN4, dont le rôle sur la dormance a été confirmé par génétique inverse. Certains motifs spécifiques semblent être impliqués dans la sélection de transcrits pour leur dégradation. / Role of mRNA post-transcriptional metabolism in the regulation of Arabidopsis thaliana dormancy.A physiological study allowed us to reveal the effect of temperature and relative humidity (RH) during Arabidopsis seed storage. Seven weeks of after ripening lead to alleviation of dormancy with different kinetics according to the conditions. Longer storage induced an induction of secondary dormancy at low RH and progressive loss of viability at high RH. Dormancy release and induction of secondary dormancy were associated with induction or repression of key genes related to abscissic acid and gibberellins pathways. We have studied the dynamics of mRNAs association with polysomes and compared transcriptome and translatome of dormant and non-dormant seeds. There was no correlation between transcriptome and translatome and germination regulation is largely translational, implying a selective and dynamic recruitment of mRNAs to polysomes in both dormant and non-dormant seeds. Some identified 5'UTR features could play a role in this selective. Dormancy is also associated with mRNA decay as demonstrated by phenotyping mutants altered in mRNA metabolism. Moreover we have shown that P-bodies were more abundant in non-dormant seeds that in dormant ones. Transcriptome analysis of xrn4-5 and vcs-8 mutants allowed us to identify several transcripts degraded via VCS ou XRN4 proteins, having a role in dormancy. This role was confirmed by reverse genetics for some of them. Some specific motifs were identified as involved in mRNA decay selection.

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Penstemon gibbensii and Penstemon haydenii are two rare, perennial forbs inhabiting remote areas of the western United States. P. gibbensii is listed as a sensitive species by the Bureau of Land Management (BLM) in Colorado, Utah, and Wyoming (Heidel, 2009). P. haydenii was designated as Endangered by the U.S. Fish and Wildlife Service in 1987 (Heidel, 2012). This thesis research was geared toward helping land managers in their efforts to protect and rehabilitate these species by providing understanding on different aspects of their seed ecology. My first study was a laboratory experiment performed on P. gibbensii and P. haydenii seed germination response to moist chilling and dry after-ripening. Wild harvested seeds were subjected to moist chilling at 2-4 °C for 0, 4, 8, 12, and 16 weeks and held in dry storage for approximately 2 years to determine effective methods for breaking primary dormancy. P. gibbensii seed germination increased consistently with increased length of chilling up to 16 weeks and exhibited habitat-correlated variation in this response. P. haydenii seed germination increased from 1 to 100% germination with 4 weeks of chilling. P. haydenii germination was greatest (96%) when incubated under a cool, diurnally-fluctuating temperature regime (10-20 °C) and responded positively to dry storage, increasing germination from 0 to 15%. My second study was an in situ field study designed to characterize the active seed bank of P. haydenii. We set up a transect line across a P. haydenii population and measured the number of seeds entering the seed bank, lost to predation post-dispersal, and persisting in the seed bank. P. haydenii does not appear to form an ecologically significant seed bank. Approximately 140 seeds/ 10m2 could potentially enter the seed bank but only 1 seed in the upper 10cm of sand persisted. Heavy post-dispersal insect predation resulted in a decrease in viability of nearly 30% in exposed P. haydenii seeds after just 12 hours. My third study explored the effects of burial by sand on P. haydenii. Wild-harvested seeds were planted in pots at 1, 2, 4, 6, 8, and 10cm deep in sand and incubated at 10-20 °C. Seed germination and mortality and seedling emergence were measured. The response of dormant seeds to post-burial incubation was determined. Burial depth decreased seedling emergence and seed germination. Shallow burial appears to induce secondary dormancy for seeds that don't germinate quickly, whereas deep burial appears to impose enforced dormancy in burial.