INTRODUCTION 331 OCCURRENCE OF DORMANCY 332 STRUCTURE AND ORGANIZATION 332 Seed Coat Efficts 332 G en eral and Mitoch ondrial Activity .... ... ... ... . .. .. ... ........ 333 Nu cleic Acids and Prot ein Syntheses . . . 334 HORMONE AND PHYTOCHROME ACTION . . . . . . . . . .. .. . . . . . . . . .. . . . . ..... . .. . . . .. .. .. . .. .. . .. .. . . . . .. .. . . 336 Abs cisic and Gibb er el ic Acids ... .. 336 Cyto kinins and Ethylene .... .... 339 Phytochrome Action ...... 341 METABOLIC REGULATION 342 APPLIED COMPOUNDS .. . . . . . . . . . .. . .. . . . . . . . . ... .... 345 TEMPERATURE EFFECTS 346 CONCLUSION 347

Dormancy in Seeds

In higher plants, seed germination is followed by postgerminative growth. One of the key developmental events during postgerminative growth is cotyledon greening, which enables a seedling to establish photosynthetic capacity. The plant phytohormone abscisic acid (ABA) plays a vital role by inhibiting seed germination and postgerminative growth in response to dynamically changing internal and environmental cues. It has been shown that abscisic acid insensitive5 (ABI5), a basic leucine zipper transcription factor, is an important factor in the regulation of the ABA-mediated inhibitory effect on seed germination and postgerminative growth. Conversely, the phytohormone cytokinin has been proposed to promote seed germination by antagonizing the ABA-mediated inhibitory effect. However, the underpinning molecular mechanism of cytokinin-repressed ABA signaling is largely unknown. Here, we show that cytokinin specifically antagonizes ABA-mediated inhibition of cotyledon greening with minimal effects on seed germination in Arabidopsis (Arabidopsis thaliana). We found that the cytokinin-antagonized ABA effect is dependent on a functional cytokinin signaling pathway, mainly involved in the cytokinin receptor gene cytokinin response1/Arabidopsis histidine kinase4, downstream histidine phosphotransfer protein genes AHP2, AHP3, and AHP5, and a type B response regulator gene, ARR12, which genetically acts upstream of ABI5 to regulate cotyledon greening. Cytokinin has no apparent effect on the transcription of ABI5. However, cytokinin efficiently promotes the proteasomal degradation of ABI5 in a cytokinin signaling-dependent manner. These results define a genetic pathway through which cytokinin specifically induces the degradation of ABI5 protein, thereby antagonizing ABA-mediated inhibition of postgerminative growth.

http://www.plantphysiol.org/content/plantphysiol/early/2014/01/17/pp.113.234740.full.pdf

Cytokinin Antagonizes Abscisic Acid-Mediated Inhibition of Cotyledon Greening by Promoting the Degradation of ABSCISIC ACID INSENSITIVE5 Protein in Arabidopsis

In higher plants, seed germination is followed by postgerminative growth. One of the key developmental events during postgerminative growth is cotyledon greening, which enables a seedling to establish photosynthetic capacity. The plant phytohormone abscisic acid (ABA) plays a vital role by inhibiting seed germination and postgerminative growth in response to dynamically changing internal and environmental cues. It has been shown that ABSCISIC ACID INSENSITIVE5 (ABI5), a basic leucine zipper transcription factor, is an important factor in the regulation of the ABA-mediated inhibitory effect on seed germination and postgerminative growth. Conversely, the phytohormone cytokinin has been proposed to promote seed germination by antagonizing the ABA-mediated inhibitory effect. However, the underpinning molecular mechanism of cytokinin-repressed ABA signaling is largely unknown. Here, we show that cytokinin specifically antagonizes ABA-mediated inhibition of cotyledon greening with minimal effects on seed germination in Arabidopsis (Arabidopsis thaliana). We found that the cytokinin-antagonized ABA effect is dependent on a functional cytokinin signaling pathway, mainly involved in the cytokinin receptor gene CYTOKININ RESPONSE1/ARABIDOPSIS HISTIDINE KINASE4, downstream histidine phosphotransfer protein genes AHP2, AHP3, and AHP5, and a type B response regulator gene, ARR12, which genetically acts upstream of ABI5 to regulate cotyledon greening. Cytokinin has no apparent effect on the transcription of ABI5. However, cytokinin efficiently promotes the proteasomal degradation of ABI5 in a cytokinin signalingdependent manner. These results define a genetic pathway through which cytokinin specifically induces the degradation of ABI5 protein, thereby antagonizing ABA-mediated inhibition of postgerminative growth.

Cytokinin Antagonizes Abscisic Acid-Mediated Inhibition of Cotyledon Greening by Promoting the Degradation of ABSCISIC ACID INSENSITIVE5 Protein in Arabidopsis 1(C)(W)

Lettuce seeds (Lactuca sativa L. cv. Grand Rapids) stimulated to germinate by gibberellin and red light produce large amounts of endo-β-mannanase. This enzyme increases markedly following radicle emergence and is capable of degrading mannose-containing polysaccharides, which are the major components of the endosperm cell wall. Non-germinated seeds contain little enzyme and under conditions where gibberellin- or red light-stimulated germination is prevented (eg. by abscisic acid or prolonged far red light) enzyme levels remain low. Cycloheximide inhibits the increase in enzyme levels when supplied to germinating seeds, but the enzyme once produced is stable in vivo in the presence of this inhibitor for at least 24h. The majority of the extractable mannanase activity is located in the endosperm and we propose that the function of this enzyme is to mobilise the endosperm cell wall polysaccharides as a nutrient source for the growing embryo.

An enzyme to degrade lettuce endosperm cell walls. Appearance of a mannanase following phytochrome- and gibberellin-induced germination

Seeds of five celery (Apium graveolens L.) cultivars germinated at 15°C in the light or dark but at 22°C only in the light. This light requirement was overcome by treatment with a mixture of the gibberellins GA4 and GA7 (GA4/7) but interactions of cytokinins, daminozide, ethephon, EDTA and N-phenyl-N′-4-pyridylurea (NC5392) with GA4/7 were observed. Varietal differences in response to GA4/7 concentration and the requirement for cytokinins were related to the upper temperature limits for germination of the different cultivars. Seeds of cultivars responding to low concentrations of GA4/7 appeared to contain less natural inhibitor than those requiring either high concentrations of GA4/7 or cytokinin in addition to low GA4/7. The cytokinin requirement for germination was partially removed by leaching the seeds with water. Interaction studies with applied hormones indicated that in seeds incubated in the light inhibition by abscisic acid was partially alleviated by N6-benzyladenine but not by GA4/7 application. The implications of these results are discussed in relation to the involvement of natural plant hormones in the dormancy mechanism of celery seeds.

Growth Regulators and the Phytochrome-Mediated Dormancy of Celery Seeds

Cytokinins overcome the inhibitory effect of abscisic acid on the germination of isolated embryos of Grand Rapids lettuce. We have used this fact to assess the content of cytokinins in embryos removed from intact seeds which have been supplied with these growth substances i.e. to determine hormonal uptake. The method has also been used to examine the hypothesis that phytochrome and gibberellin lead to increases in active cytokinin levels in seeds. Appreciable amounts of cytokinins enter intact seeds yet seem unable to promote germination. On the other hand, little or no cytokinin-like activity seems to result from light or gibberellin treatments. We conclude that there are unresolved difficulties inherent in the “cytokinin hypothesis” of phytochrome and gibberellin action.

M. Black

Papers

Lettuce seed germination and cytokinins: their entry and formation.