Roles of abscisic acid and gibberellins in maintaining primary and secondary dormancy of Korean pine seeds

Primary dormancy of seeds of Korean pine(Pinus koraiensis Sieb.et Zucc.)after dispersal in the autumn and the induction of secondary dormancy the fi rst summer following seed dispersal limit the regeneration of mixed broadleaved Korean pine forests in Northeast China.This study was to determine how changes in the levels of abscisic acid(ABA)and gibberellic acid(GA)maintain primary and secondary dormancy of Korean pine seeds under germination conditions.We transferred seeds with one of fi ve primary dormancy states or three secondary dormancy states to germination conditions and measured changes in the levels of ABA,GA 1+3(GA 1 and GA 3)and GA 4+7(GA 4 and GA 7)in the seed coat,megagametophyte and embryo during incubation.Seed coat ABA levels in primary dormant seeds(PDS)and ABA levels in various parts of secondary dormant seeds(SDS)gradually declined during incubation but were still higher than in seeds for which dormancy was progressively released.GA 4+7 and GA 1+3 levels in embryos greatly decreased 35%and 24%,respectively,during incubation of SDS,and thus,the ratio of ABA to GA 4+7 in embryos and megagametophytes signifi cantly increased.The ratio of ABA to GA 1+3 in various parts of SDS increased slightly during incubation.In contrast,in seeds for which secondary dormancy was already released,GA 4+7 and GA 1+3 levels in the embryo,GA 4+7/ABA ratio in the embryo and seed coat,and the GA 1+3/ABA in the embryo and megagametophyte signifi cantly increased during incubation.There was no trend in the changes in the levels of ABA,GA 4+7 or GA 1+3 in embryos and megagametophytes of PDS or the levels of GA 4+7 or GA 1+3 in megagametophytes of SDS during incubation.The results suggest that high ABA levels in the seed coat maintain primary dormancy of Korean pine seeds.Maintenance of secondary dormancy involves a reduction of GA 4+7,GA 1+3,GA 4+7/ABA,and GA 1+3/ABA and the retention of high ABA levels.

The coupled effect of light and temperature on dormancy release and germination of Pinus koraiensis seeds

Elucidating the regulatory mechanisms of environmental factors on seed dormancy and germination will provide guidance for tree regeneration.Toward understanding the coupled effect of light and temperature on dormancy release and germination of Pinus koraiensis seeds,we set up three light conditions(L200:200μmol m^(-2) s^(-1),L20:20μmol m^(-2) s^(-1),L0:0μm^(-2) s^(-1))and four storage temperatures[T-5:-5°C(50 days),T5:-5°C(50 days)+5°C(50 days),T25:-5°C(50 days)+5°C(50 days)+25°C(50 days),T15:-5°C(50 days)+5°C(50 days)+25°C(50 days)+15°C(50 days)]using imbibed seeds,then quantified phytohormones gibberellic acid(GA_(3))and abscisic acid(ABA)during the stratification.Germination percentage(G_(P)),mean germination time(T_(M)),and germination value(G_(V))under 25/15°C temperature and the three light conditions were then determined.Phytohormone levels and germination performances were significantly affected by light and temperature.No consistent trend was found between the phytohormone levels and G_(P) caused by light levels.Under the three light conditions,ABA concentrations in the embryo and endosperm decreased as storage temperature shifted from T-5 to T25 and increased from T25 to T15;GA_(3) decreased in nearly all four storage temperatures.G_(P) reached 40–60%in T25 storage without light irradiance.In the three light conditions,G_(P) and G_(V) were higher at T5 and T25 than at T-5 and T15;so T5 and T25 are considered as optimum storage temperatures for dormancy release and germination.At optimum temperatures,light(L200,L20)significantly increased the G_(P) and G_(V) compared with the dark(L0).At L200 and L20,significant negative correlations between G_(V) and the ABA concentrations and positive correlations between G_(V) and GA/ABA in the seed embryo were found.Temperature played a more important role in primary dormancy release and germination;light was unnecessary for primary dormancy release.Light facilitated seed germination at optimum temperatures.The dormancy release and germination of P.koraiensis seeds were controlled by a decrease in ABA concentrations or an increase in GA/ABA induced by temperature variations.

A Novel RGL2-DOF6 Complex Contributes to Primary Seed Dormancy in Arabidopsis thaliana by Regulating a GATA Transcription Factor

The DELLA protein RGA-LIKE2 （RGL2） is a key transcriptional repressor of gibberellic acid （GA） signaling that regulates seed germination. We identified GATA12, a gene encoding a GATA-type zinc finger transcription factor, as one of the downstream targets of RGL2 in Arabidopsis thaliana. Our data show that freshly harvested （unstratified） seeds of GATA 12 antisense suppression lines have reduced dormancy compared with the wild-type, while ectopic expression lines show enhanced seed dormancy. We show that GATA12 expression is negatively regulated by GA, and its transcript levels decline dramatically under dormancy- breaking conditions such as dry storage and cold stratification of seeds. GATA12 promoter has several GAMYB- and DOF-associated motifs that are known to be GA- and RGL2-responsive, respectively. Chromatin immunoprecipitation assay showed that a protein complex containing RGL2 can bind to GATA12 promoter and thereby regulate its expression. RGL2 lacks a DNA binding domain and requires a transcription factor to induce GATA12 expression. Our data show that this RGL2-containing protein complex includes DNA BINDING1 ZINC FINGER6 （DOF6）, which is a known negative regulator of germination in freshly harvested seeds. We further show that this novel RGL2-DOF6 complex is required for activating GATA12 expression, thus revealing a molecular mechanism to enforce primary seed dormancy.