G. Garello

Changes in abscisic acid biosynthesis and catabolism during dormancy breaking in Fagus sylvatica embryo

Abstract. At harvest, embryos of Fagus sylvatica are dormant. A cold pretreatment without medium at 30% moisture content allowed them to germinate. A comparison of the abscisic acid (ABA) content before and after the pretreatment has no significant relevance since dormancy is expressed during the culture at 23°C. During this culture, both de novo biosynthesis and conjugate hydrolysis contributed to maintain a high level of ABA in the dormant axis. The level of conjugates and the rate of hydrolysis were not modified substantially by the cold pretreatment. In contrast, the dormancy release was associated with a strong decrease in the capacity for ABA synthesis. Moreover, feeding (+)-[3H]ABA to untreated and pretreated embryos proved that the cold treatment also induced a hastening of ABA catabolism.

Evidence for the role of abscisic acid in the genetic and environmental control of dormancy in wheat ( Triticum aestivumL. )

Grain production of two wheat cultivars ( Triticum aestivum L. cv Recital and Scipion), known for their different germination behaviour, was studied at two different temperatures. The study of dormancy onset during grain development showed that in both cultivars, dormancy developed on the mother plant. In Recital grain, after a period of high germinability, dormancy developed for a transient period. However, full germination was obtained on medium supplemented with fluridone, showing that dormancy was associated with abscisic acid (ABA) biosynthesis inside the grain. Dormancy progressively disappeared during natural drying, at a slower rate for grains developing at 15°C than for grains developing at 25°C. However, at both temperatures, the release from dormancy was complete at maturity. In Scipion grain, dormancy was almost total throughout grain development irrespective of the temperature. However, grains could germinate in the presence of fluridone: changes in sensitivity to fluridone were observed during grain development, showing an increase in dormancy during the first half of development, followed by a progressive decrease during the second half. This decrease occurred later for grains developing at 15°C than for those developing at 25°C. In Scipion grain, unlike the cultivar Recital, release from dormancy was not completed before the end of development on the mother plant. An additional dry storage period was necessary which was shorter for grains developed at 25°C than for grains produced at 15°C. A comparison of embryo ABA levels after a 24-h culture in the presence or absence of fluridone, allowed the ABA synthesis to be estimated. It appears that the depth of dormancy was related to the estimated ABA synthesis capacity of the embryos.

Oxygen availability and ABA metabolism in Fagus sylvatica seeds

At harvest, beechnuts have a very deep dormancy whichlies both in the structures surrounding the embryo andwithin the embryo itself. Covering structures preventembryo germination by interfering with water uptakeand gaseous exchange. To understand the role of thecovering structures and oxygen availability on ABAcatabolism, (+)-[3H] ABA metabolism was studied in isolated embryos as well as in intact seeds. ABAdegradation resulted essentially in oxidative products(PA, DPA). These products were more abundant inisolated embryos than in intact seeds. Theyaccumulated mainly as alkali-nonhydrolyzableconjugates of DPA. A small amount of free andesterified forms were oftenobserved. In isolated embryos a decrease in oxidativeproducts was observed either by lowering the oxygenavailability or by feeding embryos with tetcyclasis(an inhibitor of monooxygenase). In the presence of the covering structures, these oxidative products werereduced in the same manner, indicating that coveringstructures were probably responsible for limiting theoxygen supply to the embryo and for the lowgermination percentage observed in the case of intactseeds.

Regulation of ABA levels in senescing petals of rose flowers

During the vase life of a rose flower, changes in the levels of abscisic acid (ABA) were observed: a decrease during the first 3 days, followed by a steady state at a low level, and finally a sharp increase in late senescence. Feeding [2-14C]ABA to isolated petals showed that metabolism was very active despite the age of the flower, oxidation processes increased with age, whereas conjugation decreased but the level of nonmetabolized ABA remained stable. When the isolated petal was subjected to water stress, whatever its age, the ABA level increased. Hydrolysis of ABA-GE was not involved in this phenomenon. Thus, ABA synthesis occurred in the isolated petal; it could be directly correlated to the decrease in water potential. However, the ABA increase in isolated petals was limited. Moreover, on the rose tree, increases in ABA levels were not correlated to water potential changes. ABA levels seemed, therefore, mainly regulated by changes in import from leaves and other parts of the flower.

De novo ABA synthesis and expression of seed dormancy in a gymnosperm: Pseudotsuga menziesii

Seeds of dormant Douglas-fir seeds germinated poorly when they were cultivated at 20–23 °C while isolated embryos germinated fully within two weeks. Seed dormancy was therefore imposed on the embryo by its surrounding structures. This physiological behaviour was well correlated with changes in ABA level during culture. Indeed, the ABA level decreased in isolated embryos while it increased in both embryo and megagametophyte during culture of whole seeds. The origin of this increase was analysed and the different ways by which seed coats could interfere with ABA accumulation are discussed.

The influence of light quality on rose flower senescence: involvement of abscisic acid

The aim of this work was to study the impact of light applied during preharvest culture on the subsequent senescence of cut rose flower and to analyse the possible involvement of abscisic acid (ABA). The longevity of cut rose flowers was longer when rose plants were previously grown under high pressure sodium lamps than under metal halide lamps. A change in light source did not lead to a change in leaf ABA content but significantly affected the petal ABA content. The relationship between ABA level and flower longevity, previously reported for differences of genetic origin, was again observed for culture-induced differences: the higher the ABA level at harvest, the shorter the vase-life observed.

Alteration of hormonal levels and hormone sensitivity by ri T-DNA-transformation of apple cuttings

Summary Changes in hormone levels and sensitivity to hormones were studied in the apple rootstock M 26 and in the clone transformed by Agrobacterium rhizogenes strain A4. The transformation of apple microcuttings by insertion of both TL-DNA and TR-DNA led to changes in leaf levels of all plant hormones studied (auxin, cytokinins and abscisic acid). An important increase in auxin and a decrease in cytokinin levels were observed. The ABA level was substantially lowered by transformation, whatever the water status of the leaves. The sensitivity to hormones of both transformed and untransformed cuttings was compared on the basis of their root induction efficiency. No significant difference in auxin sensitivity could be observed on this long-term response. However, the transgenic clone was less sensitive to cytokinin-induced rooting inhibition and more sensitive to ABA-induced rooting inhibition than the untransformed clone. The consequences of the changes in both hormone levels and sensitivity to hormones are discussed.

ABA Involvement in the Psychrolabile Dormancy of Fagus Embryo

Embryos of Fagus sylvatica isolated from fruits at harvest were dormant. After a cold-treatment in restricted water conditions, embryos were able to germinate at a percentage which increased with chilling duration. Embryo ABA content decreased during the dormancy-releasing treatment at 4°C; it also decreased in the same proportions during embryo culture at 23°C, a temperature allowing dormancy to be strongly expressed. It thus appears that embryo ABA content was not correlated with the physiological potentialities of the embryo. The significant decrease in ABA level observed during culture at 23°C could be explained by a very rapid ABA metabolism, revealed by the results of [3H]ABA feeding experiments. Also, when fluridone, an inhibitor of carotenoid synthesis, was applied directly to axes, dormant embryos were able to germinate after a one-week culture. The comparison between axis ABA content in the absence or in the presence of fluridone after a 6-day culture gave an estimation of the axis capacity for a de novo ABA synthesis. Consequently, it appears that the psychrolabile dormancy of Fagus is associated with the ability of the axis to synthesize its own ABA.

Onset of water stress tolerance in developing Helianthus annuus embryos

Excised Helianthus annuus embryos were capable of precocious germination as early as 7 d after pollination. At this stage they were not capable of withstanding desiccation treatment. The desiccation tolerance was acquired at a stage of development which depended on the rate of water loss to which embryos were subjected. Coincident with the acquisition of tolerance to a rapid desiccation, they gained the capacity to withstand 24 h preincubation in agitated water; this acquisition could be correlated to a sharp decrease of the leaching of electrolytes, amino acids and proteins in the incubation water, suggesting that a change in membrane properties played a key role in this transition. Moreover, the acquisition of tolerance to these stresses was coincident with the increase of endogenous abscisic acid (ABA). ABA applied in vitro to young embryos induced tolerance both to desiccation and to preincubation in water: this in vitro acquisition of tolerance was again correlated with a decrease in the leakage.