Water Content during Abscisic Acid Induced Freezing Tolerance in Bromegrass Cells

Changes in water content and dry weight were determined in control cells and those induced to cold harden in response to abscisic acid (ABA) treatment (7.5 × 10⁻⁵ molar). Bromegrass (Bromus inermis Leyss cv Manchar) cells grown in suspension culture at room temperature (23°C) for 7 days acclimated to −28°C (LT₅₀) when treated with ABA, or to −5°C when untreated. ABA significantly reduced cell growth rates at 5 and 7 days after treatment. Growth reduction was due to a decrease in cell number rather than cell size. When the cell water content was expressed as percent water (percent H₂O) or as grams water per gram dry weight (gram H₂O/gram dry weight [g DW]), the water content of hardy, ABA-treated cells decreased from 85% to 77% or from 6.4 to 3.3 g H₂O/g DW in 7 days. Control cell water content remained static at approximately 87% and 7.5 g H₂O/g DW. However, cell water content, expressed as milligrams water per million cells (milligram H₂O/10⁶ cells), did not differ in ABA-treated or control cells. The dry matter content of ABA-treated cells, expressed as milligram DW/10⁶ cells increased to 3.3 milligram/10⁶ cells in 7 days, whereas the dry weight of the control cells remained between 1.4 to 2.1 milligrams/10⁶ cells. The osmotic potential of ABA-treated cells decreased by the fifth day while that of control cells increased significantly and then decreased by day 7. Elevated osmotic potentials were not associated with increased ion uptake. In contrast to much published literature, these results suggest that cell water content does not decrease in ABA-treated cells during the induction of freezing tolerance, rather the dry matter mass per cell increased. Cell water content may be more accurately expressed as a function of cell number when accompanying changes to dry cell matter occur.     

Role of Abscisic Acid in Drought-Induced Freezing Tolerance,Cold Acclimation,and Accumulation of LT178 and RAB18 Proteins in Arabidopsis thaliana

Embryogenic tissues of Pinus caribaea Morelet var hondurensis produce extracellular proteins; among them germins have been identified. Two-dimensional electrophoresis followed by electroblotting onto a polyvinylidene difluoride membrane allowed isolation and N-terminal amino acid sequencing of extracellular GP111, which is present within the five embryogenic cell lines studied. The amino acid sequence showed strong homologies with the sequences of germins deduced from cDNA sequencing, starting at the same amino acid position but one, compared with other sequences of mature germins deduced from protein sequencing. Immunoblots of embryogenic and nonembryogenic extracellular proteins indicated that the polypeptide GP111 plus two others with similar relative molecular mass values are present in embryogenic cell lines but not in nonembryogenic ones. They were recognized by an antiserum raised against the nonglycosylated monomer of wheat germin. The cross-reaction between pine and wheat apoproteins was highly specific. An antiserum against the glycosylated pentameric germin-like protein (an oxalate oxidase) of barley cross-reacted with all three, as well as with several other glycosylated polypeptides.     

Role of Abscisic Acid in the Induction of Desiccation Tolerance in Developing Seeds of Arabidopsis thaliana

In contrast to wild-type seeds of Arabidopsis thaliana and to seeds deficient in (aba) or insensitive to (abi3) abscisic acid (ABA), maturing seeds of recombinant (aba,abi3) plants fail to desiccate, remain green, and lose viability upon drying. These double-mutant seeds acquire only low levels of the major storage proteins and are deficient in several low mol wt polypeptides, both soluble and bound, and some of which are heat stable. A major heat-stable glycoprotein of more than 100 kilodaltons behaves similarly; during seed development, it shows a decrease in size associated with the abi3 mutation. In seeds of the double mutant from 14 to 20 days after pollination, the low amounts of various maturation-specific proteins disappear and many higher mol wt proteins similar to those occurring during germination are induced, but no visible germination is apparent. It appears that in the aba,abi3 double mutant seed development is not completed and the program for seed germination is initiated prematurely in the absence of substances protective against dehydration. Seeds may be made desiccation tolerant by watering the plants with the ABA analog LAB 173711 or by imbibition of isolated immature seeds, 11 to 15 days after pollination, with ABA and sucrose. Whereas sucrose stimulates germination and may protect dehydration-sensitive structures from desiccation damage, ABA inhibits precocious germination and is required to complete the program for seed maturation and the associated development of desiccation tolerance.     

Seed Development in the Recalcitrant Species Quercus robur L: Water Status and Endogenous Abscisic Acid Levels

Water status and endogenous ABA levels were measured duringthe development of the embryonic axis and cotyledons in fruitsof Quercus robur L. As dry matter accumulated in the embryoduring development, both moisture content and osmotic potential(     

Effects of Abscisic Acid Metabolites and Analogs on Freezing Tolerance and Gene Expression in Bromegrass (Bromus inermis Leyss) Cell Cultures

Optical isomers and racemic mixtures of abscisic acid (ABA) and the ABA metabolites abscisyl alcohol (ABA alc), abscisyl aldehyde (ABA ald), phaseic acid (PA), and 7[prime]hydroxyABA (7[prime]OHABA) were studied to determine their effects on freezing tolerance and gene expression in bromegrass (Bromus inermis Leyss) cell-suspension cultures. A dihydroABA analog (DHABA) series that cannot be converted to PA was also investigated. Racemic ABA, (+)-ABA, ([plus or minus])-DHABA, and (+)-DHABA were the most active in inducing freezing tolerance, (-)-ABA, ([plus or minus])-7[prime]OHBA, (-)-DHABA, ([plus or minus])-ABA ald, and ([plus or minus])-ABA alc had a moderate effect, and PA was inactive. If the relative cellular water content decreased below 82%, dehydrin gene expression increased. Except for (-)-ABA, increased expression of dehydrin genes and increased accumulation of responsive to ABA (RAB) proteins were linked to increased levels of frost tolerance. PA had no effect on the induction of RAB proteins; however, ([plus or minus])- and (+)-DHABA were both active, which suggests that PA is not involved in freezing tolerance. Both (+)-ABA and (-)-ABA induced dehydrin genes and the accumulation of RAB proteins to similar levels, but (-)-ABA was less effective than (+)-ABA at increasing freezing tolerance. The (-)-DHABA analog was inactive, implying that the ring double bond is necessary in the (-) isomers for activating an ABA response.     

Changes in the Translatable RNA Population during Abscisic Acid Induce Freezing Tolerance in Bromegrass Suspension Culture

Abscisic acid (ABA) has been shown to increase freezing toleranceof bromegrass (Bromus in-ermis Leyss cv. Manchar) cell suspensioncultures from a LT₅₀ (the temperature at which 50% cells werekilled) of –7 to – 30?C in 5 days at 23?C. Our objectivewas to study the qualitative changes in the translatable RNApopulation during ABA induced frost tolernace. In vitro translationproducts of poly(A)⁺ RNA isolated from bromegrass cells withor without 75 µM ABA treatment for various periods oftime were separated by 2D-PAGE and visualized by fluorography.SDS soluble proteins from the same treatments were also separatedby 20-PAGE. After 5 days treatment, at least 22 new or increasedabundance SDS soluble polypeptides were observed. From fluorographs,29 novel or increased abundance in vitro translation productscould be detected. The pattern of changes between ABA inducedSDS-soluble proteins and translation products from the 2D gelswere similar. A time course study (0–7 days) showed that17 of the 29 translation products were detected after 1 dayABA treatment, and at least 14 were present after 1 h. Coldtreatment (+4?C) induced fewer changes in the pool of translatableRNA than with ABA treatment. Three translation products inducedby cold appear to be similar to 3 of the ABA induced translationproducts. The majority of the ABA inducible translatable RNAsappeared at 10 µM or higher which coincides with the inductionof freezing tolerance. Many of these ABA inducible RNAs persisted7 days after ABA was removed from the media and correspondinglythe LT₅₀ (–17?C) was still well above the control level(–17?C). The results suggest that ABA alters the poolof translatable RNAs during induction of freezing tolerancein bromegrass suspension culture cells. ¹Oregon Agricultural Experiment Station Technical Paper No.9256. (Received August 3, 1990; Accepted October 18, 1990)     

Abscisic Acid Elicits the Water-Stress Response in Root Hairs of Arabidopsis thaliana

Water stress has been shown to cause root hairs to become short and bulbous. Because abscisic acid (ABA) mediates a variety of water-stress responses, we investigated the response of Arabidopsis thaliana root hairs to ABA. When wild-type root hairs were treated with ABA, they exhibited the water-stress response. The Arabidopsis mutants abi1 and abi2, which are insensitive to ABA at the seedling stage, did not display the root hair response. These data suggest that ABA may mediate the response of root hairs to water stress. The drought response of root hairs resulting in an inhibition of tip growth will provide an easy screen to select mutations that are insensitive to ABA and/or involved in tip growth.     

Abscisic Acid Translocation and Influence of Water Stress on Grain Abscisic Acid Content

The movement of foliar applied [1-¹⁴C]abscisic acid (ABA) inwheat plants (Triticum aestivum L., cv. Kolibri) was investigatedat two stages of grain development (1000 grains, weight 19 and24 g dry matter). [1–¹⁴C]ABA seemed to be readily translocated within 12h into the developing grains as well as in other plant parts.A subsequent rapid metabolism took place leading to a decreasedactivity of the ABA-containing chromatogram fraction in theyounger plants 48 h after application. The metabolism seemodto be less intensive in the older grains, where the activityrunning with the ABA increased over 64 h. Treating the leaves of barley plants (Hordeum vulgare, L., cv.Union) 2 weeks after anthesis with a gentle stream of warm air(36° C) resulted in a significant increase in the ABA contentof all parts of the ear. The results mentioned above indicatethat this may be partially due to translocation from other partsof the plant such as the leaves.     

An Acer palmatum R2R3-MYB Gene,ApMYB77, Confers Freezing and Drought Tolerance in Arabidopsis thaliana

Journal of Plant Growth Regulation - Low temperature is one of the most prominent environmental factors affecting plant growth. As a deciduous arboreal tree, Acer palmatum has considerable...     

拟南芥ABA生物合成基因NCED6对葡萄糖诱导其种子萌发延迟的作用

抑制ABA的生物合成可以缓解葡萄糖抑制拟南芥种子萌发作用,说明ABA的生物合成参与葡萄糖诱导种子萌发的延迟。ABA生物合成基因9-顺式环氧类胡萝卜素双加氧酶6（NCED6）可以被不同浓度的葡萄糖上调表达,而nced6突变体的种子对葡萄糖不敏感。     

Interaction of Osmotic Stress, Temperature, and Abscisic Acid in the Regulation of Gene Expression in Arabidopsis

The impact of simultaneous environmental stresses on plants and how they respond to combined stresses compared with single stresses is largely unclear. By using a transgene (RD29A-LUC) consisting of the firefly luciferase coding sequence (LUC) driven by the stress-responsive RD29A promoter, we investigated the interactive effects of temperature, osmotic stress, and the phytohormone abscisic acid (ABA) in the regulation of gene expression in Arabidopsis seedlings. Results indicated that both positive and negative interactions exist among the studied stress factors in regulating gene expression. At a normal growth temperature (22°C), osmotic stress and ABA act synergistically to induce the transgene expression. Low temperature inhibits the response to osmotic stress or to combined treatment of osmotic stress and ABA, whereas low temperature and ABA treatments are additive in inducing transgene expression. Although high temperature alone does not activate the transgene, it significantly amplifies the effects of ABA and osmotic stress. The effect of multiple stresses in the regulation of RD29A-LUC expression in signal transduction mutants was also studied. The results are discussed in the context of cold and osmotic stress signal transduction pathways.     

脱落酸与Em基因的表达调控

Em基因的表达受ABA诱导,干旱和盐胁通过增加ABA含量或改变植物细胞对ABA的敏感性而诱导Em基因的表达。植物Em基因启动子存在3个功能区:5′远端AT富集区通过影响转录调节表达量,作用类似于非专一性增强子;ABA应答元件ABRE在ABA存在的情况下与转录因子EmBP1相互作用能显著增强Em基因的表达;5′UTR可能通过转录后调控而影响最终表达水平 。     

Overexpression of TaFBA-A10 from Winter Wheat Enhances Freezing Tolerance in Arabidopsis thaliana

Journal of Plant Growth Regulation - Freezing stress is the principal abiotic stress that is not conducive to plant growth and yield. Fructose-1, 6-bisphosphate aldolase (FBA; EC 4.1.2.13) is a key...     

Role of Abscisic Acid in the Induction of Freezing Tolerance in Brassica napus Suspension-Cultured Cells

Brassica napus suspension-cultured cells could be hardened in 6 days at 25°C by the addition of mefluidide or ABA to the culture medium. Cells treated with mefluidide (10 milligrams per liter) or ABA (50 micromolar) attained an LT₅₀ of −17.5°C or −18°C, respectively, while the LT₅₀ for the comparable nonhardened control (sucrose) was −10°C. The increased freezing tolerance of mefluidide-treated cells was paralleled by a 4- to 23-fold increase in ABA, as measured by gas-liquid chromatography using electron capture detection. Application of 1 milligram per liter of fluridone, an inhibitor of abscisic acid biosynthesis, prevented the mefluidide-induced increase in freezing tolerance and the accumulation of ABA. Both these inhibitory effects of fluridone were overridden by 50 micromolar ABA in the culture medium. On the basis of these results, we concluded that increased ABA levels are important for the induction of freezing tolerance in suspension-cultured cells.     

Integrin-like Protein Is Involved in the Osmotic Stress-induced Abscisic Acid Biosynthesis in Arabidopsis thaliana

We studied the perception of plant cells to osmotic stress that leads to the accumulation of abscisic acid （ABA） in stressed Arabidopsis thaliana L. cells. A significant difference was found between protoplasts and cells in terms of their responses to osmotic stress and ABA biosynthesis, implying that cell wall and/or cell wall-plasma membrane interaction are essential in identifying osmotic stress. Western blotting and immunofluorescence localization experiments, using polyclonal antibody against human integrin β1, revealed the existence of a protein similar to the integrin protein of animals in the suspension-cultured cells located in the plasma membrane fraction. Treatment with a synthetic pentapeptide, Gly-Arg-Gly-Asp-Ser （GRGDS）, which contains an RGD domain and interacts specifically with integrin protein and thus blocks the cell wall-plasma membrane interaction, significantly inhibited osmotic stress-induced ABA biosynthesis in cells, but not in protoplasts. These results demonstrate that cell wall and/or cell wall-plasma membrane interaction mediated by integrin-Iike proteins played important roles in osmotic stress-induced ABA biosynthesis in Arabidopsis thaliana.     

过量表达棉花GaSus3基因增强拟南芥的耐盐性

构建了植物过量表达载体p35S：：GaSus3,通过花序浸染法成功获得转GaSus3基因拟南芥植株。利用NaCl模拟盐胁迫处理,证实转基因拟南芥与野生型相比耐盐性明显增强。在盐胁迫下,转基因拟南芥受到的影响较小,而野生型则受盐害影响严重：转基因拟南芥具有更好的萌发率和主根长度,以保证植株正常生长;盐胁迫下转基因拟南芥能保持较多的绿色叶片,而野生型则过早黄化死亡。研究还发现,转基因拟南芥的过氧化氢酶活性在胁迫前后都高于野生型,这说明转GaSus3基因能够提高拟南芥抗氧化胁迫的能力。研究结果为进一步探讨GaSus3基因在棉花耐盐方面的功能奠定了基础。