Irreversible Effects of Water Stress on Growth and Stomatal Development in Cotyledons of Etiolated Squash Seedlings

Growth of etiolated squash cotyledons and hypocotyls was suppressedwhen the seedlings were subjected to 60 mM polyethylene glycol(PEG) in 1/5 strength of Hoagland solution. The fresh weightof the hypocotyl completely recovered when the water stresswas relieved after one day of PEG treatment. The fresh weightof the cotyledons, however, did not completely recover eventhree days after the relief of water stress. The transpiration rate of the cotyledons was substantially reducedby the water stress, and it also did not completely recoverafter the water stress was relieved. Microscopic observationof stomata of the cotyledons by a replica method revealed thatthe water stress reduced the increase in both stomatal widthand density, and this reduction did not completely recover afterthe water stress was relieved. After one day of water stress, the endogenous ABA content ofthe cotyledon was increased from 68 to 114 ng/g fr wt. Afterthe water stress was relieved, the increased ABA content decreasedlinearly over two days to 20 ng/sg fr wt, the same value asin unstressed cotyledons. These results indicate that the effect of water stress on thehypocotyl growth was reversible but that the effect on the cotyledonswas irreversible. The irreversible effect of water stress onthe growth of the cotyledons probably resulted from the inhibitionof stomatal development, but endogenous ABA did not appear toinhibit the development. (Received March 24, 1986; Accepted June 25, 1986)     

Changes in Wall Polysaccharides of Squash (Cucurbita maxima Duch.) Hypocotyls under Water Stress Condition: I. Wall Sugar Composition and Growth as Affected by Water Stress

Hypocotyl growth of dark-grown squash (Cucurbita maxima Duch.)seedlings was greatly reduced by the addition of 60 mM polyethyleneglycol (PEG) to hydroponic solution (water stress). When PEGwas removed after one day, growth promptly recovered. The contents of hemicelluloses and cellulose in the wall increasedunder unstressed condition as hypocotyls grew but these increaseswere substantially reduced by water stress. The increases inwall polysaccharide contents recovered when the water stresswas relieved. The amounts per hypocotyl of cellulose and thatof uronic acid in pectin changed in parallel with the growth(r=0.95 and 0.98, respectively). The amounts of most of thesugar components of hemicelluloses also changed in parallelwith hypocotyl growth. Pectic and hemicellulosic galactose contentof unstressed hypocotyls increased to day 2 when the hypocotylgrew at a maximum growth rate, then decreased. In contrast,galactose content of stressed hypocotyls progressively increasedto the end of the experiment. The results indicated that water stress substantially reducednet increases in most of the polysaccharides of the hypocotylcell walls when it reduced the growth, but it did not affectsyntheses of some galactosic polysaccharides in pectin and hemicelluloseB. We assume that the syntheses of non-galactosic wall polysaccharidesare associated with hypocotyl growth and the synthesis of galactose-containingpolysaccharides with preservation of the potential of the cellwall to be loosened, since hypocotyl growth promptly and completelyrecovers when water stress is relieved. (Received December 15, 1986; Accepted June 8, 1987)     

Water Deficit and Abscisic Acid Cause Differential Inhibition of Shoot versus Root Growth in Soybean Seedlings : Analysis of Growth, Sugar Accumulation, and Gene Expression

Roots often continue to elongate while shoot growth is inhibited in plants subjected to low-water potentials. The cause of this differential response to water deficit was investigated. We examined hypocotyl and root growth, polysome status and mRNA populations, and abscisic acid (ABA) content in etiolated soybean (Glycine max [L.] Merr. cv Williams) seedlings whose growth was inhibited by transfer to low-water potential vermiculite or exogenous ABA. Both treatments affected growth and dry weight in a similar fashion. Maximum inhibition of hypocotyl growth occurred when internal ABA levels (modulated by ABA application) reached the endogenous level found in the elongating zone of seedlings grown in water-deficient vermiculite. Conversely, root growth was affected to only a slight extent in low-water potential seedlings and by most ABA treatments (in some, growth was promoted). In every seedling section examined, transfer of seedlings into low-water potential vermiculite caused ABA levels to increase approximately 5- to 10-fold over that found in well-watered seedlings. Changes in soluble sugar content, polysome status, and polysome mRNA translation products seen in low-water potential seedlings did not occur with ABA treatments sufficient to cause significant inhibition of hypocotyl elongation. These data suggest that both variation in endogenous ABA levels, and differing sensitivity to ABA in hypocotyls and roots can modulate root/shoot growth ratios. However, exogenous ABA did not induce changes in sugar accumulation, polysome status, and mRNA populations seen after transfer into low-water potential vermiculite.     

表油菜素内酯对黄瓜下胚轴伸长、内源GA~3,ABA及淀粉含量的影响

1 ppm表油菜素内酯(epiBR)能显著地促进光下生长的黄瓜下胚轴伸长。其促进伸长的效应需约5h滞后期,明显超过IAA而与GA_3的滞后期接近。GA_3对epiBR促进黄瓜下胚轴的伸长具有增效作用。epiBR能提高内源GA_3,ABA的水平,处理24h后,GA_3/ABA值约为对照的两倍。经epiBR处理后的黄瓜下胚轴内淀粉含量较低,并维持在同一水平,这与GA_3的作用颇为一致。     

Longitudinal Gradients of Indole-3-Acetic Acid and Abscisic Acid in the Hypocotyl of Etiolated Bean Seedlings

The longitudinal distribution of abscisic acid (ABA) and indole-3-aceticacid (IAA) along the hypocotyl of 5-d-old etiolated Phaseolusvulgaris L. cv. Limburg seedlings was measured. IAA was analysedby the L-methyl-indole--pryone assay (2-MIP) and ABA by electroncapture gas chromatography (ECD-GC). Length and width of theinner parenchyma cells, growth rate and protein content werealso measured. Cell expansion occurred predominantly in a region20 mm below the centre of the hook where elongation rate wasmaximal and where protein concentration decreased rapidly withdistance from the hook. The ratio between ABA and IAA was constant along the lengthof the hypocotyl. On a fresh weight basis the concentrationof both growth substances was maximal in the upper (youngest)part, decreased in slightly older sections where cell expansionwas proceeding and was smallest in the basal regions where cellexpansion was complete. However, when expressed on a proteinbasis the concentration gradient of the hormones was the reverseof that described on a fresh weight basis. Key words: IAA, ABA, hypocotyl, etiolated, bean     

Blue light control of hypocotyl elongation in etiolated seedlings of Lactuca sativa (L.) cv. Grand Rapids related to exogenous growth regulators and endogenous IAA, GA3 and abscisic acid

Results of studies on the interaction of blue light (B) and exogenous applied plant hormones (IAA, GA3 and ABA) as well as inhibitors of their synthesis on the control of hypocotyl elongation in etiolated seedlings of Lactuca sativa (L.) cv. Grand Rapids are presented, and compared with endogenous GA3, IAA and ABA levels measured by capillary gas chromatography-mass spectrometry-selected ion monitoring (GC-MS-SIM). Hypocotyl elongation of etiolated seedlings was linearly inhibited by increasing the B fluence rate between 0 and 18.3 mol m–2 s–1. Both GA3 and IAA added to the incubation medium at different concentrations were able to eliminate partially the inhibition of growth caused by 7.2 mol m–2 s–1 B. When dark (D)-grown seedlings were treated with Ca-prohexadione, a specific inhibitor of gibberellin 3-hydroxylation, they showed a growth inhibition similar to under B. Also, a suppression of growth as in B was obtained when D-grown seedlings were treated with ABA 380 M. By lowering ABA levels with fluridone (an inhibitor of ABA synthesis) a partial reversion of hypocotyl growth inhibition was obtained in B-grown seedlings. While none of the growth promoters used were able to reverse completely the growth inhibition caused by B, a proper combination of GA3, IAA and (eventually) fluridone, abolished the B effects. Correspondingly, lower levels of GA3 and IAA and a higher concentration of ABA were measured by GC-MS-SIM in B-grown hypocotyls than in D-grown ones. These results support the hypothesis that hormones are implicated in mediation of B light-dependent inhibition of hypocotyl elongation, which seems to be the result of a balance among endogenous levels of growth promoting and growth inhibiting hormones.     

芦丁对绿豆幼苗营养生长的影响及其与IAA的相互作用

观察了植物体内的天然黄酮芦丁和吲哚乙酸(IAA)对绿豆幼苗营养生长的影响并测定胚轴中的芦丁和IAA含量.光照条件下芦丁(60μg/mL以下)处理对绿豆幼苗生长有一定促进作用,表现为胚轴和主根伸长加快、侧根数目增多、鲜重或干重增加;而光照条件下更高浓度芦丁(80μg/mL以上)处理及黑暗条件下芦丁(20～100μg/mL)处理对绿豆幼苗生长有抑制作用.当培养基中的芦丁浓度为60～80 μg/mL时,光照下的幼苗比暗处理的幼苗在胚轴中积累更多的芦丁;而芦丁浓度为40μg/mL以下和接近100μg/mL时幼苗在光照下累积的芦丁较暗处理的幼苗更少.0.1μg/mL以上的IAA促进芦丁的累积而进一步抑制幼苗胚轴和主根的伸长.当培养基中含有40 μg/mL的芦丁和0.5μg/mL的IAA时,胚轴中累积的芦丁达到高峰.芦丁降低黄化幼苗内源性IAA在胚轴中的累积,并抑制幼苗对IAA的吸收.     

Changes in Levels of Auxin and Abscisic Acid and the Evolution of Ethylene in Squash Hypocotyls after Treatment with Brassinolide

Brassinolide stimulated an increase in fresh weight of segmentsof hypocotyls of etiolated squash (Cucurbita maxima Duch. cv.Houkou-Aokawaamaguri). Brassinolide-treated segments containedhigher levels of IAA than water-treated segments and also showeda tendency towards decreased levels of ABA. However, treatmentwith brassinolide did not have much effect on the productionof ethylene. (Received May 30, 1988; Accepted May 22, 1989)     

Distribution of Indole-3-Acetic Acid in the Apoplast and Symplast of Squash (Cucurbita maxima) Hypocotyls

The concentration of endogenous IAA was higher in an apoplasticsolution (2.3xl0^–7M) than in a symplastic solution (0.5x 10^–7 M) obtained from segments of etiolated squash (Cucurbitamaxima Duch.) hypocotyls. Exogenously applied IAA (10^–5M) increased the level of IAA in both the apoplastic and thesymplastic solution. The concentration of IAA in the apoplasticsolution increased to 75% of the concentration of exogenousIAA in 4 h, but that in the symplastic solution increased onlyto 23% of the concentration of exogenous IAA. These resultsdemonstrate that the concentration of endogenous IAA is higherin the apoplast than in the symplast of squash hypocotyls, andthey suggest that IAA exerts its physiological effects, at leastto some extent, from the outside of cells. (Received September 20, 1996; Accepted January 10, 1997)     

Endogenous Phytohormones and Regulation of Morphogenesis of Arabidopsis thalianaby Blue Light

We studied the effects of blue light (BL) on the levels of endogenous phytohormones (IAA, ABA, gibberellins, and cytokinins) and morphogenesis of the 7-day-old Arabidopsis thaliana(L.) Heynh seedlings of wild type (Ler) and its hy4mutant with a disturbed synthesis of cryptochrome 1 (CRY1), which is a receptor for BL. In darkness, the mutant contained considerably less free IAA and zeatin, but much more ABA as compared to the wild-type seedlings. BL retarded the hypocotyl growth in the wild-type seedlings but stimulated it in the mutant. Elongation of mutant hypocotyls was accompanied by accumulation of free IAA and a decrease in the content of free ABA; the level of cytokinins did not change. We believe that the response of the hy4hypocotyls to BL is mediated by a BL receptor distinct from cryptochrome 1. The conclusion is that light and hormonal signals interact in the control of the hypocotyl growth in A. thalianaseedlings.     

Changes in the concentration of indole-3-acetic acid during the growth of etiolated lupin hypocotyls

The longitudinal distribution of unaltered radioactive indole-3-acetic acid (IAA), after application of [5-³H]-IAA to decapitated etiolated lupin hypocotyls. exhibited a wave-like pattern similar to that obtained with endogenous IAA. Waves of radioactive IAA were localizated both in the elongation zone and in the non-growing basal region of the hypocotyl. These IAA waves were transient because of basipetal polar transport and metabolism of IAA.
The level of endogenous IAA in different zones of the hypocotyl varied with age, following a wave-like pattern. During the elongation period of each zone, IAA was parallel to the bell-shaped curve of the growth rate. In addition, a role in secondary cell wall deposition is suggested for the other IAA wave that appeared after the cell elongation period, since an electron microscopic morphometric analysis of the cell wall showed that the cell wall thickness increased once the cell elongation ceased.
As the oscillation of endogenous IAA level occured in both space (distribution along the hypocotyl) and time (variation with age), it is suggested that the level of IAA really depended on the growth status of the cells. The response of the cells to the positional information submitted by the auxin waves as regards the growth status of the cell is discussed.     

The Effect of Epibrassinolide on Arabidopsis Seedling Morphogenesis and Hormonal Balance under Green Light

We studied the effect of 24-epibrassinolide (EB) on the levels of endogenous hormones and photomorphogenesis of Arabidopsis thaliana (L.) Heynh wild-type (Ler) and mutant (hy4) seedlings. This mutant is deficient in the cryptochrome 1 (CRY1) synthesis. CRY1, which is a product of the HY4 gene, is a blue light photoreceptor in wild-type plants, but is sensitive to green light as well. In dark-grown seven-day-old mutant seedlings, the ABA/zeatin ratio differed from this ratio in wild-type seedlings. Thehy4 mutant exhibited a lower zeatin and higher free-ABA contents, which could retard its hypocotyl growth in darkness. EB retarded the growth of hypocotyls in etiolated hy4 seedlings and enlarged their cotyledons more efficiently than in wild-type seedlings. Green light (GL) did not affect the growth of hypocotyls but enlarged cotyledons of hy4 seedlings, which might be associated with some increase in the level of free IAA and a considerable decrease in free ABA and also with a decrease in the cytokinin level in seedlings. The hy4 cotyledon response to GL depended evidently on photoreceptors other than CRY1. GL enhanced the effects of EB on the morphogenesis of both Ler and hy4 seedlings, which was coupled with changes in the balance of endogenous IAA, ABA, and cytokinins. We may suppose that EB is involved in the control of photomorphogenesis by interaction with endogenous hormones, which are involved in the transduction of a light signal absorbed by the GL photoreceptors.     

Water Potential and Mechanical Properties of the Cell Wall of Hypocotyls of Dark-Grown Squash (Cucurbita maxima Duch.) under Water-Stress Conditions

Hypocotyl growth of seedlings of dark-grown squash (Cucurbitamaxima Duch.) was greatly reduced by the addition of 60mM polyethyleneglycol (PEG) to hydroponic solution (water stress). Apoplastic solution (A) and cell sap (C) were separately collectedfrom the hypocotyl segments by a centrifugation method. Theosmotic potentials of A (_A) and C (_c), and (=_c–_A) ofstressed hypocotyls were always lower than those of unstressedhypocotyls. Suction force was measured by immersing the segments into solutionsof different concentrations of mannitol. Suction force was significantlycorrelated with _C (r= –0.99). The mechanical properties of the cell wall of hypocotyl segmentswere measured by stressrelaxation technique. Minimum stressrelaxation time (T_o), relaxation rate (R) and residual stressof unstressed hypocotyls were low during the growth period andincreased when the growth ceased. T_o and R of stressed hypocotylsdecreased one day after the stress treatment, but the residualstress was not decreased by the water stress throughout theexperiment. These results suggest that the suppressed growth of dark-grownsquash hypocotyls under water stress was due neither to thereduction of the osmotic potential difference between innerand outer space of the cell, nor to the decrease in suctionforce, but was partly due to the unchanged mechanical propertiesof the cell wall, as represented by one stress-relaxation parameter,residual stress. (Received February 5, 1988; Accepted September 8, 1988)     

Effect of light on peroxidase and lignin synthesis in mungbean hypocotyls

Two days of light irradiance reduced the growth of mungbean hypocotyls as well as the levels of endogenous indole-3-acetic acid (IAA). In hypocotyls, both peroxidase and laccase activities were enhanced by light. The lignin content in mungbean hypocotyls was enhanced twofold by light. The inhibition of mungbean hypocotyl growth caused by light might be due to the decline of endogenous IAA, which could be degraded by a cationic peroxidase. The higher levels of lignin were correlated with the increased anionic peroxidase activity in light-treated tissues.     

Endogenous Abscisic Acid Promotes Hypocotyl Growth and Affects Endoreduplication during Dark-Induced Growth in Tomato (Solanum lycopersicum L.)

Dark-induced growth (skotomorphogenesis) is primarily characterized by rapid elongation of the hypocotyl. We have studied the role of abscisic acid (ABA) during the development of young tomato (Solanum lycopersicum L.) seedlings. We observed that ABA deficiency caused a reduction in hypocotyl growth at the level of cell elongation and that the growth in ABA-deficient plants could be improved by treatment with exogenous ABA, through which the plants show a concentration dependent response. In addition, ABA accumulated in dark-grown tomato seedlings that grew rapidly, whereas seedlings grown under blue light exhibited low growth rates and accumulated less ABA. We demonstrated that ABA promotes DNA endoreduplication by enhancing the expression of the genes encoding inhibitors of cyclin-dependent kinases SlKRP1 and SlKRP3 and by reducing cytokinin levels. These data were supported by the expression analysis of the genes which encode enzymes involved in ABA and CK metabolism. Our results show that ABA is essential for the process of hypocotyl elongation and that appropriate control of the endogenous level of ABA is required in order to drive the growth of etiolated seedlings.     

Biosynthesis of Auxin-Induced Ethylene. Effects of Indole-3-Acetic Acid, Benzyladenine and Abscisic Acid on Endogenous Levels of 1-Aminocyclopropane-l-Carboxylic Acid (ACC) and ACC Synthase

Auxin-induced ethylene biosynthesis and its regulatory stepsin etiolated mung bean hypocotyl segments were examined. Theendogenous content of 1-aminocyclopropane- 1-carboxylic acid(ACC), an immediate precursor of ethylene, increased correspondingto the rate of ethylene production. Benzyladenine (BA), whichis a synergistic stimulator of auxin-induced ethylene production,increased the ACC content parallel to the rate of ethylene productionin the presence of IAA, but failed to increase the ACC contentin the absence of IAA while ethylene production was significantlystimulated by BA. Abscisic acid (ABA) inhibited the formationof ACC. The ACC synthase activity in the tissue was increasedby IAA, and the increase was further promoted by the presenceof BA. Cycloheximide severely inhibited the development of auxin-inducedACC synthase. The enzymatic properties of mung bean ACC synthasewere similar to those of the tomato fruit enzyme. Aminoethoxyvinylglycine(AVG) and aminooxyacetic acid, which inhibit the ACC synthasereaction, stimulated the development of ACC synthase. The regulatorymechanisms of the growth regulators are discussed in relationto ACC formation. (Received December 3, 1980; Accepted January 22, 1981)     

The interaction of light irradiance with auxin in regulating growth of <Emphasis Type="Italic">Helianthus annuus</Emphasis> shoots

Plants growing under canopy shade or in near-neighboring proximity of taller vegetation are the receivers of shade light conditions. The effect of light irradiance (photosynthetically active radiation [PAR]), one of the main components of shade light, on the growth of various tissues of sunflower seedlings and the possible role of auxin were investigated. Gradual reductions in PAR irradiance level from near-normal to low and very low result in significant and gradual increases in sunflower hypocotyl growth and endogenous auxin content. Similar reductions in PAR level resulted in significant and gradual decreases in sunflower cotyledon and leaf growth, and endogenous auxin content. Exogenously applied auxin increased hypocotyl elongation under near-normal PAR, where IAA levels are below optimum, but decreased elongation under very low PAR, where IAA levels are already at optimum. These results suggests that auxin acts as positive growth regulator of sunflower hypocotyls subjected to low light irradiance stress. This is further supported by the transfer experiments where seedlings transferred, for example, from near-normal PAR to very low PAR showed increased elongation associated with increased IAA levels. Therefore, it is reasonable to conclude that light irradiance-mediated changes in hypocotyl elongation of young sunflower seedlings are regulated by endogenous auxin levels.     

Gibberellin and auxin contents and ethylene production in the hypocotyls of green and etiolated bean plants treated with chlorocholine chloride

The effect of CCC treatment on the GA and IAA contents of the hypocotyls of green and etiolated bean plants and on their distribution was studied. In the treated plants the GA content of the hypocotyls was higher than that of the control. On the application of CCC the distribution of the hormone content within the organ altered: in the basal part of the hypocotyls of the treated plants the hormone content increased. In the hypocotyls of the treated plants the IAA content was less than in the controls and it was accumulated in the basal part. The ethylene production of the aplcal and basal parts of the hypocotyls proved to be correlated with their IAA content. Presented at the International Symposium “Plant Growth Regulators” held on June 18–22, 1984 at Liblice, Czechoslovakia.     

NaCI Reduces Indole-3-Acetic Acid Levels in the Roots of Tomato Plants Independent of Stress-Induced Abscisic Acid

Indole-3-acetic acid (IAA) was measured in leaves and roots of tomato (Lycopersicon esculentum) genotypes subjected to salt stress. An abscisic acid (ABA)-deficient mutant of tomato (sitiens), the genetic parent (Rheinlands Ruhm, RR), and a commercial variety (Large Cherry Red, LCR) of tomato were treated with 50 to 300 mM NaCl in nutrient culture. Both LCR and RR had significantly higher levels of IAA in the roots compared with that in sitiens prior to treatment. The initial levels of IAA in the roots of LCR and RR declined by nearly 75% after exposure to NaCl, whereas those in roots from the sitiens mutant remained unchanged. IAA levels in the leaves of all genotypes remained unchanged or increased slightly in response to NaCl. ABA was highest in leaves from the normal genotypes after exposure to NaCl. ABA levels in the roots of sitiens were similar to the levels in the normal genotypes, whereas levels in the leaves were only 10% of the levels found in normal genotypes regardless of the salt treatment. Treatment of LCR and sitiens with exogenous ABA increased the ABA levels in leaves and roots, but there were no measurable changes in endogenous IAA. Therefore, the reduction in IAA appears to result from an ABA-independent effect of NaCl on IAA metabolism in the roots of stressed plants.     

Changes of Endogenous ABA Content in Relevance to the Vigor of Peanut Seeds

During the development of peanut (Arachis hypogaea L. ) seed, the endogenous ABA content increased steadily'in hypocotyl, increased to a peak at 40 d after pegging with a drastic decline afterwards in testa; and in cotyledon, increased to a peak at 60 d after pegging but with a slight fall afterwards. There seemed to be a close relationship between the increose of vigor index and net loss of endogenous ABA content in the peanut seed germinating in vitro. Osmoticum (mannitol) promoted the endogenous ABA in the cotyledon and hypocotyl. and Fluridone inhibited that in the cotyledon. There were two different paths of the endogenous ABA synthesis in peanut seed, C40 in the cotyledon and C15 in the hypocotyl. When peanut seeds were put in the conditions of precocious maturation or germination tine endogenous ABA content fell down. Result from this experiment concluded that the hypocotyls played an important role in the transition from development to germination of. peanut seed.