An endogenous growth inhibitor, 3-hydroxy-β-ionone. II. Changes in its levels associated with the light-induced growth inhibition of hypocotyls of Phaseolus vulgaris

Dark-grown, 4-day-old seedlings of dwarf bean ( Phuseolus vulgaris L. cv. Morocco) and tall bean (cv. Kentucky Wonder) were transferred to conditions of continuous light at various fluence rates, and the resulting changes in growth rates and concentration of 3-hydroxy-β-ionone, a novel endogenous growth inhibitor, were monitored. The light-induced inhibition of growth and the levels of the inhibitor concentration were dependent on the fluence rate of light applied. The growth rate of hypocotyls of both cultivars decreased rapidly and reached a plateau 18 h after the onset of light. However, the more noticeable changes were the marked inhibition of growth of the hypocotyls of the dwarf cultivar. The concentration of 3-hydroxy-β-ionone in the hypocotyls of both cultivars increased after a 20-min lag and reached plateau values after 12 h. The concentration of the inhibitor in the hypocotyls of the dwarf cultivar increased to about 4 to 8 times the level in the nonirradiated controls, while that in the hypocotyls of the tall cultivar increased only about 2- to 3-fold under the same conditions. The high concentration of 3-hydroxy-β-ionone in the dwarf cultivar under steady-state conditions in continuous light, as compared with that in the tall cultivar, may determine the growth habit of these cultivars.     

An endogenous growth inhibitor, 3-hydroxy-β-ionone. III. Its longitudinal gradients in the first internode of Phaseolus vulgaris

Dark-grown, 10-day-old bean seedlings (Phaseolus vulgaris L. cv. Morocco) were transferred to continuous light, and the resulting changes in growth rate and concentration of 3-hydroxy-β-ionone, an endogenous growth inhibitor, were monitored. The growth rate of the first internodes of the seedlings decreased rapidly and leveled off 20 h after onset of light. This plateau value was about 25% of the growth rate of the non-irradiated control. The concentration of 3-hydroxy-β-ionone in the internodes of the irradiated seedlings increased rapidly and reached a plateau value after 16 h. This increased level of the inhibitor was about 5 times the level in the non-irradiated control. The changes in the levels of the inhibitor in the internodes reflected the light-induced growth inhibition of the internodes. The longitudinal distribution of the growth inhibitor along the first internodes of the seedlings was closely correlated with the light-induced growth inhibition of the corresponding region of the internodes. The present results suggest that the endogenous growth inhibitor 3-hydroxy-β-ionone may play an important role in the inhibition by light of internode growth of bean seedlings.     

The effect of cotyledons on hypocotyl elongation in light-grown bean seedlings: Comparison between dwarf and tall varieties

Hypocotyl sections with and without the cotyledons were cutfrom bean seedlings and incubated under white light of 6000lux. The cotyledons had an inhibitory effect as well as a promotiveeffect on hypocotyl growth. The former effect was more strikingin the dwarf variety, and the latter in the tall variety. Whenthe hypocotyl units were exposed to light for shorter times(6 hr or less) or incubated under weaker light (1600 and 50lux), the inhibitory effect of the cotyledons decreased greatly,and in the tall variety the presence of cotyledons producedno inhibition, but a promotion of hypocotyl growth. GA treatmentenhanced hypocotyl growth and counteracted the growth inhibitioncaused by the cotyledons. On the whole, the GA effect was moremarked in the tall variety than in the dwarf. The elongation of bean hypocotyls may be controlled by a balancebetween the inhibitory and promotive effects of cotyledons,and the predominance of the former over the latter may be oneof the causes for expressing dwarfing. (Received November 13, 1976; )     

Light growth inhibition and growth inhibitors in Sakurajima radish seedlings

Growth of Sakurajima radish seedlings (Raphanus sativus var.hortensis f. gigantissimus Makino) was studied in relation tothe effects of light, the effectiveness of plant hormones, andthe variation in endogenous growth inhibitor content Externally applied GAs and IAA had no effect on the elongationof the hypocotyl in the light as well as in the dark. BA wasslightly promotive in the light, but was not so effective asto nullify the light-induced inhibition of elongation. ABA wasstrongly suppressive in the light as well as in the dark. Growth inhibitors in seedlings were extracted, analyzed by thin-layerchromatography and the Sakurajima radish hypocotyl elongationtest, and 3 acidic and 3 neutral inhibitors were detected. Amongthem all of the acidic and one of the neutral inhibitors increasedwith the time period of illumination, whereas the other 2 neutralsubstances remained almost unchanged in the light but decreasedin the dark. Thus the levels of all 6 inhibitors was higherin light-grown seedlings than in dark-grown ones. This suggeststhat light inhibition in Sakurajima radish hypocotyls may becontrolled by the variation in the inhibitor levels in the seedlings. (Received December 1, 1977; )     

Natural growth inhibitors associated with the cotyledon effect in light-grown dwarf and tall beans

The contents of non-acidic, acidic and bound growth inhibitorsin hypocotyls and cotyledons were compared between dark- andlight-grown dwarf and tall beans by means of thin-layer chromatographyand bioassay. In the non-acidic fraction, one major inhibitoryactivity appeared on the chromatogram, but its Rf zone was differentbetween hypocotyls and cotyledons. In both the acidic and boundinhibitor fractions, one major inhibitory activity appearedat the Rf zone corresponding to ABA. The ABAIike substance whichwas the major inhibitor in the hypocotyl was more abundant inlight-gorwn than dark-grown ones, especially in the dwarf variety,but light irradiation did not cause its transport from cotyledonsto the hypocotyl. A larger amount of bound ABA-like substance,which was the major inhibitor in the cotyledon, was presentin the dwarf than the tall variety regardless of the light condition.Cotyldon-enhanced photoinhibition of hypocotyl growth couldnot be explained by the levels of the xanthoxinand ABA-likeinhibitors, or the transport of these free inhibitors from cotyledons. (Received September 8, 1977; )     

Comparison of the effects of white light and the growth retardant paclobutrazol on the ethylene production in bean hypocotyls

At a concentration of 17 µmol·L^–1, paclobutrazol (PP), a triazole plant growth retardant, effectively reduced the elongation and increased the thickness of hypocotyls in 6-day-old Phaseolus vulgaris L. cv. Juliska seedlings, both in the light and in the dark. PP treatment did not increase the cell number in transverse sections of hypocotyls. The diameter of hypocotyls was uniform from the zone of intensive elongation along the whole hypocotyl in etiolated plants, but those grown in the light exhibited an additional lateral expansion at the base. Ethylene evolution was not reduced by PP in etiolated hypocotyls, and did not differ significantly in the elongating apical and fully grown basal zones. PP reduced the ethylene release by the growing zones in green hypocotyls, but not in the basal parts, which resulted in an increasing ethylene gradient towards the hypocotyl base. The level of 1-aminocyclopropane-1-carboxylic acid (ACC), the immediate precursor of ethylene, was much higher in retardant-treated hypocotyls than in the controls, which was due in part to the reduced malonylation. The swelling of the hypocotyl bases could be eliminated by inhibitors of ethylene biosynthesis or action, or could be induced by 10 µmol·L^–1ACC in control plants in the light. None of these treatments had a significant effect on the lateral expansion of hypocotyls in etiolated seedlings. PP treatment induced a similar effect to that of white light in etiolated seedlings, and amplified the effect of light in green plants with respect to the ACC distribution, and consequently, the ethylene production in the hypocotyls of 6-day-old bean seedlings. It can be concluded that the lateral expansion of hypocotyl bases in PP-treated green plants is controlled by ethylene.     

Glycosidases and acid-invertase activities are not correlated with Phaseolus hypocotyl growth

Changes in α-galactosidase, β-galactosidase, β-glucosidase and acid invertase activities were examined in Phaseolus vulgaris hypocotyls treated with gibberellic acid (GA), naphthyl acetic acid (NAA) and distilled water (DW) (control) in light condition. The activities were estimated both in cytoplasmic and ionically wall-bound fraction. The upper segment showed considerable elongation growth while there was hardly any growth in lower segment. GA and NAA showed distinct promotion and inhibition respectively in hypocotyl growth in upper segment. The glycosidase activities were detected in both the fractions but the activity was more pronounced in cytoplasmic than in wall fraction. Acid invertase activity was present only in cytoplasmic fraction. In lower segment, in both cytoplasmic and wall fraction, the glycosidase activity, in general, showed a decreasing trend and no effect of treatment could be envisaged. In upper segment, though the trend was similar to the lower one, in α- and β-galactosidase NAA treated segment had more activity. Invertase activity also did not show a clear trend to implicate its function in hypocotyl elongation growth. The results are discussed in relation to establishing a correlation between an activity (glycosidase and invertase) and a physiological process (hypocotyl elongation). It is concluded that these wall-loosening enzymes have no role in elongation growth of Phaseolus vulgaris hypocotyls.     

Sequestration of 3-oxygenated α-ionone derivatives in the male rectal gland of the solanaceous fruit fly, Bactrocera latifrons

The solanaceous fruit fly, Bactrocera latifrons (Hendel) (Diptera: Tephritidae), infests various solanaceous fruits including eggplant and chili peppers. We found that a freshly cut fruit of an eggplant cultivar [ Solanum melongena L. cv. Long Purple Oriental (Solanaceae)] selectively attracted and provoked voracious feeding behavior in adult B. latifrons males (but not in females) in an indoor test. One of the male-specific attractants/feeding stimulants in eggplant was identified as 3-hydroxy-α-ionone. Sexually mature males that fed on the eggplant pulp selectively accumulated a series of 3-oxygenated α-ionone/α-ionol analogs (e.g., 3-oxo-α-ionol and 3-oxo-7,8-dihydro-α-ionol) in the rectal gland, a suspected pheromone reservoir in male flies. Males fed on synthetic 3-hydroxy-α-ionone, 3-oxo-α-ionol, or 3-oxo-α-ionone partially biotransformed the compounds into 3-oxo-α-ionol, 3-oxo-α-ionone, and 3-oxo-7,8-dihydro-α-ionol and sequestered substantial quantities (as high as 5 µg/gland as a mixture) in a similar ratio in the rectal gland within 6 h after ingestion. These results suggest that the rectal sequestrates may serve as a sex pheromone similar to other Bactrocera species that use phenylpropanoid volatiles to attract conspecific females.     

Aging progression involving dwarfism and its acceleration by red light in bean hypocotyls

The effect of red light on the aging progression of the bean (Phaseolus vulgaris L.) hypocotyl segment unit was examined in relation to dwarfism using Kentucky Wonder (tall) and Masterpiece (dwarf) varieties. In both plants, red light promoted the elongation of younger zones and inhibited that of mature zones. The zone exhibiting maximum elongation was shifted to the younger zones by red light irradiation regardless of the plant type, but its extent was greater in the dwarf than in the tall. Thus, red light hastens both the beginning of elongation in the younger portion and its termination in the mature portion of the hypocotyl, particularly of the dwarf plant. These red light responses in each zone of both the tall and dwarf hypocotyl units were reversed by subsequent exposure to far red light regardless of the duration and intensity of red light, thus indicating that the hastened aging progression of the hypocotyl by red light is mediated by phytochrome. However, there is no difference in the rate of decay of Pfr between the tall and dwarf hypocotyls.     

NaCl and wounding induced changes in NAD reductase in hypocotyls and root tips ofPhaseolus vulgaris L.

The effect of 300 mM NaCl and wounding on the nicotinamide adenine dinucleotide (NAD) kinase, revealed by the guanosine triphosphate-dependent NAD reductase activity, was studied in two differently resistant bean cultivars using densitometric analysis of electrophoretic gels. In the presence of NaCl the total activity of NAD reductase was increased, in hypocotyls and root tips of resistant cultivar. The contribution of each of NAD reductase isoforms to the total activity was not significantly different between cultivars. Conversely, after wounding the hypocotyl, an increase could be observed in both cultivars and differences were demonstrated in the contribution of the different isoforms.     

The Role of Various Regions of the Bean Hypocotyl on Red Light-induced Hook Opening

Measurement of various zones on the concave half of etiolated Phaseolus vulgaris L. (cv. Black Valentine) hypocotyls has shown that growth at the basal portion of the elbow and the contiguous upper portion of the shank was stimulated earliest by red light. Growth of these two zones was unaffected by the tissue of the convex half but was inhibited by tissue distal to them. The inhibition was alleviated by the continuous presence of shank tissue below the growing zones. Based on cuts made halfway through the hypocotyl at positions above, below, or between the two zones of growth, it is suggested that cells at the inner portion of the upper shank control in some way the light-induced growth of the elbow cells directly above.     

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.     

Reduced de-etiolation of hypocotyl growth in a tomato mutant is associated with hypersensitivity to, and high endogenous levels of, abscisic acid.

A recessive single gene mutant, 7B-1, in tomato was originally selected for its photoperiod-dependent male sterility. The 7B-1 mutant also has some pleiotropic effects including reduced light-induced inhibition, i.e. de-etiolation, of the hypocotyl in long days (LD), increased seed size and weight, and reduced transpiration rate. These traits led us to investigate the sensitivity of 7B-1 to exogenous hormones and the interaction of these responses with daylength. In LD, but not in short days (SD), 7B-1 was more sensitive than wild-type (WT) to exogenous abscisic acid (ABA) for inhibition of seed germination, root elongation and transpiration rate. 7B-1 mutant also exhibited reduced responses to exogenous gibberellin (GA(3)) for hypocotyl elongation, and to inhibitors of GA biosynthesis for seed germination and root and hypocotyl elongation. 7B-1 hypocotyls contained a higher level of endogenous ABA than WT in both photoperiods, although ABA levels were higher in LD than in SD. In contrast, growth-active GAs, i.e. GA(1), GA(3) and GA(4), and IAA were low in the mutant hypocotyls. The 7B-1 mutant appears to be an ABA-overproducer, and the photoperiod-regulated ABA levels may be responsible for the hypersensitivity of the mutant to exogenous ABA.     

Absence of Red Light-Induced Stimulation of Growth in the Hook of Etiolated Dwarf-Pea Seedlings

Red light inhibited the growth of the apical part of the hookin dark-grown seedlings of a dwarf variety (cv. Progress No.9) of pea (Pisum sativum L.), whereas it promoted such growthin a tall variety (cv. Alaska). In the elongation zone of theepicotyl of the dwarf variety the extent of inhibition of growthwas similar to or even smaller than that in the tall variety.Local irradiation of the apical part of the hook also causedinhibition of growth in the hook of the dwarf variety and promotionof growth in the tall variety. The inhibition of growth in theapical part of the hook of cv. Progress may be involved in thedwarfism induced by irradiation with red light of this cultivar. (Received May 15, 1989; Accepted April 27, 1990)     

Molecular weight distribution of hemicellulosic polysaccharides of the cell wall of tall and dwarf rice cultivars, and the effect of GA3

Growth rate, osmotic potential of the cell, cell wall mechanical properties, sugar composition and molecular weight (MW) distribution of water-soluble hemicellulosic polysaccharides of the second leaf sheath of one tall ( Oryza sativa L. cv. Nihonbare) and two dwarf ( Oryza sativa L. cvs. Tan-ginbozu and Waito C) cultivars of rice were compared. The effect of gibberellic acid (GA₃) on the above-mentioned parameters was also studied using the tall (Nihonbare) and one of the dwarf (Waito C) cultivar. The minimum stress-relaxation time (T₀) was higher in the cell wall from the two dwarfs than in the tall cultivar. Furthermore, in the water-soluble hemicellulosic polysaccharides the mass-average MW of β -glucan was higher and that of arabinoxylan was lower in the tall cultivar than in the dwarf ones. Thus, dwarfism of cvs Tan-ginbnozu and Waito C might be correlated with the different MW distributions of β -glucan and arabinoxylan. GA₃ induces growth in the dwarf Waito C cultivar, decreases the T₀ value of the cell wall, and also decreases the average MW of water-soluble hemicelluloses. Changes in β (1–3)(1–4)glucan or arabinoxylan or in both are proposed as part of the cell wall loosening mechanism induced by gibberellin.     

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.     

Gibberellins control Arabidopsis hypocotyl growth via regulation of cellular elongation

The gibberellins (GAs) are endogenous regulators of plant growth. Experiments are described here that test the hypothesis that GA regulates hypocotyl growth by altering the extent of hypocotyl cell elongation. These experiments use GA-deficient and altered GA-response mutants of Arabidopsis thaliana (L.) Heyhn. It is shown that GA regulates elongation, in both light- and dark-grown hypocotyls, by influencing the rate and final extent of cellular elongation. However, light- and dark-grown hypocotyls exhibit markedly different GA dose-response relationships. The length of dark-grown hypocotyls is relatively unaffected by exogenous GA, whilst light-grown hypocotyl length is significantly increased by exogenous GA. Further analysis suggests that GA control of hypocotyl length is close to saturation in dark-grown hypocotyls, but not in light grown hypocotyls. The results show that a large range of possible hypocotyl lengths is achieved via dose-dependent GA-regulated alterations in the degree of elongation of individual hypocotyl cells.Key words: Arabidopsis, cell elongation, gibberellin (GA), GA mutants, hypocotyl.      

PHYTOCHROME KINASE SUBSTRATE4 modulates phytochrome-mediated control of hypocotyl growth orientation

Gravity and light are major factors shaping plant growth. Light perceived by phytochromes leads to seedling deetiolation, which includes the deviation from vertical hypocotyl growth and promotes hypocotyl phototropism. These light responses enhance survival of young seedlings during their emergence from the soil. The PHYTOCHROME KINASE SUBSTRATE (PKS) family is composed of four members in Arabidopsis (Arabidopsis thaliana): PKS1 to PKS4. Here we show that PKS4 is a negative regulator of both phytochrome A- and B-mediated inhibition of hypocotyl growth and promotion of cotyledon unfolding. Most prominently, pks4 mutants show abnormal phytochrome-modulated hypocotyl growth orientation. In dark-grown seedlings hypocotyls change from the original orientation defined by seed position to the upright orientation defined by gravity and light reduces the magnitude of this shift. In older seedlings with the hypocotyls already oriented by gravity, light promotes the deviation from vertical orientation. Based on the characterization of pks4 mutants we propose that PKS4 inhibits changes in growth orientation under red or far-red light. Our data suggest that in these light conditions PKS4 acts as an inhibitor of asymmetric growth. This hypothesis is supported by the phenotype of PKS4 overexpressers. Together with previous findings, these results indicate that the PKS family plays important functions during light-regulated tropic growth responses.