THE DEVELOPMENTAL MORPHOLOGY OF NICOTIANA TABACUM ‘WHITE BURLEY’ AS INFLUENCED BY VIRUS INFECTION AND GIBBERELLIC ACID

Stein , Diana B. (U. Montana, Missoula.) The developmental morphology of Nicotiana tabacum ‘White Burley’ as influenced by virus infection and gibberellic acid. Amer. Jour. Bot. 49(5): 437–443. Illus. 1962.—‘White Burley’ tobacco with Severe Etch Virus (SEV) displayed a reduction of plant height which was overcome to a limited degree by spraying with gibberellic acid (GA). Spraying with GA, while hastening maturity in terms of earlier elongation and flowering, also prolonged the life of treated plants. Infection with SEV caused an increased rate of leaf production, and since flowering was also delayed, the greater number of leaves is produced by infected plants. Spraying with GA also increased the rate of leaf production but did not increase the final number of leaves produced. All groups except the unsprayed virus-infected plants showed a spurt in leaf production just prior to flowering. The pattern of internode elongation was obtained by the periodic measurement of individual internodes. In general, this pattern reflected the stunting properties of the virus and the growth-promoting properties of the GA. Internodes which were mature at time of spraying with GA were not affected. Infection with virus generally delayed elongation and shifted the internode pattern. Infection with SEV tended to reduce the size of leaves already present prior to inoculation, but some leaves produced after infection were actually 1arger than the same leaves on the controls. Spraying a healthy plant with GA made older leaves longer and wider, while less mature leaves at time of treatment tended to be longer and narrower. Spraying with GA reversed the reduction in size caused by the virus only if the leaf was a very young primordium or was formed during the course of treatment with GA.     

Effect of Gibberellic Acid on Growth, Gibberellin Content, and Chlorophyll Content of Leaves of Potato (Solanum tuberosum)

Spraying potato plants with a solution of gibberellic acid (GA)when the 15th leaf was emerging increased the area of this leafand its total gibberellin content, assayed by dwarf French beanleaf disks. GA, assayed by lettuce hypocotyls, was not detectedin leaves from untreated plants. The GA content of leaves fromGA-treated plants decreased after 2 weeks and none was detectableafter 5 weeks; apparently it was converted to another gibberellin,possibly the same as the natural gibberellin. GA increased chlorophyllcontent per leaf but increased leaf area more so that the chlorophyllper unit area decreased, and the leaves were paler than untreatedleaves.     

Growth rate and biomass partitioning of wildtype and low-gibberellin tomato (Solanum lycopersicum) plants growing at a high and low nitrogen supply

The growth-promoting effects of gibberellins (GAs) on plants are well documented, but a complete growth analysis at the whole plant level on plants with an altered GA biosynthesis has never been reported. In the present work, the relative growth rate (RGR), biomass partitioning and morphological parameters of wildtype (Wt) tomato ( Solanum lycopersicum L. cv. Moneymaker) plants were compared with those of isogenic ( gib ) mutants with a reduced biosynthesis of gibberellins. GA deficiency reduced RGR and specific leaf area (SLA, leaf area per unit leaf mass) and increased the net assimilation rate (NAR, the rate of biomass increment per unit leaf area). Despite the free access to nitrogen in the rooting medium, the low-GA mutants had a much higher root mass ratio (RMR, the root mass per unit plant biomass) than the Wt, suggesting that the mutants were disturbed in their growth response to nitrate supply. The experiment was repeated at a low exponential nitrate supply, which forced all plants to grow at the same low RGR. The persistence of the differences in RMR at low N-supply indicated that the high RMR of the mutants was a direct effect of low GA, which was independent of nitrate supply. Because the low N-supply increased the RMRs of all genotypes to the same extent, the response of RMR to N-supply does not seem to depend on GA. Although many of the traits of the slow growing GA mutants were very similar to those of inherently slow growing plant species from unproductive habitats, gibberellins are unlikely to be a main determinant of a plant's potential RGR.     

The effects of gibberellic acid and nutrient sprays on growth and yield of brussels sprout

Young plants of brussels sprout, cv. Cambridge Special, growing in pots in a glasshouse, were sprayed on ten occasions with gibberellic acid (GA) at 0, 25, 100 and 400 p.p.m. and ammonium nitrate (NH₄NO₃) at 0, 0·0125, 0·025 and 0·05 M concentrations in all combinations. In 24 days both GA and NH₄NO₃ increased leaf area, leaf number, dry weights of leaf, stem and root, and fresh weights of leaf and stem. GA increased stem height and decreased fresh weight/unit leaf area (leaf thickness), whereas NH₄NO₃ did not affect stem height and increased leaf thickness. Of the GA treatments, 100 p.p.m. gave the largest plants as judged by fresh weight of the whole plant and leaf area, and of the N treatments 0·05 M NH₄NO₃ increased growth most. The best treatment combination was 0·05 M NH₄NO₃ with 100 p.p.m. GA, which gave the greatest fresh and dry weights of the whole plant, leaf area and leaf dry weight as well as increasing leaf thickness. Significant interactions were found between GA and N for dry weight of leaf, fresh and dry weight of root, and leaf thickness but not for leaf area or stem growth (fresh weight, dry weight, height). The combination of levels of GA and N that can be expected to increase leaf area, leaf dry weight and leaf thickness simultaneously, may lie within narrow limits for a particular crop. In a second experiment plants were sprayed with all combinations of GA (0 and 100 p.p.m.) seven times under glass in pots and ten times in the field, and three levels of KH₂PO₄ on twelve occasions in the field. Sprouts (axillary buds) were harvested in October and February. At the first harvest GA did not affect fresh weight or number, but increased both the total number of sprouts picked (with GA = 80·7; without GA = 69·8 per plant) and the total fresh weight of saleable sprouts (with GA = 2·59; without GA = 2·33 lb/plant). KH₂PO₄ also increased the weight and number of sprouts at the final harvest and the number of small sprouts at the first. There were interactions between GA and KH₂PO₄ (P= < 0·001) for both tota weight and number of saleable sprouts.     

Effects of CCC and GA on internodal development of barley

Summary The effect of applying (2-chloroethyl)trimethylammonium chloride (CCC) or gibberellic acid (GA) as foliar sprays on internodal development of barley was studied. CCC applied to the whole plant at main tiller leaf stages 1, 2 or 3 decreased shoot elongation, and prevented elongation of internode 6 (internode 5 subtended the head). CCC at all stages delayed senescence of the lower leaves. CCC sprayed at all 6 leaf stages and GA sprayed at main tiller leaf stages 1, 2, 3 or 4 reduced plant height at maturity. GA treatment at leaf stages 2, 3 or 4 initially stimulated internodal elongation; elongation of later developed internodes was inhibited resulting in shorter plants at maturity. Only the treatment with GA at leaf stages 5 and 6 resulted in increased plant height.     

Endogenous gibberellins in Lolium perenne and influence of defoliation on their contents in elongating leaf bases and in leaf sheaths

Gibberellin (GA) levels in leaf sheaths and in elongating leaf bases of perennial ryegrass ( Lolium perenne L. cv. Bravo) were monitored in undefoliated and defoliated plants. Nine C-13-hydroxylated GAs (GA8, GA97, GA29, GA1, GA20, GA44, GA19, GA17, GA53) and one C-13-non-hydroxylated GA (GA9) were identified by combined gas chromatography-mass spectrometry in leaf extracts. The total level of GA8, GA29, GA1, GA20, GA44, GA19 and GA53, determined by selected ion monitoring, was 7 times higher in elongating leaf bases than in mature leaf sheaths. In both leaf tissues, defoliation induced an increase in GA53 level, while GA20 and GA1 levels decreased, suggesting that the GA53→GA44, as well as GA19→GA20, conversions were slowed down. The roles of GA1 in the control of leaf elongation and fructan mobilization following defoliation are discussed.     

Engineering of gibberellin levels in citrus by sense and antisense overexpression of a GA 20-oxidase gene modifies plant architecture

Carrizo citrange (Citrus sinensisxPoncirus trifoliata) is a citrus hybrid widely used as a rootstock, whose genetic manipulation to improve different growth characteristics is of high agronomic interest. In this work, transgenic Carrizo citrange plants have been produced overexpressing sense and antisense CcGA20ox1 (a key enzyme of GA biosynthesis) under control of the 35S promoter to modify plant architecture. As expected, taller (sense) and shorter (antisense) phenotypes correlated with higher and lower levels, respectively, of active GA1 in growing shoots. In contrast, other phenotypic characteristics seemed to be specific to citrus, or different from those described for similar transgenics in other species. For instance, thorns, typical organs of citrus at juvenile stages, were much longer in sense and shorter in antisense plants, and xylem tissue was reduced in leaf and internode of sense plants. Antisense plants presented a bushy phenotype, suggesting a possible effect of GAs on auxin biosynthesis and/or transport. The main foliole of sense plants was longer, although total leaf area was reduced. Leaf thickness was smaller in sense and larger in antisense plants due to changes in the spongy parenchyma. Internode cell length was not altered in transgenic plants, indicating that, in citrus, GAs regulate cell division rather than cell elongation. Interestingly, the phenotypes described were not apparent when transgenic plants were grafted on non-transgenic rootstock. This suggests that roots contribute to the GA economy of aerial parts in citrus and opens the possibility of using the antisense plants as dwarfing rootstocks.     

Modification of growth habit of Majestic potato by growth regulators applied at different times

The effect of gibberellic acid, CCC (2-chloroethyltrimethylammonium chloride) and B 9 (N-dimethylaminosuccinamic acid) was studied on growth of potato plants in pots. Growth was analysed on four occasions and changes in habit defined in terms of internode lengths, leaf areas and growth of lateral branches. Soaking seed pieces for 1 hr. in GA solution caused six internodes to elongate greatly, an effect not prevented by CCC applied when the shoot emerged from the soil. The effects on internode extension were determined by the length of the interval between GA treatment and CCC treatment. Treatment at emergence with CCC shortened all internodes and more CCC applied 4 weeks later had no effect. Late application of CCC or B9 shortened the top two lateral branches, usually very long in untreated plants. The regulators affected leaf growth differently from internode growth: usually growth regulators had less effect on leaf growth. Effects on growth depended on when the regulators were applied. Treatment with GA alone inhibited bud development at higher nodes than in untreated plants; when followed by late treatment with CCC, lateral growth at higher nodes was also less. CCC retarded development of lateral branches especially when applied early. B9 had a similar effect to CCC applied late. When regulators retarded growth of lower laterals, upper laterals often grew more than in untreated plants. Treatments did not affect the number of leaves on the main stem at first but later GA hastened senescence. GA increased the number of leaves on lateral stems, and the effect was enhanced by CCC. CCC alone increased the number of first- and second-order lateral leaves. GA lengthened and CCC shortened stolons. The effect of CCC persisted throughout the life of the plant. CCC or B 9 shortened stolons whenever applied. CCC hastened tuber initiation but slowed tuber growth. CCC at first retarded formation of lateral tubers but had no effect on the ultimate numbers of lateral and terminal tubers. The value of E (net assimilation rate) did not alter with time. CCC applied at emergence increased E, probably because it hastened tuber initiation and temporarily increased sink capacity. Although tubers formed earlier with CCC, their growth was slower and their demand for carbohydrate was also less. The increase in second-order laterals in CCC-treated plants indicates that they utilize carbohydrate that would normally go to tubers. This experiment also demonstrates that crowding leaves by shortening stems did not diminish E, possibly because another over-riding process (bigger sinks) offsets the effect of shading.     

TRANSLOCATION OF APPLIED GIBBERELLIN IN BEAN SEEDLINGS

The direction and extent of GA transport in Pinto beans has been studied. The increase in growth rate was used as a measure of the amount of GA which reached the stem apex. The evidence showed that a similar increase in stem growth occurred whether GA was applied to the first trifoliolate leaf or to the apex of the shoot, but considerably less elongation resulted when GA was applied to primary leaves. When leaves were treated with GA after remaining in darkness for extended times, no increase in stem elongation was observed; however, growth was promoted when the plants were returned to light. The time required for a sufficient amount of GA to be translocated from the leaf to increase stem growth is less than 1 hour. The maximum growth response was found when the treated leaf was left on the plant for 3 or more hours. A study of GA movement in two-branched plants was made. The untreated branch showed no growth response when GA was applied to the apex of the other branch, even if the dose of GA was 20 × greater than a saturating dose. Similar results were observed when GA was applied to the first trifoliolate leaf. Considerable GA moved from a mature leaf to the opposite shoot if this untreated branch had been defoliated. The pattern of GA movement to the opposite shoot was dependent on the position of the treated leaf on the shoot. It is concluded that the movement of applied GA is related to carbohydrate transport within the plant.     

Potato purple top phytoplasma‐induced disruption of gibberellin homeostasis in tomato plants

Phytoplasmas are phloem‐inhabiting, cell wall‐less bacteria that cause numerous plant diseases worldwide. Plants infected by phytoplasmas often exhibit various symptoms indicative of hormonal imbalance. In this study, we investigated the effects of potato purple top (PPT) phytoplasma infection on gibberellin homeostasis in tomato plants. We found that PPT phytoplasma infection caused a significant reduction in endogenous levels of gibberellic acid (GA₃). The decrease in GA₃ content in diseased plants was correlated with down regulation of genes responsible for biosynthesis of bioactive GAs ( GA20ox1 and GA3ox1) and genes involved in formation of GA precursors [geranyl diphosphate synthase (GPS) and copalyldiphosphate synthase (CPS)]. Exogenous application of GA₃ at 200 µmol L^?1 was able to restore the GA content in infected plants to levels comparable to those in healthy controls, and to attenuate the characteristic ‘big bud’ symptoms induced by the phytoplasma. The interesting observation that PPT phytoplasma‐infected plants had prolonged low expression of key GA biosynthesis genes GA20ox1 and GA3ox1 under GA deficiency conditions led us to hypothesise that there was a diminished sensitivity of the GA metabolism feedback regulation, especially GA biosynthesis negative feedback regulation, in those affected plants, and such diminished sensitization in early stages of infection may represent a central element of the phytoplasma‐induced disruption of GA homeostasis and pathogenesis.     

PROCERA encodes a DELLA protein that mediates control of dissected leaf form in tomato

Leaves of seed plants can be described as simple, where the leaf blade is entire, or dissected, where the blade is divided into distinct leaflets. Mechanisms that define leaflet number and position are poorly understood and their elucidation presents an attractive opportunity to understand mechanisms controlling organ shape in plants. In tomato (Solanum lycopersicum), a plant with dissected leaves, KNOTTED1-like homeodomain proteins (KNOX) are positive regulators of leaflet formation. Conversely, the hormone gibberellin (GA) can antagonise the effects of KNOX overexpression and reduce leaflet number, suggesting that GA may be a negative regulator of leaflet formation. However, when and how GA acts on leaf development is unknown. The reduced leaflet number phenotype of the tomato mutant procera (pro) mimics that of plants to which GA has been applied during leaf development, suggesting that PRO may define a GA signalling component required to promote leaflet formation. Here we show that PRO encodes a DELLA-type growth repressor that probably mediates GA-reversible growth restraint. We demonstrate that PRO is required to promote leaflet initiation during early stages of growth of leaf primordia and conversely that reduced GA biosynthesis increases the capability of the tomato leaf to produce leaflets in response to elevated KNOX activity. We propose that, in tomato, DELLA activity regulates leaflet number by defining the correct timing for leaflet initiation.     

Growth and Nitrogen Accumulation in Young Tomato Plants Treated with Gibberellic Acid

The effects of foliar sprays of gibberellic acid (GA) on thegrowth of tomato plants cv. Potentate were studied in growthrooms and a glasshouse. Four sprays of GA (5 ppm) increasedleaf area and whole plant weight relative to water controlsgrown at constant temperatures (7, 17, 22, and 27 °C) for12 days, the largest plants being obtained with 5 ppm. Experimentsmade at four photoperiods (5, 10, 15, and 20 h) and at two lightintensities (7000 and 10 750 lx) showed that GA increased leafand whole plant weight at 15 h, leaf area at 10 and 15 h andstem height at all photoperiods; area, height, and weight increaseswere obtained at both light intensities, leaf growth being increasedmore by GA at 7000 lx and stem growth more at 10 750 lx. Four foliar sprays of GA (5 ppm) were combined with N supplementsapplied via leaf and/or root to plants in sand culture. Withlow supply to the roots (20 ppm N) GA failed to increase growth,but increased it at higher levels. Total N in leaf and stemwas increased by GA or by NH₄NO₃ (10 sprays 280 ppm N) at alllevels of N supplied to roots, but when applied together theeffect on total leaf N was more than additive except at thehighest level (540 ppm) GA increased the concentration of N(as per cent dry matter) in leaf and stem at all levels of Nsupplied to roots. GA and NH₄NO₃ together resulted in a greateramount and a higher concentration of N in the shoots (and usuallyalso in roots) than did NH₄NO₃ alone. Leaf thickness (as freshweight/unit area) could only be increased appreciably by sprayingwith a complete nutrient solution which reduced leaf area butnot dry weight. Growth increases induced by GA were detectable 43 days afterthe first of four sprays in the glasshouse and after 30 daysin the growth room. The persistence of GA effects was comparedwith those induced by sprays of NH₄NO₃.     

Translocation of Photosynthates in Tall and Dwarf Varieties of Pea, Pisum sativum

Quantitative studies of the translocation of radiocarbon from a young expanded leaf of two tall varieties (Improved Pilot and Thomas Laxton) and two dwarf varieties (Little Marvel and Meteor) of Pisum sativum showed that 40 to 45 per cent of the radiocarbon was exported from the ¹⁴CO₂ treated leaf after 24 hours in all four varieties. Although substantial export to the upper shoot always occurred it was more marked in the two tall varieties. Pre-treatment with GA did not affect total fixation but increased total export from the ¹⁴CO₂ treated leaf in cv. Meteor and decreased it in cv. Improved Pilot. GA had no effect on the translocation pattern in the tall plants but modified that of the dwarf plants to correspond to that found in the tall varieties.     

Histological, Physiological and Biochemical Interactions between Vesicular-Arbuscular Mycorrhizae and Thielaviopsis basicola in Tobacco Plants

Interactions between the mycorrhizal fungus Glomus monosporum and the root rot pathogen Thielaviopsis basicola and their effects on tobacco plants were investigated over a 4 week period. Mycorrhizal tobacco plants, obtained by preinoculation with G. monosporum, showed a better tolerance to T. basicola than non-mycorrhizal seedlings. Root and leaf dry weights of mycorrhizal plants were greater than those of controls. Mycorrhizal plants inoculated with T. basicola showed higher root and leaf dry weights than non-mycorrhizal infected plants, but lower values than mycorrhizal plants which were not infected. No appreciable differences in free aminoacid composition were observed among the different treatments with two exceptions: proline content was higher in infected and mycorrhizal infected plants compared to control and mycorrhizal plants; arginine content was higher in infected and mycorrhizal infected plants compared to control and mycorrhizal plants; arginine content was higher in mycorrhizal plants than in all the other treatments. The mechanisms by which (VAM) fungi can reduce disease incidence and pathogen development are discussed.     

The Arabidopsis GIBBERELLIN METHYL TRANSFERASE 1 suppresses gibberellin activity,reduces whole‐plant transpiration and promotes drought tolerance in transgenic tomato

Previous studies have shown that reduced gibberellin (GA) level or signal promotes plant tolerance to environmental stresses, including drought, but the underlying mechanism is not yet clear. Here we studied the effects of reduced levels of active GAs on tomato (Solanum lycopersicum) plant tolerance to drought as well as the mechanism responsible for these effects. To reduce the levels of active GAs, we generated transgenic tomato overexpressing the Arabidopsis thaliana GA METHYL TRANSFERASE 1 (AtGAMT1) gene. AtGAMT1 encodes an enzyme that catalyses the methylation of active GAs to generate inactive GA methyl esters. Tomato plants overexpressing AtGAMT1 exhibited typical GA‐deficiency phenotypes and increased tolerance to drought stress. GA application to the transgenic plants restored normal growth and sensitivity to drought. The transgenic plants maintained high leaf water status under drought conditions, because of reduced whole‐plant transpiration. The reduced transpiration can be attributed to reduced stomatal conductance. GAMT1 overexpression inhibited the expansion of leaf‐epidermal cells, leading to the formation of smaller stomata with reduced stomatal pores. It is possible that under drought conditions, plants with reduced GA activity and therefore, reduced transpiration, will suffer less from leaf desiccation, thereby maintaining higher capabilities and recovery rates.     

AN ANALYSIS OF THE GROWTH RESPONSE OF YOUNG TOMATO PLANTS TO INFECTION BY VERTICILLIUM ALBO-ATRUM

Infection of seedling tomatoes with Verticillium albo-atrum checked growth but did not result immediately in leaf yellowing. Localized wilting occurred in some plants 2 weeks after the check to growth was evident. 8 weeks after inoculation, dry weights of leaf, stem and root were decreased by 72, 70, and 65% respectively.
Of the growth attributes studied, leaf area was most reduced by infection and this was due to a failure of the leaves to expand rather than to a fall in the rate of leaf production. Neither water nor nitrogen appeared to be limiting factors in this respect. The water content of infected leaves was not reduced until 6 weeks after inoculation, when leaf yellowing and necrosis had also appeared. The percentage N contents of stem, root and leaf of infected plants exceeded those of the healthy controls 24 days after inoculation. N uptake was not seriously impaired until 21 days later.
The photosynthetic efficiency of the green leaves of infected plants was reduced. The mean values for net assimilation rates were: Healthy 0.47 and infected 0.39 g./dm.²/week.
Plants, in which two-thirds of the root system had been killed by crushing, were placed in contact with mycelium in soil. This initial root injury did not significantly affect the growth of infected plants.
The data accord with a toxin theory of damage to infected plants, but the slow development of chlorosis and wilting symptoms in the young plants suggested a greater tolerance to the toxin than is found in older plants.     

Immunomodulation of gibberellin biosynthesis using an anti-precursor gibberellin antibody confers gibberellin-deficient phenotypes

Immunomodulation is a means to modulate an organism's function by antibody production to capture either endogenous or exogenous antigens. We have recently succeeded in obtaining gibberellin (GA)-deficient phenotypes in Arabidopsis thaliana by using anti-bioactive GA antibodies. In this study, a single-chain antibody (scFv) against GA(24), a precursor GA, was utilized to repress the biosynthesis of bioactive gibberellins. Stable accumulation of the scFv in endoplasmic reticulum (ER) was achieved by being produced as a fusion with GFP as well as KDEL ER-retention signal. The transgenic plants showed GFP fluorescence in the reticulate cortical ER network in epidermal cells. The GFP-scFv fusion produced in plants maintained its binding activity. The transgenic plants showed GA-deficient phenotypes, including reduced rosette leaf development, delayed flower induction and reduced stem elongation of the main culm, especially in the early stage of inflorescence growth. Contrarily, stem elongation of the main culm at a later stage, or that of lateral shoots was much less affected by scFv production. These phenotypes were different from anti-bioactive GA scFv-producing lines, whose stem elongation was continuously repressed throughout the inflorescence development. The GA-deficient phenotypes were recovered by treatment with GA(24) and bioactive GA(4), the latter being more effective. The transgenic lines contained conspicuously higher endogenous GA(24) and clearly less GA(4) than wild-type plants. The expression of GA 20-oxidase and GA 3-oxidase genes, which are feedback-regulated by GA signaling, were up-regulated in those plants. These results demonstrate that the scFv trapped GA(24) in ER and inhibited metabolism of GA(24) to bioactive GA(4).     

The influence of a reduced gibberellin biosynthesis and nitrogen supply on the morphology and anatomy of leaves and roots of tomato (Solanum lycopersicum)

We investigated how the differences in growth and morphology, between fast-growing wildtype (Wt) tomato ( Solanum lycopersicum L.) plants and slow-growing gibberellin (GA) deficient W335 mutants, were reflected in cell numbers and cell sizes. We also studied whether the differences between the Wt and the low-GA mutant would persist at a growth-limiting supply of nitrate. Both a low endogenous GA concentration and a low supply of nitrate reduced the number and size of leaf cells, whereas they increased the size and number of root cortex cells. The effects of low N-supply on the size and number of leaf and root cells did not depend on endogenous GA concentrations. The mutant's higher allocation to roots seemed to be the result of the strongly reduced growth of the shoot.     

Qualitative and Quantitative Analysis of Endogenous Gibberellins in Onion Plants and Their Effects on Bulb Development

Evidence has been reported that bulb development in onion plants (Allium cepa L.) is controlled by endogenous bulbing and anti-bulbing hormones, and that gibberellin (GA) is a candidate for anti-bulbing hormone (ABH). In this study, we identified a series of C-13-H GAs (GA₁₂, GA₁₅, GA₂₄, GA₉, GA₄, GA₃₄, and 3-epi-GA₄) and a series of C-13-OH GAs (GA₄₄, GA₂₀, GA₁ and GA₈) from the leaf sheaths including the lower part of leaf blades of onion plants (cv. Senshu-Chuko). These results suggested that two independent GA biosynthetic pathways, the early-non-hydroxylation pathway to GA₄ (active GA) and early-13-hydroxylation pathway to GA₁ (active GA), exist in onion plants. It was also suggested that GA₄ and GA₁ have almost the same ability to inhibit bulb development in onion plants induced by treatment with an inhibitor of GA biosynthesis, uniconazole-P. The endogenous levels of GA₁ and GA₄, and their direct precursors, GA₂₀ and GA₉, in leaf blades, leaf sheaths, and roots of 4-week-old bulbing and non-bulbing onion plants were measured by gas chromatography/selected ion monitoring with the corresponding [²H]labeled GAs as internal standards. In most cases, the GA levels in long-day (LD)-grown bulbing onion plants were higher than those of short-day (SD)-grown non-bulbing onion plants, but the GA₁ level in leaf blades of SD-grown onion plants was rather higher than that of LD-grown onion plants. Relationship between the endogenous GAs and bulb development in onion plants is discussed.