Effect of light and gibberellic acid on cell division in the first foliage leaf of durum wheat (Triticum durum Desf.)

The present paper is part of a research program which aims at a quantitative analysis of the effects of light and gibberellic acid (GA₃) on growth of the first foliage leaf in durum wheat (Triticum durum Desf.). Since leaf growth is the combined result of the increase in cell number (cell division) and cell enlargement, the influence of light and GA₃ treatment on cell division in the basal meristem of the first leaf in two cultivars, Cappelli and Creso, was investigated. Creso is a short-strawed cultivar carrying the Gai 1 gene which influences both plant height and insensitivity to applied GA₃. Cell division, as measured by mitotic index, was similar in darkness, continuous red light and dichromatic irradiation (far-red plus red), while lower mitotic rates were observed under continuous far-red light: this indicates that the response of cell division is modulated by a high-irradiance reaction of phytochrome in both cultivars. The two cultivars showed different responses to blue light. In Cappelli, blue light and dichromatic irradiation (blue plus red) gave lower mitotic indices than the dark control, indicating the action of a specific blue-light-absorbing photoreceptor, whereas in Creso the response kinetics to all light regimes which included blue light were more complex. On the basis also of the results obtained with GA₃ application in Cappelli, it appears that (i) the hormonal treatment is able to change the pattern of mitotic index only in the presence of the action of a blue-light receptor and (ii) the different responses of the two cultivars could be the result of different endogenous hormonal levels. The importance of the observations in relation to the data for first-leaf longitudinal growth reported in a previous paper (Baroncelli et al. 1984, Planta 160, 298–304) is discussed.Abbreviations BL blue light - D darkness - FR far-red light - GA gibberellin - GA₃ gibberellic acid - m.i. mitotic index - Norflurazon 4-chloro-5-(methylamino)-2-(,,,-trifluoro-m-totyl-3(2H)) pyridazinone - R red light - WL white light - phytochrome photoequilibrium     

Interactions of daminozide, 2, 2′-dipyridyl,and gibberellins A3, A9 and A20, in stem extension inChrysanthemum morifolium Ramat: an indication that daminozide may not inhibit the responses to GA9 or GA20 by blocking gibberellin hydroxylation

Interactions between the growth retardant daminozide (a substituted succinamic acid) and a subsequent application (1 or 10 g) of either gibberellin A₃, A₉ or A₂₀, on stem extension inChrysanthemum morifolium cv. Bright Golden Anne, indicated that pre-treatment of plants with daminozide largely prevented the response to GA₂₀ as well as to GA₉. The daminozide-GA₃ interaction on total stem length was dependent upon the dose of GA₃ such that, by flowering time, 1 g of GA₃ had virtually eliminated the retardant effect, while 10 g of GA₃ increased stem length to a value similar to that achieved by control shoots receiving 10 g of GA₃. In contrast, prior application of 2, 2-dipyridyl (an inhibitor of hydroxylation in some plant and animal systems) had no significant influence on the time courses of response to any of the GAs. In the absence of daminozide (and 2, 2-dipyridyl) all three GAs were very active in promoting internode extension soon after their application. If 2, 2-dipyridyl can block hydroxylation reactions in chrysanthemum tissues, the results do not support the hypothesis that daminozide restricts GA₉- (or GA₂₀-) induced stem elongation by preventing the hydroxylation of GA₉.     

The effect of light quality and gibberellic acid (GA3) on photosynthesis and respiration rates of pea seedlings

CO₂ exchange were measured on pea seedlings (Pisum sativum L. var. Bördi) cultivated from seeds imbibed either in water (C-plants) or in gibberellic acid (GA₃) at the concentration of 25 g/1 (GA-plants), and then grown under 17 W/m² blue light (B-plants) or 11 W/m² red light (R-plants).When measured under the same light conditions as during growth the net photosynthesis (APS) rate in B-plants was about twice higher than that in R-plants. Dark respiration (DR) rate was 70% higher in B- than in R-plants. Red light retarded the development of photosynthetic activity, but GA₃ suppressed this effect. The hormone enhanced net photosynthesis and dark respiration to the same extent.When measured under saturating white light net photosynthesis rate of C-plants was also two times higher in B-plants than in R-plants. Growth conditions had only a slight effect on the APS of GA-plants under white light. APS rates of GA-plants grown under red light were higher under white light than those of C-plants, but lower than those of plants grown under blue light.We assume that blue light induced formation of plants that were adapted to higher light intensity: red light had an opposite effect, whereas gibberellic acid induced formation of plants that were adapted to medium light intensity.     

Tissue culture ofKarwinskia humboldtiana—a plant producing toxins with antitumoural effects

Isolated embryos ofKarwinskia humboldtiana were cultured in vitro. The growth of embryos and development to plantlets on woody plant medium supplemented with indole-3-acetic acid 6.10^-2 mol l^–1, gibberellic acid (GA₃) 3.10^-2 mol l^–1, and 6-benzylaminopurine (BA) 2 mol l^–1 was obtained. Multiplication of shoots and rooting of excised shoots has been achieved. Callus formation on modified Murashige-Skoog medium supplemented with 1-naphthaleneacetic acid 10 mol l^–1, GA₃ 14 mol l^–1, and kinetin 5 mol l^–1 on hypocotyls, or on root cultures on medium supplemented with 2.4-dichlorophenoxyacetic acid 10 mol l^–1 and BA 10 mol l^–1 was induced.Abbreviations BA 6-benzylaminopurine - 2,4-d 2,4-dichlorophenoxyacetic acid - GA₃ gibberellic acid - IAA indole-3-acetic acid - NAA 1-naphthaleneacetic acid - TEM transmission electron microscopy     

Structure of the genes encoding Hordeum vulgare (1→3,1→4)-β-glucanase isoenzymes I and II and functional analysis of their promoters in barley aleurone protoplasts

Summary Barley (1 3,1 4)--glucanase isoenzyme II is synthesized in the aleurone cells during germination and secreted into the endosperm for hydrolysis of the cell walls. Its synthesis is stimulated by gibberellic acid (GA₃) and repressed by abscisic acid. The gene for isoenzyme I is expressed in the aleurone, scutellum and prominently in young leaves. Close functional relatedness between the two enzymes is attested by 92 % identity at the level of the amino acid sequence. The structural genes for the two enzymes each contain a large intron of 2505 by and 2952 bp, respectively, in the codon for amino acid 25 of the 28-residue signal peptide. During evolution, homologous regions of the two introns have changed position and orientation. Furthermore, a large palindromic sequence of 327 by in the 5 end of the intron is present only in the gene for isoenzyme II. In transient expression assays using barley aleurone protoplasts and chloramphenicol acetyl transferase as reporter the promoter of the isoenzyme I gene showed no response to GA₃. However, removal of a unique 151 by region extending from positions –402 to –552 upstream of the TATA box permitted low levels of GA₃-induced expression of the reporter gene, suggesting a silencer function for this domain. High levels of GA₃-responsive expression were obtained in aleurone protoplasts using the promoter of the gene encoding isoenzyme II. Truncation of this promoter revealed that sequences located within 253 bp upstream from the TATA box are sufficient to direct GA₃-stimulated expression. Using the homologous barley aleurone protoplast transfection assay, it was possible to reproduce the in vivo expression characteristics of the genes for the barley (1 3,1 4)--glucanase isoenzymes I and II with reporter gene constructs.     

Regulation of Medicago sativa L. somatic embryogenesis by gibberellins

The influence of exogenous gibberellic acid (GA₃) andpaclobutrazol, an inhibitor of gibberellin biosynthesis, on growth of callusandsomatic embryogenesis in petiole-derived tissue cultures of Medicagosativa L. has been investigated. GA₃ (0.5–500M) or paclobutrazol(5–100 M) were added to either an induction (with 2,4 Dand kinetin) or a differentiation medium (without plant growth regulators).Gibberellin A₃, applied during the induction as well as thedifferentiation stage, reduced the weight of callus and increased the number ofsomatic embryos in Medicago sativa L. tissue cultures.Somatic embryo production was increased more by the presence of exogenousGA₃ in the differentiation than induction medium. The inclusion ofpaclobutrazol in the induction or differentiation medium caused the inhibitionof callus growth and embryo production. Callus growth was much less affectedthan embryogenesis. These results indicate that gibberellins are beneficial forboth embryoinduction and formation. The level of endogenous gibberellins is presumablysufficient for callus induction and growth. However, it seems not optimal forthe induction and particularly for the differentiation of embryos.     

The identification and metabolism of GA9 and its metabolites in seed coats and shoots of pea,line G2

[³H]gibberellin A₉ was applied to shoots or seed parts of G2 pea to produce radiolabeled metabolites. These were used as markers during purification for the recovery of endogenous GA₉ and its naturally occurring metabolites. GA₉ and its metabolites were purified by HPLC, derivatized and examined by GC-MS. Endogenous GA₉, GA₂₀, GA₂₉ and GA₅₁ were identified in pea shoots and seed coats. GA₅₁-catabolite and GA₂₉-catabolite were also detected in seed coats. GA₇₀ was detected in seed coats following the application of 1 g of GA₉. Applied [³H]GA₉ was metabolized through both the 13-hydroxylation and 2-hydroxylation pathways. Labeled metabolites were tentatively identified on the basis of co-chromatography on HPLC with endogenous compounds identified by GC-MS. In shoots [³H]GA₅₁ and [³H]GA₅₁-catabolite were the predominant metabolites after 6 hrs, but by 24 hrs there was little of these metabolites remaining, while [³H]GA₂₉-catabolite and an unidentified metabolite predominated. In seed coats [³H]GA₅₁ was the initial product, later followed by [³H]GA₅₁-catabolite and an unidentified metabolite (different from that in shoots), with lesser amounts of [³H]GA₂₀, [³H]GA₂₉ and [³H]GA₂₉-catabolite. [³H]GA₇₀ was a very minor product in both cases. [³H]GA₉ was not metabolized by pea cotyledons.Edited by T.J. Gianfagna.Author for correspondence     

Hormonal regulation of gene expression in the “slender” mutant of barley (Hordeum vulgare L.)

The slender mutant of barley resembles a normal barley plant treated with high doses of gibberellic acid (GA₃). Expression of GA₃-regulated and abscisic acid (ABA)-regulated mRNAs was studied in the endosperm and roots of mutant and wild-type (WT) plants.Production of -amylase (EC 3.2.1.1) by WT embryoless half-grains was dependent on the presence of GA₃, and was prevented by ABA. In contrast, -amylase was produced by half-grains of the slender mutant in the absence of added GA₃, although it was still reduced by ABA. The spectrum of -amylase mRNAs in slender embryoless half-grains incubated in the absence of added GA₃ was the same as in WT endosperm half-grains incubated in the presence of GA₃. These results indicate that the endosperm of the slender mutant exhibits similar properties to WT endosperm treated with GA₃.In roots the expression of an ABA-inducible mRNA was similar in slender and WT seedlings either treated with exogenous ABA or exposed to dehydration. This result, and the effect of ABA on -amylase production by the endosperm, indicate that the slender plants retain sensitivity to ABA.Abbreviations ABA abscisic acid - AMV avian myeloblastosis virus - GA gibberellin - GA₁ gibberellin A₁ - GA₃ gibberellic acid - WT wild-type     

The role of acidification in gibberellic acid- and fusicoccin-induced elongation growth of lettuce hypocotyl sections

The roles of gibberellic acid (GA₃) and fusicoccin (FC) in the elongation growth and acidification of the medium by excised hypocotyl sections of lettuce (Lactuca sativa L.) were investigated. Hypocotyl sections incubated in buffer without GA₃ elongate optimally at pH 4.0–4.25 while sections incubated with GA₃ show the same growth between pH 4.25 and 6.0. Preincubation of sections at pH 6.0 for 6 h does not affect the subsequent elongation response to acidic medium (pH 4.25); however, the sections become refractory to further acid treatment after their initial burst of growth in response to pH 4.25. Sections made refractory to acid are responsive to GA₃ application, however, and the rate of growth in response to GA₃ of sections pretreated for 6 h at pH 4.25 is 85% of that of sections pretreated at pH 6.0. Although preincubation of sections for 48 h in medium at pH 6.0 abolishes the GA₃ response, it does not affect the response to buffer at pH 4.25. FC stimulates elongation growth in letuce hypocotyls at an optimal concentration of 1 M, and pretreatment of sections at pH 4.25 does not affect this elongation response. Although both GA₃ and FC increase elongation of the section, neither causes appreciable acidification of the medium. Addition of KCl or NaCl to FC-treated sections causes rapid medium acidification but addition of salts to GA₃-treated tissue does not cause acidification. Abrasion of the hypocotyl to remove the cuticle does not enhance acidification of the medium by the sections nor deos it affect elongation of the sections in response to GA₃ or FC. Medium acidification by the sections is not a passive process since it is abolished both by low temperature (2° C) and metabolic inhibitors (carbonyl cyanide-m-chlorophenyl-hydrazone, azide). The acidification of the medium by barley (Hordeum vulgare L.) roots in response to FC is also dependent on the presence of KCl. We conclude that the acid-growth hypothesis does not explain GA₃- or FC-induced elongation in lettuce hypocotyls.Abbreviations FC tusicoccin - GA₃ gibberellic acid - HEPES N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid - CCCP carbonyl cyanide-m-chlorophenyl-hydrazone - MES 2-(N-morpholino)ethanesulphonic acid - Tris tris-(hydroxymethyl)aminomethane     

Plant regeneration from leaf protoplasts of apple

Protoplasts were isolated from young leaves or etiolated shoot apices. For initiation of divisions the protoplasts were embedded in sodium alginate and cultivated in MS or MI medium supplemented with 2.2 M BA, 2.6 M NAA and 2.2 M 2,4-dichlorophenoxyacetic acid. The protoplasts of all seven lines tested developed to protocalluses at high frequencies. No genotypic differences were observed. When BA was used in combination with NAA in the regeneration experiments, only a few protocalluses (highest frequency 3%) exhibited shoot organogenesis. When BA was replaced with thidiazuron, the percentage of protocalluses that developed shoots increased in two of three tested lines to 7% and 56%, respectively. Shoot development was achieved under light conditions. The shoots were then rooted and transferred into soil.Abbreviations ABA abscisic acid - BA 6-benzyladenine - 2,4-d 2,4-dichlorophenoxyacetic acid - FW fresh weight - GA₃ gibberellic acid - IBA indole-3-butyric acid - MES 2-N-morpholinoethane sulphonic acid - NAA -naphthaleneacetic acid     

Total protein release from barley aleurone layers as a bioassay for gibberellins

The effect of gibberellic acid on the secretion of proteins from barley (Hordeum vulgare L.) aleurone layers has been investigated for its suitability as a gibberellin bioassay. Concentrations from 10^–4 g/ml to 100 g/ml of GA₃ resulted in the release of proportionally increasing amounts of total protein. The release of proteins is not affected by indoleacetic acid and kinetin. This method has been applied and compared with the -amylase assay for the estimation of gibberellin in extracts of tomato fruits and maize seedlings.Abbreviations GA₃ gibberellic acid - IAA indoleactic acid - K kinetin     

Effects of two growth retardants on tissue permeability in Pisum sativum and Beta vulgaris

The effects of growth retardants, 4-hydroxy-5-isopropyl-2-methylphenyltrimethylammonium chloride-1-piperidine carboxylate (AMO-1618 or AMO) and 2-chloroethyltrimethylammonium chloride (CCC), applied with and without gibberellic acid (GA₃), on -[³H]alanine uptake and leakage from pea (Pisum sativum L.) and betacyanin efflux from beetroot (Beta vulgaris L.) tissue were examined. Both compounds decreased the amount of -[³H]alanine taken up into pea leaf discs, and increased the quantity of radioactive label that subsequently leaked out of this tissue. Efflux of betacyanin from slices of beetroot was also found to be promoted by treatment with CCC or AMO-1618. In no case were these effects reversed by application of GA₃. It is concluded that the growth retardants may be altering tissue permeability by an interaction with the cell membranes, and this may account for some of the side effects of the retardants which cannot be explained on the basis of their inhibiting action on gibberellin synthesis.Abbreviations AMO-1618 4-hydroxy-5-isopropyl-2-methylphenyltrimethylammonium chloride-1-piperidine carboxylate - CCC 2-chloroethyltrimethylammonium chloride - GA₃ gibberellic acid     

Stimulatory effect of ethephon,ACC, gibberellin A3 and A4+7 on germination of methyl jasmonate inhibited Amaranthus caudatus L. seeds

Methyl jasmonate (JA-Me) inhibited or retarded germination of Amaranthus caudatus seeds in darkness at 24°C, Ethephon, ACC and gibberellins (GA₃ or GA₄₊₇) partially or completely reversed this inhibition depending on the concentration of JA-Me applied. Both ethephon and the gibberellins were more effective than ACC. Both GA₃ and GA₄₊₇ enhanced the stimulatory effect of ethephon or ACC on germination of seeds inhibited by JA-Me.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - JA jasmonic acid - JA-Me methyl jasmonate     

The influence of gibberellic acid on reassociation kinetics of DNA of Daucus carota L.

Carrot DNA, extracted from the tap root of untreated and gibberellic acid (GA₃)-treated plants and of different varieties, was analyzed by reassociation kinetics. Differences due to GA₃ treatment appear mainly in the intermediate repeated DNA region. Differences in approximately the same region are found using carrot DNA of different varieties, which also show differences in the slow reassociating sequences. By hybridizing a family of the unique DNA range with DNA obtained from GA₃-treated plants and the controls, respectively, it could be shown that changes in the composition of total DNA are the result of GA₃ treatment.Abbreviations C₀t product of DNA concentration (mol nucleotide l^-1)xtime (s) - GA₃ gibberellic acid - T_m temperature for 50% denaturation     

In vitro propagation of the Greek olive cultivar `Chondrolia Chalkidikis'

The effect of basal media (Woody Plant Medium (WPM), Quoirin and Lepoivre (QL) and Olive Medium (OM)) and of various concentrations of cytokinins (6-benzyladenine (BA), zeatin, 6-(,-dimethylallyl-amino) purine (2iP)), solely or in combinations with each other and with gibberellic acid (GA₃), on in vitro shoot proliferation of the greek olive cultivar `Chondrolia Chalkidikis' was investigated. WPM proved to be the most effective one, resulting in better morphological appearance of the microshoots produced. The highest number of new microshoots/explant (1.68), with a 3.0 cm shoot height and a 4.2 proliferation rate, was obtained when this medium was supplemented with 20 M zeatin. The number of microshoots/explant and proliferation rate increased to 1.85 and 6.8, respectively, by using the combination of 5–20 M zeatin with 1 M BA, but shoot height was reduced. 2iP was the least effective of the cytokinins tested. Combination of 20 M zeatin with 10 M GA₃ affected positively shoot proliferation resulting in 1.80 microshoots/explant, 3.0 cm shoot height and 7.0 proliferation rate. However, the same concentration of GA₃ in combination with 1 M BA reduced the number of new microshoots/explant (0.48) as well as the proliferation rate (2.4), although shoot height remained nearly the same (2.7). The effect of indole-3-butyric acid (IBA), -naphthaleneacetic acid (NAA) and putrescine on root induction was also studied. A rooting percentage of up to 70% and 2.3 roots/microshoot were achieved by the combination of 12 M IBA + 3 M NAA, however, abscission of shoot tips and leaves appeared. Both variables were increased, up to 93% and 4.0, respectively, by the addition of 30 M putrescine in the medium, without any undesired side effect. After acclimatization, survival of rooted microshoots was high (90%) while that of non-rooted ones was low (30%).     

Infloreszenz- und Blütenentwicklung bei der KugeldistelEchinops exaltatus (Asteraceae)

InEchinops the flowers are surrounded by several scales and initiated in an acropetal and spiral succession on a cone-like inflorescence axis (Figs. 1–6). The floral organs originate in the following sequence: petals—stamens—carpels—pappus. The petals arise from a meristematic rim and therefore are already interconnected when they arise as primordia. This sympetalous zone remains rather inconspicuous for a long period, but eventually, the elongated corolla tube is formed through intercalary growth in a ring zone. Thereby, the stamens are moved upwards and form ledges on the corolla tube (Fig. 34). In the inferior ovary the usual zones of the typical angiospermous gynoecium can be distinguished, namely a synascidiate, symplicate and hemisymplicate zone. The ovule is borne on carpellary tissue.

     

Production of gynogenic haploids of Hordeum vulgare L.

Haploid plants were regenerated from cultured unfertilized ovaries of Hordeum vulgare L. (barley). Optimal response was obtained by the addition of 0.6 M 4-chloro-2-methylphenoxyacetic acid (MCPA), 2.8 M indole-3-acetic acid (IAA) and 4.4 M 6-benzyladenine (BA) in the N₆ medium. Further increase in the rate of callus formation and the number of green plants produced was possible with the addition of 90 g/l sucrose and 100 g/l coconut water. The stage of development of the ovaries at the time of culture was critical; the largest number of plants being produced by ovaries from flowers at the trinucleate stage of pollen.Abbreviations (BA) 6-benzyladenine - (MCPA) 4-chloro-2-methylphenoxyaceticacid - (2,4-D) 2,4-dichlorophenoxyaceticacid - (GA₃) gibberellic acid - (IAA) indole-3-acetic acid     

Changes in photosynthetic rates and growth following root treatments of tomato plants with phytohormones

The effects of root applications of kinetin, gibberellic acid (GA₃) and indoleacetic acid (IAA) on photosynthesis was measured using an open infrared CO₂ gas-exchange system. There was a 30–35% increase in the photosynthetic rates (mg CO₂/dm²/hr) of attached leaves within 8 hr following root treatment with 0.47 M kinetin. On a short-term basis (up to 2 days) 0.47 M kinetin was shown to have the optimal stimulatory effect on photosynthesis, relative growth rate (RGR) and total plant dry weight. If the roots were in contact with 0.47 M kinetin for longer than two days there was severe branching of the root system and growth was severely decreased. When plants were left in contact with the kinetin treatment for up to 7 days the optimal stimulatory concentration was considerably lower (0.0047 M) . Plants receiving a 4, 8, or 12 hr pulse with 0.47 M kinetin to the roots exhibited higher leaf photosynthetic rates than the control. Plants receiving an 8 or 12 hr pulse with 0.47 M kinetin maintained photosynthetic rates higher than the control for the duration of the experiment (8 days) while the 4 hr pulse remained higher than the control for only 5 days. A sharp decrease in the photosynthetic rate, RGR and total plant dry weight was observed two days following continual treatments with 0.47 M kinetin to the roots. At low light levels there was approximately a 100% increase in the photosynthetic rate two days following treatment with 0.47 M kinetin while at a saturating irradiance there was a 30 to 35% increase. Indoleacetic acid either showed no effect on the photosynthetic rate, RGR and total plant dry weight or an inhibitory effect was observed. Either GA₃ or kinetin alone were shown to stimulate photosynthesis, RGR and total plant dry weight, however, when GA₃ at a 1.4 M concentration was applied in combination with kinetin at a 0.0047 M concentration to the roots of tomato plants there was no additive effect. In all cases kinetin dramatically reduced leaf resistance whereas GA₃ had no effect.By supplying either GA₃ or kinetin to the roots of tomato plants a highly reproducible stimulation in the photosynthetic rate, RGR and total plant dry weight can be achieved at physiologically relevant concentrations, whereas IAA appears to have an inhibitory effect.Approved for publication on July 29, 1981 as paper number 6281 in the journal series of the Pennsylvania Agricultural Experiment Station.Research Assistant and Assistant Professor, respectively.     

Promotion of flowering in apple trees with gibberellin A4 and C-3 epi-gibberellin A4

The proportion of spurs flowering on apple trees (Malus domestica Borkh. cv Golden Delicious) displaying a high degree of alternate-year flowering was increased in the off year by gibberellin A₄ (GA₄) and C-3 epi-GA₄ applied in the previous year. When applied 4.5 weeks after anthesis amounts of GA₄ ranging from 3 to 300 g per spur and 25 or 50 g of C-3 epi-GA₄ per spur were effective. Treatments with GA₄ made seven weeks after anthesis were less effective. A combination of 30 g GA₄ and 30 g zeatin (6-(4-hydroxy-3-methylbut-trans-2-enylamino)purine) promoted flowering at both treatment times, and tended to be more effective than GA₄ alone.Abbreviation GA gibberellin or gibberellin-like substance Contribution No. 618     

Mannanase production by the lettuce endosperm

Endo--mannanase (EC 3.2.1.78) is produced and secreted by the cells of the endosperm of lettuce (lactuca sativa L.) seeds (achenes). In imbibed intact seeds, production is prevented by inhibitors. If the endosperm is incubated alone, these inhibitors can be removed by leaching, allowing mannanase production. Abscisic acid, a component of lettuce seeds, inhibits the production of mannanase in the isolated endosperm, and may be involved in regulation of mannanase production in intact seeds. During germination the inhibition is removed, beginning 4–8 h after red-light irradiation, which was given 4 h from sowing. The cotyledons participate in this process, and are controlled by events occuring in the axis within 4 h from red-light irradiation. This control by the axis apparently depends on the exchange of diffusible substances. Both benzyladenine and gibberellic acid can replace the influence of the axis if the latter is removed, and may therefore be involved in the control by the axis of the rest of the seed.Abbreviations ABA abscisic acid - BA 6-benzyladenine - GA₃ gibberellic acid - IAA indol-3-yl acetic acid - MES 2-(N-morpholino)ethane sulfonic acid - R red light Part of this work was carried out by P. Halmer at the Department of Biology, Washington University, St. Louis, MO 63130, USA (his present address)