Relation between the level of endogenous growth regulators and the differentiation of the fungusLentinus tigrinus studied in a synthetic medium

The fungusLentinus tigrinus was cultivated in a synthetic medium. During vegetative growth of mycelium and differentiation of fruiting bodies samples serving for determination of endogenous growth regulators were collected. The material was extracted with methanol, separated by chromatography and the isolated fractions were tested. Substances of auxin type were assayed with coleoptile segments of oat plants, gibberellins by measuring the hypocotyl growth in the lettuce and cytokinins from the degree of chlorophyll decomposition in the leaf segments of barley. Very low content of auxins was found in all phases of growth and differentiation of the fungus. The content of gibberellin substances was relatively high during the whole life of the fungus. The highest content is found in the vegetative mycelium and in the differentiated fruiting body, especially in the cap; it decreases during the differentiation of fruiting bodies. The content of cytokinins is equally high and the pattern of its change resembles that of the gibberellins. Its maximum is reached in young vegetative mycelium, it decreases when fruiting bodies are formed and when they are growing. Minimum is reached in the differentiated stalk, where also the amount of inhibitions is the highest. Experimental results show that during differentiation of fruiting bodies and their morphogenesis not only gibberellins and cytokinins are of importance, but also substances of an inhibitory character. A relatively high amounts of these substances was secreted into the cultivation medium. In a synthetic nutrient solution they may be easily demonstrated.     

Endogenous Hormonal Profiles in Hop Development

Changes in gibberellins (GAs), indole-3-acetic acid (IAA), and cytokinins associated with the transition from vegetative growth to reproductive growth in Humulus lupulus L. buds and leaves harvested at fortnight intervals were studied. During vegetative growth, GA₁ increased gradually and the lowest content was observed during flower development. Both GA₃ and GA₄ showed a dramatic increase in the samples taken from the apical part of axillary branches from plants 4–5 m high, which corresponds to the maximum vegetative development prior to macroscopically visible inflorescences. Notable increases in the cytokinins trans-zeatin (t-Z), isopentenyladenine (iP), and the riboside and ribotide forms of iP were also obtained. The auxin, indole-3-acetic acid, was the most abundant plant hormone, and its content was highest during vegetative growth. These results show for the first time a relationship between endogenous hormone profiles and both vegetative and reproductive development in hop plants, which may be relevant for future research on the control of the flowering by exogenous hormone applications.     

Exogenous regulators in the mycelium ofPleurotus ostreatus after exogenous application

The mycelium of the fungusPleurotus ostreatus was cultured on a synthetic medium with growth regulators. Best growth was observed on media with 100 ppm IAA, 200 ppm GA₃ and 200 ppm of kinetin. Growth of terminal hyphae and the appearance of colonies differed characteristically. Auxin bio-assays showed that the content of substances in younger mycelium slowly increased during cultivation without conspicuous changes in the medium. After exogenous application of IAA the content of auxins in younger mycelium increased and the content of IAA decreased gradually in the culture medium. Gibberellin bio-assays showed their content in the mycelium fell during cultivation and while it increased in the medium. Application of GA₃ to the medium leads to a significant increase in these substances in the young mycelium and in the medium their level remains the same. Cytokinin bio-assays showed their content to vary during cultivation. An addition of kinetin to the nutrient medium results in an increase of cytokinins in the mycelium while in the medium the level of cytokinins is maintained. Addition of one substance thus influences the levels of other endogenous regulators.     

Hormonal Profile in Vegetative and Floral Buds of Azalea: Levels of Polyamines, Gibberellins, and Cytokinins

The floral transition includes a complex system of factors that interact and involve various biochemical signals, including plant growth regulators. The physiological signals involved in the control of the floral transition have been sparsely studied and mainly in plant species whose genetics are poorly known. In this work, the role of polyamines, gibberellins, and cytokinins was investigated by analyzing their endogenous content in vegetative and floral buds of azalea. The results showed that there is a clear distinction between floral and vegetative buds with respect to the levels of these plant hormones, with floral buds containing higher amounts of conjugated polyamines, gibberellins (GAs) from the non-13-hydroxylation pathway (GA₉, GA₇, and GA₄), and cytokinins (particularly isopentenyl-type species), and vegetative buds containing higher amounts of free polyamines and gibberellins from the early 13-hydroxylation pathway and fewer cytokinins. In conclusion, there is a specific pattern of endogenous hormone profiles in both vegetative and floral bud development in azalea, which may be relevant for future research on the control of flowering by exogenous hormone applications.     

Gibberellin-induced delay of leaf senescence of Alstroemeria cut flowering stems is not caused by an increase in the endogenous cytokinin content

The effects of various chemically pure gibberellins and cytokinins on leaf yellowing of Alstroemeria were described. The loss of chlorophyll was measured both in leaves of cut flowering stems and in a model system consisting of detached leaf tips. It was demonstrated that plant growth substances affected chlorophyll loss in both systems to the same extent. Leaf senescence was delayed by various gibberellins and cytokinins. The results demonstrated that some of the gibberellins (GA₄ and GA₇) are far more effective in delaying chlorophyll loss than GA₃, which is commonly used as a postharvest treatment for Alstroemeria cut flowering stems. Immunoassays were used to demonstrate that the effect of gibberellins on leaf yellowing does not involve an increase in the endogenous cytokinin concentrations in the leaves as an intermediate step.Abbreviations GA gibberellin A - HPLC high performance liquid chromatography - GA_3Mc GA₃-methyl ester - ZR zeatin riboside - IPAR isopentenyl adenine riboside.     

Plant Growth Regulators in Citriculture: Factors Regulating Endogenous Levels in Citrus Tissues

Since the review on endogenous growth substances of citrus tissues by Goldschmidt in 1976 (HortScience, 11: 95-99), much information regarding this topic has been published in a wide array of journals. The present review provides a comprehensive overview of published information on endogenous levels of the five classes of plant growth substances (i.e., auxins, cytokinins, gibberellins, ethylene, and abscisic acid), plus polyamines and other endogenous substances that appear to have a role in regulating citrus growth and development. It is the first in a three-part series that next examines hormonal regulation of physiological processes in citrus followed by review of current uses and commercial applications of plant growth regulators in citrus production. In this article, a brief history of the detection and characterization of each class of plant growth substances is given. Following this, variation in endogenous levels associated with different organs (and/or tissues), stages of development, species, cultivars, cultural practices, and environmental factors is reviewed. For each class, current knowledge regarding biosynthesis, metabolism and transport in citrus tissues is summarized. The concluding section deals with future research directions.     

Changes in the level of endogenous cytokinins in apical buds ofChenopodium rubrum L.

CCC (2-chloroethyl)trimethylammonium chloride applied to plants ofChenopodium rubrum during floral induction led to an increase in the level of endogenous cytokinins in the apical buds. Application of gibberellic acid or indole-3-acetic acid at concentrations reversing the effect of CCC reduced the level of cytokinins. After simultaneous treatment with both CCC and one of the growth substances this reduction was less pronounced. From the comparison bf the present results, as well as of those published in previous papers it follows that in apical buds ofChenopodium rubrum there exists a mutual interaction between gibberellins and cytokinins. Under certain conditions both these groups of hormones may substitute for each other in flowering. IAA seems to affect flowering by regulating the level of both gibberellins and cytokinins.     

Phytohormonal changes in intact shoots of wheat and oilseed rape treated with the acylcyclohexanedione growth retardant prohexadione calcium

The effects of the acylcyclohexanedione-type growth retardant prohexadione calcium on seedling growth and endogenous levels of immunoreactive phytohormone-like substances in shoots of wheat ( Triticum aestivum L. cv. Kanzler) and oilseed rape ( Brassica napus L. ssp. napus cv. Lirajet) were studied. After treatment of seedlings with increasing retardant concentrations in hydroponics, plant height and fresh weight of shoots were reduced by up to 40%. Concomitantly, the amount of immunoreactive gibberellins decreased, on a fresh weight basis, when compared with levels in the shoots of control plants. In contrast, the levels of abscisic acid and dihydrozeatin riboside and isopentenyladenosine-type cytokinins were considerably elevated by the growth retardant. The content of 3-indoleacetic acid decreased slightly. These results suggest that, in addition to its effect on gibberellin content, prohexadione calcium also influences the levels of endogenous abscisic acid and cytokinins.     

Plant growth regulator and graft control of axillary bud formation and development in the TO-2 mutant tomato

The torosa-2 tomato mutant is characterized by a strong inhibition of release of axillary shoots, that is not under the control of the main apex and IAA. Microscopic examination indicated that about 70% of leaf axils do not have axillary buds. Of the growth regulators tested, gibberellic acid and cytokinins were able to modify the to-2 phenotype: increasing bud number (GA₃ treated) and developing shoots (both substances). Sequential application of growth regulators demonstrated that bud production was only affected by treatments given between sowing time and 32 days after germination. Grafting experiments indicated that endogenous root factors have no essential role in the lateral branching of the genotypes investigated. The control of axillary bud differentiation and the branching pattern in the to-2 appears to be dependent of a complex mechanism involving gibberellins and cytokinins.     

Role of plant growth regulators in host-pathogen relationships

The effect of indol-3-ylacetic acid, gibberellie acid, kinetin, abscisic acid and Ethrel on the growth of mycelium, sporulation and germination of spores ofFusarium culmorum of different pathogenicity to wheat seedlings was studied. The production of gibberellins, auxins, cytokinins, ethylene and growth inhibitors by these isolates was determined as well. It has been found that most pronounced and explicit effect on growth and development in fungi was produced by Ethrel which strongly inhibited these processes. ABA proved to be a strong growth and development stimulator, though to a different extent in different isolates. GA₃ strongly stimulated sporulation and spore germination in some isolates. The effect of IAA and K on growth and development in fungi was slight. More sensitive to growth regulators were the fungi in earlier stages of growth. No correlation between the pathogenicity of the isolates and their ability to produce growth regulators as well as between their susceptibility to exogenous growth substances in the processes of fungal growth and development was stated. Presented at the International Symposium “Plant Growth Regulators” held on June 18–22, 1984 at Liblice, Czechoslovakia.     

Gibberellin-like Substances in the Sporophores of Agaricus bisporus (Lange) Imbach

Sporophores of cultivated Agaricus bisporus (Lange) Imbach,were shown to contain a gibberellin-like substance active inthe dwarf maize (d-5), -amylase and other bioassays. Ethyl-acetateextraction followed by paper, column, and thin-layer chromatographyrevealed the presence of one major active substance. Ficin hydrolysisof dried sporophore powder, after the complete removal of freesubstances, released more gibberellin-like substances, one ofwhich appeared identical to the free compound. The free substance was predominantly in the lamellae and residualpileus tissue. The major active substance released by ficinoccurred mostly in the lamellae but also in substantial equalamounts in both stipes and pilei. No activity was found in extractsof dikaryotic vegetative mycelium on malt agar. The level ofactivity in extracts from sporophores stored at – 20 °Cfell sharply after 7 d, and then remained constant over a periodof 6 weeks. The content of gibberellin-like substances in youngand old whole sporophores showed wide variation between experiments.In most cases young 2-d tissue had higher levels than old, 11-dtissue on a fresh-weight basis. Purified sporophore extractsand authentic gibberellins had no stimulating effect on growthof sporophores or of cultured vegetative mycelium. The inhibitorsof diterpene biosynthesis, CCC, and AMO-1618 induced a smallincrease in mycelial growth rate. Ethyl-acetate extraction ofhorse-straw compost prior to inoculation with Agaricus bisporusshowed the presence of gibberellin-like activity in significantamounts.     

Cytokinin-Regulated Mesophyll Cell Division and Expansion during Development of Cucurbita pepo Leaves

The role of cytokinins in the development of mesophyll structure was studied in developing pumpkin Cucurbita pepo L. leaves. Leaves were treated with cytokinins at different stages of growth: when they reached 25 or 50% of their final size (S _max), immediately after leaf growth ceased, and during senescence. At the early stages of leaf development, treatment with exogenous benzyladenine accelerated division of mesophyll cells. At the later stages of development, BA treatment activated expansion of growing cells and those, which have just accomplished their growth. The exogenous cytokinin did not affect the senescent leaf cells. The content of endogenous cytokinins changed during mesophyll development. The juvenile leaves (25% of S _max) were characterized by low level of these phytohormones. In the expanding leaves (50% of S _max), the content of phytohormones increased and decreased when leaf growth ceased. In the senescent leaves, the cytokinin content decreased markedly. It was concluded that the response of mesophyll cells to cytokinin depended on the cell growth phase at the moment of hormone action. Furthermore, in the young leaves, lower cytokinin concentrations were required for division of mesophyll cells in vivo than for cell expansion at the final stage of leaf development.     

The Location of Cytokinins and Gibberellins in Wheat Seeds

Wheat seeds (Triticum aestivum L. cvs. Yorkstar and Sirius) were cut transversely into embryoless and embryo-containing (embryonated) halves and the content of endogenous cytokinins and gibberellins in both halves determined before and after the seeds imbibed water for 12–15 h at 22°C in the light. Dry seeds contained little ethyl acetate-extractable gibberellin activity as measured by bioassay but n-butanol-soluble cytokinins were detected mainly in the embryoless halves. Dry, embryonated half-seeds contained water-soluble gibberellins which could be extracted into acidic ethyl acetate after treatment of the aqueous extract with either alkaline phosphatase, β-glucosidase or a crude pectolytic enzyme preparation. When half-seeds were allowed to imbibe water for 12 h and then extracted, cytokinin activity was largely lost from the embryoless halves and completely from the embryonated halves and water-soluble gibberellins were also lost from the embryonated halves. However, ethyl acetate-soluble gibberellins were present in the latter suggesting that “bound’ gibberellins were released during imbibition. The hormones present in normal and naturally embryoless dry grains of cv. Yorkstar were also determined. Both gibberellin and cytokinin activity was higher in normal grains suggesting that the presence of an embryo is essential for synthesis or accumulation of these hormones in the grain during development.     

Note on study of the sporulation of fungi: endotrophic sporulation in the genusPenicillium

Conidiophore formation and sporulation can be induced inPenicillium sp. strain P 17 by an environmental factor—carbohydrate (carbon) starvation. Both surface and submerged mycelium, when transferred from synthetic medium to glucose-free salt solution, form conidiophores and sporulate, while in the control cultures on complete medium, vegetative growth continues. The time required for the formation of conidiophores, i.e. the induction interval, is 7–14 h and its length increases with the age of both surface and submerged mycelia. During the induction phase the mycelium undergoes autolysis, associated with degradation of energy motabolism involving the comsumption of reserve substances, a rapid drop in endogenous respiration and the endogenous reducing activity of the mycelium, a decrease in the labile phosphate concentration, proteolysis, an increase in the ammonia and orthopsphate concentration and exhaustion of readily oxidized amino acids from the pool. A transient increase in respiration occurs before differentiation of the conidiophores starts. During the second half of the induction phase, polyphenol substances and polyphenol oxidase appear in the mycelium.The enzyme is not induced by exogenous phenols. Its possible role in the sporulation of fungi is considered.     

Changes in growth regulator content of radish seedlings, Raphanus sativus, infested with the aphid Myzus persicae

Following 10 days of infestation by the aphid Myzus persicae there was an increase in the amount of growth inhibiting substances and a decrease in cytokinins, gibberellins and auxins in infested as compared with similar uninfested radish seedlings. Even after previously infested seedlings have been freed of aphids for 10 days, differences in the hormone balance remained. The possible relationships between the changes in hormonal balance and the effect of the aphid infestation on growth, translocation and wilting are discussed.     

Effects of Far-red Light on the Hormonal Control of Side Shoot Growth in the Tomato

Side shoot growth in young tomato plants was almost completelysuppressed by a 5 min period of far-red light immediately followinga 16 h photoperiod from fluorescent tubes, whereas plants givenan identical photoperiod but lacking the far-red treatment branchedprofusely. The influence of far-red light on the degree of sideshoot suppression and the correlated changes in the levels ofauxins, gibberellins, cytokinins and abscisic acid is presentedand discussed in relation to current hypotheses of correlativeinhibition. It is suggested that far-red light causes increasedauxin synthesis in the apex and young leaves, which in turninduces the formation of abscisic acid in or near the axillarybuds, and it is this hormone which inhibits bud outgrowth. Therole of cytokinins and gibberellins remains uncertain but theyprobably act in a sequential manner, the gibberellins promotingbud growth following cytokinin-mediated release from apicaldominance.     

Gibberellins in Diffusates from Shoots of Apple Trees

Excised shoot apices, leaves and internodes from shoots of apple trees (Malus×domestica) give off gibberellins by diffusion on agar. A methanol extract of the agar was prepared, the extract separated on thin layer plates, and the gibberellin activity estimated by means of Rumex and lettuce hypocotyl bioassays. The largest amounts of gibberellin are found in diffusates from the shoot apex, the two upper leaves and the two upper internodes. Several promotive fractions are found on the chromatograms as well as growth inhibitors. Removal of young leaves retards elongation of the internodes. Probably gibberellins produced in young leaves exercise some control over this process. The growth regulators Alar and CCC also retard internode elongation. Diffusates from shoots treated with these substances were also examined. Preliminary results suggest that the amount of diffusible gibberellins from treated shoots is not reduced.     

Changes in endogenous gibberellins in plant organs producing and utilizing photosynthates

The distribution of endogenous gibberellin-like substances in individual organs ofZea mays L., cv. CE 250, plants was investigated during the transition from the vegetative to the generative phase of development. The gradient of the content of gibberellin-like substances and photosynthetic activity in leaf segments was followed in different parts of the Jeaves, as well as the changes in the content of gibberellin-like substances in leaf segments during an exposure in the light and in the dark. The gradient of the content of endogenous gibberellin-liko substances in the leaves, in the stem and in the spike is interpreted in terms of possible relationship of these compounds to the regulation of sink — source.     

The influence of kinetin on the endogenous content of indoleacetic acid in swelling seeds of Phaseolus, Zea and Pinus and young plants of Phaseolus

Treatment of different plant materials, seeds of Phaseolus vulgaris, Zea mays and Pinus silvestris and young plants of Phaseolus, with kinetin increased the level of extractable IAA. For seeds this increase was most pronounced in bean seeds, which contained the lowest amount of endogenous IAA and cytokinins, and lower in maize seeds with high endogenous content of IAA and cytokinins. – For young bean plants the kinetin treatment significantly increased the extractable amounts of IAA from all parts of the plant, hypocotyls, cotyledons, epicotyls and primary leaves, when the cut plants were placed for 24 h in kinetin solution. For plants sprayed with kinetin solution only the primary leaves showed a significantly higher level of extractable IAA, which could be explained by the fact that the plants were growing very close together, so that the primary leaves received most of the kinetin during spraying.     

Effect of 6-Benzylaminopurine on the Structure of the Photosynthetic Apparatus of Faba Bean (Vicia faba L.)

The level of endogenous cytokinins changed with growth and development of faba bean (Vicia faba L.) leaves. Typical of juvenile leaves, amounting to 25% of the final leaf size (S _max), was a low content of these plant hormones. The level of cytokinins increased in growing leaves (50% of S _max) and decreased in leaves that stopped growing. The content of cytokinins in senescent leaves dropped considerably. Exogenous treatment with 6-benzylaminopurine (BAP) had no effect on the structure of the terminal phloem; however, it did (1) stimulate elongation of mesophyll cells; (2) increase the area and thickness of the leaf blade, amount of photosynthetic pigments, and assimilation potential; and (3) delay senescence of the leaves and defoliation, thereby increasing the biomass of the aboveground plant part. It is inferred that BAP has the potential for inducing the development of photosynthetic apparatus and increasing the yield of the green mass of faba bean.