Studies on the physiology of hyacinth bulbs (Hyacinthus orientalis L.) the effect of plant growth regulators on metabolic activities in non-chilled hyacinth bulbs

The effect of gibberellic acid, benzyladenine and a mixture of these compounds on the activity of some hydrolytic enzymes was studied in hyacinth bulbs non-exposed to low temperatures. Plant growth regulators were applied on the heel of dormant bulbs in the middle of July. An intensive elongation of the inflorescence was observed only in the plants treated previously with GA₃ and grown in a warm greenhouse for 63 and 84 days. The activities of amylase, invertase and acid phosphatase were higher in the flower buds of unrooted bulbs treated with growth regulators than in the control plants, although growth regulators did not affect the level of extractable proteins. The elongation of the inflorescence in the plants treated with gibberellic acid was correlated with a sharp increase of invertase activity in this organ. The effect of GA₃ and BA on the activity of other enzymes in buds and scales varied with the period of plant growth.     

ABA and GA3 affect the growth and pigment composition in Andrographis paniculata Wall.ex Nees., an important folk herb

In this study, 5 μmol·L⁻¹ abscisic acid (ABA) and gibberellic acid (GA₃) were used to study the effect of both growth regulators on the morphological parameters and pigment composition of Andrographis paniculata. The growth regulators were applied by means of foliar spray during morning hours. ABA treatment inhibited the growth of the stem and internodal length when compared with control, whereas GA₃ treatment increased the plant height and internodal length. The total number of leaves per plant decreased in the ABA-treated plants, but GA₃ treatment increased the total number of leaves when compared with the control. Both growth regulators (ABA and GA₃) showed increased leaf area. ABA and GA₃ treatments slightly decreased the total root growth at all the stages of growth. The growth regulator treatments increased the whole plant fresh and dry weight at all stages of growth. ABA enhanced the fresh and dry weight to a larger extent when compared with GA₃. An increase in the total chlorophyll content was recorded in ABA and GA₃ treatments. The chlorophyll-a, chlorophyll-b, and carotenoids were increased by ABA and GA₃ treatments when compared with the control plants. The xanthophylls and anthocyanin content were increased with ABA and GA₃ treatments in A. paniculata plants.     

Stimulation of the formation of fruiting bodies of the fungusLentinus tigrinus (Bull.) Fr. by growth regulators

Growth regulators, indole-3-acetic acid (IAA), gibberellic acid (GA₃), and kinetin (KIN), were used in different concentrations to stimulate the initiation and further development of the fruiting bodies of the fungusLentinus tigrinus. Vegetative mycelium of the fungus was cultivated on cellulose cylinders soaked with a synthetic nutrient solution or with a 3% malt extract. When the mycelium covered the surface of the cylinders, further cultivation was carried out in graduated concentrations of the growth regulators mentioned above. The number of developed fruiting bodies showed that the optimum IAA and GA₃ concentrations were in both media 300 p.p.m. The optimum concentration of kinetin was 400 p.p.m. The individual growth regulators influenced characteristically also the shape of the fruiting bodies and changed in them the natural level of endogenous growth regulators. The addition of IAA into the medium raised the level of endogenous auxins in the cap. The presence of gibberellic acid and of kinetic in the medium resulted in an increase in the level of these regulators in the fruiting bodies.     

In vitro thiophene production by transformed root cultures of Tagetes laxa (Cabrera)

The effect of different concentrations of carbohydrates, nitrogen, sulphate, plant growth regulators and elicitors on growth and thiophene accumulation by transformed root cultures of Tagetes laxa (Cabrera) was studied. The combinations of sucrose (30 g/l), nitrogen (60 mM), sulphate (150 mM) and the ratio N_ox:N_red 2:1 are the most appropriate combination to support growth and thiophene accumulation, which was increased by 90% when the cultures were elicited with homogenate of Sclerotinia sclerottiorum. The plant growth regulators used produced dedifferentiation with a decrease in thiophene biosynthesis.     

A protocol for direct somatic embryogenesis of Protea cynaroides L. using zygotic embryos and cotyledon tissues

We describe a protocol for somatic embryogenesis of Protea cynaroides, with potential for high frequency production of this important horticultural species. Somatic embryos formed directly on both P. cynaroides mature zygotic embryos and excised cotyledons cultured on MS medium without growth regulators. The addition of growth regulators such as naphthalene acetic acid (NAA) (5; 13 and 27 μM) and 2,4-dichlorophenoxyacetic acid (2,4-D) (5; 11 and 23 μM), in combination with thidiazuron (TDZ) (1 μM), benzylaminopurine (BAP) (1 μM) or kinetin (1 μM) suppressed the formation of somatic embryos. After eight weeks in culture, formation of somatic embryos was observed. Zygotic explants formed the most embryos when cultured in a 12-h photoperiod in comparison to explants cultured in the dark. Up to 83% of these embryos germinated after transferal to the germination medium containing 0.3 μM GA₃. Significantly fewer embryos germinated in MS medium with no growth regulators, or supplemented with higher concentrations of GA₃, while low germination percentages were also observed in MS media containing casein hydrolysate and coconut water. The germination of normal somatic embryos (two separate cotyledons and a single radicle) was observed only in media containing either no growth regulators, 0.3 μM GA₃ or 1 μM GA₃. All embryos that germinated in high concentrations of GA₃ were malformed.     

Effect of Plant Growth Regulators on Phytoremediation of Hexachlorocyclohexane-Contaminated Soil

The influence of three plant growth regulators, indolebutyric acid (IBA), thidiazuron (TDZ) and gibberellic acid (GA₃), either individually or in pair-wise combinations, on the ability of waxy corn plant to remove hexachlorocyclohexane (HCH) from contaminated soil was studied. Waxy corn seeds were immersed for 3 h in solutions of 1.0 mg/l IBA, 0.01 mg/l TDZ, 0.1 mg/l GA₃, or a mixture of two of the growth regulators, and then inoculated in soil contaminated with 46.8 mg/kg HCH for 30 days. Pretreatment of corn seeds with the plant growth regulators did not enhance corn growth when compared with those immersed in distilled water (control), but the pretreatment enhanced HCH removal significantly. On day 30, HCH concentration in the bulk soil planted with corn seeds pretreated with GA₃ or TDZ+GA₃ decreased by 97.4% and 98.4%, respectively. In comparison, HCH removal in soil planted with non-pretreated control waxy corn seeds was only 35.7%. The effect of several growth regulator application methods was tested with 0.01 mg/l TDZ. The results showed that none of the methods, which ranged from seed immersion, watering in soil, or spraying on shoots, affected HCH removal from soil. However, the method of applying the growth regulators may affect corn growth. Watering the corn plant with TDZ in soil led to higher root fresh weight (2.2 g) and higher root dried weight (0.57 g) than the other treatments (0.2–1.7 g root fresh weight and 0.02–0.43 g root dried weight) on day 30. Varying the concentrations of GA₃ did not affect the enhancement of corn growth and HCH removal on day 30. The results showed that plant growth regulators may have potential for use to enhance HCH phytoremediation.     

The Effect of Growth Regulators on Quality Parameters and Ginsenosides Accumulation in Panax quinquefolium L. Roots

American ginseng (Panax quinquefolium L.) is a perennial medicinal herb originally grown in Canada and USA, and recently also in China, Australia, Holland and Poland. Several commercial preparations are produced from ginseng roots, that are known for their antifatigue, antitumor, antistress and immune system stimulating functions. The medicinal properties are due mainly to the active components – ginsenosides. In this work, the results of field cultivation experiments are presented that examine the effects of foliar application of several growth regulators on quality parameters and ginsenoside content of P. quinuefolium roots. The growth regulators tested, i.e., kinetin, daminozide, mixture of gibberellic acid (GA₃) with potassium salt of α-naphthalene acetic acid (kNAA) and new preparation – IPO-1 – benzimidazole derivative (obtained from the Institute of Organic Industry in Warsaw – at present during the process of patent), were applied at a concentration of 100 or 200 mg l⁻¹ in the middle of June in the 2nd year of vegetation. After 4 years of cultivation, the roots were dug up and dried, and subsequently the quantitative analysis of individual saponins (R_b1, R_b2, R_c, R_d, R_e, R_g1) by HPLC was performed. Growth regulators significantly affected quality parameters, morphological features and accumulation of individual and total ginsenosides in ginseng roots. Regardless of doses, the plant roots treated with growth regulators had a higher content of total ginsenosides in comparison to the control. The growth regulators also affected individual ginsenosides level and narrowed the ratio of R_b:R_g group. The application of kinetin, daminozide and benzimidazole derivative for foliar spray during 2nd year of American ginseng vegetation caused a significant increase in air dry weight of roots and aboveground parts whereas the mixture of GA₃ and kNAA showed a decreasing effect. An increase of roots size was observed using higher doses (200 mg l⁻¹) of kinetin and daminozide while a decreasing tendency appeared with the application of the other preparations.     

Callus culture from hypocotyls of Kosteletzkya virginica (L.) seedlings

It was shown that callus established from Kosteletzkya virginica (L.) Presl. (Malvaceae) can grow in salinities higher than 200 mM NaCl if previously accomodated stepwise. Callus lines developed from seedlings of different harvests or of the same harvest at different times, all showed the same pattern of growth and sensitiviy to salinity. The absorption of Na⁺ into the callus increased with increasing external NaCl concentration. In the callus, Na⁺ was apparently distributed outside and inside a cellular membrane (possibly the plasmalemma). This membrane was, apparently, capable of regulating the Na⁺ concentration in the protoplast. Outside this membrane Na⁺ accumulated to concentrations higher than in the external growth medium. Exogenously supplied proline or glycine-betaine did not affect the growth of the callus. Externally applied ABA stimulated growth under saline conditions and increased the accumulation of proline. Growth and proline content were positively correlated in callus exposed to salinity, but in the presence of ABA they were negatively correlated. ABA was involved in both growth and proline accumulation, but there was no clear relationship between these two effects. Both ABA and proline, if added to the growth medium, improved the appearence of the callus.Abbreviations ABA abscisic acid - B₅ Gamborg's medium - BA benzylalanine - 2,4-d 2,4-dichlorophenoxy acetic acid - FW fresh weight - G B₅ medium without growth regulators - GH B₅ medium supplemented with growth regulators - NAA naphthalene acetic acid - PGR plant growth regulators - Q _T total amount of a certain ion in the tissue - Q _s amount of the ion that has leaked out - QAC Quaternarty Ammonium Compounds - RGR mean relative growth rate - W₁ and W₂ fresh weight at times t₁ and t₂     

Plant growth regulators and adventitious root development in relation to auxin

Adventitious root formation in stem cuttings of mung bean was enhanced by ethrel, which had an additive effect when employed simultaneously with indolebutyric acid (IBA). Abscisic acid (ABA) did not influence the number of roots per cutting whereas gibberellic acid (GA₃) and kinetin were without effect on rooting at lower concentrations but were inhibitory at higher concentrations. Nevertheless, all three of these chemicals showed synergistic interactions with IBA and/or indol-3-ylacetic acid (IAA) and thereby significantly promoted root formation. A localised application of morphactin to the epicotyl of cuttings totally inhibited root production irrespective of which of the foregoing growth regulators were suppliedvia the hypocotyl. Morphactin application also prevented root formation in cuttings treated with vitamin D₂. The various growth regulators employed had differing effects on growth of roots but there was no simple relationship between their effects on root formation and subsequent root growth.     

Growth and Differentiation of Seedlings of an Aquatic Plant,Trapella sinensis OLIV II. Photosynthesis and Respiration of Seedling in Early Stages

Trapella sinensis seedlings were examined to determine whether the effect of growth regulators such as gibberellin (GA₃), kinetin and the extract of Chlorella cells on chlorophyll formation, photosynthetic activity and cytochrome oxidase activity depend upon the developmental stage. Each addition promoted the growth on both the basis of dry weight and of fresh weight. Among these regulators, only Chlorella extract promoted growth as a result of an activated rate of photosynthesis through increased chlorophyll formation. There also was an increased respiratory rate through increased cytochrome oxidase activity.     

Effects of nutritional factors on the formation of Anthraquinones in callus cultures of Rheum ribes

Rheum ribes L. (Polygonaceae) is the source of one of the most important crude drugs in Asiatic regions .The medicinal character of rhubarb is due to its anthraquinone content . Different parts of sterile seedlings were cultured on MS medium to study the generation of callus. The explants were cultured with different ranges of plant growth regulators and the best range of plant growth regulators for generation of callus was IBA (1 mg l^–1) and BA (1 mg l^–1). The content of anthraquinones were determined by HPLC . The concentration of sucrose, vitamins and Myo-inositol and ratio of NO₃ to NH₄ in the medium was changed and growth rate and content of 2 anthraquinones was determined . The growth rate of callus declined with increased rate of secondary metabolites production. Myo-inositol 100 mg l^–1 in the medium increased anthraquinone content and in medium that had NO₃/NH₄:1/1 the maximum content of anthraquinone was obtained.     

The effect of plant growth regulators and their interaction with electric current on winter wheat development

The aim of the experiments presented here was twofold. On the one hand, to study the influence of plant growth regulators, i.e. auxins (IAA and NAA), cytokinins (kinetin, zeatin and zeatin riboside), gibberellins: (GA₃) and polyamines (spermidine, spermine and putrescine) on the generative development of winter wheat plants, while on the other to study the interaction of plant growth regulators with electric current in affecting the efficiency of the flowering of wheat plants. Winter wheat var. Grana was subjected to a short vernalization (2 weeks at 5°C + 10 days at 10°C) on Murashige and Skoog medium containing growth regulators at different concentrations. For selected substances, seedlings were additionally treated with electric current by applying a constant voltage for a given period of time (30 V for 30 s or 1.5 V for 1 h) with the anode or the cathode inserted into apical leaves and the reference electrode into the media. After that, the seedlings were transferred to a glasshouse (20/17°C) where they were grown until heading was achieved. All the substances studied stimulated the generative development of winter wheat, in contrast to non-treated plants (control), especially at their highest concentrations. Spermidine, kinetin and GA₃ were the most effective in this process as they stimulated 100–80% of generative plants (whereas for the control the percentage stood at only 30% plants). The studied regulators also increased the rate of generative development, i.e. shortening the length of the vegetative phase (by about 30% in comparison with the control). Moreover, these substances treatments decreased the fresh mass of both seedlings and flowering plants and increased the number of spikelets in the ear. The electric current treatment interacted with the applied substances: the anode insertion into the leaves generally increased, whereas the cathode insertion decreased, the inductive effect of the growth regulators on generative development. However, the inhibition of the flowering process was more pronounced than the induction. Moreover, the passage of electric current shortened the time to heading. This decrease in the length of the vegetative phase was more visible after the anode was inserted into the leaves. The least noticeable effect of the electric current interaction with plant growth regulators on both the generative development and the time of heading was observed for auxins.     

Microspore culture of small grain cereals

Androgenesis of wheat, rice and triticale was studied in isolated microspore culture. It is the first publication which studies microspore culture reaction of Hungarian rice varieties. The effect of different basic media, lack and absence of growth regulators in culture media were tested on important parameters of microspore culture. Direct embryogenesis was observed in microspore culture of wheat and triticale genotypes. In the case of rice, calli were induced in isolated rice microspore culture and haploid rice plantlets were regenerated via organogenesis.In wheat, the effect of basic media (W₁₄, A2, CHB₃, P4-m) was compared and among them the W₁₄, and A2 had a superior effect on embryo production and albino and green plantlet regeneration. In rice the C, CHB₃ and MS_m media were tested in microspore culture and the significantly highest numbers of calli were achieved by using C and CHB₃ media depending on the genotypes. The lack of exogenous growth regulators was observed in isolated microspore culture of triticale and rice. Growth regulator-free medium had a positive effect on embryo production and plant regeneration of triticale genotypes, whereas in rice microspore culture multicellular structures did not continue their division without growth regulators from the third week of microspore culture. Developing of microspore-origin calli was maintained by supplement of 2,4-D and Kinetin combination in the microspore culture medium.     

Direct shoot formation in spontaneously occurring root pseudonodules of alfalfa (Medicago sativa L.)

Summary A simple and rapid procedure for direct organogenesis from root nodulelike structures of alfalfa (Medicago sativa L.) line SGg, spontaneously induced on growth regulator-free Gamborg (B₅) medium, was developed. Prolific adventitious shoot initiation was obtained using a combination of 1.0 mg/liter TIBA and 0.5 mg/liter 2iP. Transfer of shoots to a medium containing 0.5 mg/liter ABA and reduced concentration of TIBA (0.5 mg/liter) before rooting markedly stimulated shoot development. Regenerated shoots rooted easily and revealed the early appearance of nodulelike structures on basal medium (B₅) lacking growth regulators. Analysis of endogenous growth regulator levels of SGg roots maintained on growth regulators free media, showed that spontaneous shoot appearances was correlated with high cytokinin-to-auxin ratios.     

The application of growth regulators for the physiological emasculation of rye(secale cereale l.) flowers

The present paper deals with the possibility of using growth regulators for the physiological emasculation of rye flowers and utilizes our knowledge of the higher sensibility of stamens to the auxin level. By means of bioassays it has been found that a relatively low content of auxins and a definite level of gibberellins and inhibitors are characteristic for normal rye spikes at the time of stamen differentiation. The higher level of auxins and expressive inhibitions occur in the later course of pistil differentiation. Two sprayings of plants with mixed solutions of MH and NAA and two further sprayings with a solution of NAA only before the stamen differentiation change slightly the natural character of endogenous regulators in spikes and cause the destruction of mother pollen cells in anthers and the sterility of anthers in the majority of rye florets. The anthers in the basis of spikes sometimes contain pollen grains after treatment, but their fertility is substantially lowered because the spray evidently disturbed the accumulation of reserve substances in pollen grains. The development of pistil was not affected by the exogenous application of growth regulators. Normal kernels arose in the majority of flowers after supplementary pollination. Some accidental effects of exogenous application of growth regulators, as for example reduced stem growth and disorder of lodicule function were lowered to a great extent by adding GA_S to the last two sprayings of NAA.     

Evaluation of the effect of three growth regulators in the germination of <Emphasis Type="Italic">comparettia falcata</Emphasis> seeds under <Emphasis Type="Italic">in vitro</Emphasis> conditions

Summary The effect of 3-indoleacetic acid (IAA), 6-furfurylaminopurine (kinetin), and gibberellic acid (GA₃) on germination of the orchid Comparettia falcata was evaluated in a factorial experiment (4×4×4) with Murashige and Skoog (1962) basal medium. It was established that seeds of this orchid could be maintained under aseptic conditions as long as the necessary nutrients and appropriate concentrations of growth regulators were provided. Of the three growth regulators used, IAA significantly decreased seed germination of Comparettia falcata. There was a synergistic effect in the kinetin:GA₃ combination that produced a positive response in both percentage seed germination and development of seedlings. This study describes a single medium-based protocol able to achieve more than 160000 seedlings within 21 wk, starting from a single capsule, sufficient for both large-scale propagation and in vitro conservation of this threatened orchid.     

Changes in responsiveness to ethylene and gibberellin during corolla expansion ofIpomoea nil

Corolla expansion inIpomoea nil appears to be triggered by changes in gibberellin concentration and ethylene production during development. We investigated the role of responsiveness to GA and ethylene in corolla expansion. The effects of growth regulators applied in vitro were measured as a change in area of corolla segments from younger (15–17 mm) and older (18–20 mm) whole corollas. Applied gibberellic acid (GA₃) significantly (p < 0.05) promoted growth in the younger segments but was less effective in the older segments. Moreover, applications of the GA biosynthesis inhibitors, PP333 (paclobutrazol) AMO₁₆₁₈ (2-isopropyl-4-dimethylamino-5-methylphenyl-1-piperidinecarboxylate methyl chloride), chlorocholine chloride, and tetcyclasis had little effect on younger segments but inhibited growth of older segments. The older corollas have apparently synthesized and accumulated enough GA-like substances to become less responsive to additional applied GA₃. The amount of growth induced by applied or endogenous GA depended on the amount of ethylene simultaneously produced in the tissue. The younger corollas rapidly produced ethylene from endogenous 1-aminocyclopropane-1-carboxylic acid (ACC) and did not respond to applied ACC whereas the older corollas naturally produced much less ethylene and were significantly (p < 0.05) inhibited by applied ACC. When ethylene production was inhibited by applying aminoethoxyvinylglycine (AVG), growth was promoted in all segments. However, only the growth of the younger segments was further stimulated by simultaneously applied AVG and GA₃ over the GA₃ control. Thus the differential responses of segments from 15- to 20-mm long corollas to applied growth regulators reflect developmental changes in responsiveness of the developing corolla. The change in responsiveness is attributed in part to the changes in production of endogenous growth regulators and to the effect of one endogenous plant growth regulator (PGR) on the responsiveness of the corolla to another PGR.     

Somatic embryogenesis and cytoplasmic sterility inBeta vulgaris L. var.saccharifera

Embryogenic callus which has maintained its embryogenic ability on media without growth regulators for three years has been induced at the base of shoots of a genotype with CMS propagated for a long timein vitro by transferring the shoots onto media richer in inorganic and organic components. The effect of two basal media (MS and PG₀) on the intensity and completeness of the proliferation of somatic embryos was examined with different combinations of growth regulators. Pollen fertility was evaluated in 87 plants regenerated from somatic embryos. Cytoplasmic male sterility was conserved in all of them.     

Factors affecting maintenance, proliferation, and germination of secondary somatic embryos of Eucalyptus globulus Labill

The described protocol for repetitive somatic embryogenesis (SE) in Eucalyptus globulus produced more somatic embryos than the primary SE protocol. Primary somatic embryos (induced on MS_3NAA) were transferred to the same medium, leading to new cycles of somatic embryos, for at least 2 years. The influence of medium (MS and B5), plant growth regulators (auxins and cytokinins), and light on secondary SE was tested. The MS medium without growth regulators (MS_WH) was more efficient for cotyledonary embryo formation and germination than the B5 medium. Reducing auxin (NAA) levels increased the proliferation of globular somatic embryos and allowed SE competence to be maintained on medium free of plant growth regulators. The addition of two cytokinins (BAP and KIN) to the MS medium did not improve proliferation of globular secondary embryos, but was crucial during later stages of the SE process (germination and conversion). Data also show that light may influence the quality of the process, depending on its stage. Darkness should be maintained until the cotyledonary stage is reached, after which exposure to light is recommended.     

Changes in endogenous growth regulators in nasturtium leaves during senescence

Summary By use of lettuce-hypocotyl and wheat-coleoptile bioassay, the presence of both gibberellin (GA)-like and abscisic-acid(ABA)-like components in acidic ethyl-acetate extracts of fully expanded nasturtium (Tropaeolum majus) leaves has been shown. During senescence of detached leaves there was a progressive decline in GA-like components and an increase in ABA-like components. Pretreatment of detached leaves with GA₃ or kinetin prevented changes in the levels of endogenous growth regulators and delayed senescence. The observations provide experimental verification for the concept that senescence is associated with changes in endogenous growth regulators.