The Effect of Growth Hormones on Cell Division and Expansion in Liquid Suspension Cultures of Acer pseudoplatanus

The effects on cell division and cell size of indole-3-aceticacid (IAA), gibberellic acid (GA), and kinetin were studiedin liquid suspension cultures of cambial cells derived fromAcer pseudoplatanus. It was shown that all three hormones promotecell division and that the effects of both GA and kinetin areadditive to those of IAA, but the effects of GA and kinetintogether are not additive. Treatment with IAA resulted in anincrease of mean cell size (indicating that cell expansion ispromoted), but after GA or kinetin treatment the mean cell sizewas smaller, indicating that little cell expansion had takenplace after each division. The results are discussed in relationto previous work on the effects of hormones in the intact cambiumand to current theories on the interactions of growth hormones.     

Quantitative Relationship between Various Growth Substances and Bud Production in Funaria hygrometrica. A bioassay for abscisic acid

Eleven growth substances were tested for their bud producing capacity in Funaria hygrometrica. IAA and gibberellic acid showed no response, neither did inositol, abscisic acid and adenine. The cytokinins: benzylaminopurine, kinetin and mercaptopurine showed a quantitative relationship while methylaminopurine did not. When different amounts of abscisic acid were added together with 10 μg kinetin (which gave maximal production of buds), there existed a linear relationship between reduction in buds and abscisic acid added. Results are obtained in two weeks and it is therefore a fairly quick method for assaying certain cytokinin and also abscisic acid.     

Growth Hormones and Propagation of Cymbidium in vitro

Protocorms of Cymbidium (Orchidaceae) were grown on solid or liquid medium with macro-nutrients according to Wimber (van Raalte 1967) and iron, micro-nutrients and vitamins according to Nitsch (1968) the medium also contained 2% sucrose. The effects of 1) the auxins; indol-3yl-acetic acid (IAA), α-naphthaleneacetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D); 2) the cytokinins; 6-furfurylaminopurine (kinetin) and benzyladenine (BA) and 3) the gibberellin; gibberellic acid (GA) were examined alone or in combinations. IAA had no effect alone. NAA resulted in optimal fresh weight at 10 μM and the protocorms were vigorous, but lighter green than usual. 2,4-D caused a high weight increase at 1 μM, but the protocorms were abnormal. Higher concentrations of NAA and 2,4-D inhibited chlorophyll synthesis. On solid medium kinetin (100 μM) induced a growth of many small shoots, but had no effect on the fresh weight. In liquid medium, kinetin promoted a callus formation and fresh weight increase. BA had effects similar to kinetin, but at lower concentrations. GA alone promoted shoot and leaf growth. Combinations of kinetin and NAA resulted in a maximal fresh weight increase at kinetin concentrations one tenth of the NAA concentrations. The optimal growth and the best development occurred at 10 μM NAA and 1 μM kinetin. NAA and kinetin together could limit the shoot and leaf growth induced by GA.     

The Effects of Growth Substances on the Growth of Tapinanthus bangwensis (Loranthaceae) in vitro

The growth response of Tapinanthus bangwensis (Engler and K.Krause) Danser to exogenous growth substances, indole-3yl-aceticacid (IAA), kinetin, and gibberellic acid (GA₃), not only varieswith the concentrations of each of these substances in the mediumbut follows a similar trend in each case. Kinetin, IAA, andGA₃ were found to enhance growth but not by very appreciableamounts, kinetin being the most effective growth promoter. Highconcentrations of these substances (5.0 and 10.0 ppm) exceptGA₃ were toxic to the parasite. Gibberellic acid induced theproduction of accessory leaves at all the concentrations used.Kinetin was also effective in inducing accessory leaves butonly at a concentration of 1.0 ppm. Some of the lower concentrationsof the growth substances affected significantly some aspectsof the parasite's growth and development. Nevertheless, thegrowth in vitro was still slower than that in vivo.     

EFFECTS OF GROWTH SUBSTANCES ON EXCISED FLORAL BUDS OF AQUILEGIA

Buds at various stages of development were grown for 3 weeks on solid media containing coconut milk, minerals, vitamins, sucrose, and varying quantities of gibberellic acid, indoleacetic acid, and kinetin. The best average growth was obtained on media containing GA at 2.0 mg/liter, IAA at 1.0 mg/liter, and kinetin at 0.05 mg/liter. When results were compared for buds explanted at very young stages and at older stages, however, young buds attained better average growth on media with 0.5 mg/liter of IAA. Evidence is presented for interaction between IAA and kinetin. With buds explanted at young stages, as the kinetin concentration was increased, optimum growth occurred on media with increasing concentrations of IAA. With older buds, on the other hand, as the kinetin concentration was increased, optimum growth occurred on decreasing concentrations of IAA. Bud growth was compared on media with growth substances sterilized by autoclaving with growth on similar media with these substances filter-sterilized. Better growth occurred generally on the media with filter-sterilized ingredients. A long-range objective of this research is the development of a system that would make possible quantitative measurements of floral development in vitro.     

Responses of tobacco pith nucleic to growth substances

Summary A cytological study was made of excised segments of pith of tobacco (ev. Wisconsin 38) grown in vitro on media containing various growth substances. Explants were cultured on a basal medium and on media containing kinetin, indoleacetic acid, gibberellic acid, and combinations of these three growth substances. Controls consisted of pith segments chemically fixed immediately after excision. The nuclei of explants on the basal medium and those on media containing the growth substances showed a response as early as 1 day after the beginning of the experiment. Nucleoli were indicators of initial stimulation in that they became swollen and displayed internal vacuolation, a known characteristic of active cells. These fatures were more pronounced by the second day for cultures grown on media containing growth regulators. Nucleoli were centers of nucleohistone staining as shown by the ammoniacal silver method. Continued and sustained growth of explants was possible only on media containing combinations of IAA and kinetin, or a combination of kinetin, IAA and GA. The greatest initial nucleolar stimulation occurred with IAA alone, even though subsequent growth did not take place. There was no nucleolar response of explants grown on abscisic acid alone.Abbreviations ABA abscisic acid - GA gibberellic acid - IAA indole-3-acetic acid The senior author is most grateful for support by fellowships from the Fulbright Commission and the John Simon Guggenheim Memorial Foundation. The authors wish to thank Mrs. Colette Nitsch, Miss Simone Hamon and Miss Linda Robinson for their valuable technical assistance.     

Growth Regulators and Wood Formation in Pinus silvestris

The formation of new xylem in the spring is preceded by bud development. In decapitated pine stem the formation of xylem is arrested until the outgrowth of interfascicular buds takes place. When indole-3yl-acetic acid (IAA) is applied to the cut surfaces of decapitated stems it induces the formation of a xylem ring on the whole length of 5-ycar old trees. Naphthaleneacetic acid (NAA) causes the formation of xylem; however, the width of the growth ring is several times broader at the point of application than at the base of the leader. Cis- and trans-cinnamic acids, coumarin, L-tryptophan, kinetin (Kin), benzylaminopurine (BAP) and gibberellic acid (GA) alone do not induce cambial divisions; however, GA and the cytokinins given jointly with IAA or NAA accelerated the basipetal stimulus which has been induced by the auxins, resulting in normal xylem formation. 2,3,5-Triiodobonzoic acid (TIBA) given jointly with IAA-induced formation of compression wood in the apical part of the stem and narrow diameter tracheids at the base. When carboxyl labelled IAA or NAA are applied to pine segments it is found that the basipetal movement of IAA is much quicker than that of NAA. GA and the cytokinins increase the rate of transport of both auxins, whereas TIBA arrests the bulk of auxin in the apical part of the stem.     

Application of the disk method: responses to growth regulators

The paper disk method of screening several plant growth regulators was evaluated. Leaf explants ofVigna unguiculata (L) Walp. were placed on solidified Murashige and Skoog's minimal organics medium containing 0.5 mg/l nicotinic acid. Hormones were tested, singly and in combinations, on paper disks in large Petri plates (150×20 mm). Hormones tested were 2,4-D (2,4-dichlorophenoxyacetic acid), 2,4,5-T (2,4,5-trichlorophenoxyacetic acid), IAA (indole-3-acetic acid), IBA (indole-3-butyric acid), picloram (4-amino-3,5,6-trichloropicolinic acid), dicamba (3,6-dichloro-2-methoxybenzoic acid), BA (6-benzyladenine), 2iP (2-isopentenyl adenine), and kinetin [6-(furfurylamino)-purine]. Root formation was stimulated by IAA and IBA; dicamba, picloram, 2,4-D, and 2,4,5-T stimulated callus formation. All cytokinins tested suppressed root formation. Dicamba in combination with either 2iP or kinetin induced the greatest callus formation. Root formation was optimal with kinetin and either IAA or IBA. The disk method provided a rapid, nonquantitative evaluation of callus and root formation from leaf disks.     

Recovery of loss in chlorophyll and 2,6-dichlorophenol indophenol Hill reaction of isolated chloroplasts during dark induced aging of intact maize seedlings

Loss in the content of pigments and decline in the efficiency of thylakoid membranes to reduce 2,6-dichlorophenol indophenol (DCPIP) have been investigated during dark induced senescence of attached leaves of maize seedlings. The chlorophyll degradation during senescence is differentially inhibited by indoleacetic acid (IAA), gibberellic acid (GA) and kinetin. IAA and GA behave as mild senescence inhibitors in comparison to kinetin. However, in comparison to light, kinetin is relatively less efficient in counteracting senescence. Dark-induced loss in chlorophyll content is fully recovered by light when the dark incubation period is relatively short. The pattern of light recovery of loss in photoelectron transport during dark-aging is similar to the recovery kinetics of chlorophyll. Continuous kinetin treatment of dark-incubated seedlings inhibits the chlorophyll degradation but with decreased duration of kinetin treatment, the efficiency of the hormone to inhibit chlorophyll loss is reduced. The kinetin-induced inhibition of pigment loss is small in comparison with the effect of light.     

Effects of gibberellic Acid, calcium, kinetic, and ethylene on growth and cell wall composition of pea epicotyls

The influence of gibberellic acid (GA), calcium, kinetin, and ethylene on growth and cell-wall composition of decapitated pea epicotyls (Pisum sativum L. var. Alaska) was investigated. Calcium, kinetin, and ethylene each caused an inhibition of GA-induced elongation of pea stems. Gibberellic acid did not reverse the induction of swelling by Ca²⁺, kinetin, or ethylene. Both Ca²⁺ and ethylene significantly inhibited the stimulatory effects of GA on the formation of residual wall material. Although GA promoted the development of walls relatively low in pectic substances and pectic uronic acid, Ca²⁺, kinetin, and ethylene favored the formation of walls rich in these constituents. Calcium, kinetin, and GA, alone or in combination, had no effect on the production of ethylene by pea epicotyls.     

Effect of Different Cytokinins on Ethylene Production by Mung Bean Hypocotyls in the Presence of Indole-3-Acetic Acid or Calcium Ion

Kinetin has been shown to act synergistically with indole-3-acetic acid (IAA) or calcium ion (Ca²⁺) to stimulate ethylene production. Several commercially available cytokinins (kinetin, kinetin-riboside, benzyladenine, benzyladenine-riboside, isopentenyladenine, isopentenyladenine-riboside, and zeatin) as well as noncytokinin bases (adenine and xanthine) were administered to mung bean (Phaseolus aureus Roxb.) hypocotyls to study their effects, alone or in combination with IAA or Ca²⁺, on ethylene production. In the presence of IAA or Ca²⁺, all cytokinins tested synergistically stimulated ethylene production and were as effective or nearly as effective as kinetin. Noncytokinin bases (adenine and xanthine) were, however, inactive in this system.     

The Effects of Cytokinins on Unfolding of Cotyledons and Opening of Hypocotylar Hooks of Cucumber

Benzyladenine (BA), zeatin and kinetin caused the unfolding of cucumber (Cucumis sativus L. Var. Jing-ye No. 4) cotyledons. But the other plant hormones e.g. gibberellic acid (GA3), indoleacetic acid(IAA), ethylene (ethrel), abscisic acid (ABA) and other related compound e.g. KCl, adenine were not so. This response was caused by even lower concentration (0.01 ppm). Therefore, it could be used as a bioassay for cytokinins. KCl affaeted synergistically on cytokinin-induced unfolding. Basally applied BA (10 ppm) stimulated the opening of the intact hypocotylar hook. The light affacted synergistically on BA-induced opening. However, basally applied BA prevented the opening of decapitated hooks, especially in the light. The cotyledons were found to play a part of effects in maintaining the hook.     

THE INFLUENCE OF GROWTH SUBSTANCES ON THE INDUCTION OF APOGAMY IN PTERIDIUM GAMETOPHYTES

Experiments employing sequences of three media demonstrated the effect of growth substances on the induction of apogamy. The most effective sequence was 4% sucrose 1–14 days, 4% sucrose plus growth substance 15–28 days, and 0.1% sucrose 29–56 days. In this sequence concentrations of NAA, IAA, and GA promoted apogamous shoot formation. A higher NAA concentration than optimal for shoot formation stimulated apogamous root formation in all medium sequences. Kinetin was without effect or inhibitory to apogamy. Combination of kinetin/GA or GA/NAA concentrations did not increase the apogamous response. One combination of the kinetin/NAA concentrations had a synergistic effect on the apogamous shoot formation. Additions of GA to the synergistic kinetin/NAA combination had an antagonistic effect on the apogamous response.     

Efficient propagation and rooting of three citrus rootstocks using different plant growth regulators

The influence of various basal medium and plant growth regulators on the efficient micropropagation of nodal explants from mature trees of alemow, sour orange, and ??Cleopatra?? mandarin citrus rootstocks was studied. All three citrus rootstock shoot cultures showed a preference for high-salt media, like Murashige and Skoog or Driver and Kuniyuki Walnut medium. Several combinations of N ⁶-benzyladenine (BA) and adenine (AD), kinetin (KIN) or gibberellic acid (GA) were tested to optimize the shoot proliferation phase. BA/GA combinations improved the proliferation of all the rootstocks studied, especially alemow. The addition of BA and AD to the culture medium improved shoot proliferation in sour orange and ??Cleopatra?? mandarin in the same way as BA and GA. The addition of different combinations of BA/KIN did not result in further improvement of any of the studied variables. The transfer of in vitro shoots to rooting media, containing different concentrations of indolebutyric acid (IBA) and indoleacetic acid (IAA), resulted in regeneration of complete plantlets. Alemow and ??Cleopatra?? mandarin shoots rooted well using these plant growth regulators; however, all combinations of IBA and IAA tested resulted in very low rooting percentages in sour orange. To improve rooting in sour orange and ??Cleopatra?? mandarin, different combinations of naphthaleneacetic acid (NAA) and IBA were tested. All NAA/IBA combinations produced higher rooting percentages than did the IBA/IAA combinations, and in sour orange nearly 100 % of explants developed roots. An efficient and simple protocol for the micropropagation of three citrus rootstocks, alemow, ??Cleopatra?? mandarin, and sour orange, by culturing nodes from mature plants, has been established.     

A Comparative Study of the Changes in Root Growth, Induced by Coumarin, Auxin, Ethylene, Kinetin and Gibberellic Acid

The effect of coumarin, IAA, ethylene, kinetin and gibberellic acid on roots of maize and wheat was investigated. Sterile attached and detached roots and isolated elongation zones were used. In some experiments a semi-sterile procedure was followed. The effects of the different regulators separately or in various combinations together with coumarin were studied on the root growth with regard to division, elongation and swelling of the cells. The ethylene production in isolated elongation zones was measured after treatment with coumarin, IAA, PCIB, kinetin, colchicine and dinitrophenol. The results show the following: 1) Each substance produces a specific morphologic pattern. 2) Changes in polarity were demonstrated for auxin-induced swelling in cell divisions and cell expansion and for coumarin-induced swelling in cell divisions. Other cell expansion in swollen parts was due to cylindric cells increasing in width while retaining their cylindric form. 3) Coumarin-induced inhibition could not be counteracted by IAA, PCIB, carbon dioxide, kinetin, gibberellic acid or Cycocel. 4) The ethylene production in isolated elongation zones increases noticeably after kinetin treatment, less strongly after auxin treatment and least after coumarin treatment. The production of ethylene does not seem to be correlated with the morphogenetic effect of the different substances. 5) The isolated elongation zones reacted to a) IAA and kinetin with an increase in length in some cases and b) gibberellic acid with a reduction of their width. 6) The inhibitory effect of coumarin on the growth in length of the elongation zones was diminished by kinetin. The swelling produced by coumarin in these zones was reduced by gibberellic acid. The effects just mentioned of kinetin and gibberellic acid were considered as indirect ones. - From the present findings it was concluded that concomitant effects of auxin, ethylene, cytokinins and gibberellins are not obligatory for coumarin to exert its morphogenetic effects on root growth. In discussing the results some pitfalls in studies of growth reactions after application of hormones to roots containing meristem were emphasized.     

Changes in the level of endogenous gibberellins and auxins in apical buds ofChenopodium rubrum L. after application of growth substances reversing the effect of (2-chlorethyl)-trimethylammonium chloride (CCC) on flowering

The application of CCC at concentrations inhibiting flowering ofChenopodium rubrum reduces the level of endogenous gibberellins in the apical buds of the plants. The effect of CCC may be reversed by appropriate concentrations of gibberellin (GA-), indole acetic acid (IAA) or kinetin. Kinetin applied to the apical bud during floral induction reduced the level of endogenous gibberellins similarly as CCC and if both CCC and kinetin were applied simultaneously their action was additive. On the other hand IAA applied under the same conditions increased the level of endogenous gibberellins and after joint application of CCC and IAA their level was the same as in untreated control plants. After application of CCC during floral induction the level of endogenous auxins did not change markedly but an active substance “x” appeared on the chromatograms of indole compounds. This substance was found also after simultaneous application of GA- and CCC but not after joint application of CCC and kinetin. If follows from our results that the same morphological phenomenon (flowering) can take place in plants considerably differing as to their level of endogenous growth substances. The ratio of different growth substances is obviously more important than the actual level of the single substances.     

Correlative Growth in Seedlings of Phaseolus vulgaris L. : Inhibition of Stem Growth by the Primary Leaves

Primary leaves of one-week-old seedlings of dwarf beans effectivelyinhibit stem elongation either in natural daylight or in continuousdarkness. Stem elongation is promoted by exogenous gibberellicacid and kinetin and inhibited by indol-3-ylacetic acid (IAA)or abscisic acid, and it is suggested that IAA is the inhibitorysubstance emanating from the primary leaf blades which affectsthe growth of the stem.     

Short-term Effects of Some Chemicals on Cambial Activity

Aqueous solutions of indol-3yl-acetic acid (IAA), 1-naphthyl-aceticacid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D), gibberellicacid (GA), 6-furfuryl-aminopurine (FAP), myo-inositol, and sucrosewere applied singly and in mixtures to the apical ends of disbuddedstem segments of willow. After 4 weeks all substances had hadsome effect on differentiation of xylem from cambial derivatives.The production of potential xylem cells, as well as their differentiationwere most markedly enhanced when IAA, GA, and FAP were appliedtogether, although the response was further augmented by additionof inositol or sucrose. The action of the substances when appliedas mixtures was often synergistic. This means that it is difficultto assess the role of different chemicals in xylem productionby extrapolation from experiments involving the applicationof single substances.     

春萝卜抽薹过程中内源激素含量变化分析

采用酶联免疫吸附法(ELISA)测定了春萝卜抽薹过程中顶端生长点处嫩叶的赤霉素(GA₄)、生长素(IAA)、细胞分裂素(iPA)和脱落酸(ABA)等含量的变化情况。研究结果表明,在萝卜抽薹过程中,GA₄含量在现蕾点存在明显的峰值,表明GA₄与现蕾抽薹密切相关,起着主导作用;内源ABA含量变化与GA₄变化趋势基本一致;与GA₄和ABA含量相比,抽薹过程中叶片IAA含量相对较低;iPA含量在现蕾期逐步增加,至抽薹中后期出现一高峰值。研究发现iPA/ABA值增大,GA₄/iPA和GA₄/IAA值变小有利于花芽分化后萝卜植株抽薹开花,表明内源激素之间的平衡对萝卜抽薹开花也起着重要作用。     

Development of Seeded and Seedless Hypanthium of Rosa Canina After Application of Growth Substances

Dog rose (Rosa canina L.) plants in the bloom stages of flowering were sprayed by indole-3-acetic acid (IAA) in concentrations of 0.06 and 0.60 mM and gibberellic acid (GA₃) in concentrations of 0.60 and 1.50 mM. Ascorbic acid, total sugar, reducing sugar and carotenoid contents gradually increased, while the protein content remained unchanged and the content of phenolic substances decreased during hypanthium development. Ascorbic acid, total sugar, reducing sugar and carotenoid contents increased in hypanthium sprayed by GA₃ and IAA. However, IAA and GA₃ applications (except low concentrations) decreased contents of phenolic substances. IAA and GA applications might be a good way to produce the high quality hypanthium in R. canina.