CONTROL OF IN VITRO SEPAL DEVELOPMENT OF EXCISED FLORAL BUDS OF AQUILEGIA (RANUNCULACEAE)

Excised floral buds of Aquilegia formosa Fisch. were grown on a coconut-milk medium containing the minerals and vitamins of Murashige and Skoog, sucrose, and kinetin. The plant growth regulators indoleacetic acid (IAA, 0.5 mg/liter) and gibberellic acid (GA, 2.0 mg/liter) were added singly and in combinations; both were omitted from the control medium. The addition of GA to the basal medium was required to support sepal development on flowers at all phases of development. The formation of stomatal complexes in the epidermis of the sepals occurred only in the presence of GA. Sepals grown in the presence of GA also contained trichoblasts and developing trichomes, while none were formed in the absence of GA. The role of IAA in the development of these idioblasts was not clear but it appeared to have no effect. The hormones GA and IAA had different effects on the growth of the sepals. In the presence of GA the sepals increase in length until comparable with sepals grown in vivo. However, the sepals remained small when GA was omitted from the medium. Upon closer examination of this effect, it was determined that there was a direct proportionality between an increase in cell number in the epidermis and an increase in sepal length. The role of the two hormones in increasing epidermal cell length in sepals was distinct and separate. Exogenous IAA had no effect upon cell division but was required for cell elongation, while GA was required for cell division and had no effect on cell elongation. The GA effect in promoting cell division in the sepals was substantiated by use of autoradiography. If the buds were grown on media with GA, twice as many epidermal cells along the central file incorporated significant amounts of tritiated thymidine. The cell cycle of the epidermal cells of the GA-treated sepals was ca. 8.7 hr in duration and ca. 13.0 hr if GA was deleted from the medium.     

The Effects of Kinetin, Gibberellic Acid, and Light on Expansion and Cell Division in Leaf Disks of Dwarf Bean (Phaseolus vulgaris)

The effects of kinetin (Kn), gibberellic acid (G), and light(L) on cell expansion and division in disks from 6-day-old etiolatedprimary leaves of dwarf bean are described. Cell number wasdetermined by direct counting after disks were digested in apectinase/EDTA mixture to separate the cells. Kn increased leafexpansion wholly by increasing cell size. G also increased cellsize; it increased cell division in the dark but not in thelight. Light also increased cell size and cell division; iteliminated the effect of G on cell division but enhanced theeffect of G on cell expansion.     

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.     

Involvement of wall microtubules in gibberllin promotion and kinetin inhibition of stem elongation

The elongation of light-grown azuki bean (Azukia angularis =Vigna angularis) epicotyl segments was promoted by indoleaceticacid (IAA) and this IAA-induced elongation was inhibited byboth kinetin and benzimidazole (BIA). Increased stem thickeningwas observed with kinetin- or BIA-treated segments, but thiswas not accompanied by incresed cell number in the transversedirection, suggesting that both kinetin and BIA promoted lateralcell expansion. Colchicine at a concentration with no effecton IAA-induced elongation reversed both the kinetic- and BIA-inducedinhibition. Electron-microscopic examination revealed that wall microtubulesin cells treated with kinetin together with IAA ran parallelto the cell axis, while wall microtubules in cells treated withonly IAA were randomly oriented. In the cell treated with gibberellintogether with IAA, wall microtubules ran tranverse to the cellaxis. (Received July 13, 1973; )     

Involvement of cellulose synthesis in actions of gibberellin and kinetin on cell expansion. Gibberellin-coumarin and kinetin-coumarin interactions on stem elongation

In azuki bean (Azukia angularis = Vignia angularis) epicotylsections, 5 ? 10^–4 M coumarin inhibited the incorporationof radioactivity from [U^–14C]glucose into the cellulosefraction by 35% in the absence of indole-3-acetic acid (IAA)and by 40% in the presence of 1 ? 10^–4 M IAA. There wasno inhibitory effect on the incorporation of radioactivity intothe other fractions. Coumarin at 5 ? 10^–4 M reversed thepromoting effect of 1 ? 10^–5 M gibberellin A₃ (GA) andthe inhibitory effect of 1 ? 10^–5 M kinetin on IAA-inducedelongation of sections with no significant effects on IAA-inducedelongation. Neither GA nor kinetin had any appreciable effectson cellulose synthesis. No inhibition of cellulose syntheiswas observed with 1 ? 10^–3 M colchichine, which has beenreported to have effects similar to those of coumarin on GA-or kinetin-affected stem elongation. Coumarin at 5 ? 10^–4M was ineffectual in breaking up wall microtubules, while adisrupting effect on wall microtubules was clearly demonstratedwith 3 ? 10^–4M colchicine. From these results, the possible involvement of cellulose synthesisin cell expansion controlled by GA or kinetin was suggested. (Received August 3, 1973; )     

Control of Hypocotyl Hook Angle in Phaseolus mungo L. Role of Growth Substances

Effects of growth hormones on the hook angle and light responseof Phaseolus mungo L. hypocotyl hooks are described and theresults are discussed with reference to the functions of otherparts of the seedling in controlling the growth and shape ofthe hook. Apically applied IAA (indolyl acetic acid) prevented hook openingin decapitated seedlings in the dark and in all the red-irradiatedseedlings. [¹⁴C]IAA experiments showed that only a small quantityof IAA (2–6 ?g per hook) was required to produce theseeffects, and that transport of IAA through the hook was negligibleand unaffected by red irradiation. ABA (abscisic acid) had little effect on the hooks or theirlight response. Applied ethylene and IAA-induced ethylene slightly closed thehooks, but only slightly reduced light-induced opening. IAAreduced the effect of ethylene in the dark, but after irradiationthe hooks appeared more sensitive to the ethylene in the presenceof IAA, resulting in light-induced hook closure. Basally applied kinetin (6-furfurylaminopurine) prevented decapitatedhooks from opening in the dark, especially when GA₃ (gibberellicacid) was also present. Some combinations of kinetin and GA₃(with high kinetin concentrations) also prevented light-inducedopening, but combinations with lower kinetin concentrationsallowed almost as much opening as was found in intact hooks. It is proposed that the terminal parts act by regulating thesupply of cytokinins and gibberellins from the basal parts,and that IAA does not mediate this funotion in this species. The results are compared with those reported for other species.     

Habituation in Nicotiana bigelovii tissue cultures: Different behaviour of two varieties

The exceptionally high capacity for transformation to autotrophyfor hormones (habituation) discovered in Nicotiana bigeloviiled us to a comparison of the responses of 2 varieties (quadrivalvisand bigelovii) to different hormone treatments applied to theirseeds. Hormones used were: kinetin (0.8, 1.6, 3.2 ppm), 2,4-D(0.1, 0.4 ppm), IAA (2 ppm) and NAA (1, 2, 4 ppm). Callus formation and habituation were scored after transferon minimal medium (without hormones) 3 months after sowing.Var. quadrivalvis showed a much higher ability to form callusbut the callus formed was autonomous in a lower percentage ofcases when IAA or kinetin treatment was considered. Analysisof auxin-like substances in leaves of the 2 varieties showeda higher content in N. bigelovii var. quadrivalvis. Data arediscussed with particular regard to their relevance to tumorousphenomena in plants. ¹ Publication No 52 of the Laboratorio di Mutagenesi e Differenziamento,C. N. R. (Received March 30, 1971; )     

Comparative Effects of Indoleacetic Acid and Gibberellic Acid on Growth of Decapitated Etiolated Epicotyls of Pisum sativum cv. Alaska

Measurements were made of the growthof the sub-apical region of decapitated, etiolated epicotyls of Pisum sativum L. cv. Alaska after treatments with indoleacetic acid (IAA), gibberellic acid (GA) and triiodobenzoic acid (TIBA). Growth was measured either at the end of a 2-day period, at short intervals during growth, or was monitored continuously for 2–3 h using a position-sensing transducer. In experiments measuring growth after 2 days, high levels (0.1–10 μg/plnat) of IAA caused expansion, whereas similar levels of GA caused elongation. When both hormones were applied together, the effects of IAA were dominant and expansion ensued, even when GA was present at 100 times the amount of IAA. Very low amounts of IAA (0.5–5 ng/plant), however, caused elongation. The elongation elicited by high GA or low IAa was inhibited to a similar extent by TIBA and this inhibition of elongation was associated with an increased expansion at the extreme tip. When application of the hormones was delayed, GA-induced elongation was reduced considerably, IAA-induced elongation was lessened somewhat and IAA-induced expansion was partially converted into elongation. In experiments measuring elongation at short intervals, high levels of IAA caused rapid elongation followed after 3 to 6 h by expnasion. Both GA and low levels of IAA extended the duration of elongation with little apparent effect on the rate of growth. In fast-growth experiments, low, intermediate and high levels of IAA doubled the rate of elongation with a lag period of about 20 min, whereas GA had at most a very slight stimulatory effect on the growth rate. It is concluded that the main role of GA in this system is to maintain physiological levels of IAA in the growing zone and that the level of IAA present determines whether elongation or expansion will take place.     

Alleviation of Seed Dormancy in the Desert ForbZygophyllum simplexL. from Pakistan

Zygophyllum simplexL. is a succulent annual that grows on thecoastal and inland saline flats around Karachi, Pakistan. Theseeds are moderately salt tolerant during germination. GerminationofZygophyllum simplexseeds under various salinity, proline,betaine, GA and kinetin treatments was determined. Proline (0.1and 1 mM) and betaine (0.1 and 1 mM) alleviated the innate dormancyof seeds, and germination reached 60–70% compared to 12%in the control set. At low salinity compatible osmotica alleviatedsome effects of salinity, but at higher NaCl concentrationsboth proline and betaine were ineffective. Gibberellic acid(0.3 and 3 mM) and kinetin (0.05 and 0.5 mM) substantially alleviatedboth innate as well as salinity-induced seed dormancy. At highersalinity (125 mM), low concentrations of kinetin (0.05 mM) andhigh concentrations of GA (3 mM) were more effective. GA completelyalleviated the effect of salinity at all concentrations used. Betaine; desert; dormancy; forb; GA; germination; halophyte; kinetin; proline; seeds; Zygophyllum simplex     

Effect of phytohormones on chlorophyll degradation during aging of chloroplastsin vivo andin vitro

Summary Phytohormones like IAA and kinetin inhibit chlorophyll loss during aging of wheat chloroplasts duringin vivo andin vitro. GA, on the other hand, stimulates the pigment degradation during aging of attached leaves in contrast to its senescence inhibiting action in detached leaves and isolated chloroplasts. A shift in optimum concentration of hormone in inhibiting chlorophyll degradation suggests a differential pool size of endogenous hormone regulating aging of chloroplastsin vivo andin vitro. The retardation of chlorophyll loss by kinetin, IAA and GA during aging of chloroplastsin vitro would indicate that the effect of hormones in preventing yellowing of senescing leaves may be mediated through their direct action on chloroplasts.     

Changes in enzyme activity during elongation and tuberization of stolons of Solanum tuberosum L. cultured in vitro

Isolated stolons of Solanum tuberosam L. were cultured in vitroin the presence of kinetin, which induced tuber initiation orgibberellic acid which inhibited initiation. Progressive changes in enzyme activity, at the locus of tuberinitiation, were monitored at specified intervals. In the presenceof kinetin soluble invertase activity was decreased with timewhereas gibberellic acid (GA) evoked substantial increases inactivity. Acid phosphatase activity was enhanced by GA but changedonly slightly under tuber inducing conditions. Enzymic hydrolysisof glucose-6-phosphate and fructose-6-phosphate decreased duringthe course of tuber induction but increased in the presenceof GA. In contrast hydrolysis of 3'AMP was stimulated undertuber inducing conditions. GA evoked substantial increases in peroxidase activity duringthe initial stages of incubation while under tuber inducingconditions increased activity was only observed after 5 days.Substantially higher levels of IAA oxidase activity were associatedwith tuber initiation. RNase activity decreased with time undertuber inducing conditions but showed an initial stimulationin the presence of GA. These results are discussed with referenceto the role of these enzymes in carbohydrate metabolism andthe regulation of hormone levels. (Received February 29, 1972; )     

Hormonal Control of Xylogenesis in Pith Parenchyma Explants of Lactuca

The time course of xylem differentiation was determined in explantsof lettuce pith parenchyma (Lactuca sativa L. cv. Romana) culturedon Murashige and Skoog (1962) medium using different concentrationsof auxin (IAA) and one cytokinin (zeatin or kinetin). Increasinglevels of auxin from I mg 1^–1 to 15 mg 1^–1 in thepresence of a constant level of a cytokinin (zeatin or kinetin)yielded up to 10 mg 1^–1 IAA, an increase in the numberof tracheary element formations. Cytokinin concentrations aboveand below o.1 mg 1^–1 interacting with an optimal xylogenicamount of auxin inhibited xylogenesis. The IAA (10 mg 1^–1)-zeatin(0.1 mg ^1–1) treatment produced the greatest number oftracheids, while kinetin compared to zeatin did not producesuch an effect. The different effectiveness of zeatin and kinetinin inducing tracheary element formations was not due to a differentcapacity of the two cytokinins to stimulate cell division butit seems likely that zeatin, because of interaction with IAA,is more active than kinetin in the determination of the dividingcells in a specific type of cytodifferentiation. The IAA (10mg 1^–1)-zeatin (0.1 mg 1^–1) treatment produced about6.9 per cent tracheids with respect to cell division while IAA(10 mg 1^–1)-kinetin (0.1 mg 1^–1) produced 4.2 percent. These results are discussed with reference to the problemsof hormonal control of xylem differentiation.     

Developmental changes in the gibberellin-induced growth response in stem segments of light-grown pea genotypes

The effects of GA on stem elongation were studied using segments from one tall and three dwarf light-grown pea genotypes varying in endogenous hormone content. Stem segments were cut at two distinct ages: when the fourth internode was at about 6–13% of full expansion (early-expansion) or at 18–25% of full expansion (mid-expansion). Light microscopy and flow cytometry were used to demonstrate that GA does not induce cell division in excised pea stem segments. The growth studied here was strictly elongation. Measurement of final segment length after 48 hours and high resolution measurement of growth kinetics over 20 hours using an angular position transducer were done on segments treated with hormone solutions. Our data indicate that the action of GA on stem elongation can be classified into two distinct modes. The first, apparent in early-expansion stem segments, shows distinct growth kinetics and is independent of the endogenous IAA concentration of the segments. Quantitation of IAA by GC/MS in early-expansion segments of wild type pea incubated with gibberellin shows that an increase in IAA concentration is part of the GA response in such segments. The second mode of GA action is evinced in mid-expansion segments. Whereas there is no short term (<20 h) response to GA alone (as determined by growth kinetics), there is a long term (48 h) response whose magnitude decreases across the genotypes with decreasing endogenous hormone content. Growth responses indicate that in mid-expansion segments exogenous GA acts by enhancing IAA action but appears to be unable to augment endogenous IAA content. Contradictory reports of the response of excised stem segments to GA can be reconciled when tissue genotype and developmental stage are considered.     

Effects of N-1-naphthylphthalamic acid on the growth and bud formation of tobacco callus grown in vitro

The effect of N-1 -naphthylphthalamic acid (NPA), indole-3-aceticacid (IAA) and kinetin on callus growth and bud formation wasstudied mainly by a tobacco callus culture method. Callus producedfrom Nicotiana tabacum var. Wisconsin 38 was used as the testplant material. Callus growth on nutrient agar containing 2mg/liter of IAA was promoted by NPA added at a concentrationof 0.5 mg/liter with 0.4 mg/liter of kinetin or by NPA addedat 5 mg/liter in the absence of kinetin. At a high concentrationof 50 mg/liter, however, NPA inhibited growth on the mediumcontaining 2 mg/liter IAA and no kinetin. Kinetin reduced thisNPA inhibition. In the presence of 0.4 mg/liter kinetin and2 mg/liter IAA, when the concentration of NPA was 50 mg/liter,buds were initiated after calluses were grown on the test mediumfor 7 weeks in dim light, but no buds formed when NPA was omittedfrom the above medium. The control of callus growth and bud initiation is based onthe active ratio of auxin (IAA) to cytokinin (kinetin) in themedium and NPA added to the medium can promote or inhibit callusgrowth and induce bud formation. Therefore, it is proposed thatNPA can itself reduce auxin activity or enhance cytokinin activityand hence change the active ratio of the two regulators. NPAmay enhance the activity of cytokinin (here supplied as kinetin)but cannot substitute for it. ¹Present address: Department of Biology, Wisconsin State University,Oshkosh, Wisconsin 54901, U. S. A. (Received March 10, 1969; )     

The Effect of Growth Substances on Growth of Excised Embryo Shoot Apices of Wheat in Vitro

Excised embryo shoot apices of wheat were grown on media containingeither indolylacetic acid (IAA), kinetin, gibberellic acid (GA),or adenine. GA and adenine stimulate the activity of the apicalmeristem whereas IAA and kinetin do not. No combination of anytwo substances significantly stimulated growth when comparedwith the most active substance present. GA can overcome theinhibitory effect of IAA or kinetin. The possible function ofthese substances in the apical meristem is discussed.     

Studies on seasonal variation of plant hormones,nitrogen flixation and indole acetic acid metabolism in root nodulus ofButea monosperma (Lam.) Taub.

Summary Mature nodules ofButea monosperma (Lam.) Taub. (B. monosperma) contained higher amounts of indole acetic acid-like (IAA), cytokinin-like (CK), gibberellic acid-like (GA) and abscisic acid-like (ABA) substances, and were more active in IAA metabolism and nitrogen fixation. Roots contained lower amount of all the hormones. The higher level of IAA in the nodules than the roots was due to more synthesis from tryptophan and simultaneously less destruction by IAA oxidase. IAA oxidase, in turn, was regulated by phenols.Regardless of the hormone levels, the nodules of a particular age attained a fixed size indicating that regulation of formation and growth of the nodules are not the sole function of the nodular hormones. Seasonal studies on the mature nodules showed positive relation of nitrogenase (N₂ase) with IAA and CK, and negative relation with GA and ABA.     

Influence of phytohormones on morphology and chlorophyll a fluorescence parameters in embryos of Fucus vesiculosus L. (Phaeophyceae)

While a variety of plant hormones from brown algae were described, there were few studies that examined the combined effects of these hormones on morphogenesis and photosynthetic physiology in developing fucoid embryos. We evaluated the effects of phytohormones to determine the extent, to which responses were similar to those of terrestrial plants. Kinetin, IAA, ABA, GA₃, and kinetin + IAA were added to seawater at a physiological concentration (1 mg/L), and embryos of Fucus vesiculosus L. were grown for 10 days. Photosynthetic activity of single embryos or embryo cells were characterized using the following fluorescence parameters: minimum fluorescence yield (F ₀), maximum quantum yield (F _v/F _m), relative maximum rate of electron transfer to photosystem II under saturation irradiances (rETR_max), photosynthetic efficiency under non-saturating irradiances (αETR) and saturation irradiance (E _k). In addition, embryo length and diameter and apical hair length and number were determined. Morphological changes associated with hormone treatments included an increase in the embryo length in the presence of IAA, an increase in the embryo diameter in the presence of IAA, kinetin, and kinetin + IAA, an increase in the maximum hair length and number in the presence of kinetin + IAA, and a decrease in the hair length and number in the presence of ABA. With respect to fluorescence parameters, significant effects of phytohormones included an increase in the F ₀ and F _v/F _m at kinetin treatment, a synergistic effect of kinetin + IAA on F _v/F _m, rETR_max, and αETR, a promotion of F _v/F _m by GA, and a decrease of the parameters by ABA. These results are consistent with the data on responses of land plants to the same hormones and suggest that brown algae have evolved regulatory mechanisms for morphogenesis and photosynthetic regulation similar to plants.     

Hormonal control of endoreduplication in sugar beet (Beta vulgaris L.) seedlings growing in vitro

The effect on endoreduplication in sugar beet (Beta vulgaris L.) seedlings of five plant hormones in MS medium, ethylene, 24-epibrassinolide (EBR), gibberellic acid (GA(3) ), kinetin and 1-naphthaleneacetic acid (NAA), as well as a combination of kinetin and NAA at two different concentrations, was studied using flow cytometry. Analyses of DNA content in nuclei of the root, hypocotyl and cotyledons of seedlings growing in vitro were performed during their early development, starting from when the root was 0.5-1.0 cm long until expansion of the first pair of leaves. The proportions of nuclei with different DNA contents were established and the mean C-value calculated. The presence of exogenous plant hormones changed endoreduplication intensity, although to different extents, depending on the organ and developmental stage. Ethylene and NAA stimulated the process, while EBR and kinetin suppressed it and GA did not clearly affect it.     

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.     

Hormone Interaction in Apical Dominance in Phaseolus vulgaris L.

Gibberellic acid (GA₃), kinetin, and indole-3yl-acetic acid(IAA) were applied to roots of Phaseolus vulgaris under twodifferent light intensities and when either young or old leaveswere removed In all cases GA₃, promoted stem and lateral growth,especially when light intensity was reduced. Promotion by GA₃,of stem growth under reduced light was reduced if IAA and kinetinwere present; promotion of lateral growth under reduced lightwas reduced if IAA was added and eliminated if kinetin or kinetinplus IAA were added to GA₃. Removal of young and mature leavesreduced main stem growth; removal of young leaves promoted,and of mature leaves reduced, lateral shoot growth. We suggestthat shoot growth and apical dominance are governed by the balanceof hormones present in elongating internodes. There may be twoways of modifying this balance; firstly by altering light, temperature,or nutrients, or by applying hormones generally to the plant.Secondly, local modifications can be made by removing apicesor young leaves, or applying hormones in lanolin to specificareas. Knowledge of both the general and local conditions maybe necessary for a complete understanding of apical dominance.