Interaction of paclobutrazol and indole-3-butyric acid in relation to rooting of mung bean (Vigna radiata) cuttings

Paclobutrazol (PB) only slightly stimulated the rooting of mung bean cuttings but, interestingly, the number of adventitious roots formed was dramatically increased when PB was used together with indole-3-butyric acid (IBA). Application of PB in the first phase of root formation, when root initials are induced, caused the greatest enhancement of the promotive effect of IBA on rooting. Investigation of the effect of PB on uptake, transport and metabolism of [5^-3H]-IBA in mung bean cuttings revealed some changes in the rate of metabolism of IBA in comparison with control cuttings. PB was found to be involved in the partitioning of carbohydrates along the cuttings. Application of sucrose, like PB to the base of IBA-treated cuttings enhanced the effect of IBA. The patterns of the effects of PB and IBA, separately and together, on rooting were similar in defoliated and intact cuttings, however the number of roots was much lower in the defoliated cuttings, which lacked a source of assimilates. PB counteracted the effect of GA₃ in the upper regions of the cuttings and seemed to increase the sink capacity at the base of the cuttings. The results of the present study clearly demonstrated the enhancing influence of PB on IBA stimulation of the rooting of mung bean cuttings. It is suggested that PB may affect the rate of metabolism of IBA during rooting and the status of the local sink, in the base of the cuttings, thus partially contributing to the enhancement of the rooting-promotive effect of IBA.     

Interaction of auxins with growth-retarding, -inhibiting and ethylene-producing chemicals in rooting of cuttings

The growth retarding chemical Cycocel significantly synergizedIAA- and IBA-induced rooting of mung bean and IBA-induced rootingof tomato cuttings. In marigold, Cycocel and B-Nine promotedrooting when applied alone but did not synergize auxin-inducedroot formation. Chlorflurenol, a growth-inhibiting morphactin,antagonized auxin-induced rooting of all three species; inhibitionwas especially marked in the presence of IBA. The ethylene-producingchemical Amchem promoted rooting of mung bean and marigold cuttingswhen applied alone. In tomato cuttings, Amchem significantlysynergized IAA-induced root formation. (Received July 5, 1972; )     

Stimulation of adventitious root formation in non-woody stem cuttings by uridine

Uridine strongly stimulated adventitious root formation in stem cuttings of sunflower (Helianthus annuus L.), mung bean (Vigna radiata L.) and common bean (Phaseolus vulgaris L.). A dose response curve of uridine induced rooting showed that the optimum concentration of uridine was 0.1 µM. At all concentrations employed, uridine had no significant effect on root elongation. The rooting response of stem cuttings to the optimal concentration of indole-3-butyric acid (10 µM) in combination with 0.1 µM uridine did not significantly differ from their response to either of these compounds when applied alone. However, the rooting response of the cuttings to sub-optimal IBA (0.01 µM) was significantly stimulated by uridine. These findings suggested that uridine may have stimulated rooting by increasing the sensitivity of the rooting tissue to auxin.     

Ethylene and auxin-ethylene interaction in adventitious root formation in mung bean (Vigna radiata) cuttings

The role of ethylene in adventitious root formation and its involvement in auxin-induced rooting were investigated in cuttings ofVigna radiata (L.). Treatment with 30 M indole-3-acetic acid (IAA) for 24 h slightly inhibited rooting, whereas the same concentration of indole-3-butyric acid (IBA) significantly stimulated it. Ethylene derived from 1-aminocyclopropane-1-carboxylic acid (ACC) increased the number of adventitious roots but inhibited their emergence and elongation. Endogenous levels of ethylene, ACC, and malonyl-ACC (MACC) were initially higher in cuttings treated with IAA. This trend was quickly reversed, and cuttings, particularly hypocotyls, treated with IBA produced higher levels of ethylene and had more ACC and MACC during most of the rooting process. Aminoethoxyvinylglycine significantly inhibited rooting, but its inhibitory effect could not be reversed by ACC. The data suggest that the stimulating effect of IBA on rooting is closely associated with its induction of ACC and ethylene biosynthesis.     

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.     

Isolation, Purification, and Characterization of an Endogenous Root-promoting Factor Obtained from Basal Sections of Pear Hardwood Cuttings

Basal segments taken from Old Home and Bartlett pear hardwood cuttings collected at intervals during the rooting period in September were extracted with ethanol and fractionated by paper chromatography in different solvent systems. Different zones on the chromatograms were bioassayed by the mung bean rooting test, which showed high levels of promotion in Old Home basal extracts when the cuttings were obtained during the period of maximum rooting. Extracts from Bartlett cuttings, however, showed considerably less promotion activity in the bioassay but did show high levels of inhibitory activity.

After the easily-rooted Old Home cuttings had been in the rooting medium for 10 days, a highly active endogenous root-promoting material was found in extracts from basal segments of cuttings having buds and which had been treated with indolebutyric acid. Similar extracts obtained from disbudded cuttings, or from cuttings with buds but not treated with indolebutyric acid, lacked this rooting-factor. Extracts obtained from all types of the difficult-to-root Bartlett cuttings also lacked this rooting-factor. The latter is believed to be produced by physiologically active Old Home buds, and is very effective in the mung bean bioassay, even at extremely low concentrations.

From paper chromatographic studies, tests with spray reagents, solubility determinations, biological tests, UV spectrum analysis, and infrared spectroscopy, it is believed that this rooting factor could be a condensation product between exogenous auxin (indolebutyric acid) and a phenolic compound produced by physiologically active Old Home pear buds.

     

N-phenyl indolyl-3-biityraitiide and phenyl indole-3-thiolobutyrate enhance adventitious root primordium development

N-phenyl indoryl-3-butyramide (NP-IBA) and phenyl indole-3-thiolobutyrate (P-ITB) are new synthetic auxins that enhance rooting of bean ( Phaseolus vulgaris cv. Top Crop) hypocotyl cuttings and jack pine ( Pinus banksiana Lamb.) seedling cuttings. In comparison with an equal concentration of indole-3-butyric acid (IBA), NP-IBA treatment resulted in greater rooting of bean and jack pine seedling cuttings. IBA and P-ITB treatment at equal concentration were equally effective in inducing rooting of bean cuttings. However, in comparison with an equal concentration of IBA, P-ITB treatment promoted greater rooting of jack pine seedling cuttings, but P-ITB treatment was less effective than NP-IBA treatment. Application of crystalline chemical to jack pine seedling cuttings indicated lesser toxic effects and earlier and greater root-promoting effects of P-ITB and NP-IBA treatments, in comparison with IBA treatment. The earlier rooting of NP-IBA-treated jack pine seedling cuttings was positively related to increase in fresh weight of basal 1 cm stem before primordia were initiated. Weight of basal steins of NP-IBA-treated cuttings initially increased during propagation at day 2 in comparison with the control, and at day 3 in comparison with IBA-treated cuttings. Results that were obtained with seedling jack pine cuttings were generally confirmed for cuttings from 7-year-old trees.     

Characterization and Rooting Ability of Indole-3-Butyric Acid Conjugates Formed during Rooting of Mung Bean Cuttings

Indole-3-butyric acid (IBA) is rapidly metabolized by mung bean cuttings during rooting. Twenty-four hours after application, less than 20% of the applied IBA remained in the free form and its level decreased continuously in the later stages of rooting. Indole-3-butyrylaspartic acid (IBAsp) and at least two high molecular weight conjugates were the major metabolites in IBA-treated cuttings. In the latter conjugates, at least part of the IBA moiety is attached to a high molecular weight constituent in an amide linkage. IBAsp level peaked 24 hours after application of IBA to the cuttings and then declined. The level of the high molecular weight conjugates increased continuously throughout the rooting process. The conjugates were active in inducing rooting of cuttings, with IBAsp being superior to free IBA. It is suggested that IBA conjugates, and particularly IBAsp, serve as the source of auxin during the later stages of rooting.     

The effect of 4-chlororesorcinol on the endogenous levels of IAA,ABA and oxidative enzymes in cuttings

Treatment of bean cuttings with 4-chlororesorcinol (4-CR), known to increase the number of roots and extend their distribution, prevented the accumulation of free indol-3-yl-acetic acid (IAA) in the hypocotyls within 24 h after cutting preparation. In mung bean there was no change in the distribution (upper half vs. 1 ower half of the hypocotyl) of IAA within the hypocotyl as a result of the treatment. In bean cuttings the treatment with 4-CR prevented the accumulation of IAA in the bottom of the cutting. Oxidation of IAA as a measure of IAA oxidase activity in bean was enhanced appreciably by 4-chlororesorcinol. The level of abscisic acid in mung bean, on the other hand, remained 3–4 fold higher than in the control, yet still about 50% lower than the zero time level. In untreated mung bean cuttings the activity of peroxidase increased after cutting preparation. In contrast, the activity of peroxidase in 4-Cr-treated cuttings was consistently lower. In order to relate to the effect of exogenously applied auxin the level of peroxidase was measured also in indol-3-yl-butyric acid-treated cuttings. The overall peroxidase activity in IBA-treated cuttings was not affected. However, when assaying for the different isozymes the drop in peroxidase activity was most evident in the inducible basic isoperoxidases both in 4-CR and IBA treatments. It appears that the exposure to 4-CR exerts an effect that is similar to that of exogenously applied auxin, affecting the activity of basic peroxidases and enhancing the oxidation of endogenous IAA, thus allowing the organization of the primordia.Abbreviations ABA - abscisic acid - 4-CR - 4-chlororesorcinol - IAA - indol-3-yl-acetic acid - IBA - indol-3-yl-butyric acid     

Promotion of adventitious root formation by 4-chlororesorcinol: A polyphenol oxidase inhibitor

The extent of rooting in cuttings of Phaseolus vulgaris L., and Vigna radiata Wilcz. was affected by 4-chlororesorcinol, a polyphenol oxidase inhibitor. More root primordia and more roots were formed after 4-chlororesorcinol treatment both with and without 10^-5M Indole butyric acid. Promotion of rooting was observed also in cuttings of Elaeagnus pungens, Gypsophilia elegans and Kalanchoe blossfeldiana. The enhancement in bean and mung bean was accompanied by a concomitant wider spatial distribution of the primordia and the resulting adventitious roots. The formation of primordia in the treated cuttings was delayed by 12–24 hours, compared to untreated cuttings. The treatment was effective only when given during the first hours after the preparation of the cutting of bean and mung bean, suggesting involvement in the initiation stage. Hypocotyl extracts of mung bean cuttings, pretreated with 4-chlororesorcinol, exhibited reduced polyphenol oxidase activity. The inhibition was not reversed by washing of the treated extract in 50% acetone or by an overnight dialysis, suggesting tight or maybe even irreversible binding of the inhibitor to the enzyme.Abbreviations 4-CR 4-chlororesorcinol - IBA Indole butyric acid - PPO polyphenol oxidase     

l-DOPA (l-3,4-dihydroxyphenylalanine) affects rooting potential and associated biochemical changes in hypocotyl of mung bean, and inhibits mitotic activity in onion root tips

A study was undertaken to explore the effect of l-DOPA (l-3,4-dihydroxyphenylalanine) on the rooting potential of hypocotyl cuttings of mung bean (Phaseolus aureus Roxb. var. SML-32) and related biochemical changes at the post-expression phase. At lower concentrations of (0.0001–0.1 mM) l-DOPA, there was no change in rooting potential, though the average number of roots per cutting and root length were significantly decreased (except at 0.0001 mM). However, at 1.0 mM concentration, a 50% inhibition in rooting potential was noticed and the root number and length were severely reduced. In contrast, at 2.5 mM l-DOPA, none of the hypocotyl cutting rooted. The decrease in rooting potential was associated with a significant effect on the biochemical changes measured in terms of protein and carbohydrate metabolism and activity of peroxidases. In the l-DOPA treated hypocotyl cuttings, there was a significant reduction in the protein and carbohydrate content, whereas activities of associated enzymes proteases and amylases decreased, particularly at higher treatment concentration (>1.0 mM). It indicated negative effect of l-DOPA on these two important metabolic processes. Likewise, activity of peroxidases also decreased in the l-DOPA treated hypocotyl mung bean cuttings thereby indicating its role in suppressing rhizogenesis as the enzyme is involved in lignification process during cell division. l-DOPA suppressed mitotic activity in the root tip cells of onion indicating thereby its interference with the cell division, which is required for the formation of new meristematic tissue during rhizogenesis. Based on the obtained results, it is concluded that l-DOPA interferes with the various biochemical processes in the mung bean hypocotyl cuttings thereby affecting their rooting potential.     

The Effect of the Ethylene Action Inhibitor 1-Cyclopropenylmethyl Butyl Ether on Early Plant Growth

Increased levels of ethylene in plants are responsible for many deleterious effects such as early senescence, fruit deterioration and inhibition of root elongation. Several cyclopropene derivatives have previously been studied as inhibitors of ethylene action in plants. This study focuses on one such compound, 1-cyclopropenylmethyl butyl ether and its effect on the growth of roots and shoots of canola plants as well as rooting of mung bean seedlings 1-cyclopropenylmethyl butyl ether increased root length in canola plants, but had no significant effect on shoot length. In rooting studies, mung bean seedlings treated with 1-cyclopropenylmethyl butyl ether prior to root excision had fewer numbers of roots than control plants that were not treated with the ethylene action inhibitor. The same rooting study, when repeated in the presence of 1-aminocyclopropane-1-carboxylic acid (ACC), demonstrated an overall increase in the number of roots of inibitor-treated and non-treated plants, however, the inhibitor was still effective in decreasing the number of roots, compared to its non-treated conterpart. Online publication: 7 April 2005     

The effect of ethylene on adventitious root formation in mung bean (Vigna radiata) cuttings

A promotive effect of ethylene on the formation of adventitious roots by mung bean cuttings was demonstrated using a recirculating solution culture system to apply dissolved ethylene. The number of roots increased in proportion to the length of exposure to the gas. Mean root numbers per cutting for a 4-day exposure to ethylene and an air control were 45 and 19, respectively. The tissue was most sensitive to a 24-h ethylene “pulse” 2–3 days after taking cuttings. Rooting was maximal at a concentration of 13 μl 1^?1 ethylene. The ethylene treatment inhibited the growth of roots and terminal buds. Application of Ag⁺, as silver thiosulfate, reversed the effect of ethylene on the two growth responses but had no effect on root numbers. Norbornadiene, another inhibitor of ethylene action, reversed all three ethylene responses.     

Ethylene and rooting of mung bean cuttings. The role of auxin induced ethylene synthesis and phase-dependent effects

We have re-examined the role of ethylene during rooting of mung bean cuttings. Cuttings were treated for 5 days with a low or a high concentration of NAA (naphthaleneacetic acid). During this 5 days period, we also applied STS (silverthiosulfate, an inhibitor of ethylene action) or ACC (1-aminocyclo-propane-l-carboxylic acid, a direct precursor of ethylene). At high NAA concentration, STS promoted and ACC inhibited rooting, respectively. At low NAA concentration, the effects were opposite, STS being inhibitory and ACC promotive. AVG (aminoethoxyvinylglycine, an inhibitor of ethylene synthesis) gave similar results as STS. Together, these data suggest supraoptimal and suboptimal ethylene levels in the tissue at high and low NAA concentration, respectively. We also examined whether the effect of ethylene varied during the successive phases of the rooting process. Thus, we gave 24 h pulses with either STS or ACC during the rooting treatment. During the first two days (0–48 h), ACC-pulses were promotive and STS-pulses inhibitory. Later on (48–168 h), the ACC-pulses were inhibitory and the STS-pulses promotive. Whether this effect was observed or not was dependent on the NAA concentration. These data indicate that ethylene promotes or inhibits rooting depending on the stage in the rooting process. When ACC was added only during the initial period, rooting was increased at all NAA concentrations in a NAA dose-response curve and the optimal NAA concentration remained the same. This suggests that ethylene renders more cells responsive to NAA.     

The influence of plant growth regulators and storage on root induction and growth in <Emphasis Type="Italic">Pelargonium zonale</Emphasis> cuttings

The effects of post harvest application of ethylene, abscisic acid (ABA), indole-3-butyric acid (IBA) treatments or dark storage on root induction and continued growth of regenerated roots in Pelargonium cuttings were investigated using hydroponics in the greenhouse. Ethylene markedly increased rooting percentage in ‘Greco’ and ‘Surfing’, reduced the number of roots per cutting in ‘Surfing’ and had no effect on the total root lengths in the two cultivars. Ethylene treatment reduced fresh root mass in ‘Surfing’, increased dry root mass and reduced root water content in both cultivars. ABA (50 μM) enhanced rooting percentage in ‘Greco’, reduced the number of roots per cutting, reduced total root lengths and fresh root mass in both cultivars. ABA increased dry root mass and reduced root water content in ‘Surfing’ but this effect was not apparent in ‘Greco’. Storing cuttings in the dark for 4 days had no effect on rooting percentage and number of roots per cutting in ‘Greco’ and ‘Surfing’. However, dark storage reduced total root lengths in ‘Surfing’ and reduced fresh root mass in ‘Greco’. Dark storage had no effect on dry root mass and water content in both cultivars. Applying 4 μl l⁻¹ IBA in the rooting solution induced maximum (100%) root induction in ‘Surfing’. However, IBA reduced the number of roots per cutting in ‘Greco’, reduced total root lengths and fresh root mass in the two cultivars. IBA treatment profoundly increased and reduced dry root mass and root water content, respectively, in ‘Greco’ and ‘Surfing’. The enhanced root induction observed after IBA and ABA applications could be ascribed to their influence on ethylene biosynthesis, since ethylene treatment increased rooting percentage in both cultivars. However, high ABA (100 μM) and IBA (12 μl l⁻¹) levels or dark storage reduced the ability of induced roots to continue growth. We attribute our results to plant stress-response mechanism and ethylene appears to play an important role in the process of root initiation and root growth in Pelargonium cuttings.     

Initiation of roots on hypocotyl cuttings of Pinus contorta in vitro

The origin of roots and wound tissue after treatments for induction of roots on hypocotyl cuttings of three-week-old Pinus contorta Dougl. ex Loud, is discussed. The cuttings were cultured in vitro and treated with 1.2 μ M to 1.5 M IBA (indole-3-butyric acid) for 6 h to 10 days. The control, which was not treated with IBA developed a wound tissue from which roots formed. Cuttings treated with IBA developed roots directly from the hypocotyl. Direct rooting was faster than indirect rooting via a wound tissue. Rooting was considered to be optimal if more than 80% of the cuttings rooted within 19 days and half of the cuttings which possessed roots after one month had acquired them within 14 days. This type of rooting was obtained after treatment with either 80 μ M IBA for 4 to 6 days or 1.25 to 5.0 m M IBA for 6 h. Suboptimal treatments gave lower rooting percentages and superoptimal treatments resulted in delayed rooting. In IBA-treated cuttings, large increases in mitotic activity (number of mitoses per mm hypocotyl) were found in the pericycle and parenchyma inside endodermis. However, the control also had similar mitotic activities as the IBA-treated cuttings but closer to the cut surface. This led us to the conclusion that similar tissues may produce either wound tissue or roots. Almost all roots obtained through direct rooting originated outside resin ducts.     

Effects of combinational treatment with ethephon and indole-3-butyric acid on adventitious rooting of <Emphasis Type="Italic">Pinus thunbergii</Emphasis> cuttings

Adventitious rooting is essential for cutting propagation of pine wilt-resistant Pinus thunbergii. To examine a variety of adventitious rooting potentials among donor plants, cuttings were taken from 31 seedlings within a half-sib family. Rooting abilities of cuttings from each seedling ranged from 0 to 100%. When 11 ortets and 11 ramets (clonally propagated from each ortet) were used as donor plants, there was a positive correlation between rooting abilities of cuttings from ortets and ramets, suggesting that adventitious rooting is dependent on genetic factors in the donor plants. To promote adventitious rooting of cuttings by hormonal treatment, we examined the effect of soaking time in Oxyberon (19.7 mM indole-3-butyric acid (IBA) solution) on rooting. Ten minutes was the best soaking time for rooted cuttings to produce more adventitious roots without impairing normal growth. When cuttings were soaked in Ethrel diluent (69.2 μM ethephon) for 24 h before soaking in Oxyberon for 10 min, a significantly higher rooting ability was observed than those soaked in Oxyberon alone. Ethrel on its own barely affected rooting ability. The positive effect of the combinational treatment was confirmed in a similar experiment using authentic ethephon and IBA instead of Ethrel and Oxyberon. When cuttings were soaked in a mixture of ethephon and silver thiosulfate (STS), an ethylene action inhibitor, before IBA-soaking, the effect was partially diminished compared with combinational treatment without STS. These findings suggest that ethylene action caused by ethephon treatment promotes IBA-mediated adventitious rooting of P. thunbergii cuttings.     

Influence of ethylene on adventitious root formation in mung bean hypocotyl cuttings

The relationship between ethylene and adventitious root formation in mung bean hypocotyl cuttings was studied.Ethephon, an ethylene-releasing compound, at 5 x 10 -5 M increased root number and root dry weight on hypo-cotyl cuttings. When ethephon was applied to hypocotyl at different times after excision, there were two effectivetimes for root production i.e. between 06 h and 18-24 h. These two time periods correspond to the induction phase and the late initiation phase of root development, respectively. After excision, three peaks of ethylene productionwere observed. The first peak commencing at 6 h started the sequence of reactions leading root formation, the second peak appearing at 12 h coincided with the beginning of the increase of the IAA level during primordia initiation, and the third peak showing at 48 h played a role in root differentiation and growth. Ethylene stimulated rooting by enhancing the increase in auxins. Thus it appears that the IAA-induced ethylene production may be a factor involved in the stimulation of adventitious root formation.     

Effect of abscisic acid on endogenous IAA, auxin protector levels and peroxidase activity during adventitious root initiation in Vigna radiata cuttings

Endogenous levels of free and conjugated IAA, auxin protectors (Prs) and peroxidase (PER) activity and their relation to adventitious root initiation (ARI) were investigated at the potential sites of adventitious rooting in relation to exogenous application of 250 μM ABA during the first 120 h after treatment. Cuttings from 7-day-old mung bean [Vigna radiata (L.) Wilcz.] seedlings were treated with 125, 250, and 500 μM ABA for 24 h. ABA significantly stimulated ARI but extremely inhibited epicotyl growth as compared to control. Free and conjugated IAA were measured by reversed-phase high performance liquid chromatography while Prs and PER activities were measured spectrophotometrically. The present results also indicate that endogenous free IAA levels peaked later in ABA-treated cuttings than that in control, suggesting that ABA extended the length of the induction phase of rooting process in treated cuttings and that might explain the significant delay of the appearance of roots at the treated cuttings. Higher level of IAA conjugates was found in ABA-treated cuttings than that in untreated ones. Pr level also peaked later in ABA-treated cuttings than that in control, indicating that ABA extended the period of Pr activity. An initial temporary decrease of PER activity was found in associating with high levels of free IAA and Prs during most of the primary events, while the opposite occurred during the secondary events of adventitious rooting process in both treated and untreated cuttings. Thus, ABA may stimulate ARI in mung bean Vigna radiata cuttings by regulating the concentration and /or activities of endogenous IAA, Prs, and PER activity in favor of inducing a large number of adventitious roots at their potential sites of adventitious rooting.     

Abscisic-acid-stimulated rooting of stem cuttings

Summary Abscisic acid (ABA) has been found to stimulate rooting of stem cuttings of mung beans and English ivy. ABA partially overcame the inhibitory effect of gibberellic acid on root formation of mung bean cuttings but at the concentrations used did not overcome the inhibitory effect of kinetin on root formation.