Effect of Reciprocal Bud-Graft Transfers Between Old Home and Bartlett Pears and Centrifugatiou on Trans-location of Endogenous Growth Substances in Hardwood Cuttings

An investigation was made of the effect of bud-graft exchanges between two pear varieties—the easily-rooted Old Home and the hard-to-root Bartlett—on the root-promoting activity of extracts obtained from basal segments of hardwood cuttings of each variety. The effect of a centrifugal force obtained from 2000 rpm for 30 minutes on the root-promoting activity of diffusates and extracts from bases of centrifuged cuttings of these two varieties was also studied. Bud exchange treatments showed no appreciable influence on the root-promoting or inhibiting activity of basal extracts unless the transbudded shoots (used for making cuttings) were girdled one month prior to the sampling date; in such cases an influence did appear. The centrifugation treatment increased the number of buds sprouting on cuttings. It also increased the root-promoting activity of extracts from the basal parts of cuttings and of their growth-active components (separated by paper chromalography) when bioassayed by the mung bean rooting test. Apparently some inhibitory materials from the buds were removed through the bases of the cuttings by centrifuging.     

Evidence for the presence of a native,non-auxinic rooting promoter in avocado (Persea americana Mill.)

The presence of a rooting promoter in paratially purified extracts of avocado (Persea amricana Mill.) organs has been demonstrated using the mung bean rooting bioassay. Extraction with 80% methanol was followed by partition into diethyl ether, paper chromatography (PC) and 3 steps of thin layer chromatography (TLC). The number of roots induced by the rooting promoter in the absence of exogenous auxin was 5 to 7 times higher than that of the water control and 50% higher than by 4-(indol-3-yl) butyric acid (IBA) at its reported optimum concentration. Rooting of tomato, Coleus and young avocado cuttings was also enhanced by the rooting promoter. The rooting promoter was inhibitory in the wheat coleoptile section elongation bioassay for auxins and had slight inhibitory activity in the split pea stem curvature test.The biological properties of the avocado rooting promoter may be comparable to those of -(p-chlorophenoxy) isobutyric acid (PCIB) which acts as an anti-auxin in certain bioassays and, nevertheless, promotes the rooting of mung bean cuttings.     

Treatment of bean cuttings by chestnut extracts during rooting modifies their IAA-oxidase activity

The IAA-oxidase activity in bean cuttings ( Phaseolus vulgaris L. cv. Contender) treated with extracts from juvenile and adult chestnut ( Castanea sativa Mill.) was studied in the light of the effect of the extracts on the rooting activity of some auxins. Extracts from adult chestnut increased IAA-oxidase activity in bean cuttings. Extracts from juvenile chestnut inhibited IAA-oxidase activity only slightly, but substantially reduced the IAA-oxidase activity of bean cuttings pre-treated with adult chestnut extracts. These findings provide evidence that there is a relationship between the IAA-oxidase system and the effect of chestnut extracts on IAA-induced rhizogenesis in bean cuttings.     

Effect of Ethephon, Indole Butyric Acid, and Treatment Solution pH on Rooting and on Ethylene Levels within Mung Bean Cuttings

Light-grown mung bean (Phaseolus aureus Roxb.) cuttings were treated with buffered and nonbuffered solutions of Ethephon, indole butyric acid (IBA), and the combination of both. Ethephon treatment resulted in increased tissue ethylene levels with increasing solution pH, but had no effect on rooting. IBA treatment had no effect on tissue ethylene levels, but strongly promoted rooting. Combinations of Ethephon and IBA had no effect on rooting of mung bean cuttings beyond that obtained by IBA alone.     

Enhancement of adventitious root formation in mung bean cuttings by 3,5-dihalo-4-hydroxybenzoic acids and 2,4-dinitrophenol

3,5-Dihalo-4-hydroxybenzoic acids enhanced adventitious root formation in mung bean (Vigna radiata L.) cuttings. 3,5-Diiodo-4-hydroxybenzoic acid was more active than 3,5-dichloro-4-hydroxybenzoic acid, increasing the number of roots formed by about 4-fold. 2,4-Dinitrophenol also enhanced significantly adventitious root formation in mung bean cuttings. The phenolic compounds were active with or without indole-3-acetic acid. The possible mechanism by which these phenolic compounds enhance rooting is discussed.Abbreviations CCCP carbonyl cyanide 3-chlorophenylhydrazone - DIHB 3,5-diiodo-4-hydroxybenzoic acid - DNP 2,4-dinitrophenol     

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.     

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     

Vegetative Propagation of Pinus sylvestris

Methods for the vegetative propagation of Pinus sylvestris L. from interfascicular shoots are described. Using 5-year-old plants the outgrowth of interfascicular shoots was promoted by removal of terminal and lateral buds; this response was augmented substantially by application of cytokinin, tri-iodobenzoic acid, alar and morphactin alone and especially in combination. The rooting capacity of shoot cuttings from interfascicular shoots appeared to be largely determined by the state of growth of the stock plant. Cuttings from dormant stock plants subjected to short-day treatment followed by a period of low temperature gave the best rooting, especially when the cuttings themselves had been cold-stored prior to planting. Rooting was optimal when such cuttings were treated with a mixture of 25 mg/l of indolebutyric acid and 25 mg/l of napthaleneacetic acid as a 48 h basal soak, and were planted on a heated mistbench under extended illumination; over 90% of such cuttings could be rooted. These results are discussed in relation to bud activity, endogenous hormone levels and promoting tissue extracts also tested.     

Changes in the rooting inhibitory effect of chestnut extracts during cold storage of the cuttings

Extracts of cold stored chestnut cuttings ( Castanea sativa Mill.) were examined for the inhibitory effect on the stimulation of rooting by 1AA which has been detected previously in extracts from freshly collected cuttings. The extracts were fractionated by paper chromatography and the different zones of the chromatograms were bioassayed together with 1AA by the bean rooting test. The bean rooting test showed that the inhibitory effect decreased with the length of cold storage period, so that after 5 months of storage, the inhibitory effect had disappeared, and a root promoting zone was found on the chromatograms. A comparative study of phenolics in this zone, before and after cold storage, revealed the formation of vanillyl and salicyl alcohols in the chilled material. Vanillyl and salicyl alcohols are rooting stimulators and increase the effect of 1AA on rooting in bean cuttings.     

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; )     

Caffeic acid affects early growth, and morphogenetic response of hypocotyl cuttings of mung bean (Phaseolus aureus)

Caffeic acid (CA) is one of the most common cinnamic acids ubiquitously present in plants and implicated in a variety of interactions including allelopathy among plants and microbes. This study investigated the possible interference of CA with root growth and the process of rhizogenesis in hypocotyl cuttings of mung bean (Phaseolus aureus=Vigna radiata). Results indicated that CA (0-1000 microM) significantly suppressed root growth of mung bean, and impaired adventitious root formation and root length in the mung bean hypocotyl cuttings. Further investigations into the role of CA in hampering root formation indicated its interference with the biochemical processes involved in rooting process at the three stages - root initiation (third day; RI), root expression (fifth day; RE), and post-expression (seventh day; PE) - of rhizogenesis. CA caused significant changes in the activities of proteases, peroxidases (PODs), and polyphenol oxidases (PPOs) during root development and decreased the content of total endogenous phenolics (TP) in the hypocotyl cuttings. The enhanced activity of PODs and PPOs, though, relates to lignification and/or phenolic metabolism during rhizogenesis; yet their protective role to CA-induced stress, especially during the PE phase, is not ruled out. At 1000 microM CA, where rooting was significantly affected, TP content was very high during the RI phase, thus indicating its non-utilization. The study concludes that CA interferes with the rooting potential of mung bean hypocotyl cuttings by altering the activities of PODs and PPOs and the endogenous TP content that play a key role in rhizogenesis.     

Comparison of cytokinin- and auxin-like activity in some commercially used seaweed extracts

Six commercially used seaweed extracts were tested for cytokinin- and auxin-like activity using the soybean callus bioassay and the mung bean rooting bioassay respectively. All the seaweed extracts tested showed cytokinin-like activity. Seamac caused the most callus growth with activity being concentrated at R _f 0.9 which co-chromatographed withiso-pentenyladenine and its riboside. The other five seaweed extracts yielded activity at R _f 0.7–0.8. This co-chromatographed with zeatin and zeatin riboside. All the seaweed extracts tested improved rooting of mung beans. Kelpak made fromEcklonia maxima (Osbeck) Papenf. gave the best rooting response.     

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.     

Effects of autoclaving and charcoal on root-promoting substances present in water extracts made from gelling agents

The root-promoting ability of water extracts made from gelling agents (agar and Gelrite) was investigated using the mungbean rooting bioassay. Autoclaving these water extracts decreased the number of roots in mungbean cuttings compared to the controls. The addition of activated charcoal to the water extracts from Agar Bacteriological and Agar Commercial Gel had no effect on their root-promoting ability. Extracts with exogenous indole-3-butyric acid (IBA) which were treated by autoclaving or via a freeze-thaw cycle, significantly increased rooting. However, incorporation of activated charcoal to similar IBA-containing extracts reduced rooting. Our results indicate that more attention should be given to the choice of gelling agent and its interaction with other additives in the media used during tissue culture.     

脱落酸对绿豆下胚轴不定根发生的影响

以10^-4 mol/L脱落酸(ABA)处理绿豆种子24 h,在幼苗下胚轴长6 cm时,切除根部作为插条,研究ABA对插条不定根发生及插条基部细胞周期时相的影响。结果表明,ABA可促进下胚轴插条不定根发生,增加生根数和生根范围;ABA提高插条基部细胞色氨酸转氨酶、吲哚丙酮酸脱羧酶和吲哚乙醛脱氢酶的比活性,增加吲哚乙酸含量,同时进入细胞周期S期的基部细胞数目增加,促进DNA合成,有利于不定根的发生。     

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.     

Initiation of Flowering and Changes in Endogenous Inhibitors and Promoters in Olive Buds as a Result of Chilling

Olive trees require a period of chilling temperatures, either diurnally fluctuating, or at a constant—but definite—level before inflorescences develop. In this study, bud and leaf samples were taken at weekly intervals during this critical period from trees receiving temperatures favorable for flowering and from trees under temperature conditions known to prevent flowering. Extracts were obtained from these samples, separated by paper chromatography and the different growth-active materials were bioassayed by the mung bean rooting test for the presence of promoters and inhibitors. Extracts from buds on trees under conditions unfavorable for flowering generally consisted only of promoters. Extracts from buds on trees placed under conditions favorable for flowering showed a definite and consistent pattern for the presence of rooting inhibitors and promoters. Within one or two weeks after being placed under such conditions a strong inhibitor band appeared from R_f 0.50 to 0.70. This persisted for 6 to 7 weeks, gradually decreasing in intensity, followed by promoters at these bands. During this period all other bands, at different R_f values, showed strong promoters. Extracts from leaves on shoots under temperature regimes favorable for flowering showed a pattern like the buds in the appearance and disappearance of inhibitors and at the same R_f values but with about one week lag in time. Bud extracts from defoliated shoots on trees under temperature conditions known to result in inflorescence production failed to show the characteristic inhibitor pattern associated with flowering. Such defoliated shoots do not form inflorescences.     

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.     

Root-promoting Substances in Salix alba

Root-promoting substances were extracted from softwood cuttings of Salix alba L. by centrifuging them with water or by shaking the ground freeze-dried stems with water. Rooting substances were partitioned by paper chromatography or chemical fractionation and their rooting activity was tested by mung bean cuttings. Both extracts indicated three major root -promoting fractions at Rf 0-0.1, 0.7-0.8, and 0.3-0.4 in a decreasing order of their activities when paper chromatographed with isopropanol:ammonia:water 8:1:1 v/v. The strongest one indicated an apparent synergistic rooting effect with indol-3yl-acetic acid (IAA) regardless of the extraction method. These results indicate that water can extract from freeze -dried sample the similar rooting substances found in the centrifugal diffusates. The Rf 0–0.1 fraction consisted of at least four fractions and the strongest one did not move from the starting line on the chromatogram when isopropanol:ammonia:water 8:1:1 was used. This starting line fraction was extremely strong in rooting activity and its highest concentration resulted in 8.7 times as many roots as controls. More thain additive rooting effect between IAA and the fraction was found only at the highest concentration. The fraction was very soluble in water but insoluble in chloroform or ethyl ether and only stimulated rooting of mung bean cuttings when it was applied within 3 days after cuttings were made. It had no effect in lengthening roots. The starting line fraction was further found to have four root-promoting subfractions at Rf 0.05, 0.35, 0.65, and 0.85 when it was chromatographed in 60 % isopropanol. Among these four, the subfractions at Rf 0.65 and 0.35 were strongly root promotive and displayed more than additive root promotion with IAA at the highest concentrations studied.     

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.