Adventitious rooting: examining the role of auxin in an easy- and a difficult-to-root plant

Therooting responses of cuttings of difficult-to-root lilac (Syringavulgaris) and easy-to-root forsythia(Forsythia×intermedia)were compared. The rooting ability of lilac cuttings declined over the growingseason (May–June). There was also a decline in the initial concentrationof free IAA at the base of the cuttings, but there was not a tight relationshipbetween basal IAA concentration and rooting ability. Polar auxin transportability was measured in lilac and forsythia during the period of maximum growthby [³H]IAA application to stem internodal tissue. Transport abilitydeclined in lilac over this time period, particularly in terms of transportintensity and percentage of [³H]IAA transported. In contrast thechanges in polar auxin transport ability in forsythia were less marked. Thisdifference between species was maintained in winter hardwood cuttings, withforsythia tissue showing greater polar auxin transport ability than lilac. Theimportance of polar auxin transport for adventitious rooting was demonstratedinboth lilac and forsythia softwood cuttings by use of the polar transportinhibitor 2,3,5-triiodobenzoic acid (TIBA). Overall the results indicate thatdifferences in polar auxin transport ability between lilac and forsythiacontribute to differences in rooting ability.     

Chemical induction of adventitious root formation in Taxus baccata cuttings

The effect of some auxins (IBA and NAA), phenolic compounds (phloroglucinol, gentisic acid and coumarin), a combination of auxins and phenolics, and a systemic fungicide (Bavistin) have been examined for stimulatory effects on adventitious root formation in stem cuttings (current season's growth) of Taxus baccata L. In general lower concentration (0.25 mM) of both IBA and NAA was more effective in inducing rooting of cuttings taken from both male and female trees. The combined treatment of IBA+NAA (0.25 mM each) showed some success in cuttings from male trees only (55%, compared to 15% rooting in cuttings from female trees). Generally, the callus formation was quite high (70%) in all auxin treatments (alone or in combination). Among the phenolics, 40% rooting success was achieved with phloroglucinol only, while coumarin and gentisic acid were ineffective. The combined treatment of auxins and phenolics also failed to promote rooting. On the other hand, Bavistin was extremely effective for callusing (90%) as well as rooting (80%). The effectiveness of various compounds tested for rooting of young stem cuttings declined in the order: 0.25 mM IBA>0.05% Bavistin>0.25 mM NAA>1.25 mM IBA>15 mM phloroglucinol>IBA+NAA (0.25 mM each). In addition to the auxins, IBA and NAA that are widely used for commercial propagation, the auxin-like properties of the fungicide Bavistin could be exploited for adventitious rooting in T. baccata, and in other plant species.     

Carbohydrates as regulatory factors on the rooting of <Emphasis Type="Italic">Eucalyptus saligna</Emphasis> Smith and <Emphasis Type="Italic">Eucalyptus globulus</Emphasis> Labill

Comparisons between related species with different rooting capacities can provide insights into the mechanisms controlling adventitious root development. The availability of carbohydrates is often considered exclusively as an energetic requirement to drive root development; the major regulatory role in the process is often attributed to phytohormones, particularly auxin. The roles of light quantity (irradiance) and carbohydrate supply available to young aseptic donor-plants on the adventitious rooting response of Eucalyptus globulus (rooting recalcitrant) and Eucalyptus saligna (easy-to-root) were examined. The effects of the type of carbohydrate supply (sucrose or glucose) on the rooting response of cuttings was also evaluated. Light intensity supplied to mother-plants (30 or 60 mol m^–2 s^–1) had limited influence on the rooting response of both species, whereas dark periods were detrimental, particularly for E. globulus. In E. globulus, rooting was promoted by the absence of sucrose in donor-plant media. Presence of sucrose in donor plant medium promoted root number but did not affect rooting percentage of E. saligna. A positive effect of glucose on cutting rhizogenesis was found if this hexose was supplied during the root induction phase, followed by sucrose in the root formation step, especially for E. globulus. The same effect was not seen with fructose. The beneficial effect of glucose in the induction phase on root number was also evident under suboptimal auxin concentrations.     

A simple and efficient method for clonal propagation of Casuarina sumatrana (de Vriese) L. Johnson

A new and efficient method for clonal propagation of Casuarina sumatrana by rooting stem cuttings is described. High percentage (about 60–70%) of rooting was achieved with mature softwood stem cuttings. A quick-dip of 5 s in NAA (1–10mM) solution followed by sand culture under high humidity were required for a high rate of survival and rooting of stem cuttings. A simple, closed chamber propagation system, using fluorocarbon polymer (tetrafluoroethyleneperfluoroalkyl vinyl ether) film (Neoflon PFA film), was successfully developed for the rooting of stem cuttings without mist. Rooted cuttings inoculated with Frankia were easily transplanted and established in field conditions with very low (about 3%) mortality. The significance of these findings for mass clonal propagation of C. sumatrana is discussed.Abbreviations IBA indole-3-butyric acid - NAA -naphthaleneacetic acid     

The influence of leaf area on the rooting physiology of leafy stem cuttings of Terminalia spinosa Engl.

Summary The effect of different leaf areas on the rooting of Terminalia spinosa Engl. cuttings in an non-mist propagation system in glasshouses at Edinburgh was investigated by trimming the leaves to 0, 7.5, 15 and 30 cm² before cuttings were severed from stockplants. Cuttings were taken to a standard length of 5 cm from the lateral shoots of previously pruned stockplants grown in a tropicalised glasshouse. During the rooting period, photosynthetic rate, stomatal conductance, water potential and relative water content of the cuttings were assessed at regular intervals. It was found that (i) removal of the entire leaf area prevented rooting; (ii) cuttings with a 7.5 cm², 15 cm² and 30 cm² leaf all achieved 80% rooting after 3 weeks; (iii) an increase in leaf area from 7.5 cm² to 30 cm² increased the rate of rooting and the length of the longest root after 2 weeks, but also increased the number of original leaves abscised after 6 weeks; and (iv) the greatest number of new leaves were produced by cuttings with 7.5 cm² and 15 cm² leaf area per cutting. All leafy cuttings actively photosynthesized during the propagation period, with a mean rate of 2 mol CO₂ m^-2 s^-1 with an irradiance of 100 mol m^-2 s^-1. Cuttings with 30 cm² leaf area had lower relative water contents, lower stomatal conductances and lower photosynthetic rates per unit leaf area than those with a 7.5 cm² and 15 cm² leaf. It was concluded that T. spinosa cuttings are easy to root, provided the cuttings have leaves to produce current assimilates.A member of the Edinburgh Centre for Tropical Forests     

The role of light and polar auxin transport in root regeneration from hypocotyls of tomato seedling cuttings

A relationship between light conditions, auxin transport and adventitious root formation by hypocotyls of tomato seedling cuttings was demonstrated. Effective rooting of tomato seedling cuttings was observed under continuous white light (WL) irradiation. However, root formation was reduced in darkness or under red (RL) or blue light (BL). At least 3/4-day-long irradiation treatment with (WL) was necessary to increase the number of roots formed in comparison with control cuttings grown in darkness. Light was most effective if applied during the first half of the 13-day-long rooting period. The role of photoreceptor-dependent light perception in the light-regulation of rooting was tested using tomato photomorphogenic mutants: aurea (au) and high pigment (hp). When exposed to WL both mutants generated fewer roots then their isogenic wild type (WT). In darkness or under BL and RL less roots were formed on all plants and no difference was observed between mutants and WT plants. TIBA (2,3,5-triiodobenzoic acid) inhibited rooting in a dose-dependent manner both in darkness and under WL. However, although rooting was suppressed by 0.75 M TIBA in the dark, 8 M TIBA was necessary to block root formation in continuous WL. Inhibition of rooting by TIBA was most efficient when applied at the initial period of rooting, a 1-day-long treatment with TIBA being sufficient to suppress rooting if given during the first 2 days of culture. Later treatment had much less effect on the root formation.     

Adventitious Root Formation in Young Shoots of Cedrus Deodara

The effect of auxins [indole-3-butyric acid (IBA) and -naphthaleneacetic acid (NAA)], phenolics (phloroglucinol and coumarin), a combination of auxins and phenolics, and a systemic fungicide (Bavistin, containing 50% carbendazim) on adventitious root formation in stem cuttings (current season's growth) of Cedrus deodara L. during winter and monsoon (rainy) seasons has been examined. Significant stimulation of rooting due to treatments was observed in cuttings planted in winter in the following order: 0.25 mM IBA (87.5% rooting) > 5 mM coumarin (70.8%) > IBA (0.5 mM) + coumarin (5 mM) (50.0%). In cuttings planted in monsoon only 0.05% Bavistin was found to be effective in inducing rooting (83.3%). Other treatments were ineffective and in some treatments drying of cuttings was noticed.     

Effect of salinity and humidity on δ13C value of halophytes—Evidence for diffusional isotope fractionation determined by the ratio of intercellular/atmospheric partial pressure of CO2 under different environmental conditions

Summary Seedlings of two mangrove species, Avicennia marina and Aegiceras corniculatum, were grown in a range of salinities and humidities in controlled environment chambers, and Phaseolus vulgaris plants were grown in the glasshouse. The fractionation of carbon isotopes in the three species was correlated with the ratio of intercellular and ambient partial pressures of CO₂. The results are consistent with fractionation being due both to diffusion in air and to carboxylation in the leaf. It was concluded that the latter process discriminates against ¹³CO₂ relative to ¹²CO₂ by about 27.     

The distribution and state of mangroves along the coast of Transkei,Eastern Cape Province,South Africa

The mangrove communities along the coastline of the former Transkei, now part of the Eastern Cape Province, have not been looked at in detail since Ward and Steinkes survey in 1982. Mangroves previously occurred in 17 estuaries but were now found in only 14 of the 76 estuaries visited, with a complete loss of mangroves evident in the Mnyameni, Mzimvubu and Bulungula estuaries amounting to 7.5 ha. Total mangrove loss amounted to 17.6 ha which represents a 6.5% loss over 17 years or 1.04 ha per annum. Tree cover had increased by 16.15 ha in eight other estuaries. This increase could be attributed to the inaccessibility of mangrove stands or to protection afforded by provincial nature reserves and hotel resorts. No new mangrove stands were recorded, although Steinke (pers. comm.) has recently recorded mangroves along the north bank of the Kei River. There has been little change in mangrove species composition in the different estuaries over the past 17 years. Total mangrove loss amounted to 17.6 ha which represents a 6.5% loss over 17 years or 1.04 ha per annum and the species recorded included Avicennia marina, Bruguiera gymnorhiza and Rhizophora mucronata. The mangrove fern Acrostichum aureum L. was recorded for the first time in the Mkozi estuary. Tree density for all estuaries was between 10 and 2594 trees ha^–1. The Mngazana and Mntafufu estuaries had the highest tree densities of 2594 and 1402 trees ha^–1 respectively, typical of riverine mangrove forests. Fringe mangroves were evident in most other systems. Removal of trees for wood has the greatest impact on mangrove cover. Fringe mangrove stands are particularly accessible to harvesters. Only 6% of the current area of trees is afforded some protection in conservation areas. The Mdumbi, Mzamba and Kobonqaba estuaries receive no conservation protection and harvesting has resulted in more than 50% of the trees being removed. The density of dead tree stumps was greater than the number of living trees and no seedlings or juvenile trees were found. Further removal of mangroves within the estuaries south of the Mzimvubu River is expected in the fringe mangroves, as most are unprotected and easily accessible.     

小果核果茶扦插繁殖技术研究

小果核果茶为山茶科石笔木属常绿乔木,树形美观,叶泽光亮,洁白亮丽的花朵和茂密的果实具有较高的观赏价值;而且其种子含油量高,是一种具有潜在开发价值的新型生物能源植物。该研究采用4因素3水平的正交设计,分析了激素种类、处理浓度、浸泡时间和扦插基质对小果核果茶扦插生根的影响,并以生根率、生根数和平均根长为扦插生根效果的评价指标,构建生根效果指数,总体评价小果核果茶的生根效果。结果表明:对生根效果指数影响最大的是扦插基质,激素种类次之,影响最小的是处理时间。其中,扦插基质对生根效果指数的影响达到显著水平(P0.05);当激素种类为NAA、处理浓度为500mg·L~(-1)、处理时间为12 h、扦插基质为泥炭土时,小果核果茶扦插的生根效果最好。该研究结果为小果核果茶的栽培生产、育种以及资源开发和利用提供了科学依据。     

Aeroponics for adventitious rhizogenesis in evergreen haloxeric tree <Emphasis Type="Italic">Tamarix aphylla</Emphasis> (L.) Karst.: influence of exogenous auxins and cutting type

Tamarix aphylla (L.) Karst., a drought resistant halophyte tree, is an agroforestry species which can be used for reclamation of waterlogged saline and marginal lands. Due to very low seed viability and unsuitable conditions for seed germination, the tree is becoming rare in Indian Thar desert. Present study concerns the evaluation of aeroponics technique for vegetative propagation of T. aphylla. Effect of various exogenous auxins (indole-3-acetic acid, indole-3-butyric acid, naphthalene acetic acid) at different concentrations (0.0, 1.0, 2.0, 3.0, 5.0, 10.0 mg l^?1) was examined for induction of adventitious rooting and other morphological features. Among all three auxins tested individually, maximum rooting response (79%) was observed with IBA 2.0 mg l^?1. However, stem cuttings treated with a combination of auxins (2.0 mg l^?1 IBA and 1.0 mg l^?1 IAA) for 15 min resulted in 87% of rooting response. Among three types of stem cuttings (apical shoot, newly sprouted cuttings, mature stem cuttings), maximum rooting (~ 90%) was observed on mature stem cuttings. Number of roots and root length were significantly higher in aeroponically rooted stem cuttings as compared to stem cuttings rooted in soil conditions. Successfully rooted and sprouted plants were transferred to polybags with 95% survival rate. This is the first report on aeroponic culture of Tamarix aphylla which can be utilized in agroforestry practices, marginal land reclamation and physiological studies.     

Use of carbon dioxide enrichment to obtain adult morphology of grapevinein vitro

A procedure has been developed forin vitro propagation ofVitis vinifera Pinot noir from lateral-bud cuttings under high CO₂ concentration (1200 µmol mol^–1). Because of inhibition of rooting by CO₂, this procedure requires a rooting pre-culture of explants on medium with sucrose before the CO₂-enriched culture on sucrose-free medium. Shoot growth was enhanced by CO₂ enrichment as a result of both a higher rate of leaf production and greater internode elongation. Leaf expansion and tendril growth were promoted and better rooting was obtained. The more significant effect of CO₂ enrichment was to promote adult morphology with, in particular, the tendril pattern. Thus, for the first time, grapevine plants have been producedin vitro without typical juvenile characteristics. CO₂ enrichment appears to be an interesting process to improve thein vitro propagation of grapevines.     

Effects of temperature at constant air dew point on leaf carboxylation efficiency and CO2 compensation point of different leaf types

The effect of temperature on photosynthesis at constant water-vapor pressure in the air was investigated using two sclerophyll species, Arbutus unedo and Quercus suber, and one mesophytic species, Spinacia oleracea. Photosynthesis and transpiration were measured over a range of temperatures, 20–39° C. The external concentration of CO₂ was varied from 340 bar to near CO₂ compensation. The initial slope (carboxylation efficiency, CE) of the photosynthetic response to intercellular CO₂ concentration, the CO₂ compensation point (), and the extrapolated rate of CO₂ released into CO₂-free air (R _i) were calculated. At an external CO₂ concentration of 320–340 bar CO₂, photosynthesis decreased with temperature in all species. The effect of temperature on was similar in all species. While CE in S. oleracea changed little with temperature, CE decreased by 50% in Q. suber as temperature increased from 25 to 34° C. Arbutus unedo also exhibited a decrease in CE at higher temperatures but not as marked as Q. suber. The absolut value of R _i increased with temperature in S. oleracea, while changing little or decreasing in the sclerophylls. Variations in and R _i of the sclerophyll species are not consistent with greater increase of respiration with temperature in the light in these species compared with S. oleracea.Abbreviations and symbols A net photosynthetic rate - C and C _i CO₂ concentration in the air and in the intercellular airspace of the leaf, respectively - CE carboxylation efficiency - E transpiration rate - R _i CO₂ release into CO₂-free air estimated from extrapolation to 0 bar CO₂ - T _i leaf temerature - VPD difference in water-vapor pressure between mesophyll and air - CO₂ compensation point     

Regeneration of whole plants from apical meristems of Pinus radiata

A methodology to regenerate whole plants of Pinus radiata from apical meristems of 3- and 7-year-old trees was developed. Meristematic domes with two or three leaf primordia were excised from surface-sterilized branch tips of field-grown plants and cultured in LP medium with half strength macronutrients (1/2 LP) and full strength micronutrients. The early growth of meristems required approximately 12 weeks, including a recovery stage during the first 2 weeks. After 8 weeks, some meristems developed abnormal phenotypes and died during the subsequent stages of development. However, healthy meristems elongated and formed shoots when they were transferred to LP medium supplemented with MS vitamins, 30 mg l^–1 casein hydrolysate, and 0.4 g l^–1 agar plus 2.85 g l^–1 Gelrite. Meristems that developed vigorous shoots were used for rooting experiments when they were 2 cm in length. Whole plants were obtained after 5 days of root induction in water-agar medium containing 8.2 M IBA and 5.4 M NAA and 1 month culture in LP medium with 10 g l^–1 sucrose. Plants regenerated from meristems were further propagated by rooting of cuttings. Of the rooted cuttings, 10% were morphologically juvenile.     

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.     

The effects of a fogging system on the physiological status and rooting capacity of leafy cuttings of woody species

The early responses of leafy stem cuttings of Prunus and Castanea species with differing rooting abilities were assessed in a fog system using fluorescence measurements. Different types of cuttings of each species were used: cherry Prunus (‘GiSelA 5’, Prunus cerasus × Prunus canescens—148/2) and chestnut Castanea (‘Marsol’ and ‘Maraval’, Castanea crenata × Castanea sativa). The physiological status of cuttings in the early initiation phase was compared to the rooting results. For all cuttings, fluorescence measurements revealed a close-to-optimum photochemical efficiency, indicating that physiological stress (severance, water, etc.) was minimal. In cherry, the potential photochemical efficiency (Fv/Fm) differed slightly between terminal and basal cherry cuttings, being lower in the basal ones at the time of severance. Later in the propagation process, the differences were smaller. The photochemical efficiency did not differ between two ‘difficult-to-root’ Castanea clones, nor was it dependent on the length of the cuttings. The high rooting capacity of long Castanea cuttings (50 cm) indicated that physiological stress could be minimized under a fogging system. An erratum to this article can be found at     

Stock plant etiolation and stem banding effect on the auxin dose-response of rooting in stem cuttings of Carpinus betulus L. ‘Fastigiata’

Experiments were undertaken to determine the effect of stock plant etiolation and stem banding, prior to cutting propagation, on the auxin dose-response of rooting in Carpinus betulus L. Fastigiata stem cuttings. Stock plants were forced in a greenhouse, etiolated for 10 days and banded with black, light-tight Velcro for 8 weeks. Indole-3-butyric acid was applied to cuttings at concentrations ranging from 0 to 79 mM. Rooting percentages and numbers increased to a peak reponse at 20 mM in light-grown and 40 mM in etiolated shoots, followed by an inhibition at higher concentrations for all except etiolated and banded shoots. Cuttings prepared from shoots which had been etiolated or banded rooted better than controls at low and optimal IBA concentrations. Cuttings from shoots receiving both etiolation and banding also yielded higher rooting percentages and more roots per rooted cutting. Furthermore, etiolation and banding reduced the sensitivity of cuttings to supra-optimal auxin-induced inhibition of adventitious root initiation.     

The influence of medium aeration on in vitro rooting of Australian plant microcuttings

Media with different air filled porosity were compared with standard agar medium for root induction and root elongation for two Australian plants Grevillea thelemanniana and Verticordia plumosa×Chamelaucium uncinatum. Microcuttings from shoot cultures were pulsed for 7 days on a high auxin (40 M IBA), agar-solidified medium in the dark. The rooting of the microcuttings was then compared on standard agar medium (M1, 1/2 MS, no hormones) and on three experimental treatments: – porous-agar medium (1/2 MS, no hormones, 30 g agar l^–1, solidified then blended to provide aeration); – white sand, or white sand wet with M1 medium; and – a sterile propagation mix. The protocol using the propagation mix is referred to as IVS (In Vitro Soil). A separate experiment involved flushing the IVS soil profile with low or normal oxygen. The controls on M1 medium showed low and variable rooting percentages. The rate of root induction and the average total root length per microcutting at final harvest was significantly higher using the IVS protocol, porous-agar or white sand, while addition of agar medium to sand suppressed the percentage rooting and elongation as did flushing the air space in the IVS rooting medium with low oxygen. Other species tested on M1 medium and IVS including Pimelea physodes, Conospermum eatoniae, Verticordia grandis, and a Chamelaucium megalopetalum×C. uncinatum hybrid all showed a significant improvement on the IVS system. The IVS culture technique reduces plant-handling costs.     

Metabolic changes during rooting in stem cuttings of five mangrove species

Vegetative propagation through rooting in stem cuttings in five tree mangroves namely Bruguiera parviflora, Cynometra iripa, Excoecaria agallocha, Heritiera fomes, and Thespesia populnea using IAA, IBA and NAA was reported. Spectacular increase in the root number was noted in the cuttings of H. fomes and C. iripa treated together with IBA (5000 ppm) and NAA (2500 ppm). The highest number of roots was obtained with IBA (2500 ppm) and NAA (500 ppm) in E. agallocha. B. parviflora and T. populnea responded better to IAA and IBA treatment. The species specific variation in the rooting response to exogenous application of auxins was reflected in the metabolic changes during initiation and development of roots in cuttings. Biochemical analysis showed increase of reducing sugar in the above-girdled tissues at initiation as well as subsequent development of roots which was further enhanced by the use of auxins. Decreases in the total sugar, total carbohydrate and polyphenols and increase in total nitrogen were recorded in the girdled tissues and the high C/N ratio at the initial stage helped in initiation of roots in all the species. Interaction of IBA and NAA promoted starch hydrolysis better than IAA and IBA during root development and subsequently reduced the C/N ratio and increased the protein-nitrogen activity during root development which suggest the auxin influenced mobilization of nitrogen to the rooting zone.Abbreviations IAA Indole-3-acetic acid - IBA Indole-butyric acid - NAA A-naphthalene acetic acid     

Establishment of willow cuttings grown in porous membrane root envelopes

Small diameter fine, fibrous roots of willow are exceedingly difficult to remove from most soils. Where field retrieval of entire plants including clean roots may be important, porous membrane rooting envelopes are a method of choice.Purple osier willow (Salix purpurea L.) stem cuttings (clone SP3) were field rooted in buried porous membrane envelopes permeable to water and nutrients. Ramets were grown for two and four months, then separated into component parts for dry weight analysis.At two months, cuttings grown in rooting envelopes were no different in shoot dry weight than plants grown in clay pots or in soil. Tests indicated that rooting envelopes may be reused at least once without affecting shoot or total plant dry weight. Likewise, membrane pore sizes of 0.45, 1.2 and 3 m did not affect SP3 dry weight in four month tests. The importance of matching plant material and experiment duration to envelope size was illustrated by the limitation of growth by envelope edges at four months compared to two months.