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Carlos A. Lazcano, Fred T. Davies Jr., Andrés A. Estrada-Luna, Sharon A. Duray, and Victor Olalde-Portugal

Mature cladodes of prickly-pear cactus (Opuntia amyclaea Tenore. `Reina') were treated with five wounding methods and four concentrations of potassium salt indole-3-butyric acid (K-IBA) to stimulate adventitious root formation. The wounding method and K-IBA had highly significant effects on root number and root dry mass of cladodes. Interaction between K-IBA and wounding methods showed that greater root number was obtained at the higher auxin concentrations and with wounding methods that had the greatest cut surface area. K-IBA concentrations from 4,144 to 41,442 μm (1,000 to 10,000 mg·L-1) increased root dry mass. Only the wounding method affected rotting of cladodes. Treatments allowing suberization had a higher percentage of nonrotted cladodes. This research validates the commercial practice of allowing cladodes to suberize early in the propagation cycle. K-IBA altered rooting polarity and stimulated adventitious root formation along the wounded cladode surfaces. The vertical nonsuberized wounding methods and auxin treatments are an excellent classroom demonstration for manipulating rooting polarity. Auxin application and wounding could be of commercial benefit for enhanced rooting in the clonal regeneration of new selections for prickly-pear cactus orchards.

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Kourosh Vahdati, James R. McKenna, Abhaya M. Dandekar, Charles A. Leslie, Sandie L. Uratsu, Wesley P. Hackett, Paola Negri, and Gale H. McGranahan

Walnuts (Juglans spp.) are difficult-to-root woody plants. The rolABC genes (rolA + rolB + rolC), derived from the bacteria Agrobacterium rhizogenes, have been shown to increase the rooting potential of other difficult-to-root woody plants. We inserted the rolABC genes into somatic embryos of a `Paradox' hybrid (J. hindsii × J. regia) clone PX1 using the A. tumefaciens gene transfer system. A transgenic sub-clone, designated PX1 rolABC 2-2 was selected and compared to the untransformed clone for a variety of phenotypic characteristics, including rooting potential. Transformed and untransformed shoots were budded onto seedling J. regia rootstock in the greenhouse and established in the field. Transformed trees displayed reduced internode length, an increase in lateral branching, and wrinkled leaves. In another test, a commercial persian walnut cultivar J. regia `Chandler' was grafted onto rooted cuttings of both the untransformed and transformed plants. The presence of the rolABC genes in the rootstock had no visible effects on the grafted scion. Several of these trees were excavated from the field and the root systems of each genotype were examined for root number, diameter, and biomass. Trees with the rolABC rootstock had significantly more small diameter roots compared to the controls and less recovered biomass. Tests of the rooting potential of leafy semi-hardwood cuttings for two years resulted in 14% to 59% rooting of the transformed cuttings compared to 51% to 81% rooting of the control. Both transformed hardwood cuttings and microshoots in tissue culture also rooted significantly less (52% and 29% respectively) than untransformed hardwood cuttings and tissue cultured shoots (82% and 54% respectively). Thus, although the rolABC genes induced a shorter internode length and a more fibrous root system (typical of rol-tranformed plants), they were not useful for increasing the rooting potential, and as rootstock they did not affect the phenotype of the scion.

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Robert L. Geneve, Wesley P. Hackett, and Bert T. Swanson

Exogenous ethylene could not substitute for NAA to induce adventitious root initiation in juvenile petiole explants of English ivy (Hedera helix L.), indicating that the action of auxin-stimulated root initiation was not directly mediated through ethylene production. Mature petioles did not initiate roots under any auxin or ethylene treatment combination. Ethephon or ACC supplied at 50 or 100 μm was inhibitory to NAA-induced root initiation in juvenile petioles. The pattern of ethylene production stimulated by NAA application was significantly different in juvenile and mature petioles. Ethylene evolution by juvenile petioles declined to near control levels during from 6 to 12 days after NAA application. Reduction in ethylene production was due to reduced availability of ACC in juvenile petioles. Mature petioles continued to produce ethylene at elevated levels throughout the course of the experiment. Ethylene does not appear to play a significant role in the differential root initiation response of juvenile and mature petioles treated with NAA. However, ethylene appeared to have an inhibitory effect during root elongation stages of adventitious root development in juvenile petioles. Chemical names used: 1-aminocyclopropane-1-carboxylic acid (ACC); 1-napthaleneacetic acid (NAA); 2-chloroethylphosphonic acid (ethephon).

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Mateja Štefančič, Franci Štampar, and Gregor Osterc

The influence of two exogenously applied auxins (IAA and IBA) on the root and shoot development of leafy cuttings was analyzed at 'GiSelA 5', the dwarfing cherry rootstock. IBA (indole-3-butyric acid) hindered the callus formation in the early period of root development and it was more successful than IAA (indole-3-acetic acid) in promoting earlier root development. IBA also influenced the stronger shoot growth and the development of acrobasal type of the rooting system, and induced higher number of roots. Those parameters are very important for the quality and survival of the new plants and they are not the consequence of the higher IAA content in the rooting zones of cuttings in the first days of root development. Both auxin treatments had no effect on the final percent of the rooted cuttings neither on the survival of cuttings, but they increased the percent of rooted cuttings without callus. The root system with callus proved less qualitative, because the cuttings with such root system developed significantly less roots per rooted cutting and their shoot length was shorter than those of the cuttings without callus at both auxin treatments. Exogenously applied auxins were not crucial for root formation, however their application resulted in higher percent of more qualitative 'GiSelA 5' leafy cuttings. IBA proved as the most efficient treatment and it additionally induced earlier root formation.

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James F. Harbage, Dennis P. Stimart, and Carol Auer

The influence of root initiation medium pH on root formation was investigated in relation to uptake and metabolism of applied IBA in microcuttings of Malus ×domestica Borkh. `Gala' and `Triple Red Delicious'. Root formation and uptake of H 3-IBA were related inversely to root initiation medium pH. Maximum root count (10.3 roots) and IBA uptake were observed at pH 4.0. Regardless of pH, overall root count of `Gala' was higher (13.5 roots) than `Triple Red Delicious' (4 roots). Uptake of IBA was highest at pH 4.0 for `Gala' (1.7% uptake) and at pH 4 and 5 for `Triple Red Delicious' (0.75% uptake). Metabolism of IBA was the same regardless of root initiation medium pH or cultivar examined. One-half of the IBA taken up was converted to a compound that coeluted with IBAsp during high-performance liquid chromatography. Apparently, pH regulates root formation by affecting IBA uptake but not metabolism. The level of auxin in tissue appeared unrelated to root formation between genotypes. Chemical names used: 1H-indole-3-butyric acid (IBA); 5-H 3-indole-3-butyric acid (H 3-IBA); indole-3-butrylaspartic acid (IBAsp).

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Wen-Quan Sun and Nina L. Bassuk

Single-node `Royalty' rose (Rosa hybrida L.) cuttings were used to examine the relationship between adventitious root formation, budbreak, and ethylene synthesis following IBA treatment. IBA was applied as a lo-second basal quick dip before rooting, and AIB, GA3, STS, and ethephon were applied either as basal dips or foliar sprays. IBA application increased rooting and inhibited budbreak of cuttings. IBA 2 600 mg·liter-1 greatly inhibited budbreak during 4 weeks of rooting. IBA treatment stimulated ethylene synthesis, which was inversely correlated with budbreak of cuttings. Ethephon also significantly inhibited budbreak. Budbreak of rose cuttings was completely prevented by repeated ethephon sprays used to maintain high endogenous ethylene levels during the first 10 days. Treatment with STS, an ethylene-action inhibitor, improved budbreak. The inhibition of budbreak by IBA treatment resulted primarily from elevated ethylene levels. Root initiation and root elongation of cuttings initially inhibited budbreak, but later promoted budbreak. Chemical names used: indole-3-butyric acid (IBA); gibberellic acid (GA3); silver thiosulfate (STS); AIB, aminoisobutyric acid (AIB); (2-chloroethyl)-phosphoric acid (ethephon).

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Mehmet Nuri Nas and Paul E. Read

Microshoots of four hazelnut genotypes grown in vitro on Nas and Read medium (NRM) containing various combinations of CuSO4 • 5H2O and myo-inositol were successfully rooted and acclimatized ex vitro without any need of in vitro hardening treatments. Dipping of shoot bases in 1000 ppm indole-3-butyric acid (IBA) solution for 5 or 10 seconds followed by placement of shoots in plant growth regulator free NRM gave rise to formation of roots as early as 8 days. Shoots treated for 5 and 10 seconds rooted similarly, and depending on genotype, 88% to 98% rooting was observed within 15 days after treatment with IBA. Ex vitro survival of shoots three months after in vitro-root induction was 73% when shoots were treated with IBA for 5 seconds and 66% when shoots were treated for 10 seconds. The highest ex vitro survival rate (97%) 3 months after root induction was observed when shoots were treated with IBA solution for 10 seconds, and then cultured directly in peat pellets. Shoots developed good roots, and grew up to 70 cm in height 3 months after root induction. The potential use of rooting and acclimatization protocol for commercial micropropagation of hazelnut is presented.

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Robert L. Geneve, Wesley P. Hackett, and Bert T. Swanson

Several inhibitors of ethylene biosynthesis and action, as well as an atmospheric ethylene scrubber, were used to investigate the role of ethylene in adventitious root initiation in de-bladed petioles from the juvenile and mature phase of English ivy (Hedera helix L.). Induction of root primordia required NAA regardless of the inhibitor treatment. Difficult-to-root mature petioles have been shown to produce higher amounts of ethylene than easy-to-root juvenile petioles. However, mature petioles failed to root under any combination of NAA and inhibitor treatment, indicating that the continued evolution of ethylene in NAA-treated mature petioles was not responsible for the absence of a rooting response. Root initiation in juvenile petioles was not affected by treatment with the ethylene action inhibitors STS and NDE, nor by removal of atmospheric ethylene with KMnO. Inhibition of ethylene biosynthesis using AVG or AOA reduced root initiation in juvenile petioles, but this response was not well-correlated to the observed reduction in ethylene evolution. The inhibitory action of AVG could not be reversed by the addition of ethylene gas or ACC, which indicated that AVG could be acting through a mechanism other than the inhibition of ethylene biosynthesis. Chemical names used: 1-naphthalene acetic acid (NAA); l-aminocyclopropane-l-carboxylic acid (ACC); silver thiosulfate (STS); 2,5-norbornadiene (NDE); aminoethyoxyvinyl-glycine (AVG); aminooxyacetic acid (AOA).

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Zhaohui Li, Yan Ma, Wanyuan Yin, Dekui Zang, and Xianfeng Guo

determine the cutting type by testing the effects of gradient concentrations of indole-3-butyric acid (IBA) and cutting positions on rooting performance and, second, to investigate the effects of the collection times of cuttings on rooting performance and to

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David Jespersen and Brian Schwartz

stomatal aperture, density of stomata, as well as canopy characteristics such as leaf texture, length, and density all play important roles in regulating transpiration rates. Rooting characteristics are another major aspect affecting drought avoidance