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Carole H. Saravitz, Frank A. Blazich, and Henry V. Amerson

Adventitious shoots developed on cotyledons of Virginia pine (Pinus virginiana Mill.) excised from seeds subjected to H2O2 treatment for 3, 6, or 9 days and cultured on media containing 0.5 to 10 mg BA/liter. Shoot regeneration was greatest (42 shoots per embryo) on cotyledons from seeds treated with H2O2 for 6 days and placed on medium containing BA at 10 mg·liter-1. Excised shoots elongated on medium lacking BA. Following elongation, shoots were placed on media containing IBA at 0 to 40 mg·liter-1 for 14 days followed by transfer to the same medium lacking auxin. Without IBA treatment, rooting was 3%, and increased to 50% for 5 to 40 mg·liter-1. Rooted shoots averaged 2.0 roots per shoot without auxin incorporation, 3.3 roots when treated with 5 mg IBA/liter, and the number of roots increased linearly with increased IBA concentration up to 40 mg·liter-1 (4.5 roots). Plantlets were transferred to growing medium and acclimated successfully to greenhouse conditions. Chemical names used: N- (phenylmethyl)-1 H- purine-6-amine (BA), 1 H- indole-3-butyric acid CBA).

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Michael A. Arnold and Eric Young

Bare-root Malus × domestica Borkh. seedlings were chilled for 0, 600, 1200, or 1800 hours at 5C (CH). Seedlings were then placed with roots and/or shoots in all combinations of 5 and 20C forcing conditions (FC) for up to 21 days. Virtually no growth occurred at 5C FC. When the whole plant was forced at 20C, all measures of root and shoot growth increased in magnitude, occurred earlier and at a faster rate with increasing CH. Thus, roots and shoots responded similarly to chilling. When shoots or roots were subjected to 20C FC, while the other portion of the plant was at 5C, the responses were reduced in magnitude and delayed. However, the overall growth enhancement by chilling was not negated. Root and shoot growth enhancement by chilling appeared to be increased if the other portion of the plant was actively growing also, but not dependent on it. Growth of adventitious shoots on roots (root suckers) was greatly enhanced with increasing CH on plants subjected to 5C shoot and 20C root FC. While total root and shoot bark protein levels on a per-seedling basis were similar, protein concentrations were lower in root bark than in shoot bark. During chilling, total protein per seedling generally increased until just before the time that chilling requirements for vegetative budbreak were satisfied. Protein degradation then began, resulting in lower protein levels through 2300 CH. Rapid protein breakdown (1200 to 1800 CH, roots; 1000 to 1800 CH, shoots) occurred at about the same time that root (1000 to 1800 CH) and shoot (800 to 1800 CH) growth responses to chilling were increasing. Warm FC resulted in increased protein breakdown with increased CH and forcing time.

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J.T.A. Proctor, T. Slimmon, and P.K. Saxena

Ginseng is an herbaceous perennial that grows in the understorey of deciduous hardwood forests and is also cultivated for its highly valued root. The primary method of propagation of ginseng is by seed which requires the breaking of dormancy by stratification, a process which takes 18–24 months. Investigation of factors controlling the growth and development of ginseng plants is a prerequisite to the development of a more efficient system of ginseng propagation. We have recently modulated the morphogenetic potential of geranium roots and stimulated de novo development of shoots and embryo-like structures which later formed whole plants using thidiazuron (TDZ). Our objective was to investigate the morphological changes in seedling and mature ginseng plants induced by TDZ, particularly in relation to root and shoot morphogenesis and economic yield. Applications of TDZ (0.22 and 2.20 ppm), either as foliar sprays or soil watering to greenhouse-grown seedlings over 18 weeks (2 weeks after sowing to 20 weeks when plants were harvested) induced similar effects. These responses included increased stem length and diameter, and shoot and root weight (economic yield). Single foliar applications of TDZ at 62.5 and 125 ppm to 3-year-old field-grown ginseng plants 3 months before harvest increased root biomass (economic yield) by 19% to 23%. Roots of TDZ-treated seedlings and 3-year-old field-grown plants developed thickened secondary roots on the upper part of the taproot. The root-like structure of these secondary roots was confirmed by histology. In addition, TDZ treatments induced adventitious buds on the shoulder of 3-year-old roots. These buds developed into shoots to give multi-stem plants following a period of dormancy, which was overcome with GA3 (gibberellic acid) treatment before planting.

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Carole H. Saravitz, Frank A. Blazich, and Henry V. Amerson

Adventitious shoots developed on cotyledons of Virginia pine (Pinus virginiana Mill.) excised from seeds germinated for 3, 6, or 9 days and cultured on media containing 0.5 to 10 mg/liter benzyladenine (BA). Shoot regeneration was greatest (46 shoots per embryo) on cotyledons from seeds germinated for 6 days and placed on medium containing 10 mg/liter BA. Shoots were excised and elongated on medium lacking BA. Following elongation, shoots were placed on media containing 0 to 40 mg/liter indolebutyric acid (IBA) for 14 days followed by transfer to the same medium lacking auxin. Without IBA treatment, percent rooting was 3% and increased to 50% for concentrations of 5 to 40 mg/liter. Rooted shoots averaged 2.0 roots per shoot without auxin treatment, 3.3 roots when treated with 5 mg/liter IBA and root number increased linearly with increased IBA concentration up to 40 mg/liter (4.5 roots). Plant lets were transferred to growing medium and acclimated successfully to greenhouse conditions.

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Carole H. Saravitz, Frank A. Blazich, and Henry V. Amerson

Adventitious shoots developed on cotyledons of Virginia pine (Pinus virginiana Mill.) excised from seeds germinated for 3, 6, or 9 days and cultured on media containing 0.5 to 10 mg/liter benzyladenine (BA). Shoot regeneration was greatest (46 shoots per embryo) on cotyledons from seeds germinated for 6 days and placed on medium containing 10 mg/liter BA. Shoots were excised and elongated on medium lacking BA. Following elongation, shoots were placed on media containing 0 to 40 mg/liter indolebutyric acid (IBA) for 14 days followed by transfer to the same medium lacking auxin. Without IBA treatment, percent rooting was 3% and increased to 50% for concentrations of 5 to 40 mg/liter. Rooted shoots averaged 2.0 roots per shoot without auxin treatment, 3.3 roots when treated with 5 mg/liter IBA and root number increased linearly with increased IBA concentration up to 40 mg/liter (4.5 roots). Plant lets were transferred to growing medium and acclimated successfully to greenhouse conditions.

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Christopher L. Rosier, John Frampton, Barry Goldfarb, Frank A. Blazich, and Farrell C. Wise

Seven concentrations of IBA and seven concentrations of NAA plus a nonauxin control were tested over three growth stages to determine their effectiveness in promoting adventitious root formation on stem cuttings taken from 3- and 4-year-old stock plants of Fraser fir [Abies fraseri (Pursh) Poir.]. Cuttings were prepared in March (hardwood), June (softwood), or November (semi-hardwood) 2001, treated with auxin concentrations ranging from 0 to 64 mm, and placed under mist. Rooting percentage, percent mortality, number of primary roots, total root length, root system symmetry, and root angle were recorded after 16 weeks. Growth stage and auxin concentration significantly affected every rooting trait except root angle. NAA significantly increased the number of primary roots and total root length. However, auxin type did not significantly affect rooting percentage or percent mortality. The highest rooting percentages (99%) occurred when softwood cuttings were treated with 5 mm auxin, however, semi-hardwood cuttings also rooted at high percentages (90%) and had no mortality when treated with 14 mm auxin. Regardless of auxin type, the number of primary roots and total root length varied in similar patterns across concentration, although, NAA tended to induce a greater response. To root Fraser fir stem cuttings collected from 3- and 4-year-old stock plants, it is recommended that a concentration of 5 mm NAA should be used on softwood cuttings and 14 mm IBA on semi-hardwood cuttings. Chemical names used: indole-3-butyric acid (IBA); 1-naphthaleneacetic acid (NAA).

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Ramon Dolcet-Sanjuan, Elisabet Claveria, Robert Gruselle, Adreas Meier-Dinkel, Christian Jay-Allemand, and Thomas Gaspar

Various factors were found to influence the in vitro induction and elongation of adventitious roots from walnut shoot microcuttings. Diverse walnut genotypes (Juglans regia, J. nigra × J. regia hybrids) and selected elite J. regia clones were micropropagated throughout the establishment of in vitro shoot-tip cultures. New evidence is presented here that demonstrates the importance of the genotype and juvenility of the plant material on the in vitro rooting ability. Selection of the best adapted genotypes to multiplication and rooting, and rejuvenation of mature clones through repetitive subcultures or micrografting were examined. Adult J. regia clones were rejuvenated through subsequent subcultures and their rooting was consequently improved. The same results were not accomplished by micrografting on juvenile shoots. A differential response to auxin type and concentration was observed for Juglans regia or J. nigra × J. regia clones. A short prerooting culture in multiplication medium, lowering the sucrose concentration in the root elongation medium and increasing the atmospheric carbon dioxide during the root elongation phase affected the number of shoots forming roots as well as the quality of plantlets and roots.

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Brian A. Kahn and Peter J. Stoffella

Field experiments were conducted in 1985 at Fort Pierce, Fla., and Bixby, Okla., to quantify and describe the distribution of nodules among root morphological components of cowpea [Vigna unguiculata (L.) Walp.]. Plants of `Knuckle Purplehull', `Mississippi Cream', and `White Acre' were sampled by cultivar on separate dates at three growth stages: pre-anthesis, seed initiation, and harvest, when most pods were dry. Root masses were partitioned into adventitious, basal, lateral, and taproot components. Nodules were removed from roots, grouped according to root morphological component of origin, and weighed. No linear correlation was found between the weight of a particular root morphological component and the nodule weight associated with that component. Total root weight and total nodule weight also were not strongly correlated. Nodule weights usually were lower at harvest than at earlier stages of ontogeny, especially for nodules from taproots. Although ≈70% of the root mass was in the taproot and its associated laterals at both locations, the taproot per se was not the primary locus of nodulation. Instead, most nodules generally were located on the basal and lateral roots. When percentage distribution of total nodule weight was examined, neither growth stage nor cultivar was found to affect nodulation of basal or lateral roots.

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Steven T. McNamara and Cary A. Mitchell

The relative contributions of auxin and ethylene (C2H4) in stimulating the initiation of adventitious root primordia (ARP) and their subsequent development into adventitious roots (ARs) by flooded tomato (Lycopersicon esculentum Mill. PI 406966) seedlings were evaluated using TIBA and STS. Flooded plants treated with STS (F + STS) produced ≈ 40% as many emerged ARs as plants that were flooded only (F). Only 7% of the ARP initiated by F + STS plants developed enough to emerge through the epidermis by 120 hours of treatment compared with 95% emerged for F plants. A band of TIBA applied below the lowest leaves of flooded plants (F + TIBA) virtually eliminated AR formation. Plants with two or four leaves below the TIBA band produced 16- and 35-fold more ARs, respectively, than those with no leaves below the TIBA band. Relative to nonflooded (NF) plants, F + STS plants exhibited a nearly 40-fold increase in C2H4 evolution, while F and F + TIBA plants exhibited about a 5-fold increase in C2H4 production. These results suggest that auxin accumulation at or above the floodline is essential for ARP initiation and that auxin action is not mediated through C2H4. Ethylene may be required for elongation of flood-induced ARP leading to their emergence as ARs. Chemical names used: 2,3.5 -triiodobenzoic acid (TIBA): silver thiosulphate (STS).

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Dennis P. Stimart and James F. Harbage

The role of the number of adventitious roots of Malus domestics Borkh. `Gala' microcuttings in vitro on ex vitro root and shoot growth was investigated. Root initiation treatments consisted of IBA at 0, 0.15, 1.5, 15, and 150 μm in factorial combination with media at pH 5.5, 6.3, and 7.0. IBA concentrations significantly influenced final root count and shoot fresh and dry weights, but not plant height, leaf count, or root fresh and dry weights at 116 days. Between 0 and 0.15 μm IBA, final root counts were similar, but at 1.5, 15, and 150 μm IBA, root counts increased by 45%, 141%, and 159%, respectively, over the control. The pH levels did not affect observed characteristics significantly. There was no significant interaction between main effects. A significant positive linear relationship was found between initial and final root count. The results suggest a limited association between high initial adventitious root count and subsequent growth. Chemical name used: 1 H -indole-3-butyric acid (IBA).