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Jenny B. Ryals, Patricia R. Knight, and Eric T. Stafne

), and 1000 ppm IBA + 250 ppm 1-naphthaleneacetic acid, potassium salt (K-NAA) (Hortus IBA Water Soluble Salts; Phytotronics, Earth City, MO). The control was water only (i.e., 0 ppm IBA, 0 ppm NAA). We combined 1000 ppm IBA with 250 ppm K-NAA to see an

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Eugene K. Blythe, Jeff L. Sibley, Ken M. Tilt, and John M. Ruter

In five experiments, singlenode cuttings of `Red Cascade' miniature rose (Rosa) were treated with a basal quick-dip (prior to insertion into the rooting substrate) or sprayed to the drip point with a single foliar application (after insertion) of Dip `N Grow [indole-3-butyric acid (IBA) + 1-naphthaleneacetic acid (NAA)], the potassium salt of indole-3-butyric acid (K-IBA), or the potassium salt of 1-naphthaleneacetic acid (K-NAA); a single foliar spray application of Dip `N Grow with and without Kinetic surfactant; or multiple foliar spray applications of Dip `N Grow. Spray treatments were compared with their respective basal quick-dip controls {4920.4 μm [1000 mg·L-1 (ppm)] IBA + 2685.2 μm (500 mg·L-1) NAA, 4144.2 μm (1000 mg·L-1) K-IBA, or 4458.3 μm (1000 mg·L-1) K-NAA}. Cuttings sprayed with 0 to 246.0 μm (50 mg·L-1) IBA + 134.3 μm (25 mg·L-1) NAA, 0 to 207.2 μm (50 mg·L-1) K-IBA, or 0 to 222.9 μm (50 mg·L-1) K-NAA resulted in rooting percentages, total root length, percent rooted cuttings with shoots, and shoot length similar to or less than control cuttings. Exceptions were cuttings sprayed with 0 to 2.23 μm

(0.5 mg·L-1) K-NAA, which exhibited shoot length greater than the control cuttings. Addition of 1.0 mL·L-1 (1000 ppm) Kinetic organosilicone surfactant to spray treatments resulted in greater total root length and shoot length. Repeated sprays (daily up to seven consecutive days) had no or negative effects on root and shoot development.

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James Robert Ault and Sandy S. Siqueira

supplemented with 0.0, 1.0, 2.0, 4.0, or 8.0 μ m dicamba, picloram, K-NAA, or 2,4-D. The PGRs and concentrations were selected based on previous reports of their use for other Lilium taxa ( Famelaer et al., 1996 ; Haensch, 1996 ; Okazaki and Koizumi, 1995

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Bruce W. Wood

control included Silwet L-77 at 0.025% v/v used in all plant growth regulator treatments at this rate); 2) 6-BA) [or N-(phenyl-methyl)-1H-purine-6-amine, at 50 mg a.i./L]; 3) NAA (K-Salt Fruit Fix ® at 10 mg a.i./L); 4) TIBA (at 100 mg a.i./L); and 5) and

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James R. Ault and Kayri Havens

Shoot explants from actively growing, greenhouse-maintained plants of Baptisia `Purple Smoke' were cultured in vitro for shoot initiation on Murashige and Skoog (MS) basal medium containing vitamins and supplemented with 30 g·L–1 sucrose, 8.87 μm BA, and 4.14 μm K-IBA. All subsequent media were supplemented with 2.47 mm NaH2PO4 to enhance shoot growth. Single-node explants were subcultured for shoot multiplication on MS medium with either no plant growth regulator or with 2.22, 4.44, 8.87, 17.74, or 35.48 mm BA in combination with 0.0 or 4.14 μm K-IBA. Explants produced a maximum of 4.1 shoots on the medium with 2.22 μm BA. Shoots rooted on all concentrations of K-IBA (2.07, 4.14, 10.36, or 20.72 μm) and K-NAA (2.23, 4.46, 11.15, or 22.29 μm) tested. Maximum rooting was 100% on MS medium with 11.15 μm K-NAA; however, this treatment induced copious stem callusing. Rooted shoots were greenhouse-acclimatized for 2.5 weeks. Overall survival was 86%. For optimal rooting and subsequent acclimatization, treatment with 2.23 μm K-NAA is recommended; this resulted in 83% rooting and 87% acclimatization. Chemical names used: N 6 benzyladenine (BA); potassium salt of indole-3-butyric acid (K-IBA); potassium salt of 1-naphthalene acetic acid (K-NAA).

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James R. Ault

Shoot formation was obtained from Lachenalia arbuthnotiae W.F. Barker, L. bulbifera (Cyrillo) Engl., and L. purpureo-coerulea Jacq. leaf tissue explants cultured on Murashige and Skoog (MS) medium supplemented with sucrose at 30 g·liter–1, 8.87 μm BA, and 0.44 μm K-NAA. Shoots of all three species rooted on subculture to MS medium supplemented with 0.0, 4.14, or 8.29 μm K-IBA or 0.0, 4.46, or 8.92 μm K-NAA. Maximum percent rooting was ≈81% from treatment with 4.14 μm K-IBA for L. arbuthnotiae and with 8.29 μm K-IBA for L. purpureo-coerulea; it was 59% from treatment with 8.92 μm K-NAA for L. bulbifera. Rooted and nonrooted shoots were acclimatized in a greenhouse. Survival of rooted plants was 93% for L. arbuthnotiae, 95% for L. bulbifera, and 94% for L. purpureo-coerulea. Survival of nonrooted shoots was 71% for L. arbuthnotiae and 91% for L. bulbifera. Chemical names used: 6-benzyladenine (BA); potassium salt of indole-3-butyric acid (K-IBA); potassium salt of 1-naphthaleneacetic acid (K-NAA).

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James R. Ault

Shoot initiation and multiplication were obtained in vitro from immature flower bud and leaf explants of Veltheimia bracteata Bak. `Lemon Flame' and from leaf explants of V. bracteata `Rosalba' cultured on a Murashige and Skoog (MS) medium supplemented with sucrose at 30 g•L–1, and either 8.87 μm BA plus 0.54 μm NAA or 8.87 μm BA plus 5.40 μm NAA. Shoot initiation and multiplication was obtained from a single leaf explant of Veltheimia capensis (L.) DC. on MS medium with 8.87 μm BA plus 0.54 μm NAA. Shoots of the three genotypes rooted on subculture to medium with 0.0, 4.14, or 8.29 μm K-IBA or 0.0, 4.46, or 8.92 μm K-NAA. Maximal rooting was 98% for V. bracteata `Lemon Flame', 95% for V. bracteata `Rosalba', and 98% for V. capensis, from medium with 4.46 μm KNAA. Rooted shoots were acclimatized for 3 to 4 weeks. Overall survival percentage was 69% for V. bracteata `Lemon Flame', 65% for V. bracteata `Rosalba', and 83% for V. capensis. Chemical names used: 6-benzyladenine (BA); potassium salt of indole-3-butyric acid (K-IBA); potassium salt of 1-naphthaleneacetic acid (K-NAA); 1-naphthaleneacetic acid (NAA).

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Jim Ault* and Sandy Siqueira

Shoot, root, and callus induction were examined in the North American lily, Lilium michiganense, in response to treatment with four auxins. Seed from controlled crosses were aseptically excised from slightly immature capsules and cultured in vitro on Murashige and Skoog basal medium and vitamins with 30 g/l sucrose, 7.0 g/l agar, and a pH = 5.7. Seed were maintained at 20 °C with a 14-h photoperiod. After 5.0-5.5 months, leaves and roots were removed from seedlings, the bulbs transversely sectioned, then the bulb sections cultured cut-surface down on the identical medium supplemented with 0.0, 1.0, 2.0, 4.0, or 8.0 μm dicamba, picloram, K-NAA, or 2,4-D. PGRs were added to medium prior to autoclaving except dicamba which was dissolved in 50% ethanol and added after medium autoclaving. 16 explants were utilized for each treatment. The experiment was conducted three times. Morphogenetic response (# of shoots produced, % of explants forming roots, and % of explants forming callus) was tabulated 4 months after treatment. Shoot formation was promoted by treatment with dicamba, picloram, and K-NAA in comparison to the control (2.5 shoots/explant). Shoot formation varied significantly in response to individual dicamba, picloram, and 2,4-D concentrations. A maximum of 7.9 shoots per explant was promoted by 4.0 μm K-NAA and 1.0 μm dicamba, respectively. Root and callus formation also varied significantly between auxin treatments. Root formation was inhibited by dicamba, picloram, and 2,4-D treatments in comparison with the control (100% rooting); callus formation was promoted by dicamba, picloram, and K-NAA treatments in comparison with the control (15% callusing).

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Leticia Santos-Hernández, Martha Martínez-García, Jorge E. Campos, and Ernesto Aguirre-León

The orchid Laelia albida is an important cultural and religious plant resource of the Tehuacan-Cuicatlan Valley Biosphere Reserve in south-central Mexico. It is gradually becoming scarce due to overcollecting and habitat perturbation. For this reason, and for preservation purposes, the aim of the present work was to obtain the conditions to successfully propagate this species, through the use of mature seeds stored at 4 °C, and basal buds following in vitro techniques. Seeds with different storage periods (lot 1 and lot 2) were analyzed for seed viability as determined by the TTC method, germination percentage and germination index were analyzed on a monthly basis for 11 months on two Knudson C (K-4003, K-4128; Sigma). Seedlings were initially grown on the same culture media variants, but 30 days later they needed to be supplemented with potato starch (20 g·L-1). Basal buds were cultured on Knudson C containing nine BAP and NAA combinations two of which were also supplemented with potato starch (20 g·L-1) and coconut water (10%). Seed viability ranged from 78% to 98% throughout the 11 months and germination percentage was 70% to 90% without significant differences between the two lots. K-4003, either alone or supplemented with potato starch, was the best culture medium used to achieve all the development stages as well as seedlings with rhizoids. Bud proliferation was also successful on K-4003 added with BAP, NAA, potato starch and coconut water. The BAP (7.4 μm) and NAA (5.3 μm) combination promoted shoots and protocorm-like bodies (PLB). The results obtained with basal buds, make its preservation through in vitro culture possible and give the possibility to obtain available plantlets for its cultivation in regional nurseries. Storage conditions tested here may be useful for seed bank management for this species. Chemical names used: 6-benzylaminopurine (BAP), 1-naphthaleneacetic acid (NAA).

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Hamid Elhag, Mahmoud M. El-Olemy, and Mansour S. Al-Said

Somatic embryogenesis of Nigella sativa was investigated with the objective of inducing and isolating somatic embryos for biosynthetic studies. Callus cultures were initiated from leaf, stem, and root explants of axenic seedlings on MSB5 basal medium supplemented with kinetin (0.46 μm) and 2,4-D (4.5 or 13.5 μm) or NAA (5.4 or 16.2 μm) in the dark. Cultures initiated and subcultured on medium containing NAA produced friable callus with numerous roots regardless of explant type. Cultures initiated, subcultured, or both, on medium with low 2,4-D concentration produced shiny embryogenic masses. These cultures differentiated into somatic embryos on medium containing NAA. The embryos developed into leafy structures on basal medium devoid of growth regulators. When the embryogenic callus was transferred to liquid medium containing NAA, numerous embryos and clusters of embryos were released into the liquid medium but, in contrast to solid medium, development remained arrested at the early embryonic stages. The developmentally arrested embryos were tested for production of active constituents of N. sativa oil. Chemical names used: 2,4-dichlorophenoxyacetic acid (2,4-D); α-naphthaleneacetic acid (NAA); kinetin (K).