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Ravindra K. Hajela, Neerja Hajela, Mark G. Bolyard, Wayne M. Barnes, and Mariam B. Sticklen

A simple gene transfer method based on Agrobacterium -mediated transformation of adventitious multiplication of Juneberry (Amelanchier laevis L.) basal shoots is described. Evidence is presented for successful integration and expression of a transformed gene in greenhouse-grown transgenic plants. This method can transform woody perennials that are difficult to regenerate from leaf disks, protoplasts, or other tissue culture regimens.

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Bruce J. Parliman, Phillip T. Evans, and Earlene A. Rupert

Abstract

A single rhizome explant of the Venus fly-trap has the potential to produce 14 or more rooted plantlets in 40 to 60 days when cultured on a medium containing half strength Murashige and Skoog salts, organic components, naphthaleneacetic acid (NAA) at 1.9 mg/liter and 6-benzylamino purine (BA) at 0.2 mg/liter. Cultures were grown in 16 hour cycles of Cool White fluorescent light at 23° to 26°C. Explants derived from either lateral buds or adventitious buds from leaf cuttings have equal potential for rapid multiplication. This same medium produced optimum plantlet size and quality. Supplementing the basal medium with 0.3 or 1.0 mg/liter of GA3 decreased the number of explants and increased the size of plantlets prior to acclimatization. Media containing higher and lower salt concentrations and higher and lower IAA, NAA, 2,4-dichlorophenoxyacetic acid (2,4-D), BA, or 6(γ,γ, dimethylallylamino)-purine (2ip), produced fewer plantlets while increasing deleterious effects. The rapid plantlet multiplication procedure described will increase commercial availability of the plants while decreasing collection pressures on wild germplasm pools.

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R.C. Apter, E.L. McWilliams, and F.T. Davies Jr.

One-node explants and one-node stem cuttings of Asian jasmine [Trachelospermum asiaticum (Siebold & Zucc.) Nakai] were rooted, respectively, in vitro [tissue culture (TC)] or by conventional macropropagation (MACRO). The TC and MACRO stem bases were then analyzed for differences in the time-course sequence of 1) root primordia initiation and development and 2) adventitious root xylem development and root-to-shoot xylem connections. Early root primordia were observed at Day 3, and, by Day 7, root-to-shoot xylem connections were equally developed in TC and MACRO systems. Continued development and emergence of adventitious roots were observed at Days 8 to 10. At Days 13 and 18, when viewed using scanning electron microscopy, TC root hairs were morphologically thicker and one-third to one-half the length of MACRO root hairs. There was no apparent difference in root-hair density. Inferior TC root-hair length may be a factor in the acclimation of TC-generated plantlets.

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Hazel Y. Wetzstein and Harry E. Sommer

Abstract

The acclimatization or hardening-off of in vitro-cultured sweetgum (Liquidambar styraciflua L.) plantlets was studied using scanning electron microscopy. Comparisons were made among leaves of plantlets differentiated in culture, plantlets acclimatized after transfer from in vitro conditions, greenhouse seedlings, and mature trees. Leaves of plantlets directly from tissue culture had superficial, circular stomata and epidermal cells with irregular, sinuous undulations in the anticlinal walls. Leaves from acclimatized plantlets had ellipsoid, depressed stomata and irregularly shaped epidermal cells. Seedling and field-grown leaves had depressed, ellipsoid stomata and well-defined isodiametric epidermal cells. Stomata in all cases were confined to the abaxial surface, with densities significantly greater in leaves of in vitro plantlets than in acclimatized plantlets or greenhouse-grown plants. Epicuticular wax was generally smooth and absent of waxy outgrowths in all conditions.

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Freddi A. Hammerschlag, Ghazala Hashmi, Robin Huettel, Dennis Werner, and David Ritchie

134 WORKSHOP 20 Reevaluating Tissue Culture Techniques for Generating Useful Variation and for in Vitro Screening

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G.R. de L. Fortes, A.M. R. Vieira, and D.L. Leite

139 POSTER SESSION 21 Cell & Tissue Culture/Cross-Commodity

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U.L. Yadava and S.K. Dhir

139 POSTER SESSION 21 Cell & Tissue Culture/Cross-Commodity

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Yasseen Mohamed-Yasseen and Walter E. Splittstoesser

134 ORAL SESSION (Abstr. 637-644) CROSS-COMMODITY TISSUE CULTURE V/MICROPROPAGATION

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S. Latha Kancherla and Prem L. Bhalla

Pandoreas, Australian natives of horticultural significance, were successfully propagated using tissue culture. A protocol for rapid in vitro multiplication of commercial cultivars was developed using nodal segments cultured on Murashige and Skoog medium containing either BA or kinetin. Maximum shoot induction and number of shoots per explant for P. pandorana (Andrews) Steenis and P. jasminoides (Lindley) Schumann were on 8.8 μm BA and 4.6 μm kinetin. Higher levels of cytokinin in the medium inhibited shoot formation. Tissue-cultured shoots were rooted with IBA. This study demonstrates that Pandoreas can be successfully micropropagated. Chemical names used: 6-benzylaminopurine (BA); 3-indole butyric acid (IBA).

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Yousef I. Dlaigan, A.E. Said, and M.A. El-Hamady

139 POSTER SESSION 21 Cell & Tissue Culture/Cross-Commodity