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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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U.L. Yadava and S.K. Dhir

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

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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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Yasseen Mohamed-Yasseen and Walter E. Splittstoesser

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

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

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

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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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Karim H. Al-Juboory and Jabar H. Al-Niami

108 POSTER SESSION (Abstr. 362–374) Cell and Tissue Culture I

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S.A. Merkle and B.A. Watson-Pauley

Low conversion rates of somatic embryos and poor early growth of somatic embryo-derived plantlets of some forest trees may be related as much to prolonged maintenance in vitro as to basic developmental problems with the embryos. We tested ex vitro conversion as an alternative method for producing the rare North American pyramid magnolia (Magnolia pyramidata Bartram) plantlets from somatic embryos. Tissue cultures were initiated from immature seed explants of pyramid magnolia. Immature seeds collected from each of three trees formed proembryogenic masses (PEMs) following 7 to 10 weeks of continuous culture on semisolid medium containing 9.0 μm 2,4-D, 1.1 μm BA, and 1 g casein hydrolysate/liter. PEMs transferred to semisolid medium without plant growth regulators produced somatic embryos that germinated following transfer to the same medium without casein hydrolysate. Conversion frequency to plantlets was higher and plantlets were more vigorous when germinants were transferred directly to potting mix and grown in a humidifying chamber instead of being maintained in plantlet development medium in test tubes. Chemical names used: 2,4-dichlorophenoxyacetic acid (2,4-D); N-(phenylmethyl)-1H-purine-6-amine (BA).

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A. Raymond Miller, Joseph C. Scheereus, Patricia S. Erb, and Craig K. Chandler

A tissue culture protocol was developed that increased the germination percentage and decreased the lag time to germination for strawberry (Fragaria x ananassa Duch.) achenes. This technique involved cutting surface-sterilized achenes across the embryo axis then placing the shoot apex/radicle-containing sections on semisolid Murashige and Skoog medium lacking hormones. Cut achenes began germinating 5 days after culture and achieved maximum germination (97% to 100%) in less than 2 weeks, compared to whole achenes, which began to germinate 7 to 10 days after sowing and required more than 7 weeks for maximum germination (<50%). Enhanced germination of cut achenes was a general phenomenon since achenes from 231 hybrid crosses responded similarly. Following placement on culture medium, cut achenes could be stored up to 8 weeks at 4C then removed to 27C, where germination and seedling development occurred at percentages and rates comparable to freshly cut achenes. Achenes did not require stratification before cutting to exhibit increased germination. Nearly 100% of the achenes from freshly harvested red-ripe, pink and white strawberries germinated after cutting and culture, although cut achenes from white and pink berries germinated more slowly than those from red-ripe berries. Achenes from green berries, whether whole or cut, did not germinate. This method of “embryo rescue” could be used to generate more seedlings from poorly germinating hybrid crosses, would considerably decrease the time from sowing to seedling production compared to traditional means, and would produce seedlings of uniform age for subsequent field evaluation.