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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.

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James R. Ault

1 To whom reprint requests should be addressed; e-mail . I thank volunteer Ling-Ling Wei for conducting much of the tissue culture work.

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Carol Auer, Jerry D. Cohen, and Todd Cooke


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Abba Upadhyaya, Tim D. Davis, Daksha Sankhla, and N. Sankhla

Both kinetin and BA promoted in vitro shoot formation from hypocotyl explants of Lupinus texensis Hook. placed on Murashige and Skoog (MS) medium. With either cytokinin, shoot formation was best at ≈4.5 μm. Adventitious root formation was observed only on tissue culture-derived shoots placed in MS media containing 5.4 to 54 μM NAA. IAA and IBA, at concentrations ranging from 5 to 55 μm, failed to stimulate rooting. Even at the optimal concentration of NAA, only 14% of the shoots produced roots. Thus, although hypocotyl explants readily produced shoots, adventitious root formation on these shoots occurred with relatively low frequency. Chemical names used: 6-benzylaminopnrine (BA); indole-3-acetic acid (IAA); indole-3-butyric acid (IBA); 6-furfurylaminopurine (kinetin); 1-naphthaleneacetic acid (NAA).

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Takashi Ikeda, Yukihiro Fujime, Satoshi Terabayashi, and Shuichi Date

Garlic (Allium sativum L.) calli in vitro were evaluated over a range of salt concentrations and by adding mannitol to culture medium with reduced salt to provide equivalent osmoticum. The water potential of the medium ranged from -0.27 to -0.73 MPa under the various salt and osmotic stress conditions. The percent increase in calli was highest in standard Murashige & Skoog (MS) medium and was reduced when MS salts were reduced but the water potential of medium was adjusted to that of standard MS medium by addition of mannitol. The water potential of callus tissue was similar to that of tissue culture media over a 20-fold range (10% to 200%) of MS concentrations. Turgor of callus tissue was not influenced by any stress conditions. These results indicate that the optimum concentration of salt and water status of medium for formation of garlic calli was provided by standard MS medium.

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Lisa J. Rowland and Elizabeth L. Ogden

Conditions for improving the efficiency of shoot regeneration from leaf sections of highbush blueberry (Vaccinium corymbosum L.) were investigated. Effectiveness of tissue culture medium supplemented with the cytokinin conjugate zeatin riboside or the cytokinin zeatin at 10, 20, or 30 μm was compared with medium supplemented with the optimum 2iP concentration of 15 μm. Use of 20 μm zeatin riboside resulted in the most shoots per leaf section, » 6-fold higher than the number of shoots produced on 2iP medium. The number of shoots produced on medium supplemented with zeatin was not significantly higher than the number of shoots produced on 2iP medium. Consequently, we concluded that the cytokinin conjugate zeatin riboside was more effective than either of the free cytokinins, 2iP or zeatin, in promoting shoot regeneration from leaf sections of highbush blueberry. Chemical names used: 6-(y,y-dimethylallylamino)-purine (2iP); 6-(4-hydroxy-3-methyl-but-2-enylamino)purine (zeatin).

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Dao-Shun Huang, Jingkun Ho, and Ralph Backhaus


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Cynthia Zurinsky, Michael E. Kane, and Nancy Philman

24 ORAL SESSION 6 (Abstr. 040-046) Cross-commodity: Cell and Tissue Culture

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Hyo-Geur. Park, Kyung-Young Choi, and Do-Hyun Lee

164 ORAL SESSION 46 (Abstr. 326–332) Cross-commodity: Cell and Tissue Culture

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Juan C. Díaz, Kenneth Shackel, and Ellen Sutter

30 ORAL SESSION 3 (Abstr. 020–026) Cell and Tissue Culture: Fruits and Nuts