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A. Raymond Miller and Craig K. Chandler

A protocol was developed for excising and culturing cotyledon explants from mature achenes of strawberry (Fragaria × ananassa Duch.). Cotyledon explants formed callus with multiple shoot buds on agar-solidified Murashige and Skoog media containing several combinations of hormones (1 μm 2,4-D; 10 μm 2,4-D; 1 μm BA + 1 μm 2,4-D; 1 μm BA + 10 μm 2,4-D; 5 μm BA; 5 μm BA + 1 μm 2,4-D; 5 μm BA + 10 μ m 2,4-D; 5 μ m BA + 5 μm NAA; 5 μ m BA + 15 μ m NAA). After three subcultures, only tissues maintained on the medium containing 5 μm BA + 5 μm NAA continued to form shoots. Tissues transferred to other media eventually died (1 μm 2,4-D; 1 μ m BA + 10 μ m 2,4-D; 5 μ m BA; 5 μ m BA + 1 μ m 2,4-D), became unorganized (1 μm BA + 1 μm 2,4-D; 5 μm BA + 10 μm 2,4-D; 5 μm BA + 15 μm NAA), or formed roots (10 μm 2,4-D). Whole plantlets were produced by transferring callus with buds to medium lacking hormones. The rapid regeneration of clonal plantlets from cotyledon explants may be useful for reducing variability in future developmental studies. Chemical names used: N-(phenylmethyl)-1H-purin-6-amine (BA); (2,4-dichlorophenoxy) acetic acid (2,4-D); and 1-naphthaleneacetic acid (NAA).

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Earl E. Albregts and Craig K. Chandler

Four strawberry (Fragaria ×ananassa, Duch.) cultivars were grown in a winter strawberry fruiting study using the annual hill cultural system and polyethylene-mulched beds during two seasons. Plants were set on 15, 30, 45, and 60 cm in row-plant spacing with two rows per bed spaced at 45 cm. Increasing plant density in the fruiting field generally increased early fruit yield and sometimes total fruit yield during two seasons. Yields of cull fruit were also increased with increased plant density. Daughter plant production decreased with increased plant density. Growers should consider planting costs, fruit rot, and harvesting problems when selecting the plant density for fruit production.

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Michael E. Kane and Craig K. Chandler

Many horticultural crops are infected with bacterial, fungal, or viral pathogens that reduce yield and/or quality. Recovery and maintenance of pathogen eradicated crops, such as strawberry (Fragaria × ananassa Duch.), have been possible following the isolation and culture of apical meristems or meristem-tips in vitro. A laboratory exercise has been developed to provide experience in the procedures required for the isolation, surface disinfection, and in vitro establishment of meristem-tip explants excised from strawberry stolons. Stolons are obtained from greenhouse-grown strawberries (`Sweet Charlie') maintained in hanging baskets under a 14-h photoperiod. Stolons are cut into single-node segments and terminal tips. The leaf blades are removed and the nodal sections are rinsed and then surface-disinfected by successive agitation in 70% ethanol and 1.05% sodium hypochlorite, followed by three rinses in sterile deionized water. In the transfer hoods, each student attempts to isolate meristem-tips and shoot tips of various sizes under high magnification provided by a stereomicroscope. Explants are inoculated onto Murashige and Skoog basal medium (Murashige and Skoog, 1962) supplemented with 30 g/liter sucrose, 80 mg/liter adenine sulfate, 1.0 mg/liter benzyladenine, 1.0 mg/liter indole-3-acetic acid, and 0.01 mg/liter gibberellic acid (GA3) and solidified as 45°slants with 1.25 g/liter Phytagel and 3.0 g/liter TC agar. Growth responses are monitored weekly. After 6 weeks, students record the percentage of visibly contaminated cultures and number shoots produced per explant. The relationship between initial explant size and in vitro growth is also determined. Students index their cultures for the presence of cultivable bacteria and fungi using sterility test media.

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Eric B. Bish, Daniel J. Cantliffe, and Craig K. Chandler

Bare-root strawberry transplants have been conventionally used for establishment of strawberry fruiting fields. These bare-root transplants have variability in vegetative vigor that results in irregular flowering patterns. We have been experimenting with a containerized transplant system to produce uniform transplants. Increasing transplant container volume by increasing perimeter, rather than depth, has resulted in increased plant size, but also increases transplant production costs. This study evaluated three container perimeters (17, 25, 32 cm) and three container shapes (circular, elliptical, and biconvex) such that different cell perimeters had the same greatest diameter. All containers had a depth of 3.5 cm. Root imaging analysis (MacRHIZOTM) was used to measure root growth in the container as well as root growth 3 and 6 weeks after transplanting. Increasing container perimeter led to increased plant growth before and after transplanting, but did not affect fruit production. Transplant container shape did not significantly alter plant growth or fruit production. Biconvex and elliptical containers required 25% and 15% less surface area, respectively. Therefore, a biconvex shaped container can be used to increase plant density during transplant propagation, decreasing surface area needed and reducing production costs.

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Julia Reekie*, Peter Hicklenton, John Duval, Craig Chandler, and Paul Struik

Our previous work on modifying strawberry plant morphology used either mowing to remove the leaf laminas and part of the petioles on `Camarosa', or a new reduced-risk gibberellin synthesis inhibitor, Prohexadione-Ca (ProCa), to restrict cell elongation in `Sweet Charlie'. These early studies showed promising results in acheiving desirable plant size and increasing fruit yield in annual hill plasticulture. Therefore, in the growing seasons of 2001 and 2002, we used `Camarosa' to explore the possibility of combining mowing and ProCa as a means of modifying strawberry transplant morphology in the nurseries, and studied its effect on fruit production in annual hill plasticulture. Plants were mowed and treated with 62.5 μL·L-1 of ProCa in a nursery field in Nova Scotia (45°26'N, 63°27'W). Treatments consisted of either mowing, the application of ProCa, or a combination of mowing and ProCa on one of two dates, 5 or 19 Sept. ProCa application early in the growing season had increased the production of daughter plants in the nursery. All plants were harvested in early October, and immediately transplanted in Dover, Fla. (28°00'N, 82°22'W). Fruits were collected twice weekly from late November to February or March. At time of harvest, both mowing and ProCa reduced plant height and total leaf area; plants which were treated with ProCa and mowed were the shortest. On average, treated plants had higher fruit yield as compared to untreated plants. In 2001, early fruit production in December was increased significantly in treated plants.

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Leighan Howard, Philip Stewart, Amit Dhingra, Craig Chandler, and Kevin Folta

Cultivated strawberry (Fragari×ananassa) is a valuable crop, yet has benefitted little from recent advances in biotechnology and genomics. A high-throughput system for transformation and regeneration would hasten elucidation of gene function for strawberry and possibly the Rosaceae in general. In this report, a protocol for high-frequency octoploid strawberry transformation and regeneration is presented. The protocol uses leaf, petiole, and stolon as explants from a newly selected genotype, `Laboratory Festival #9'. This genotype was selected from progeny of a `Strawberry Festival' self-cross exclusively for its rapid regeneration and robust growth in culture. Direct organogenesis has been achieved from the leaf or from prolific callus with multiple shoots being visible in as few as 14 days. Over 100 viable regenerants may be obtained from a single leaf explant of about 3-cm2 size. This laboratory-friendly genotype allows high-throughput, statistically relevant, studies of gene function in the octoploid strawberry genetic background as well as generation of large transgenic populations.

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Craig K. Chandler, Diane Doud Miller, and David C. Ferree

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Craig K. Chandler, Daniel E. Legard, and Charles A. Sims

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Craig K. Chandler, T. E. Crocker, and E. E. Albregts

During the past 10 years, the Florida strawberry growers, through the Florida Strawberry Growers Association, have made a serious commitment to fund university research on strawberries. They have purchased equipment and donated monies for facilities at Dover. They have also helped support a new faculty position in breeding and genetics. During this same period, the University of Florida has made an equally strong commitment to support strawberry research and extension. These commitments are beginning to pay significant dividends for industry and the University. Cultural and pest management information has been generated that is saving the industy money, and the breeding program is developing new cultivars that will keep the industry competitive in the marketplace. The University has benefitted through the acquisition of new facilities, equipment, and faculty and graduate student support.