Rockwool plugs were placed in Magenta G-7 boxes (Sigma) and then autoclaved at 121°C for 20 min. Fifty milliliters of cool autoclaved liquid medium was poured into Magenta G-7 boxes in aseptic conditions before microcuttings of Amelanchier, Cercis canadensis, cherry, and apple were transferred. Murashige and Skoog medium (MS, M-5519, Sigma) containing 30 g·L–1 sucrose, and with/out 1 ppm of NAA, pH 5.5 were used in all experiments. All cultures were incubated at 23 ± 1°C under a 16-hour lighting period with a light intensity of about 4000 lux of white fluorescent light. Microcuttings of Amelanchier, Cercis, Apple, and cherry rooted in rockwool plugs in 3 weeks after transfer and were ready to be out-planted in 6 weeks. Out-planted plantlets were leached with tap water and potted in 4-inch pots with Metrolite mix, then, placed in mist bench under 50% shade for 2 weeks before taking to bench with full sun light. The survival was 100%. Conditions and growth rate of rockwool-plug-rooted plantlets were much better than those plantlets rooted in agar medium. Rockwool plug plantlets had 2–3 flushes of growth before dormancy in greenhouse and were ready to be planted in the field or garden in 8 months after out-planting. Using a rockwool plug system simplifies out-planting procedure, produces better plantlets, increases out-planting survival, and greatly shorten time needed from out-planting to field-plantable size. This system is a very useful system for difficult-to-root woody ornamentals.
Chin-Chang Chu and Kenneth W. Mudge
Chin-Chang Chu and Kenneth W. Mudge
Seeds from mature seed pods of Cypripedium calceolus var. parviflorum were germinated on 1/4 MSMO (Sigma) + 100ml/l coconut water + 1% sucrose +/- 8g/l agar (pH 6.0), and with or without prechilling at 5C for 8 weeks. Protocorm with apex (stage 3) was use as an index of germination. Seeds sown on agar medium withou chilling treatment resulted in a 40% germination rate in 120 days but the germination was very uneven. Seeds germinated on agar medium with prechilling developed more synchronously with 92% germination in 60 days (ie. about 120 days after sowing). Suspension culture of seeds without prechilling resulted in 85% germination after 90 days. The synchronization of seed germination in suspension culture was intermediate between that on agar with and without prechilling. Protocorms germinated in suspension culture appeared morphologically identical to those germinated on agar medium. All stage 3 protocorms developed further on the same agar medium in darkness.
Both agar and suspension culture in media containing coconut water provided reliable seed germination methods for this orchid species.
Kenneth W. Mudge and Chin-Chang Chu
In vitro asymbiotic seed germination, subculture, and outplanting of orchids is presented as a laboratory exercise suitable for students of plant propagation or tissue culture. Dendrobium antennatum (Lindley), Phalaenopsis (Blume) white hybrid, or both, are used in this exercise because they flower predictably in the greenhouse, are reliable for seed production, and germinate and grow rapidly in vitro. The exercises can be used to instruct students in the skills involved in orchid seed sterilization, sowing, and culture, as well as instruct students in the unique features of orchid reproductive biology and symbiosis. A schedule is suggested for stock plant flower pollination, capsule harvest, seed sowing, and seedling subculture so that the necessary plant material is available for students to sow, subculture, and outplant seedlings during a single laboratory session.