Traditional and biotechnological breeding techniques are being united to develop exciting new plants and to improve existing cultivated plants by introducing natural variability from germplasm resources. Intervarietal, interspecific and intergeneric crosses can be accomplished by using plant embryo culture techniques, sometimes also referred to as embryo rescue. Embryo culture involves the isolation and growth of immature or mature zygotic embryos under sterile conditions on an aseptic nutrient medium with the goal of obtaining a viable plant. The technique depends on isolating the embryo without injury, formulating a suitable nutrient medium, and inducing continued embryogenic growth and seedling formation. The culture of immature embryos is used to rescue embryos from hybrid crosses that were once thought to be incompatible because they would normally abort or not undergo the progressive sequence of ontogeny. The culture of mature embryos from ripened seeds is used to eliminate seed germination inhibitors, to overcome dormancy restrictions, or to shorten the breeding cycle. New and exciting cultivars of Alstroemeria, also known as Lily-of-the-Incas, Inca Lily, or Peruvian Lily, have been bred by using zygotic embryo culture; these techniques and applications will be discussed.
In vitro growth and development of Alstroemeria `Cornell Pink' were evaluated on media containing different amounts of CaCl2, MgS O4, FeSO4, NO3, or NH4. Six levels of calcium chloride were originally examined (from 0 to 75 mM); the low levels proved to be most beneficial. Subsequent experiments used CaCl2 levels from 0 to 3.0 mM. Again, the low levels were most productive. Two experiments, with different gelling agents, were designed for MgSO4. The levels ranged from 0 to 15 mM. The 15 mM level produced explants with the greatest fresh weight. Three experiments were used to study the effect of FeSO4. The range was the same in all of the experiments (0 to 1 mM), but the increments and the gelling agents differed. In all three experiments, the 1 mM level proved to be toxic. The group with treatments from 0.01 to 0.5 mM had the best response over time. Both experiments with nitrogen found no response to different NO3:NH4 ratios. A positive linear response to rate was found within the range studied (20 to 80 mM).
Red- and pink-flowering cultivars of Alstroemeria were grown with and without cooling tubes placed in the growing medium. Cooling tubes were placed immediately below, 5 cm below, and 10 cm below the surface of the medium. Cooling was obtained by circulating well water (10 to 15C) through polyethylene tubing. Air and media temperatures were recorded every 3 to 10 days in the morning and in the late afternoon to determine if positioning of the cooling tubes had an effect on the medium temperature. The air temperature was warmer than the noncooled medium, which, in turn, was significantly warmer than the cooled medium. There were no significant differences in media temperatures among the three cooling treatments. Flowering of cooled Alstroemeria cultivars continued for 2 months longer at the end of the summer and reinitiated 1 month earlier than the noncooled control. Both cultivars produced significantly more total flowers when grown in cooled medium. Flower production was greatest in the summer for plants with tubes 10 cm below the surface and least for plants in noncooled medium. This trend continued when flowering was reinitiated; however, by April of the following year, all treatments were equal in flower production.
Presowing treatments and temperature regimes were tested to improve germination of Alstroemeria hybrids 3 to 12 months following harvest. In addition, seeds from 20 intraspecific F1 hybrids of five selections were also tested 3 to 7 or 8 to 12 weeks following harvest. Seeds were pretreated by chipping the seedcoat above the embryo, general abrasion of the entire seedcoat, or soaking 12 hours in distilled water, GA, (0.029, 0.29, 2.9 mm), or KNO3 (0.5 and 1.0 m). Pretreatments were evaluated under three environmental regimes: 8 weeks at a constant 18-25C (warm), 4 weeks at 18-25C followed by 4 weeks at 7C (warm-cold), or 4 weeks at 7C followed by 4 weeks at 18-25C (cold-warm). There was an interaction between pretreatment and environmental regime for percent germination. Germination percentages for the water soak and GA, at 0.29 or 2.9 mm were significantly higher than for the other pretreatments, but were not significantly different from one another. The warm-cold environment yielded higher germination percentages than the other environments. The time to germination was longest for the cold-warm regime. This response depended on the genotype and the age of the seed. Chemical name used: gibberellic acid (GA3).
Three Alstroemeria genotypes—A30, ER292, and 16-1-2—developed at the Univ. of Connecticut were grown in vitro on two media: Murashige and Skoog (MS) basal medium and Alstroemeria medium (ALA). Each medium had four levels of 6-benzylaminopurine (BA) added: 0, 9, 18, and 36 μm. Alstroemeria rhizomes initially containing one bud were cultured on the 24 treatments for 12 weeks with transfers onto fresh medium at 4 and 8 weeks. At each transfer and at the end of the experiment plants were scored for the number of shoots produced, number of new buds, fresh weight, and presence of roots. No difference was observed between the MS and ALA media. No interaction was found between medium and hormone concentration. BA inhibited the formation and growth of roots that were observed only in the control without BA. The control was different from 9, 18, and 36 μm BA for the number of buds produced, number of shoots and fresh weight while no differences were observed between the various concentrations of BA. ER292 gave the highest yields in number of buds, shoots and fresh weight of all the genotypes.
Mussaenda, a tropical ornamental shrub developed in the Philippines is being examined as a potential greenhouse potted crop in the United States. Showy sepals of white, picotee, pink or red and fragrant, yellow flowers make Mussaenda an attractive patted plans however, the profuse upright growth habit of some Mussaenda cultivars is undesirable for pot plant culture. With this in mind experiments were conducted to determine the effects of three growth regulators at two concentrations each, as well as the application method and the number of applications on Mussaenda plant height.
Three growth regulators, daminozide (B-Nine), ancymidol (A-Rest), and paclobutrazol (Bonzi) were applied at two commercially recommended rates and two application methods (spray or drench). The treatment were daminozide at 2500 ppm and 5000 ppm (spray), ancymidol at 33 and 66 ppm (spray) and at 0.25 and 0.50 mg/pot (drench), and paclobutrazol at 25 and 50 ppm (spray) and at 0.125 and 0.25 mg/pot (drench). In subsequent experiments, the same growth regulators were applied with an increase in concentration and either two or three applications. The treatments were daminozide at 5000 ppm (spray), ancymidol at 66 and 132 ppm (spray) and at 0.50 and 1.0 mg/pot (drench), and paclobutrazol at 50 and 100 ppm (spray) and at 0.25 and 0.50 mg/pot (drench).
The most attractive potted plants were produced with two applications of daminozide at 5000 ppm or two applications of ancymidol at 0.5 mg/pot (drench). Higher concentrations or additional applications excessively reduced plant height. Three spray applications of 132 ppm ancymidol also produced an attractive potted plant. Paclobutrazol sprays or drenches at any concentration or application number were ineffective for reducing Mussaenda `Queen Sirikit' plant height.
Disinfected midrib sections of Mussaenda `Queen Sirikit' ≈3 to 4 mm in size were cultured on a basal medium of Murashige and Skoog salts and vitamins, 87.7 mm sucrose, and 5 g Sigma agar/liter supplemented with several concentrations of indole-3-acetic acid (IAA) (0, 5.0, 10.0, 20.0 μm) and 6-benzylaminopurine (BAP) (0, 0.5, 1.0, 2.5, 5.0, 10.0, 25.0, 50.0 μm). Cultures were subculture onto the same treatment after 5 weeks and observed weekly for 15 weeks for the presence of somatic embryos. As somatic embryos were produced, they were subculture onto basal medium supplemented with 0.5, 1.0, 2.5, or 25.0 μm BAP. Callus was first observed at 2 weeks in cultures grown on basal medium supplemented with 5.0–20.0 μm IAA and 0–50.0 μm BAP. Somatic embryos were observed at 8 weeks on basal medium supplemented with 5.0–10.0 μm IAA and 2.5–5.0 μm BAP. Callus cultured on 0–10 μm IAA and 5.0–10.0 μm BAP produced the greatest number of somatic embryos by 15 weeks. Somatic embryos subculture to basal medium supplemented with 25.0 μm BAP proliferated shoots, while eliminating BAP from the medium resulted in root and callus production. Shoots and entire plants were removed from in vitro conditions and successful] y acclimated to greenhouse conditions. Somatic embryo-derived plants flowered sporadically 25 to 35 weeks after removal from in vitro conditions. Variations in sepal number and leaf number per node were observed at 1% to 5%.
Mussaenda, a tropical, hybrid ornamental plant from India and the Philippines, is being evaluated as a potential greenhouse ported crop in the united States. Showy sepals of white, picotee (White with rosy edges), light pink, dark pink, or red complemented by fragrant, yellow flowers and dark green, pubescent foliage make Mussaenda a very attractive potted plant. However, sometimes the height of Mussaenda is unsuitable for pot plant culture. With the use of chemical growth regulators. plant height is reduced thus making Mussaenda a more feasible potted crop.
In the summer of 1992, a growth regulator study was conducted to evaluate three growth regulators and concentrations capable of reducing plant height in Mussaenda. Daminozide (B-Nine SP), ancymidol (A-Rest), or paclobutrazol (Bonzi) was applied at two concentrations each. Daminozide was tested as a spray at 2500 ppm and 5000 ppm. Ancymidol was applied as a spray at 33 ppm and 66 ppm or as a drench at 0.25 mg/pot and 0.50 mg/pot. Paclobutrazol was tested as a spray at 25 ppm and 50 ppm or as a drench at 0.125 mg/pot and 0.25 mg/pot. Growth regulators were applied as a single application or a double application with two weeks separating applications.
Daminozide at 2500 ppm and 5000 ppm was most effective in controlling plant height. Ancymidol as a drench at 0.25 mg/pot and 0.50 mg/pot was also effective in plant height control. Two applications of these growth regulators were more effective in controlling plant height than a single application.
Meristems from three different positions were excised from in vitro plants of Alstroemeria genotype A30. Explants were removed from the most-distal vegetative shoot apical meristems, rhizome tip apical meristems, and rhizome tip axillary meristems. Meristems were cultured on four different media to compare the effect of meristem position and medium on the ability to produce Alstroemeria rhizomes from meristems. The meristem culture media were Murashige & Skoog salts plus 8.39 μM pantothenic acid, 1.19 μM thiamine, and 0.55 mmmyo-inositol (MSM), MSM plus 8.88 μM of 6-benzylaminopurine (BA), MSM plus 8.88 μM BA, and 0.72 μM gibberellic acid (GA3), and MSM plus 0.72 μM GA3. Meristems that were removed from the vegetative shoot apices did not develop rhizomes on any medium. Rhizome tip apical meristems developed less than 10% rhizomes when subcultured on media containing BA and GA3. However, rhizome tip axillary meristems developed rhizomes on all media with best results achieved when the medium was supplemented with BA.