No abstract

Tillandsia eizii is an epiphytic bromeliad that due to over-collection, habitat destruction, and physiological constraints has declined to near threatened status. This species exhibits high mortality in the wild, and seed are characterized by low percentages of germination. As a means to conserve this species, in vitro culture protocols were developed to enhance seed germination and seedling growth. A sterilization protocol using 70% ethanol for 2 minutes followed by 2.6% NaOCl for 40 minutes disinfested seed and promoted seedling growth. Sucrose incorporated into the culture medium had no effect on germination or growth, while NAA inhibited growth, but not germination. Cultures maintained under a 16-hour photoperiod at 22 °C exhibited greater growth than those grown at 30 °C. Seed that germinated in the dark remained etiolated and failed to develop even after transfer to light conditions. Plants grown in vitro were successfully acclimatized and transferred to the greenhouse. Over 86% survival and rapid growth were obtained with either an all-pine-bark medium, or a mixture of 2 redwood bark: 2 fir bark: 2 potting mix: 1 perlite. This demonstrated that in vitro culture of seed may be used to rapidly produce large numbers of T. eizii, and thus can be used for the conservation and reintroduction of this species.

SummaryThe bromeliad Tillandsia eizii is a striking species with large, colorful, and persistent inflorescences that can reach 1 m in length. The value of this plant as an ornamental and its importance in cultural and religious activities has led to its overcollection in the wild. Clonal propagation via tissue culture may be a means to repopulate native stands while meeting the demands for this species as an ornamental and ceremonial plant. Adventitious bud proliferation was induced from axenically germinated seedling material. Parameters evaluated were the age of explant material at the time of transfer onto bud-induction medium, the concentration of plant growth regulators, and the period of exposure to induction medium. Light and scanning electron microscopy (SEM) established the origin and development of buds. Twelve-week-old seedling explants rapidly initiated adventitious buds after a 30-d induction period on shoot-initiation medium. Adventitious buds were induced in 40% of the explants placed on media with 2 mg l 21 6-benzylaminopurine (BA) (8.88 mM) plus 0.1 mg l 21 a-naphthaleneacetic acid (NAA) (0.54 mM) with some cultures becoming highly prolific after repeated subculture. Shoots elongated in proliferating cultures, and plants were successfully acclimatized and planted into the greenhouse. The results indicate that tissue culture may be used as a means to propagate this epiphytic bromeliad species, which is being seriously affected by deforestation and habitat destruction. In addition, adventitious bud proliferation can provide a means to propagate superior genotypes.

The mitotic inhibitors, colchicine and oryzalin, were evaluated for their effects on callus, adventitious shoot formation, and tetraploid induction of Euphorbia pulchurrima `Winter Rose'. In vitro grown leaf sections were placed on various media supplemented with either colchicine or oryzalin at various concentrations for 1 to 4 days. Colchicine was less damaging to leaf tissues than oryzalin. On various colchicine-containing media, prolific calluses were produced and adventitious shoot formation was observed. Regenerated shoots were found to be diploid as determined by flow cytometry. On media supplemented with oryzalin (28.9 μm to 144 μm), leaf tissues produced callus but failed to form adventitious shoots. Samples of calluses produced on oryzalin-containing media were subject to analysis using flow cytometry and were found to be diploid. These results suggest that the colchicine is less toxic on poinsettia tissues and shoot induction than oryzalin. Additional experiments are needed to establish a protocol for in vitro induction of poinsettia tetraploid with colchicine and oryzalin.

Poinsettia, Euphorbia pulcherrima, is an important holiday symbol and is the number one flowering potted plant in the United States. The technique of chromosome doubling has been utilized to increase size of flowers, stems, and leaves of many species, and has been used in poinsettia breeding to obtain new cultivars. Application of colchicine or oryzalin to in vitro tissues may be used to enlarge the inflorescences and brackets and reduce the height of `Winter Rose'™ poinsettias, reduce the likelihood of chimeric tetraploids, and provide a rapid means for producing many tetraploid plants. The purpose of this research was to evaluate the effect of colchicine and oryzalin on callus and adventitious shoot formation of `Winter Rose'™ poinsettia with in vitro grown leaf tissues and its potential for tetraploid induction. In vitro grown leaf midvein sections were placed on various media supplemented with either colchicine or oryzalin at various concentrations for 1–4 days. Colchicine was least damaging to leaf tissues at concentrations of 0.25 or 250.4 μm. A large amount of callus, as well as adventitious shoots, were produced. Regenerated shoots were found to be diploid, determined by flow cytometry. On media with oryzalin (28.9–144 μm), leaf tissues produced callus, but not adventitious shoots. Calluses produced on oryzalin-containing media were tested using the flow cytometer and were found to be diploid.