In vitro flowering of orchids.

Flowering is the most elusive and fascinating of all plant developmental processes. The ability to induce flowering in vitro in orchids would reduce the relatively long juvenile phase and provide deeper insight into the physiological, genetic and molecular aspects of flowering. This review synthesizes all available studies that have been conducted on in vitro flowering of orchids with the objective of providing valuable clues as to the mechanism(s) that is possibly taking place.

Photoperiod and temperature effects on in vitro growth and flowering of P. pusilla, an epiphytic orchid.

Psygmorchis pusilla Dodson and Dressler, an epiphytic orchid, has been shown to be an interesting model to study in vitro flower formation. In the present study, the effects of photoperiod and temperature on vegetative and reproductive development were investigated. Although photoperiod had limited effects on leaf number, an etiolating process was verified in darkness and a higher growth was detected under long days. A positive relationship was observed between long days and floral spike formation. However, plant incubation under 20 h photoperiod or longer days negatively affected floral bud development, inhibiting anthesis and reducing flower longevity. Higher soluble sugar and starch levels were detected in plants cultivated under long days, while chlorophyll and carotenoids contents were negatively affected under these conditions. Plants showed great sensitivity to temperature variations; 27 degrees C being the most adequate for growth, leaf and floral spike formation. Temperatures of 22 and 32 degrees C were not appropriate for in vitro development of P. pusilla.

Endogenous hormonal levels and growth of dark-incubated shoots of Catasetum fimbriatum.

Apical shoots and Lateral buds of the epiphytic orchid Catasetum fimbriatum give rise to rootless etiolated stolons, when cultured in the presence of light and then transferred to the dark. The stolons are characterized by fast and continuous apical longitudinal growth. Measurements of four endogenous cytokinin, indole-3-acetic acid (IAA) and abscisic acid (ABA) levels in etiolated shoots and light-grown plants were low. However, after transfer of green plants to the dark, cytokinin Levels increased 3- and 7-fold by 10 and 30 days of incubation, respectively. IAA levels also increased significantly, but the increase was not as great as for cytokinins. A similar trend was observed in the roots. A close relationship seems to exist between both cytokinin accumulation and the formation of etiolated stolons. Variations in ABA levels were practically inconspicuous. The presence of paclobutrazol in the medium, a potent inhibitor of gibberellin synthesis, strongly inhibited etiolated and non-etiolated longitudinal shoot growth, although no apparent effect was observed on apical meristem activity.

Dark-induced hormone changes coincide with the resumption of light-inhibited shoot growth in Catasetum fimbriatum (Orchidaceae).

Catasetum fimbriatum plants cultivated in the absence of light exhibit continuous shoot growth leading to the formation of nodes and internodes. On the other hand, when these plants are incubated in the presence of light, shoot longitudinal growth is inhibited and pseudobulbs develop just below the shoot apical meristem. These facts provide evidence of a possible influence of light on mitotic cell division in the shoot apex as well as on pseudobulb initiation. The effects of light and dark on the interruption and/or maintenance of shoot apex mitotic activity and the subsequent formation of pseudobulbs in the sub-meristematic regions were investigated by means of histological and hormonal studies. The interruption of shoot apex development occurred around the 150th d of light incubation and seems to have resulted from the establishment of a strong storage sink in the region of the future pseudobulb, in detriment to the continuous activity of the shoot apical meristem. The reduced total cytokinins/IAA ratio in the apex, mainly due to high levels of IAA, could be a key factor in the interruption of cell divisions. Transfer to the dark brings about the resumption of shoot apex development of plants through the re-entrance of cells in the cell cycle which coincides with a significant increase in the total cytokinins/IAA ratio.