Conservation Horticulture: In Vitro Micropropagation and Acclimatization of Selected Florida Native Orchids

Abstract

Florida is home to 106 native orchid species, of which 77 are listed as endangered or threatened by the State of Florida. The Institute for Regional Conservation (IRC) has classified 62 of these species as either critically imperiled, imperiled, or rare in South Florida. Because of lack of endosperm, orchid germination rates are very low in nature, as they depend on an obligate relationship with mycorrhizal fungi for nutrients. Most orchid seeds can be germinated in vitro without the need for specific mycorrhizal fungi. This study aims are to establish a fast and efficient protocol for in vitro seed germination using different nutrient media and plant growth regulator (PGR) combinations, and to optimize seedling acclimatization protocols using different greenhouse media. To determine germination preferences, three different in vitro seed germination media were tested supplemented with PGRs, including 1) Murashige & Skoog (MS) (control), 2) MS supplemented with 1.5 mg/L 6-benzylaminopurine (BAP), and 3) MS supplemented with 1 mg/L BAP and 0.5 mg/L 1-Naphthaleneacetic acid (NAA) on Cyrtopodium punctatum, a state-listed endangered, IRC critically imperiled epiphytic orchid. There was no significant difference amongst the treatments after 2 and 6 weeks of seed sowing culture. To understand post-culture survivorship, two epiphytic and one terrestrial native species (Trichocentrum undulatum, Encyclia tampensis, and Oncidium ensatum) were chosen for the acclimatization study with two commercially available potting substrates (coir, sphagnum). To measure seedling growth rates, phenotypic measurements [leaf number, leaf length, root length, plantlet height, light intensity, pH, and electrical conductivity (EC)] and Soil Plant Analysis Development (SPAD) and Normalized Difference Vegetation Index (NDVI) values were recorded monthly for five months. All media/PGR combinations resulted in an initial high percentage of stage I growth but inhibited Protocorm-like bodies (PLBs) development, suggesting more research is necessary to determine later improvements or detriments to MS basal media with BAP and NAA. Comparing acclimatization media with the three species of orchid chosen for this experiment, neither O. ensatum nor E. tampensis showed a marked preference for sphagnum moss or coir. However, T. undulatum did perform better with coir compared with sphagnum. This research will help botanical gardens and commercial plant tissue culture laboratories to have a better understanding on selection of PGR combinations for in vitro cell culture and acclimatization media on increasing the viability and plant health and decreasing the mortality of endangered plants.