Abstract
Many Disa species (80%) have not been germinated in vitro, fuelling conservation fears. Winter-rainfall species are typically germinable, whilst summer-rainfall species are exclusively intractable in vitro. We aimed to establish the in vitro germination requirements for previously ungerminated Disa species. Conventional asymbiotic seed culture protocols were reviewed and their efficacy in summer-rainfall Disa established. Mature seed was subjected to manipulations of media composition and viscosity, incubation temperature and illumination. Immature cultures were also established across four seed maturity classes. Four first-time germinations (D. cooperi, D. nervosa, D. pulchra and D. woodii) resulted from media with increased water availability. Germination rate (>12 weeks), percentage ( 80% germination in 8 weeks). Germination of immature seed under control conditions was similar to mature seed germination under modified conditions, but neither percentage approached the calculated germination potential (∼viability). Germination control in Disa is proposed as a trade-off between water availability and the presence of phyto-inhibitors in the environment of the embryo—two features typical of seeds exhibiting water-impermeable dormancy.
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