In vitro Pollen Performance of Pinus densiflora and P. rigida: Temperature and Medium Nutrient Effects

Effects of environmental factors on in vitro pine pollen performance were investigated. Pinus densiflora and P. rigida pollen grains collected at Mt. Kwanak, Korea were used. Three environmental factors, such as pollen storage temperature, pollen culture temperature and nutrient condition in medium, were tested. To determine the storage temperature effects on pollen viability, pine pollen was stored at $-70^{\circ}C$, $-12^{\circ}C$, $4^{\circ}C$ and $22^{\circ}C$. Pollen viability was substantially extended at the storage temperatures of $-12^{\circ}C$ and $4^{\circ}C$ for more than 300 days. To elucidate the culture temperature effects on pine pollen germination and tube growth, pollen grains were cultured at the temperatures from $5^{\circ}C$ to $40^{\circ}C$ at $5^{\circ}C$intervals. The germination rate and tube growth were highest at $25^{\circ}C$ and decreased above $30^{\circ}C$. To investigate boron and sucrose effects on pollen tube growth, the pollen was cultured at different sucrose and boric acid concentrations. Germination rate was optimal in germination medium containing 3 or 5$\%$ sucrose with 0.01 $\%$ boric acid. These results indicate that the pine pollen can be stored for considerable length of time without noticeable loss of viability at storage temperature below or near $0^{\circ}C$. Optimal germination medium conditions were established for pine pollen. Therefore, pine pollen can be used for many biological and environmental monitoring researches.