Quick Detection of Pollen Developmental Stages of Hamelia patens Jacq.

DAPI is a DNA-specific fluorochrome that facilitates observation of nuclei during pollen development and can confirm the developmental stages of pollen grains. However, the nuclei in Hamelia patens Jacq. pollen grains are little stained by DAPI due to the resistance of the pollen wall. An effective "water-bath heating and oxidation" method was used to treat the pollen wall of H. patens. The results showed that the: (1) best incubating conditions for uninucleate microspores and bicellular pollen grains were 20~50 min at 65℃ and 20~40 min at 55℃, respectively; and, (2) lengths of flowers and anthers were 0.90~1.00 cm and 0.50~0.60 cm at the tetrad stage, 1.10~1.60 cm and 0.60~0.85 cm at the uninucleate microspore stage, and 1.80~2.70 cm and 0.91~1.01 cm at the bicellular pollen stage, respectively.