J Integr Plant Biol. ›› 1998, Vol. 40 ›› Issue (4): -.

• Research Articles •    

Isolation of Viable Male and Female Gametes in Cunninghamia lanceolata

CHENG Xiao-Fei, LI Wen-Dian and HONG Ju-Sheng   

Abstract: Viable male and female gametes were isolated from pollinated ovules of Cunninghamia lanceolata (Lamb.) Hook. Prior to their penetration into the female gametophyte, the pollen tubes were drawn out from the nucetlus. The isolated pollen tubes were branched and one of them became swollen. An enlarged spermatogenous cell and subsquently a pair of sperm cells were formed as the pollen tube reached the regions over and against the archegonia. The sperm cells were released from the pollen tubes manually with the use of a stereomicroscope. The positive FDA reaction gave evidence of the sperm cells viability, and the Fluorescent Brightener 28 positive demonstrated the presence of cell wall. The egg cells were enzymatically isolated from the female gametophytes. The isolated egg cells were spherical, contained 1 to 2 large and many small vacuoles. FDA test showed the egg cells were viable, and the viability sustained for 8 days in 2 to 4 ℃ without any protectants. Fusion between single pair of male and female gametic protoplasts was attempted with PEG method, but only adhesion of the two was obtained.

Key words: Isolation, Male gamete, Female gamete, Cunninghamia lanceolata

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