百合生殖细胞和精细胞的大量制备及其蛋白组成的比较

MASS ISOLATION AND PURIFICATION OF VIABLE GENERATIVE CELLS AND SPERM CELLS FROM LILIUM DAVIDII DACH AND COMPARISON BETWEEN THEIR PROTEIN COMPOSITION

新鲜的或低温贮存的兰州百合(Lilium davidii Duch.)花粉,在BKS15中萌发,分别收集萌发5h和30h的花粉管,采用低渗冲击胀破和percoll密度梯度离心纯化等方法,获得批量纯化的生殖细胞和精细胞。用10%三氯乙酸和丙酮沉淀制备生殖细胞和精细胞的蛋白质,用SDS-PAGE对两种细胞蛋白质进行比较。结果表明,二者在蛋白质组成成分上没有明显差异;但在蛋白质的含量上有两种成分差异显著,即:精细胞中的40KD蛋白多于生殖细胞;而98KD蛋白又少于生殖细胞。对40KD和98KD蛋白质的可能生理功能进行了讨论。

Fresh or pollen stored at low temperature was germinated in BKS 15 medium. After 5 h and 30 h, pollen tubes were collected respectively. Then the tubes were subjected to hypotonic shock treatment. The generative cells and sperm cells were collected and purified by density gradient centrifugation. Large amount of purified, viable generative cells and sperm cells were obtained. The total protein of generative cells and sperm cells was precipitated by 10% trichloroacetic acid and actone. Protein of both kinds of cells was electrophoretically analyzed with SDS-PAGE. Results show that there is no significant difference between their protein composition, but in sperm cells it is noticed that 40 KD protein exists in larger amount, while 98 KD protein in smaller amount than in generative cells. The possible physiological function of the 40 KD and 98 KD protein is discussed.