莴苣胚囊细胞分离

用酶解和解剖方法分离了莴苣的卵细胞,助细胞,中央细胞和合子。莴苣子房先在酶液中酶解40～50min,然后在不含酶的分离液中用解剖针解剖子房。在解剖出的胚囊中,可看到卵细胞,两个助细胞和中央细胞的轮廓。将胚囊的合点端切破,轻轻挤压胚囊的珠孔端,四个细胞即可逸出。在最佳条件下,90min可从40个子房中分离出29个胚囊,进一步从中分离出11个卵细胞。分离出的胚囊细胞用显微操作仪收集备用。莴苣卵细胞的成功分离为进行离体受精探索创造了条件。

Isolation of Embryo Sac Cells of Lettuce (Lactuca sativa)

Viable egg cells,synergids,central cells and zygotes of lettuce （Lactuca sativa L.） were isolated using enzymatic digestion and mechanical dissection. The ovaries were digested in enzymatic solution for 40-50 min,and then transported into the isolation solution without enzymes to dissect. In the dissected embryo sacs,the outline of an egg cell,two synergids and a central cell could be observed. When dissected embryo sacs were sliced from the chalazal end,four cells could be released by gently pushing its micropylar end. In optimal condition,29 embryo sacs and 11 egg cells could be isolated from 40 ovaries during 90 min. The isolated egg cells could be collected using micromanipulator for preparation of molecular biology of egg cell of lettuce. The isolation of egg cell of lettuce will make a great chance for in vitro fertilization in a dicot plant.