酶法分离胚囊的荧光显微观察

用荧光显微技术对酶法分离的胚囊进行了三方面的研究:(1)利用与DNA结合的荧光染料Hoechst 33258对金鱼草、蚕豆、芝麻、烟草、龙葵等植物的分离胚囊??染色,可以观察胚囊中各组成细胞的核以及合子和胚乳的核。(2)用金胺O和樱草黄两种荧光染料染色,证明胚囊壁中含有耐果胶酶与纤维素酶作用的成分,可能是角质,因而虽经上述酶处理仍能保持胚囊的整体性。(3)用脱色苯胺蓝染色观察了被果胶酶分离的泡桐大孢子发生的各期材料,对这一过程中胼胝质的动态作了描述。以上各项实验表明:酶法分离技术与荧光显微技术相结合,是研究胚囊细胞生物学的一种简便有效的方法。

FLUORESCENCE MICROSCOPIC OBSERVATIONS ON ENZYMATICALLY ISOLATED EMBRYO SACS

The enzymatic isolation of embryosac combined with various histochemi-cal assays could offer a new means forstudying the female gametophyte ofangiosperms.In this paper some re-sults of fluorescence microscopic obser-vations on enzymatically isolated emb-ryo sacs are reported.Ovules were ma-cerated in pectinase-cellulase solutionin the same manner as previously de-scribed and naked embryo sacs wereliberated out.Using a DNA-bindingfluorescent dye Hoechst 33258,cell nu-clei within the embryo sacs of Antir-rhinum majus,Vicia faba,Sesamum indicum,Nicotiana tabacum and Solanum nigrumwere stained well.This not only servesas a means for identification of nucleiin embryo sac cells,but also may pro-vide a basis for further fluorimetric mea-surement of DNA content in thesenuclei.Auramine O and primulinstaining revealed the cutin,or/and spo-ropollenin,nature of the boundary wallof snapdragon embryo sac.This mayexplain the tolerance of this wall to thepectinase-cellulase action,resulting inmaintenance of the intactness of the iso-lated embryo sacs.Fluorescence micro-scopic observations on the megasporo-genesis in enzymatically isolated materi-als of Paulownia stained by leuco anilineblue showed some interesting behaviorof callose depositon in this species.These preliminary results indicate thatthe enzymatic isolation method combin-ed with fluorescence microscopy is use-ful for studying the biology of embryosac.