铁皮石斛种子共生萌发形态学观察

Morphologic Techniques for Observing Seed Symbiotic Germination of Dendrobium Officinale

本研究以铁皮石斛种子-胶膜菌属菌根真菌共生体为实验材料,利用4种真菌特异性染剂,包括苯胺蓝(aniline blue)、氯唑黑E(chlorazol black E)、台盼蓝(trypan blue)和麦胚凝集素-异硫氰酸荧光素(wheat-germ agglutinin-fluorescein isothiocyante,WGA-FITC)染剂,整体透明试剂(Hoyer′s溶液)和多种显微观察仪器对共生萌发种子和原球茎进行形态学观察.研究结果显示4种真菌特异性染剂能有效检测菌根真菌在共生萌发种子和原球茎中的定殖,共生原球茎经过整体透明试剂处理后,最大程度展示了菌根真菌在皮层细胞内的结构和真实定殖情况.本研究为兰科植物种子共生萌发检测提供了多种简单有效的形态学方法.

In this study, dendrobium officinale seed-tulasnella sp were used as experimental materials. Four fungal specific dyes, including aniline blue, chlorazol black E, trypan blue and WGA-FITC dyes, Hoyer′s solution and various microscopic observation instruments were used to study the morphology of symbiotic germinated seeds and protocorms. The study showed that four fungal specific dyes could effectively detect the colonization of mycorrhizal fungi in symbiotic germinating seeds and protocorms. After the treatment of symbiotic protocorms with whole transparent reagents, the structure and true colonization of mycorrhizal fungi in cortical cells were displayed to the greatest extent. This were an efficient way to analyse the infected protocorm development without sectioning. These findings would provide a variety of efficient morphological methods for detection of symbiotic germination of orchid seeds.