论文部分内容阅读
为了建立一种甘蔗根系内生真菌最佳的染色方法,便于更好观察甘蔗根际土壤丛枝菌根(Arbuscular mycorrhiza,AM)真菌和深色有隔内生真菌(Dark septate endophytes,DSE)对甘蔗根系的侵染情况。将甘蔗根样置于20%的KOH溶液中90℃水浴透明90~120 min;接着加入碱性H_2O_2脱色60 min;5%的乙酸酸化5 min后,用5%的墨水醋染液(Quink牌纯黑墨水、北京牌蓝黑墨水)、酸性品红、苏丹红Ⅳ、台酚蓝、苯胺蓝为染液,在66℃水浴染色30 min,后用清水浸泡(12 h)脱色后即可镜检。经过碱性H_2O_2脱色60 min后Quink牌纯黑墨水染色的根系,AM真菌的菌丝和泡囊以及DSE微菌核能够清晰可见。此实验建立的墨水醋染法安全性高而且操作简便,成本低廉。相对于其他染色方法毒性低,对环境的污染小,而且染色效果稳定,更适于观察和镜检拍照。
In order to establish an optimum staining method for endophytic fungi in sugarcane root, it is convenient to observe the effects of arbuscular mycorrhiza (AM) fungi and dark septate endophytes (DSE) on rhizosphere soil of sugarcane Sugarcane root infection. The sugarcane roots were placed in 20% KOH solution for 90-120 min in a 90 ° C water bath, followed by alkaline H 2 O 2 decoloration for 60 min. After 5 min of 5% acetic acid acidification for 5 min, Pure black ink, Beijing brand black ink), acid fuchsine, Sudan IV, phenolic blue, aniline blue dye solution, dyed in 66 ℃ water bath for 30 min, after soaked in water (12 h) Check Quink black dye-stained roots, AM fungi mycelium and vesicles, and DSE micro-sclerotia were clearly visible after 60 min of alkaline H 2 O 2 decoloration. The ink vinegar method established in this experiment is safe and easy to operate with low cost. Relative to other staining methods, low toxicity, less pollution to the environment, and the dyeing effect is stable, more suitable for observation and microscopic examination of photographs.