An Improved Ink-acetic Acid Technique for Staining Arbuscular Mycorrhizas of Citrus

作者：Cao, Ming-Qin; Wu, Qiang-Sheng^*; Zou, Ying-Ning

来源：International Journal of Agriculture and Biology, 2013, 15(2): 386-388.

• Arbuscular mycorrhizal fungi
• Blue ink
• Citrus
• Mycorrhizal staining
• Root mycorrhizal colonization

Trypan blue is usually applied to stain arbuscular mycorrhizas of higher plant roots, whilst it is harmful for human health. The objective of the study was to exploit a safe, non-toxic, and cheap technique for staining mycorrhizal structure of plants such as citrus (Citrus unshiu grafted on Poncirus frifoliata). The 1-2 cm root segments were cleared with 10% KOH for 90 min at 95 degrees C, blanched with H2O2 (10%) for 15 min, acidified with 0.2 mol/L HCl for 10 min, stained with 5% black, blue or red ink prepared with 5% acetic acid for 3-8 min, and destained with 5% acetic acid. It is clear that both 5% blue ink-acetic acid solution and 0.05% frypan blue solution showed better color contrast and visible mycorrhizal structures. The mycorrhizal colonization of roots stained with 5% blue ink-acetic acid solution for 3-5 min was not significantly different, compared with 0.05% trypan blue staining for 5 min On the other hand, the mycorrhizal colonization was higher in the roots stained with the 5% blue ink-acetic acid or 0.05% frypan blue than with the other ink-acetic acid staining The results suggest that use of frypan blue for arbuscular mycorrhizal staining could be replaced with 5% blue ink-acetic acid solution for 3-5 min, at least in citrus roots.

• 出版日期2013
• 单位长江大学