TY  - JOUR
TI  - Fast, Easy, and Comprehensive Techniques for Microscopic Observations of Fungal and Oomycete Organisms Inside the Roots of Herbaceous and Woody Plants
AU  - Toma, Tomáš 
AU  - Kováč, Ján 
AU  - Ďurkovič, Jaroslav 
VL  - 14
IS  - 11
PY  - 2024
DA  - 2024/06/05
SP  - e5013
C1  - Bio-protocol 2024;14:e5013
DO  - 10.21769/BioProtoc.5013
UR  - https://doi.org/10.21769/BioProtoc.5013
AB  - The roots of herbaceous and woody plants growing in soil are complex structures that are affected by both natural and artificial fungal colonization to various extents. To obtain comprehensive information about the overall distribution of fungi or oomycetes inside a plant root system, rapid, effective, and reliable screening methods are required. To observe both fine roots, i.e., a common site for penetration of fungi and oomycetes, and mature roots, different techniques are required to overcome visual barriers, such as root browning or tissue thickening. In our protocol, we propose using fast, cost-effective, and non-harmful methods to localize fungal or oomycete structures inside plant roots. Root staining with a fluorescent dye provides a quick initial indication of the presence of fungal structures on the root surfaces. The protocol is followed by clearing and staining steps, resulting in a deeper insight into the root tissue positioning, abundance, and characteristic morphological/reproductive features of fungal or oomycete organisms. If required, the stained samples can be prepared by using freeze-drying for further observations, including advanced microscopic techniques.
KW  - Fungi staining
KW  - Methyl blue staining
KW  - NaOH/KOH clearing
KW  - Oomycetes staining
KW  - Plant root microtechniques
KW  - Root clearing
KW  - Roots of herbaceous plants
KW  - Roots of woody plants
KW  - Solophenyl flavine staining
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False