Advances in AMF Research: Isolation, Histochemical Staining, Enumeration, Morphological and Molecular Techniques

The mutualistic association of arbuscular mycorrhizal fungi (AMF) is an important symbiotic association between the roots of plant and fungi that belongs to Glomeromycota order. The AM fungal isolation involves various techniques such as wet sieving and decanting and gradient centrifugation methods which are discussed here. For detection and quantification of AMF in roots, various histochemical staining and light microscopy-based techniques have been widely used. These techniques include trypan blue and DAB (3, 3′-diaminobenzidine) staining methods. The traditional system of identification of AM fungi mainly relied on morphological characterization of spore regarding color, size, and wall structure, etc. Molecular techniques for identification of AM fungi by SSU rDNA analysis give more accurate and scientific results and also used extensively in AMF taxonomic and diversity analysis. In this chapter, we reviewed the progress made in the traditional methods of mycorrhizal research and discussed recently developed techniques which are being utilized extensively in the field of AMF research. The chapter specifically focuses on the isolation and identification of AMF, providing a comprehensive summary of the diverse techniques employed throughout this process.