A Simple and Nontoxic Ink and Acetic Acid Staining Technique for Arbuscular Mycorrhizal Structures

Abstract:

Trypan blue is generally used to stain mycorrhizal hyphae, but trypan blue is also possible carcinogenic to humans. The present research conducted to develop a simple and nontoxic mycorrhizal staining technique with red, blue and black inks and acetic acid solution combined with different staining times in roots of Prunus salicina grafted on Amygdalus persica. After cleared with 10% KOH and acidified with 0.2 M HCl, the root segments were stained with 0.05% trypan blue or 5% ink-acetic acid solution. We found that root mycorrhizal colonization was no significantly different between 0.05% tryblune staining for 5 min and 5% blue ink-acetic acid staining for 3-8 min, and mycorrhizal colonization stained with other red and black inks-acetic acid solutions was significantly lower than those with 0.05% trypan blue. In addition, using 5% blue ink-acetic acid staining for 3-5 min could exhibit better contrast and clearly visible intraradical and extraradical hyphae, vesicles and arbuscules. It suggests that a simple and safe technique for staining arbuscular mycorrhizal structures with 5% blue ink-acetic acid solution for 3–5 min could potentially substitute trypan blue staining and exhibit good contrast and visible mycorrhizal structures.

• Periodical: Advanced Materials Research (Volumes 518-523)
• Pages: 679-682
• DOI: https://doi.org/10.4028/www.scientific.net/AMR.518-523.679
• Online since: May 2012
• Authors: Qiang Sheng Wu, Ming Qin Cao, Ying Ning Zou
• Keywords: Arbuscular Mycorrhiza, Blue Ink, Mycorrhizal Colonization, Trypan Blue

References

[1] F. Maillet, V. Poinsot, O. André, V. Puech-Pagés, A. Haouy, M. Gueunier, L. Cromer, D. Giraudet, D. Formey, A. Niebel, E.A. Martinez, H. Driguez, G. Becard and J. Dénarié: Nature Vol. 469(2011), p.58

DOI: 10.1038/nature09622

[2] Q.S. Wu and Y.N. Zou: Plant Soil Environ. Vol. 55 (2009), p.436

[3] Q.S. Wu, Y.N. Zou and X.H. He: Acta Physiol. Plant. Vol. 32 (2010), p.297

[4] Q.S. Wu, Y.N. Zou and G.Y. Wang: Commun. Soil Sci. Plant Anal. Vol. 42 (2011), p.1825

[5] J.M. Phillips and D.S. Hayman: Trans. Br. Mycol. Soc. Vol. 55 (1970), p.158

[6] A. Mauler-Machnik A and P. Nass: Gesunde Pflanze Vol. 42 (1990), p.130

[7] H. Vierheilig, A.P. Coughlan, U. Wyss and Y. Piche: Appl. Environ. Microbiol. Vol. 64 (1998), p.5004

[8] Q.S. Wu, R.X. Xia and Y.N. Zou: Eur. J. Soil Biol. Vol. 44 (2008), p.122

[9] C. Grace and P. Stribley: Mycol. Res. Vol. 95 (1991), p.1160