Published article

From Darkness to Light: Unlocking the Secrets of Ectomycorrhiza with Fluorescent Proteins in Laccaria

Figure: Fluorescence microscopy of wild type, pHTB16202GFP and pHTB16202mCherry transformed Laccaria confirmed specific nuclear localization of H2B-FP. a Fluorescence signals detected in DAPI stained wild type (wt), pHTB16202mCherry (H2B-mCherry), and pHTB16202GFP (H2B-GFP) strains. b A magnification of (a) and shows DAPI staining of nuclei in the wild type fungus. c Specific nuclear localization in a selected pHTB16202mCherry strain. d Demonstrates specific nuclear localization in a selected pHTB16202GFP strain. a ×400 total magnification, rest of the panels ×1000 magnification. bf bright field.

Deep in the forest, a tiny fungus known as Laccaria bicolor forms a symbiotic relationship with the roots of trees. This relationship, known as ectomycorrhiza (ECM), is vital to the health of the forest ecosystem, but until now, scientists had struggled to study it in detail especially .at the molecular level. The problem? The fungal partner of the ECM, Laccaria, was notoriously difficult to study using traditional methods.

But now, we have cracked the code- at least partially. We have successfully used fluorescent proteins (FPs) to study Laccaria in a new way. These FPs, called eGFP and mCherry, act as markers, lighting up specific parts of the fungus, making it much easier for scientists to study its behavior and interactions with trees.

But getting these FPs to work wasn't easy. We had to figure out the specific requirements for FP expression in Laccaria bicolor, and then design a set of tools to make it easy for other researchers to use FPs in their studies as well. We discovered that FPs needed specific sequences, known as Kozak sequences, to work efficiently in Laccaria and that we could use specific genes from Laccaria bicolor to control where the FPs were localized within the fungus, making it possible to study specific parts of the fungus in greater detail. This discovery can finally shed light on the mysterious world of the Laccaria bicolor fungus and its relationship with trees. And it's not just Laccaria that will benefit, but other biotrophic or saprotrophic basidiomycete species susceptible to genetic transformation.

Author: Jamil Chowdhury

1 April 2020.

Check out the article published in Current genetics.