# Protocol for conditional gene expression in Magnaporthe oryzae via fungal nitrate reductase promoter replacement

## TL;DR

This paper describes a method to control gene expression in a rice blast fungus using a nitrogen-dependent promoter, enabling easy observation of genetic effects.

## Contribution

A new protocol for conditional gene expression in Magnaporthe oryzae using a nitrogen-responsive promoter replacement.

## Key findings

- Replacing the BUF1 promoter with a nitrogen-responsive promoter allows conditional gene expression.
- Nitrogen source-dependent expression leads to observable pigmentation changes in M. oryzae.

## Abstract

Functional genomic studies rely on controlled gene expression, which is challenging for essential genes or those active only under specific conditions. We provide a protocol for generating Magnaporthe oryzae (synonym of Pyricularia oryzae) with conditional BUF1 gene expression. We describe steps for using targeted promoter replacement to substitute the native BUF1 promoter with the nitrogen-responsive promoter. This approach enables nitrogen source-dependent control of expression in M. oryzae, leading to observable pigmentation changes that facilitate easy phenotypic screening.

•Protocol for conditional gene expression in Magnaporthe oryzae•Details replacement of native promoter with nitrogen-responsive pMoNIA1•Enables observable phenotypic changes based on nitrogen availability

Protocol for conditional gene expression in Magnaporthe oryzae

Details replacement of native promoter with nitrogen-responsive pMoNIA1

Enables observable phenotypic changes based on nitrogen availability

Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Functional genomic studies rely on controlled gene expression, which is challenging for essential genes or those active only under specific conditions. We provide a protocol for generating Magnaporthe oryzae (synonym of Pyricularia oryzae) with conditional BUF1 gene expression. We describe steps for using targeted promoter replacement to substitute the native BUF1 promoter with the nitrogen-responsive promoter. This approach enables nitrogen source-dependent control of expression in M. oryzae, leading to observable pigmentation changes that facilitate easy phenotypic screening.