Klebsiella michiganensis: A nitrogen-fixing endohyphal bacterium from Ustilago maydis

Author:

Liang Pengyu1,Jiang Jianwei1,Sun Zhengxiang1,Li Yanyan2,Yang Chunlei2,Zhou Yi1

Affiliation:

1. Yangtze University

2. Tobacco Institute Inc

Abstract

Abstract Ustilago maydis is a pathogenic fungus in Basidiomycota causing corn smut disease. A strain of U. maydis YZZF202006 was isolated from the tumor of corn smut collected from Jingzhou city in China. The intracellular bacteria were confirmed inner hyphal of the strain YZZF202006 by PCR amplification and fluorescence in situ hybridization (FISH) and SYTO-9. An endohyphal bacterium YZUMF202001 was isolated from the protoplasts of the strain YZZF202006. It was gram-negative, short rod-shaped with smooth light yellow colony. The endohyphal bacterium was genomic evidenced as Klebsiella michiganensis on the basis of average nucleotide identity (ANI) analysis and the phylogram. And then K. michiganensis was GFP-Labeled and reintroduced into U. maydis, which confirmed the bacterium can live in hpyphal of U.maydis. The bacterium can grow on N-free culture media. Its nitrogenase activity was reached av. 646.25 ± 38.61 nmol·mL−1·h−1 C2H4 by acetylene reduction assay. A cluster of nitrogen fixation genes (nifJHDKTXENXUSVWZMFLABQ) was found from its genome. The endohyphal K. michiganensis may play an important role to help nitrogen fixation for fungi in the future.

Publisher

Research Square Platform LLC

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