Kosakonia cowanii, a new bacterial pathogen affecting foxtail millet (Setaria italica[L.]P. Beauv.) in China

Abstract

Foxtail millet (Setaria italica [L.] P. Beauv.) is an important cereal worldwide. From 2021 to 2022, stalk rot disease of foxtail millet was identified in Shanxi province, northern China, with an 8% and 2% field incidence rate in Xinzhou (2 different locations), respectively. It caused necrosis, decay, stem lodging, and sometimes death. This study aimed to identify the causal agent of the disease through morphophysiological and molecular identification of the isolates. Stalk rot specimens were collected in Xinzhou, from foxtail millet plants exhibiting typical symptoms, and the pathogen was isolated with dilution plating. It was cultured at 28 °C for 48 h on nutrient agar, revealing circular, convex, and pale-yellow colonies, with a smooth surface and an entire edge. Scanning electron microscopy showed that the pathogen is rod shaped, round ended and has an uneven surface ranging from 0.5 to 0.7 μm in diameter and 1.2-2.7 μm in length. It is a motile gram-negative facultative anaerobic bacterium that can reduce nitrate and synthesize catalase but cannot hydrolyze starch. It also shows a negative reaction in the methyl red test and optimum growth at 37 °C. The pathogenicity test was performed on foxtail millet variety 'Jingu 21' stem to confirm Koch's postulates. The biochemical tests were done in the Biolog Gen III MicroPlate, revealing 21 positive chemical sensitivity tests, except those for minocycline and sodium bromate. Furthermore, among 71 carbon sources, the pathogen utilized 50 as the sole carbon source, including sucrose, d-maltose, α-d-lactose, d-galactose, D-sorbitol, D-mannitol, glycerol, and inositol. Finally, molecular characterization of the pathogen using 16S rRNA and rpoB gene sequencing and subsequent phylogenetic analysis identified the strain as Kosakonia cowanii. This study is the first to report K. cowanii as a stalk rot-causing pathogen in foxtail millet.

Keywords: Foxtail millet; Koch's postulates; Molecular identification; Pathogenicity; Stalk rot.