New emerging Pseudomonads as causal agents of bean blight disease

Common bean (Phaseolus vulgaris) production is negatively affected by several bacterial plant diseases. A severe outbreak of bean blight was observed in the nine major bean-producing provinces of Iran during 2021–2022. Necrotic spots were observed on leaves which become necrotic with chlorosis beyond it. One hundred-sixty bacterial strains with a greyish-white colony appearance were consistently isolated from the one-hundred eighteen symptomatic bean leaf, pod, seed and root samples on nutrient agar. All strains were screened for their ability to cause disease bean leaves among which 134 strains induced chlorosis three days after inoculation on bean leaves of cv. Yaghoot followed by necrotic spots on the inoculated sites one week after inoculation. The pathogenic strains clustered in three groups based on phenotypic characters and multilocus sequencing analyses. Three housekeeping genes including gyrB, rpoB, and rpoD of representative strains were partially sequenced. In the phylogenetic tree based on concatenated sequences of these three genes or each gene individually, the strains were divided into three clusters each with high bootstrap support. The first group of the strains clustered with Pseudomonas simiae CCUG 50988^T. This is the first report of this species as causal agent of bean blight. The second group clustered close to Pseudomonas alloputida NMI5768^T which apparently belong to a new species. The third group clustered in a separate clade in Pseudomonas chlororaphis species which belongs to a new subspecies based on pathogenicity on bean, phenotypic characters, sequences of five housekeeping genes which named as subsp. phaseoli subsp. nov. with the type strain UT-M315.