First Report of Pantoea agglomerans Causing Leaf Blight and Bulb Rot of Onion in Chile

Onion (Allium cepa L) is an important vegetable crop in Chile with 3.675 ha (ODEPA, www.odepa.gob.cl). Since 2017, a series of symptoms has been increasingly observed, mainly in mature bulbs which include water-soaked lesions on mature leaves and external and central cataphylls, which evolve to necrotic lesions and rot, reducing yields and affecting the postharvest life. As the season progresses over 90% incidence has been estimated in severely affected fields. Symptomatic plants were collected for disease identification in summer (February 2022) in Santa Cruz, O´Higgins, Chile, (34º41'32"S and 71º27'17"W). Isolations were done on semi-selective OEM medium (Zaid et al. 2012) and isolated colonies were transferred and maintained on King's B (KB) medium. Colonies on OEM medium were yellow slightly green in the center, circular to irregular shape, elevated, smooth margin, and mucoid texture (Tho et al. 2015). On KB medium colonies were yellow cream, slightly darker in the center, with smooth margins and circular or slightly irregular. Two isolates (CHS B3, CHS B1), obtaining in three different onion fields, were further identified by PCR amplification of partial 16S rRNA using primers 27F/1492R (Frank et al., 2008). The resulting sequencing were deposited in GenBank under accession numbers OP934962.1 and OP934961.1. A BLAST search of GenBank showed that these sequences exhibited 99,89 to 100 % nucleotides similarities with multiple Pantoea agglomerans sequences. Three isolates were used to test pathogenicity on detached cataphylls and onion plants tests in the laboratory and greenhouse, respectively. Cataphylls (n=10) were surface disinfected with 2% NaOCl for 2 min, placed in sterile Petri dishes, and injected with 50 µL (106 UFC/mL) inoculum suspension with the aid of a hypodermic syringe. Cataphylls were incubated in humid chamber at 22 + 2ºC for 14 days. Additionally, three-month-old onion plants (n=10), growing in pod, were inoculated by injecting leaves with 1 mL inoculum suspension (106 UFC/mL). Plant were incubated under greenhouse for 30 days to 28ºC. An equal number of cataphylls and onion plants, injected with sterile water, were left as controls. Water soaked lesions on inoculated cataphylls appeared five days after inoculation. While onions plants develop water soaked lesion 1- 4 days after inoculation. Blight and bulb rot were observed only on inoculated plants. Symptoms were identical to those previously observed in the field. Reisolations were accomplished for 100% of the inoculated cataphylls and leaves fulfilling Koch's postulates (Biochemical characterization and PCR amplification of partial 16S rRNA). Previously, Pectobacterium carotovorum was reported causing a similar disease in Chile (Acuña 2008). However, to our knowledge this is the first report of P. agglomerans causing onion leaf blight and bulb rot in O'Higgins Region, Chile. In agreement with other reports (De Armas et al. 2015, Tho et al. 2015), leaf blight and bulb has been a very aggressive disease in Chile. These results do not exclude the possibility that other Pantoea species (Armas et al. 2015) may be involved in the leaf blight and bulb rot of onions.