First Record of Bacterial Crown Rot of Papaya (Carica papaya) Caused by an Erwinia papayae-Like Bacterium in the Kingdom of Tonga.

Symptoms resembling papaya bacterial crown rot (1,3) attributed to Erwinia papayae were first observed on 'Waimanalo' and 'Solo Sunrise' papaya on the island of Tongatapu, Kingdom of Tonga in July 2009. Spreading, dark green, water-soaked lesions formed on juvenile stem tissue and developed into a foul-smelling, wet rot that destroyed large sections of the stem. Coalescing, brown, angular, marginal, and intercostal lesions killed large areas of the lamina. Elongated lesions on petioles resulted in breakage and leaf death. Symptoms on stems typically moved toward the crown with the growing point being killed or the whole crown breaking off at a canker below. Isolations at 28°C on King's medium B (KB) yielded slow-growing, raised, white, mucoid colonies that produced a conspicuous, nondiffusable blue pigment in the medium. Two-day-old suspensions (1 × 10⁸ CFU/ml) of two cultures were injected into juvenile stem tissue, petioles, and laminae of four healthy papaya seedlings of 'Solo Sunrise' with a sterile 1-ml insulin syringe. Sterile water was used as a negative control. Typical water-soaked lesions appeared at all bacterial inoculation sites on all plants within 5 days but not on controls. Pigment-producing colonies similar to those used for inoculation were reisolated from four different stem lesions. Bacteria isolated from diseased tissues were gram negative, facultative anaerobes, oxidase negative, nonfluorescent on KB, induced a hypersensitive reaction on tobacco leaves, but could not cause soft rot on potato slices. Those characteristics, combined with blue pigment production, are consistent with the bacterium E. papayae. A partial sequence of the 16S rDNA gene of ~804 bp was amplified from four Tongan isolates (ICMP18248-18251) using primers 27f and 1492r (4). Sequences of these strains were 100% identical to each other (GenBank Nos. HQ286366-HQ286369), 99 and 98% identical to the 16SrDNA sequences of E. mallotivora strains LMG2708 (Z96084.1) and DSM4565 (AJ233414.1) respectively, and 97% identical to the 16SrDNA sequence of E. papayae strain NCPPB 4294 (AY131237.1). E. mallotivora and E. papayae cause different diseases, a leaf spot on Mallotus japonicus (2) and bacterial canker on papaya, respectively. They are closely related and in the laboratory are distinguished by only very few biochemical characteristics (1). E. papayae produces a blue pigment on KB and can utilize arabinose but not mannitol. E. mallotivora does not produce a blue pigment and can utilize mannitol but not arabinose. The four Tongan strains produced a blue pigment and could utilize mannitol and arabinose. Symptoms caused by the strains isolated from Tonga are typical of those caused by E. papayae and the strains identified share most of the characteristics of E. papayae. Because the Tongan strains were able to utilize mannitol as well as arabinose and their 16S rDNA was only 97% similar to E. papayae, these strains are referred to as an E. papayae-like bacterium. The taxonomic position of these isolates will be resolved with techniques such as Multilocus Sequence Typing analysis. To our knowledge, this is the first report of this highly destructive papaya disease in the Kingdom of Tonga and of a pathogen closely related to E. papaya in the country. References: (1) L. Gardan et al. Int. J. Syst. Bacteriol. 54:107, 2004. (2) M. Goto. Int. J. Syst. Bacteriol. 26:467, 1976. (3) N. H. Maktar et al. New Dis. Rep. 17:4, 2008. (4) F. Martin-Laurent et al. Appl. Environ. Microbiol. 67:2354, 2001.