Identification and Enumeration of Small-Spored Alternaria Species Associated with Potato in the US Northwest

A number of Alternaria spp. have been isolated from potato worldwide but only Alternaria solani and A. alternata have been described as pathogenic to this host in the United States. These taxa are easily differentiated based on conidial morphology but species delimitation among the small-spored Alternaria spp. associated with potato are much more challenging. Accurate identification methods for small spored Alternaria spp. are necessary so that a more thorough understanding of Alternaria epidemiology can be obtained. Isolations of Alternaria fungi from lesions on potato leaves collected in the U.S. Northwest were made between 2008 and 2011. Large-spored taxa (putatively A. solani), were isolated less frequently than small-spored taxa (putatively A. alternata sensu lato), except in 2010. Colletotrichum coccodes was isolated from necrotic lesions in 2008 to 2010 but not in 2011. Frequency of isolation ranged from 0.05 (5%) to 0.11 (11%) during the 3 years the fungus was detected. Anonymous genomic region OPA1-3, previously used for Alternaria systematics, allowed for the discrimination of phylogenetic lineages among 210 small-spored isolates. When OPA1-3 was restricted using enzyme ApaI, 65 isolates (31%) displayed a restriction banding pattern consistent with previously characterized morphospecies A. alternata and A. tenuissima and 145 (69%) displayed a restriction banding pattern consistent with the previously characterized morphospecies A. arborescens. Morphological characterization of a subsample of 59 small-spored Alternaria isolates randomly selected with each restriction pattern was compared with phylogenetic lineage. In all, 54 (92%) isolates were consistently assigned to the same group by both methods. Three isolates exhibited conidial morphologies that were inconsistent with any described morphospecies. A small number of isolates were identified as A. arbusti (infectoria group) via sequencing of the glyceraldehyde-3-phosphate-dehydrogenase locus and BLAST searches.