First Report of "Candidatus Liberibacter solanacearum" in Pepper Plants in México.

Bell pepper (Capsicum annuum) plants exhibiting symptoms that resembled those of potato psyllid (Bactericera cockerelli Sulc) damage and "Candidatus Liberibacter solanacearum" infection (2) were observed in a pepper field in La Cruz de Elota, Sinaloa, México in March 2009, with an infection rate of 1.5%. Plants exhibited chlorotic or pale green apical growth and leaf cupping, sharp tapering of the leaf apex, shortened internodes, and an overall stunting (2). Total DNA was extracted from the top whole leaf tissue of nine symptomatic and five asymptomatic pepper plants with cetyltrimethylammoniumbromide (CTAB) buffer (3,4). Seven and eight of the nine selected symptomatic pepper plants yielded the expected 1,168-bp 16S rDNA and the expected 669-bp rplJ/rplL ribosomal protein gene amplicons with the "Ca. L. solanacearum" specific OA2/OI2c and CL514F/CL514R primer pairs, respectively, indicating the presence of liberibacter (2,4). Nucleic acid from asymptomatic pepper plants yielded no products with these primers. Three amplicons generated from symptomatic pepper plants with each primer pair were cloned into pCRII-TOPO plasmid vectors (Invitrogen, Carlsbad, CA) and three clones of each amplicon were sequenced in both directions (ACGT, Inc., Wheeling, IL). BLAST analysis of the 16S rDNA consensus sequence (GenBank Accession No. FJ957896) showed 100% identity to 16S rDNA sequences of "Ca. L. solanacearum" amplified from Solanum betaceum (EU935004) and S. lycopersicum (EU834130) from New Zealand (2), and "Ca. L. psyllaurous" from potato psyllids (EU812559) (1). The ribosomal protein gene consensus sequence (GenBank Accession No. FJ957894) was 100% identical to the analogous rplJ and rplL "Ca. L. solanacearum" ribosomal protein gene sequence amplified from S. lycopersicum (EU834131) from New Zealand (2) and to 'Ca. Liberibacter' sp. sequence amplified from zebra chip-infected potato tubers from Lancaster, CA (FJ498803). To our knowledge, this is the first report of "Ca. L. solanacearum" associated with bell pepper in México. "Ca. L. solanacearum" was first reported in tomato and pepper plants in 2008 in New Zealand, where it has resulted in plant decline and significant yield loss, resulting in millions of dollars in losses to the commercial glasshouse tomato and pepper industry (2). Zebra chip, a new and emerging potato disease associated with 'Ca. Liberibacter' sp., was first identified in México in 1994, where it has caused significant economic damage, often leading to abandonment of entire potato fields (3,4). References: (1) A. K. Hansen et al. Appl. Environ. Microbiol. 74:5862, 2008. (2) L. W. Liefting et al. Plant Dis. 93:208, 2009. (3) J. E. Munyaneza et al. J. Econ. Entomol. 100:656, 2007. (4) J. E. Munyaneza et al. Plant Dis. 93:552, 2009.