RESUMEN
In Oklahoma, during the late summer of 2004, an elm tree (Ulmus americanus L.) located in the Oklahoma Botanical Gardens near Stillwater showed symptoms of marginal leaf scorch bordered by a yellow band between necrotic and green tissues, indicating possible Xylella fastidiosa infection. Three leaves from the symptomatic tree and one leaf from an asymptomatic nearby elm were sampled. DNA was extracted with the Extract-N-Amp kit (Sigma, St. Louis, MO). Samples were tested for X. fastidiosa using real-time polymerase chain reaction (PCR) with Xylella genus specific primers XfF1/XfR2 and dual-labeled TaqMan probe XfP2 (2). Infected oleander from California was used as a positive control. All three samples from symptomatic leaves and the positive control were PCR positive, and the sample from the asymptomatic tree was PCR negative. Attempts to culture an isolate of the bacteria from petioles and branch tissues on PD3 and PW, media selective for X. fastidiosa, failed. For more detailed molecular characterization of the putative pathogen, DNA from additional symptomatic petioles from the same tree was isolated using the cetyltrimethylammoniumbromide (CTAB) extraction. X. fastidiosa specific primers BBXFOUTF1 (5'-AAGCGCCTCCGTGAGTTATC-3') and BBXFOUTR1 (5'-CCTTCACGCATATCATCACC-3') were used to PCR amplify the gyrB gene. The amplification product was recovered after gel electrophoresis with QIAquick gel extraction kit (Qiagen, Valencia, CA) and was subjected to automated sequencing (Oklahoma State University Recombinant DNA/Protein Resource Facility). BLASTN alignment (1) of the obtained 381 bp sequence revealed 100% identity with the gyrB gene from elm (GenBank Accession No. AF534966) and mulberry (GenBank Accession No. AF534965) isolates of X. fastidiosa. During 2005, petiole samples from the tree were collected and serological diagnosis was confirmed using enzyme-linked immunosorbent assay (Agdia, Inc., Elkhart, IN). Some strains of X. fastidiosa have very wide host ranges and many of the hosts may be asymptomatic. Therefore, the economic importance and implications of the detection of X. fastidiosa in the state of Oklahoma remain to be determined. To our knowledge, this is the first report of X. fastidiosa in Oklahoma. References: (1) S. F. Altschul et al. J. Mol. Biol. 215:403, 1990. (2) N. W. Schaad et al. Phytopathology 92:721, 2002.
RESUMEN
Downy mildew, incited by Phytophthora phaseoli Thaxt., is the most important disease of lima bean (Phaseolus lunatus L.) on the east coast of the United States. It has been a serious threat to commercial lima bean production in Delaware, Maryland, and New Jersey for the past 5 years. Growers have attempted to manage this disease using resistant cultivars and copper hydroxide fungicides. In August and September 1995, a new pathogenic race of P. phaseoli was isolated from infected pods of the lima bean cv. Packer in a production field near Milton, DE. Races of P. phaseoli are determined using a modification of a cultivar differential developed by Wester (3). The cv. 184-85, which is resistant to races A, B, C, and D (1), is susceptible to the new race, designated as E. In August 2000, another new pathogenic race of P. phaseoli was isolated from infected pods of cv. 184-85 near Middletown, DE. The lima bean line BG2-408, which is resistant to races A, B, C, D, and E, is susceptible to the new race, designated as F. Symptoms produced on lima bean plants infected by races E and F are similar to each other, and to those produced by all other races. All races of P. phaseoli have the same cultural characteristics on lima bean pod agar. Evaluations of in field weather station data and disease occurrence indicate that races E and F may have temperature maxima greater than 32°C, whereas race D has a maximum of less than 32°C (2). During the 2000 growing season, 118 isolates of P. phaseoli were collected from 44 production fields in Delaware and the eastern shore of Maryland, with 86% characterized as race E and 5% as race F. References: (1) C. R. Davidson et al. Biol. Cult. Tests 2001:V80. (2) R. A. Hyre and R. S. Cox. Phytopathology 43:419, 1953. (3) R. E. Wester. Phytopathology 60:1856, 1970.