Biology and Management of Bacterial Spot of Peppers in Oklahoma.

Bacterial spot, an important disease of bell and chili peppers grown in Oklahoma, is caused by Xanthomonas euvesicatoria. We evaluated isolates from 1995 to 2015 (n = 72) for avirulence alleles and race by assessing hypersensitive responses (HRs) on differentials with resistance genes Bs1, Bs2, Bs3, or Bs4. Most isolates (96%) expressed AvrBs2 (races 1, 3, 7, 8), and only three were virulent on Bs2 (race 6). Chili cultivars, grown locally for capsaicin production, were susceptible to all races. Copper-based spray programs were evaluated on bell pepper hybrids with and without Bs2 resistance from 2008 to 2010 and on bell and jalapeño hybrids with Bs2 or Bs1-2-3 from 2017 to 2018. Bs2 and Bs1-2-3 hybrids generally had lower disease and higher yields (≤21 t/ha) than susceptible entries. Copper reduced disease by 20 to 40% and increased yield by an average of 4 t/ha, but yield responses were not always significant (P = 0.05). In August 2018, disease increased to >50% on Bs2 hybrids but remained low on the Bs1-2-3 hybrid. Despite the breakdown of Bs2 resistance, yields of the Bs2 hybrids were not reduced. Avirulence alleles and race of isolates from susceptible, Bs2, and Bs1-2-3 hybrids at the end of the 2018 trial depended on source plant genetics. AvrBs2 was expressed in 86% of isolates from a susceptible hybrid but not in any isolates from the Bs2 and Bs1-2-3 hybrids. HR resistances effectively protected yield, but their deployment may not be sustainable without cultural practices such as crop rotation that limit pathogen survival and transmission to subsequent crops.

Characterization of the Race Structure of Leptosphaeria maculans Causing Blackleg of Winter Canola in Oklahoma and Kansas.

Blackleg, caused by the fungus Leptosphaeria maculans, is a widespread disease of winter canola (Brassica napus) in Oklahoma and Kansas. Deployment of genetic resistance is the primary strategy for managing blackleg. Resistance genes (Rlm) in canola interact with avirulence genes in the fungus (AvrLm) in a gene-for-gene manner. Little is known about the diversity and frequency of avirulence genes and the race structure in the region. Isolates of Leptosphaeria spp. were collected from diseased leaves in nine counties in Oklahoma and one county in Kansas from 2009 to 2013. Based on pathogenicity and PCR amplification of mating type and species-specific internal transcribed spacer loci, most isolates (n = 90) were L. maculans. The presence of avirulence genes was evaluated using phenotypic interactions on cotyledons of differential cultivars with Rlm1, Rlm2, Rlm3, and Rlm4 and amplification of AvrLm1, AvrLm4-7, and AvrLm6 by PCR. The avirulence alleles AvrLm6 and AvrLm7 were present in the entire L. maculans population. AvrLm1 was found in 34% of the population, AvrLm2 in 4%, and AvrLm4 in only 1%. A total of five races, defined as combinations of avirulence alleles, were identified that included AvrLm1-2-6-7, AvrLm2-6-7, AvrLm4-6-7, AvrLm1-6-7, and AvrLm6-7. Races virulent on the most Rlm genes, AvrLm1-6-7 at 32% and AvrLm6-7 at 62%, were predominant. Defining the avirulence allele frequency and race structure of L. maculans should be useful for the identification and development of resistant cultivars and hybrids for blackleg management in the region. The results suggest that Rlm6 and Rlm7 would be effective, although their deployment should be integrated with quantitative resistance and cultural practices, such as crop rotation, that limit selection pressure on Rlm genes.

Survival and growth of foodborne pathogens in pesticide solutions routinely used in leafy green vegetables and tomato production.

BACKGROUND: The consumption of fresh produce has increased tremendously in the past few years as have outbreaks of foodborne illnesses associated with these commodities. Pesticides routinely used in crop production could influence the outcomes of foodborne pathogen contamination of fresh produce. Experiments were performed to determine the effects of pesticides on the survival and growth characteristics of Escherichia coli O157:H7 and Salmonella spp. Eight commercial fungicides and insecticides commonly used for disease and insect pest control on leafy green vegetables and tomatoes were evaluated. RESULTS: Among the pesticides tested, copper hydroxide, acetamiprid, cypermethrin and permethrin were found to be significantly (P < 0.05) inhibitory to pathogens while no effect was observed for chlorothalonil, flonicamid and methoxyfenozide. At the highest concentration tested (2.66%), azoxystrobin had a significant (P < 0.05) stimulatory effect on the growth of E. coli O157:H7 after 24 h incubation. The results indicated that some pesticides can stimulate the growth of human pathogens if contaminated water is used in their preparation, whereas others were likely to inhibit or reduce pathogen populations. CONCLUSION: This information is helpful in mitigating the risk of microbial contamination in fresh produce, which is critical to public health and safety.

Detection, Survival, and Sources of Inoculum for Bacterial Diseases of Leafy Crucifers in Oklahoma.

Xanthomonas campestris pv. campestris, X. campestris pv. armoraciae, and Pseudomonas syringae pv. maculicola are bacterial pathogens that cause leaf spot diseases on leafy crucifers in Oklahoma. Polymerase chain reaction (PCR) amplification of the cfl gene from the gene cluster encoding the phytotoxin coronatine was used to identify coronatine-producing strains of P. syringae, and the expected 0.65-kb PCR product was detected in 19 strains of P. syringae pv. maculicola originating from diseased crucifers in Oklahoma. A simple, rapid PCR method based on primers from the cfl gene was developed to detect coronatine-producing strains of P. syringae in planta. Pathogenicity tests confirmed the cfl-positive strains to be P. syringae pv. maculicola. To monitor the survival of X. campestris pv. armoraciae and P. syringae pv. maculicola in the field, turnip and collards were inoculated with rifampicin-resistant strains and were buried beneath the soil or left on the soil surface. Both pathogens were recovered from turnip and collard debris up to 2 months following burial, but neither pathogen was recovered from soil after the debris had decomposed. However, both pathogens were recovered from plant debris left on the soil surface for up to 5 months. Four production fields were surveyed for sources of inoculum of the bacterial pathogens from October 1999 to May 2000. X. campestris pv. campestris was isolated from the weed shepherd's purse (Capsella bursa-pastoris) in all fields, and from volunteer turnip and kale in three fields. X. campestris pv. campestris and P. syringae pv. maculicola were isolated from surface debris and regrowth from crop stubble left in one field after harvest in the fall. X. campestris pv. campestris was detected in 6 of 51 lots of crucifer seed assayed. X. campestris pv. armoraciae and P. syringae pv. maculicola were not recovered from weeds, volunteer plants, or seed lots.

Bacterial Leaf Spot Diseases of Leafy Crucifers in Oklahoma Caused by Pathovars of Xanthomonas campestris.

Fields of kale, spinach mustard, and turnip were severely damaged by bacterial leaf spots during 1994 to 1996. Symptoms included circular to angular necrotic lesions with yellow halos and water-soaking on the abaxial leaf surface. Yellow, mucoid strains isolated from leaf spots were identified as Xanthomonas campestris using Biolog. Four strains caused black lesions on stems of cabbage seedlings in an excised cotyledon assay, leaf spots and sunken dark lesions on petioles of spray-inoculated crucifers, and leaf spots on spray-inoculated tomato. These strains were classified as X. campestris pv. armoraciae. Most other strains from leafy crucifers and all strains from a cabbage field caused black rot in the cotyledon assay and in spray-inoculations. Many of these strains also caused leaf spots on collard and kale but not stem and petiole lesions. The strains causing black rot were classified as X. campestris pv. campestris. Cluster analysis of Biolog profiles yielded a small group that contained local strains of both pathovars, and a large group comprised of reference and local strains of each pathovar, and some local, nonpathogenic strains. Five fingerprint groups were identified by rep-polymerase chain reaction using the BOXA1R primer. Local and reference strains of each pathovar occurred in two of the groups. Two pathovars of X. campestris are involved in the leaf spot diseases. Both pathovars were recovered within several fields, and also were recovered along with Pseudomonas syringae pv. maculicola. This is the first report of Xanthomonas leaf spot caused by X. campestris pv. armoraciae in Oklahoma.

Bacterial Leaf Spot of Leafy Crucifers in Oklahoma Caused by Pseudomonas syringae pv. maculicola.

During 1995 and 1996, bacterial leaf spots severely damaged fields of kale, spinach mustard, and turnip in Oklahoma. Symptoms were small, brown, necrotic spots with irregular edges surrounded by chlorotic halos. Lesion margins were often water-soaked on the abaxial surface. The spots enlarged and coalesced, causing extensive leaf yellowing and necrosis. Nineteen strains of a fluorescent Pseudomonas spp. were isolated from symptomatic plants. LOPAT tests and carbon source oxidation using Biolog GN MicroPlates were used to classify the strains as P. syringae. Cluster analysis of carbon source oxidation profiles for the local strains and selected reference strains of P. syringae pv. maculicola and pv. tomato produced one group with 79.5% similarity. In spray inoculations, all local strains caused chlorotic or water-soaked lesions on collards, kale, cauliflower, and tomato. A few local strains caused necrotic lesions on mustard. Most local strains caused one of the three lesion types on turnip and spinach mustard. Reference strains of P. syringae pv. maculicola caused similar symptoms. All but three of the local strains produced coronatine in vitro. The local strains were thus classified as P. syringae pv. maculicola, the cause of bacterial leaf spot of crucifers. Two distinct groups of P. syringaepv. maculicola were identified by repetitive sequence-based polymerase chain reaction (rep-PCR) with both REP and BOXA1R primers. Three subgroups within each group were further identified using the BOXA1R primer. Except for two strains of P. syringae pv. tomato which were pathogenic on crucifers, the pathovars maculicola and tomato had different genetic fingerprints. The pathogen was recovered from seven of ten fields sampled during 1994 to 1996. In five of the fields with P. syringae pv. maculicola, pathovars of Xanthomonas campestris were also isolated from lesions forming a bacterial disease complex. This is the first report of bacterial leaf spot caused by P. syringaepv. maculicola on leafy crucifers in Oklahoma.