Tar Spot: An Understudied Disease Threatening Corn Production in the Americas.

Tar spot of corn has been a major foliar disease in several Latin American countries since 1904. In 2015, tar spot was first documented in the United States and has led to significant yield losses of approximately 4.5 million t. Tar spot is caused by an obligate pathogen, Phyllachora maydis, and thus requires a living host to grow and reproduce. Due to its obligate nature, biological and epidemiological studies are limited and impact of disease in corn production has been understudied. Here we present the current literature and gaps in knowledge of tar spot of corn in the Americas, its etiology, distribution, impact and known management strategies as a resource for understanding the pathosystem. This will in tern guide current and future research and aid in the development of effective management strategies for this disease.

First Report of Downy Mildew Caused by a Peronospora sp. on Basil in Florida and the United States.

Basil is grown as a specialty crop in greenhouse and field production in Florida and other regions of the United States. Downy mildew on basil (Ocimum basilicum) was detected from four production sites (Collier, Hendry, Miami-Dade, and Palm Beach counties) in south Florida in the fall of 2007, and within months, it was also found in west-central north Florida (Hillsborough County). Incidence reached nearly 100% on some of the affected crops and caused complete yield losses on basil grown both in the field for fresh market and potted herbs market. Symptoms developed during transit on basil that appeared symptomless at harvest. Symptoms initially appeared as yellowing on the lower leaves that was typically delineated by the veins, although in some cases the entire leaf area of the leaf surface was affected. A gray, fuzzy growth was apparent on the abaxial leaf surface. Microscopic observation detected dichotomous branching, hyaline sporangiophores (220 to 750 × 4 to 9 µm) bearing single sporangia. Sporangia were light brown, ovoid to slightly ellipsoid, and measured 14 to 15 × 15 to 18 µm. Oospores were not observed. Leaves of potted basil plants and coleus (Solenostemon scutellarioides) were inoculated with a suspension containing 1 × 105 sporangia/ml and sprayed till runoff (approximately 15 ml per plant) with a hand-held pressurized aerosol canister. Plants were covered with a plastic bag for 24 h and maintained in the greenhouse under ambient conditions. Noninoculated plants served as controls. After 7 days, symptoms typical of downy mildew occurred only on the inoculated basil plants and sporulation was confirmed microscopically. The internal transcribed spacer regions of an isolate collected in Hendry County were sequenced bidirectionally. The consensus sequence was deposited into GenBank (Accession No. FJ346561). Sequence data matched (100% homology) with a Peronospora sp. reported on sweet basil in Switzerland (GenBank Accession No. AY884605) and was similar (99% homology) to an isolate (GenBank Accession No. DQ523586) reported on coleus, although inoculation to coleus failed to confirm pathogenicity on this host. The sequence data also distinguished the isolate from P. lamii (87% homology) previously reported to occur on basil. The pathogen was identified as a Peronospora sp. based on morphological characteristics and sequencing homology (1-3). References: (1) L. Belbahri et al. Mycol. Res. 109:1276, 2005. (2) S. Francis. CMI Descriptions of Pathogenic Fungi and Bacteria. No. 688. CMI, Kew, England, 1981. (3) A. McLeod et al. Plant Dis. 90:1115, 2006.

First Report of Brown Ring Patch Caused by Waitea circinata var. circinata on Poa trivialis in Florida.

Brown ring patch is a newly described disease of cool-season turfgrass first reported in Japan on creeping bentgrass (Agrostis palustris) (2) and later reported in California on annual bluegrass (Poa annua) (1). The disease is characterized by either patches or rings of discolored to blighted turfgrass that can range from a few centimeters to a meter in diameter. Affected turfgrass plants turn chlorotic and can be blighted from the crown to the leaf tips. Blight symptoms have been associated with fluffy white-to-cream aerial mycelium after extended incubation of the sample. Symptoms including patches of blighted turfgrass approximately 10 cm in diameter were observed on roughstalk bluegrass (Poa trivialis) that had been overseeded onto a dormant 'Tifdwarf' bermudagrass (Cynodon dactylon) putting green in Palatka, FL. A sample was submitted by the superintendent in June 2005 because symptoms were confused with dollar spot and a fungicide resistance issue was suspected. The sample produced abundant aerial mycelium after incubation. The pathogen was isolated on potato dextrose agar amended with rifampicin (100 ppm) and streptomycin (100 ppm) from Poa plants surface disinfested with 70% ethanol for 30 s. Colony and sclerotia morphology were consistent with Waitea circinata var. circinata as previously described (1,2). The teleomorph W. circinata var. circinata was not observed on plant material or culture plates. Amplified fragments of rDNA including internal transcribed spacers from the isolate were sequenced bidirectionally from four bacterial clones. The consensus sequences (GenBank Accession Nos. FJ029103, FJ029104, FJ029105, and FJ029106) matched with 99% homology (99% sequence overlap) isolate TRGC1.1 of W. circinata var. circinata described by Wong, NCBI Accession No. DQ900586 (1). Pots of 'Cypress' roughstalk bluegrass that were 1 week postemergence were inoculated with the pathogen using 10 infested wheat grains. Plants were incubated at 25°C in a sealed plastic bag with a moist paper towel in the bottom. Hyphae grew from the grains and colonized the grass. Individual plants began to turn chlorotic within 3 days and greater than 90% of the turf in pots was dead after 1 week. The fungus was reisolated from affected plants. Control pots were inoculated with uninfested wheat grains and showed no disease symptoms after 1 week. Inoculations were repeated twice more with the same results. To our knowledge, this is the first report of brown ring patch on P. trivialis in Florida. References: (1) K. A. de la Cerda et al. Plant Dis. 91:791, 2007. (2) T. Toda et al. Plant Dis. 89:536, 2005.

Bidens mottle virus Identified in Tropical Soda Apple in Florida.

Tropical soda apple (TSA) (Solanum viarum Dunal), a plant native to South America, was first identified in Florida in 1988 (4). It rapidly became a noxious weed in pastures throughout the state and it is known to be a reservoir for Cucumber mosaic virus, Potato leafroll virus, Potato virus Y (PVY), Tobacco etch virus (TEV), Tomato mosaic virus, and Tomato mottle virus, viruses that infect important vegetable crops in Florida (3). During a routine survey of Florida weeds during May of 2004, a TSA plant with chlorotic, young leaves found near Okeechobee, FL was determined to be infected with a potyvirus by using a commercially available enzyme linked immunosorbent assay kit (Agdia, Elkhart, IN). The results of a host range study indicated this potyvirus was neither PVY nor TEV. The virus caused local lesions in Chenopodium amaranticolor and systemic symptoms in C quinoa, Coreopsis sp. (C. A. Baker, unpublished), Helianthus annus, Nicotiana benthamiana, Petunia × hybrida, Verbena hybrida, and Zinnia elegans. It did not infect Gomphrena globosa, N. glutinosa, Pisum sativum, or Phaseolus vulgaris (1). Cylindrical inclusions consistent with those observed in plants infected with Bidens mottle virus (BiMoV) were observed in Z. elegans. Immunodiffusion tests with antiserum to BiMoV (Department of Plant Pathology, University of Florida) gave a reaction of identity with leaf extracts of the symptomatic zinnia, a known sample of BiMoV originally isolated from Bidens pilosa and a recent isolate of BiMoV from lettuce in Belle Glade, FL (C. A. Baker and R. Raid, unpublished). A partial polyprotein gene fragment (GenBank Accession No. EF467235) was amplified from total RNA of an inoculated C. quinoa plant by reverse transcription (RT)-PCR with previously described degenerate potyvirus primers (2). Analysis of the RT-PCR product sequence confirmed the host range results and indicated that the potyvirus infecting TSA was neither PVY nor TEV. However, the nucleotide and deduced amino acid sequences of a 247-bp portion of the RT-PCR product were 94 and 98% identical, respectively, with the coat protein sequence (GenBank Accession No. AF538686) of Sunflower chlorotic spot virus (SCSV). SCSV is a tentative potyvirus species described from Taiwan that is not yet recognized as an accepted species by the International Committee on Taxonomy of Viruses. On the basis of our concurrent host range, inclusion body, and serological data, it is likely that SCSV is in actuality the previously described and currently accepted potyvirus species BiMoV, for which no previous sequence data existed. As part of a comprehensive viral disease management plan, it is recommended that TSA plants growing in and around lettuce-production areas be controlled along with other weed hosts of this virus. References: (1) A. A. Brunt et al., eds. Plant Viruses Online: Descriptions and Lists from the VIDE Database. Version: 20 at http://biology.anu.edu.au/Groups/MES/vide/ , 1996. (2) A. Gibbs and A. J. Mackenzie. Virol. Methods 63:9, 1997. (3) R. J. McGovern et al. Int. J. Pest Manag. 40:270, 1994. (4) J. J. Mullahey et al. Weed Technol. 7:783, 1993.

First Report of a Leaf Spot Disease of Wild Rocket (Diplotaxis tenuifolia) in Florida Caused by Xanthomonas campestris pv. raphani.

Wild rocket, also known as wall rocket or sand rocket (Diplotaxis tenuifolia (L.) DC, family Brassicaceae), is grown in Florida as a salad green and herb, especially for addition to "spring mix" and other bagged salad blends. It is similar in texture and flavor to the more widely known garden arugula (Eruca vesicatoria (L.) Cav. subsp. sativa). During the winter vegetable season of 2006-2007, a leaf spot disease causing severe economic loss was seen in commercial fields of wild rocket near the town of Sebastian in Indian River County, FL. Discrete water-soaked to greasy appearing spots were observed on leaf blades that rarely exceeded 1 mm in diameter with some surrounded by a narrow, yellow halo. There was no evidence of marginal V-shaped lesions suggestive of vascular black rot disease caused by Xanthomonas campestris pv. campestris. A bacterium that formed yellow colonies on nutrient agar was consistently isolated from these lesions. Four strains were isolated, purified, and characterized. All strains were strictly aerobic, gram-negative rods. Strains were positive for esculin hydrolysis and gelatin liquefaction and negative for oxidase, nitrate reduction, urease production, fluorescence on King's B medium, and utilization of asparagine as a sole source of carbon and nitrogen. Proteolysis and an alkaline reaction were observed in inoculated tubes of litmus milk. Colonies were bright yellow and mucoid on plates of yeast extract-glucose-calcium carbonate agar. Carbon source utilization, as revealed by the Biolog system, indicated a match to X. campestris. Fatty acid methyl ester analysis indicated a match with Florida library strains of X. campestris pv. raphani (similarity indices of 0.737 to 0.779). Suspensions (2 × 107 CFU/ml in sterile phosphate-buffered saline) of the four wild rocket strains and a strain isolated in 2003 as a cause of a discrete leaf spot disease of cabbage in southern Florida (1) were sprayed onto plants until runoff with a hand-held plastic mist bottle. Pathogenicity of strains was tested in the greenhouse on seven replicate plants of green cabbage cv. Copenhagen Market, salad arugula cv. Rocket, and wild rocket (an unnamed selection propagated from seed provided by a Florida grower). Symptoms appeared within 6 to 7 days. The wild rocket strains and the cabbage strain were pathogenic on all wild rocket and cabbage test plants, producing small, nonvascular leaf spots. No symptoms were seen on salad arugula or on control plants of wild rocket, cabbage, or arugula sprayed with phosphate-buffered saline. The bacterium was reisolated from infected plants and identified as X. campestris pv. raphani, thus completing Koch's postulates. We have classified the bacterial leaf spot pathogen of wild rocket as X. campestris pv. raphani using the nomenclature of Vincente et al. (2) for X. campestris strains producing nonvascular leaf spots on brassicas. This disease problem seems to be related to widespread use of overhead irrigation in the fields where the disease is prevalent. We have not observed the disease in seepage-irrigated fields of wild rocket. References: (1) K. Pernezny et al. Plant Dis. 87:873, 2003. (2) J. G. Vincente et al. Phytopathology 96:735, 2006.

First Report of Powdery Mildew of Parsley Caused by Erysiphe heraclei in Florida.

Parsley (Petroselinum crispum (Mill.) Nym. ex A.W. Hill) is an important leaf crop in the Everglades Agricultural Area of southern Florida. During the spring of 2005 and 2006, disease signs and symptoms resembling those incited by powdery mildew were observed on parsley at a commercial vegetable farm located 15 km east of Belle Glade. Symptoms consisted of leaf chlorosis, particularly in the dense lower canopy, and desiccation of affected tissue. A dense, white-to-light gray fungal growth was visible macroscopically on the surface of affected leaf tissue. Microscopic examinations revealed ectophytic hyphae with lobed appressoria and hyaline, straight conidiophores bearing single conidia. Conidia were short-cylindrical to cylindrical, measured 33 to 44 µm long and 13 to 16 µm wide, and lacked fibrosin bodies. Conidiophore foot cells were also cylindrical, straight, and measured 27 to 37 × 9 to 10 µm. Ascocarps of the teleomorph were not observed. The fungus closely matched the description of Erysiphe heraclei DC, a pathogen previously reported as attacking parsley on the U.S. West Coast (1,2). Pathogenicity was verified by inoculating adaxial leaf surfaces of 12 plants (cv. Dark Green Italian) with conidia collected from infected tissue by using a small brush. Inoculated plants and 12 noninoculated plants were lightly misted, held in a moist chamber for 48 h (22°C), and then incubated in a growth chamber for 4 weeks at 22°C with a photoperiod of 16 h. Symptoms that developed on inoculated plants were similar to those observed in the field, with no symptoms evident on the controls treated in a similar manner. To our knowledge, this is the first report of powdery mildew on parsley in Florida, even though parsley has been grown in the area for at least six decades. Noted as being somewhat unique among fungal pathogens because it favors dry rather than moist climatic conditions, it is probably no coincidence that powdery mildew was observed both years during the month of April, the height of Florida's dry season. The fact that monthly rainfall totals of 22 and 35 mm were recorded during April of 2004 and 2005, respectfully, well below the historical average of 72 mm, may have been a contributing influence. Glawe et al. (1), in issuing a first report of E. heraclei on carrots and parsley in the state of Washington and observing ascocarps on carrot tissue, mentioned the prospect of contaminated seed serving as a potential source of dissemination. Although they did not observe the teleomorph on parsley, prospects for its occurrence seem likely. With the bulk of parsley seed planted in Florida being produced in Washington, Oregon, or California, the observations reported herein may provide credence to such a hypothesis. References: (1) D. A. Glawe et al. Online publication. doi:10.1094/PHP-2005-0114-01-HN. Plant Health Progress, 2005. (2) S. T. Koike and G. S. Saenz. Plant Dis. 78:1219, 1994.

First Report of Peronospora farinosa f. sp. spinaciae Race 5 Causing Downy Mildew on Spinach in Florida.

Downy mildew, caused by Peronospora farinosa f. sp. spinaciae, is an economically important disease in most areas where spinach is grown. This disease has become increasingly important in production fields for prepackaged salad mixes where plant densities typically are very high. In Florida, spinach production for these markets has reached approximately 200 ha. Currently, seven physiological races of the downy mildew pathogen have been described (1). Downy mildew was observed in several commercial spinach fields in the Everglades agricultural area of Palm Beach County, Florida in January 2003 on cvs. Unipak 151 and Merlo Nero. Symptoms appeared as chlorotic and necrotic leaf spots. Disease incidence reached approximately 25% in some field locations. Economic losses were significant, since entire plantings in several fields were not harvested as a result of diminished quality. The race of a field isolate recovered from the cv. Unipak 151 was determined following greenhouse inoculation procedures and using differentials outlined by Irish et al (1). Greenhouse inoculation tests were conducted twice. Disease reactions on a U.S. and international set of differentials indicated that the isolate was race 5. To our knowledge, this is the first report of race 5 occurring outside of the California/Arizona spinach production area in the United States. There are commercial spinach lines with resistance to race 5, as well as the other described races (1). References: (1) B. M. Irish et al. Plant Dis. 87:567, 2003.

In situ hybridisation of chick embryos with p53-specific probe and their immunostaining with anti-p53 antibodies.

Tumor-suppressor protein p53 is an important regulator of cell cycle and apoptosis. On the level of embryo extracts it has been shown earlier that both p53 protein and mRNA are expressed in developing chicken. Here we describe the expression patterns of p53 mRNA and protein in developing chicken embryos (stages 2-12) using in situ hybridisation and immunostaining with p53-specific monoclonal antibody Mab421. p53 mRNA is equally localised all over the embryo in the stages observed. According to electron microscopy data a subfraction of p53 mRNA is bound to dissolving yolk granules expressing acid phosphatase activity characteristic for lysosomes. Protein p53 is synthesised starting from the medium primitive streak stage (stage 3) and reaches its maximum level at the full primitive streak stage. During these stages protein p53 is distributed evenly across the embryos. After gastrulation p53 protein remains visible at higher levels only in certain anlages and areas. In developing nervous system the expression is observable in neuroectoderm, during the closure of the neural tube and in mesenchyme in the area of migrating neural crest cells. In cardiogenesis protein p53 is expressed during formation of tubular heart in the epimyocardium, endocardium and cardiac jelly. p53 protein localises in the neurocoele (obviously connected with cellular debris) and cardiac jelly. Our data support the role of p53 in early development, especially during embryo gastrulation, the development of central nervous system, neural crest and heart. In some cases increased p53 amounts colocalise with the areas of intensive epithelium-mesenchyme transition.

Occurrence of Bacterial Leaf Spot of Escarole Caused by Pseudomonas cichorii in the Everglades Agricultural Area of Southern Florida.

In the fall of 1997, 1998, and 2000, a leaf spot disease of escarole (Cichorium endivia L.) was widespread among commercial plantings in the Everglades Agricultural Area (EAA), south and east of Lake Okeechobee, FL. Symptoms consisted of dry, dark gray-to-black lesions that expanded to ≈4 cm in diameter. Concentric rings were often observed in mature lesions. Growers and scouts in the area consistently identified this disease as Alternaria leaf spot, because the symptoms closely resembled Alternaria leaf spots seen on a number of other vegetables. Prolific bacterial streaming occurred when cut portions of lesions were observed microscopically. A fluorescent bacterium was consistently isolated when a sterile inoculation needle was pushed through lesions. Eight bacterial strains were isolated, restreaked to obtain pure cultures, and characterized. All strains were aerobic, gram-negative rods that were oxidase positive and arginine dihydrolase negative. Negative reactions were recorded for levan formation and rotting of potato slices. All strains utilized glucose, mannitol, and m-tartrate and were negative for sucrose, sorbitol, benzoate, d-arabinose, l-rhamnose, and cellobiose. Results for utilization of D-aspartate were variable. Based on these results, the causal agent of bacterial leaf spot of escarole was identified as Pseudomonas cichorii. Greenhouse-grown plants of escarole, cv. Full Heart, and Cos lettuce, cv. Tall Guzmaine, were mistinoculated with a suspension (107 CFU/ml) of each test strain from escarole and P. cichorii strain Pc28, originally isolated from celery (1). Plants were bagged for 3 days after inoculation. Symptoms characteristic of this disease were evident on escarole inoculated with all test strains and Pc28 6 days after inoculation. Pure cultures of P. cichorii were recovered from lesions on King's B medium. Three test strains produced mild leaf spot symptoms in Cos lettuce, but the symptoms were distinctly different from those associated with the common bacterial leaf spot of lettuce in Florida caused by Xanthomonas campestris pv. vitians (2). To our knowledge, this is the first report of P. cichorii causing this unusual target spot symptom on escarole in the EAA. References: (1) K. Pernezny et al. Plant Dis. 78:917, 1994. (2) K. Pernezny et al. Plant Dis. 79:359, 1995.

Natural Infection of Vicia faba by Bidens mottle virus in Florida.

In a study to evaluate the potential of Vicia faba (faba bean) as a cover and forage crop for Florida, 60 accessions of faba bean with diverse genetic backgrounds and geographic origins were acquired from the USDA Germplasm Repository in Pullman, WA. The beans were grown south of Lake Okeechobee in Belle Glade, FL, from December 2000 to April 2001. Reddish-brown elliptical lesions first appeared on the leaflets of two of the faba bean plants 10 to 12 weeks after planting. Within 2 weeks of initial symptoms, a mosaic pattern was expressed on the newly emergent leaves of the same plants. After disease expression, new pods aborted, while developing pods became stunted, distorted, and blistered. Potyvirus cylindrical inclusions (CI) were found in leaf strips (1) of the original plants. Viral symptoms were expressed in manually inoculated plants of Chenopodium amaraticolor, C. quinoa, Lactuca sativa, Nicotiana benthamiana, Petunia × hybrida, Verbena × hybrida, Vicia faba, and Zinnia elegans. Inoculated species of Phaseolus and Pisum were not infected. The virus causing the disease was identified as Bidens mottle virus (2) based on host range, characteristic CI in Z. elegans, and homologous lines of precipitation in SDS-immunodiffusion using antiserum to Bidens mottle and a known antigen. Both the primary host of this virus Bidens mottle virus and its aphid vectors are ubiquitous throughout Florida. To our knowledge, this is the first report of Bidens mottle virus infecting a member of the Leguminosae. References: (1) R. G. Christie and J. R. Edwardson. Light and Electron Microscopy of Plant Virus. Monogr. 9, IFAS, University of Florida, 1994. (2) D. E. Purcifull et al. Bidens mottle virus. Descriptions of Plant Viruses. No. 161. CMI/AAB, Surrey, England, 1976.

Dorsoventral compartmentalization of mesoderm in heart-forming area of chick embryo.

In early chick development (stages 5-8) the seemingly homogeneous mesoderm in the heart-forming area splits to somatic and splanchnic cardiogenic layers. Little is known about dorsoventral compartmentalization before splitting. Electron microscopic analysis shows the early dorsoventral polarization of precardiomyocytes. The dorsal compartment has epithelial and the ventral compartment mesenchymal features with numerous protrusions. At stage 5+-6 staining for wheat germ agglutinine (WGA) transiently demarcates the ventral part of mesoderm. The glycosomes (beta-glycogen) show a dorsoventral gradient in the mesoderm of the cardiogenic field during the initial step of the compaction. The differential expression of glycosomes depends on the activity of glycogen synthase kinase 3-beta, a component of the wnt-signaling pathway, and might in this spatiotemporal developmental window be involved in the commitment of presumptive cardiogenic and somatic cells. To verify this hypothesis simulation experiments with LiCl in vitro were carried out. The normal splitting of the mesoderm and the development of heart primordia were disturbed. Blocking the receptors of WGA by WGA in vitro at stage 5-5+ perturbs the migration of mesoderm to anterio-medial direction. It appears that early specification of dorsal and ventral compartments of the mesoderm in the heart-forming area correlates with the gradient of glycosomes. Our results suggest that the target of LiCl action (glycogen synthase kinase 3-beta) might be involved in the specification of heart primordia and that WGA receptors mediate the migration of mesoderm to the anteriomedial direction.

Disease Severity and Yield of Sweet Corn Hybrids with Resistance to Northern Leaf Blight.

Reactions of supersweet (sh2) sweet corn to northern leaf blight (NLB) and associated yields were evaluated in Belle Glade, Florida and Urbana, Illinois in yield-loss trials, hybrid evaluations, and evaluations of breeding materials. Hybrids differed significantly for NLB in all trials. Severity of NLB ranged from 0 to 66% on 35 sh2 hybrids in yield-loss trials, and from 0 to 60% on 80 sh2 hybrids in hybrid evaluations. NLB ratings ranged from 1 to 9 (approximately 0 to 80% severity) on 375 hybrids and 186 inbred lines in evaluations of breeding materials. Various methods of rating NLB and ratings from multiple dates were highly correlated, with correlation coefficients ranging from 0.76 to 0.98. Yield, measured as weight of ears and number of marketable ears from inoculated plots as a percentage of that from control plots, decreased as disease severity increased. Linear or quadratic regression models explained 31 to 70% of the variation in percent yield as a function of disease severity at harvest. The effects of NLB on yield were limited by NLB-resistance in several hybrids, including CCO 3268, Chieftain, Crisp N Sweet 710A, Day Star, Envy, Forever, GSS 1526, Jupiter, Midship, Prime Plus, Sch 5005, and SummerSweet 7630. Although high levels of partial resistance to NLB were prevalent among 375 new experimental sh2 hybrids and 186 sh2 inbred lines evaluated in 1995, use of the gene HtN may increase in the near future as breeders are incorporating this resistance into new inbreds and hybrids. Breeders and plant pathologists would be wise to continue improving partial resistance to NLB without using the gene HtN in genotypes with adequate levels of partial resistance, because the widespread use of the gene HtN will select for virulent races of Exserohilum turcicum which occur in Florida, or for races with new combinations of virulence.

Bacterial Leaf Spot of Cilantro in Florida.

In the 1995 to 1996 winter vegetable season, a leaf spot disease of cilantro (fresh coriander) (Coriandrum sativum L.) was widespread throughout commercial plantings in the Everglades Agricultural Area south and east of Lake Okeechobee. Symptoms first appeared as water-soaked spots ≤1 mm in diameter. These spots became dark brown to black and enlarged up to 2 mm in diameter. No chlorotic haloes developed around the lesions. The disorder was observed in numerous plantings of the cultivar Longstanding at four separate locations. Symptoms were apparent throughout each planting, with disease severity ranging from a few individual spots to numerous lesions covering nearly entire leaflets. Severe outbreaks were correlated with heavy precipitation events. In several instances, disease levels were great enough to render entire plantings totally unmarketable. A non-fluorescent bacterium was consistently isolated on King's medium B when a cooled inoculation needle was pushed through lesions. Six representative strains were chosen for further characterization. All strains were aerobic, gram-negative rods, and were oxidase and arginine dihydrolase negative. Levan was produced, but potato slices were not rotted. Tests for utilization of l-tartrate, l-lactate, and erythritol were negative. Biolog analysis identified all strains of the bacterium as Pseudomonas syringae. Highest similarity indices (0.52 to 0.81) were with P. syringae pv. pisi for four of the six strains. Suspensions of each strain were swab inoculated onto leaves of 4-week-old Longstanding cilantro seedlings in the greenhouse. Control plants were swabbed with sterile water only. Plants were covered with clear polyethylene bags for 72 h. Watersoaked spots were evident on test plants when bags were removed. Typical brown, greasy-looking leaf spots were seen by 6 days after inoculation. Control plants were symp-tomless. In a host-range study, cilantro and the following plants were mist-inoculated with a 107 CFU/ml suspension of each of the six test strains: carrot (Daucus carota L. 'Fancy Pack'), celery (Apium graveolens L. var. dulce (Mill.) Pers., 'June Belle'), garden pea (Pisum sativum L. 'Melting Sugar'), snap bean (Phaseolus vulgaris L. 'Pod Squad'), and onion (Allium cepa L. 'Evergreen Bunching'). Six days after inoculation, characteristic symptoms were evident on the cilantro. Four of the six strains produced a few (less than 10 per plant) bacterial leaf spot symptoms in carrot from which P. syringae was readily recovered. Some necrosis was observed on young, emerging leaves of snap bean. No symptoms were recorded for the other host species or the controls. This disease is similar to one reported on cilantro in California (1) and Germany (2). References: (1) D. A. Cooksey et al. Plant Dis. 75:101, 1991. (2) H. M. Toben and K. Rudolph. J. Phytopathol. 144:169, 1996.

[Changes in the distribution of adenylate cyclase activity in PC 12 cells under the influence of nerve growth factor]. / Izmenenie v raspredelenii adenilattsiklaznoi aktivnosti v kletkakh PC 12 pod vliianiem faktora rosta nervov.

Changes in distribution of adenylate cyclase in PC 12 cells under the influence of nerve growth factor (NGF) have been studied using cytochemical methods. The adenylate cyclase activity was predominantly associated with the plasma membrane. In cell cultures without NGF the activity was revealed on the contacting surfaces of cell aggregates; single grains of reaction product were revealed on exposed cell surface only in cultures with a high cell density. One day after administration of NGF, the adenylate cyclase activity on exposed cell surface increased, and three days later the whole cell surface was covered with lead sediment. The enzyme activity was also revealed in growth cones, filopodia and microcytospheres. The role of adenydlate cyclase system in neuron-like differentiation of PC 12 cells is discussed.