MGIS: managing banana (Musa spp.) genetic resources information and high-throughput genotyping data.

Database URL: https://www.crop-diversity.org/mgis/.

Characterization of a resistance locus (Pfs-1) to the spinach downy mildew pathogen (Peronospora farinosa f. sp. spinaciae) and development of a molecular marker linked to Pfs-1.

Downy mildew is a destructive disease of spinach worldwide. There have been 10 races described since 1824, six of which have been identified in the past 10 years. Race identification is based on qualitative disease reactions on a set of diverse host differentials which include open-pollinated cultivars, contemporary hybrid cultivars, and older hybrid cultivars that are no longer produced. The development of a set of near-isogenic open-pollinated spinach lines (NILs), having different resistance loci in a susceptible and otherwise common genetic background, would facilitate identification of races of the downy mildew pathogen, provide a tool to better understand the genetics of resistance, and expedite the development of molecular markers linked to these disease resistance loci. To achieve this objective, the spinach cv. Viroflay, susceptible to race 6 of Peronospora farinosa f. sp. spinaciae, was used as the recurrent susceptible parent in crosses with the hybrid spinach cv. Lion, resistant to race 6. Resistant F(1) progeny were subsequently backcrossed to Viroflay four times with selection for race 6 resistance each time. Analysis of the segregation data showed that resistance was controlled by a single dominant gene, and the resistance locus was designated Pfs-1. By bulk segregant analysis, an amplified fragment length polymorphism (AFLP) marker (E-ACT/M-CTG) linked to Pfs-1 was identified and used to develop a co-dominant Sequence characterized amplified region (SCAR) marker. This SCAR marker, designated Dm-1, was closely linked ( approximately 1.7 cM) to the Pfs-1 locus and could discriminate among spinach genotypes that were homozygous resistant (Pfs-1Pfs-1), heterozygous resistant (Pfs-1pfs-1), or homozygous susceptible (pfs-1pfs-1) to race 6 within the original mapping population. Evaluation of a wide range of commercial spinach lines outside of the mapping population indicated that Dm-1 could effectively identify Pfs-1 resistant genotypes; the Dm-1 marker correctly predicted the disease resistance phenotype in 120 out of 123 lines tested. In addition, the NIL containing the Pfs-1 locus (Pfs-1Pfs-1) was resistant to multiple races of the downy mildew pathogen indicating Pfs-1 locus may contain a cluster of resistance genes.

First Report of Phytophthora palmivora, Causal Agent of Black Pod, on Cacao in Puerto Rico.

Black pod or Phytophthora pod rot is the most economically important and widespread disease of cacao, Theobroma cacao L. Total losses due to Phytophthora exceed $400 million worldwide (1), and several species are known to attack cacao with P. palmivora (E.J. Butler) E. J. Butler as the most common. All plant parts are infected, but pod infections are particularly damaging. Symptoms resembling those of black pod disease were observed at the National Plant Germplasm Collection System of cacao at the USDA-ARS Tropical Agriculture Research Station (TARS) in Mayaguez, Puerto Rico for a number of years. During May of 2005, to determine the etiology of the disease, small, surface disinfested sections of pod lesions were placed on water agar and incubated for 4 days. The formation of papillate, deciduous, ellipsoidal to ovoid sporangia produced on sympodial sporangiophores on fruits, fit the description of P. palmivora and the identification was confirmed on cultures on water agar (2). Chlamydospores were readily observed in diseased pods and observed in pure cultures on V8 agar (2). Eight, single hyphal tips were transferred to potato dextrose agar (PDA) (Sigma-Aldrich, St. Louis, MO) and maintained as stock cultures. For pathogenicity tests, healthy mature pods were surface disinfested and placed in a humidity chamber lined with moist paper towels. Eight isolates were tested on four fruits per isolate and the pathogenicity test was repeated once. Inoculum was prepared by growing each isolate on PDA for 5 days with irradiation at 24°C, adding approximately 3.0 ml of water to each plate, dislodging the sporangia with a glass rod, mixing the suspension, estimating spore numbers with a hemacytometer, and adjusting to 104 sporangia per ml. A small, sterile scalpel was used to make an approximately 20.0 mm cut on the fruit epidermis, and approximately 0.2 ml of inoculum was placed on the wound. Pods were evaluated daily for 2 weeks. For molecular analysis, each of the eight cultures were grown in 50% potato dextrose broth to produce mycelia for DNA extraction using the FastDNA kit (Q-Biogen1, Irvine, CA). The internal transcribed spacer (ITS) region of the ribosomal RNA gene cluster was amplified, purified, and sequenced for all eight isolates. The ITS sequences of GenBank Accession Nos. DQ987915 to DQ987922 were identical and exhibited strong similarity (>99% identity) to that of three previously described isolates of P. palmivora from cacao (GenBank Accession Nos. AF 228097, AF467093, and AF467089). P. palmivora has been reported on citrus, coconut, black pepper, and Arracacia xanthorrhiza in Puerto Rico (2,3) and inoculum may have originated from these host or imported on cacao planted into the cacao collection before 2000. USDA-ARS-TARS is the official site for the cacao germplasm collection, thus, a detailed integrated pest management plan that includes the evaluation for resistance, sanitation measures, and use of fungicides to reduce disease levels has been implemented. Decreasing incidence and severity of this disease is a top priority. To our knowledge, this is the first report of P. palmivora on cacao in Puerto Rico. References: (1) M. C. T. Braga et al. Agrotropica 1:108, 1989. (2) D. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society. St. Paul, MN, 1996. (3) E. Rosa-Marquez. J. Agric. Univ. P. R. 84:53, 2000.

Three New Races of the Spinach Downy Mildew Pathogen Identified by a Modified Set of Spinach Differentials.

Spinach downy mildew, caused by Peronospora farinosa f. sp. spinaciae, is the most economically important disease of spinach worldwide. During the past few years, spinach cultivars resistant to the seven previously described races of P. farinosa f. sp. spinaciae were observed to be severely affected by downy mildew in both the United States and the European Union. Four new isolates of P. farinosa f. sp. spinaciae were collected from California and The Netherlands and characterized based on disease reactions on two modified sets of spinach differentials. The results led to the description of three new races of the downy mildew pathogen, designated races 8, 9, and 10. Four differential cultivars with resistance to races 1 to 7 were used to distinguish the three new races. Dolphin was susceptible to races 8 and 10 but resistant to race 9; Lion was susceptible to race 10 but resistant to races 8 and 9; Lazio was resistant to races 1 to 7 as well as races 8, 9, and 10; and Tarpy was susceptible to all three new races. The three new races also were used to evaluate the disease reactions on 43 contemporary commercial spinach cultivars in greenhouse trials. A survey of 58 isolates of P. farinosa f. sp. spinaciae collected in California and Arizona between 2004 and 2006 revealed that race 10 predominated in the areas sampled.

Mycosphaerella fijiensis, Causal Agent of Black Sigatoka of Musa spp. Found in Puerto Rico and Identified by Polymerase Chain Reaction.

Black Sigatoka, also known as black leaf streak, is caused by Mycosphaerella fijiensis Morelet (anamorph Pseudocercospora fijiensis (Morelet) Deighton). It is the most significant disease of bananas and plantains (Musa spp.) because most of the economically important cultivars of exported and staple commodities are highly susceptible. The Caribbean is one of the few regions of the world where black Sigatoka is not widespread. Black Sigatoka has been reported in the Bahamas, Cuba, Hispaniola, and Jamaica (2). Yellow Sigatoka, caused by M. musicola Leach (anamorph P. musae (Zimm.) Deighton), has been recognized in Puerto Rico since 1938-1939 (3). In August 2004, symptoms resembling black Sigatoka were first observed in Añasco, Puerto Rico by extension personnel from the University of Puerto Rico. Since black and yellow Sigatoka produce similar disease symptoms, a survey was conducted in the western banana- and plantain-production region of Puerto Rico to confirm the presence of black Sigatoka. Leaf samples were collected from production fields near the towns of Las Marias, Maricao, and Añasco. Single-ascospore isolates were recovered using the discharge technique from moistened pseudothecia in necrotic lesions that were inverted over water agar, and ascospores were transferred to potato dextrose agar. The isolates were subcultured in potato dextrose broth for mycelium production. DNA was isolated from mycelium with the FastDNA kit (Q-Biogen, Irvine, CA) for 19 isolates. Internal transcribed spacer as well as the 5.8s rDNA regions were polymerase chain reaction amplified with primers specific to M. fijiensis or M. musicola (1). Amplification products (˜1,100 bp) were observed for 18 of the 19 isolates, 6 of which were M. fijiensis and the remaining 12 were M. musicola, while the positive controls for both species were also amplified with the respective primer pairs. M. fijiensis was recovered from production fields close to all three towns. The source of M. fijiensis in Puerto Rico is unclear, but it may have originated from introduced leaf material and/or wind dispersed ascospores from neighboring countries. The presence of black Sigatoka in Puerto Rico will most likely increase production costs where fungicide applications will be needed to maintain yields. The USDA-ARS, Tropical Agriculture Research Station is the official Musa spp. germplasm repository for the National Plant Germplasm System. As such, efforts are underway to introduce and evaluate black Sigatoka disease-resistant clones that can satisfy local and export market criteria. References: (1) A. Johnasen. Detection of Sigatoka leaf spot pathogens of banana by the polymerase chain reaction. Chatman, UK, Natural Resource Institute, 1997. (2) R. C. Ploetz. Plant Dis. 88:772, 2004. (3) R. H. Stover. Trop. Agric. Trinidad. 39:327, 1962.

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.

Identification and Cultivar Reaction to Three New Races of the Spinach Downy Mildew Pathogen from the United States and Europe.

Since 1996, commercial spinach cultivars with resistance to four previously described races of Peronospora farinosa f. sp. spinaciae (races 1, 2, 3, and 4) were observed with high incidences of downy mildew both in California and Europe. Isolates of P. farinosa f. sp. spinaciae collected in California between 1997 and 2001, Arizona in 1999, and a single isolate collected in the Netherlands in 1996 were examined for their disease reaction on differential spinach cultivars and a set of commercial spinach cultivars. Disease reactions on the differential cultivars indicated the occurrence of three new races of P. farinosa f. sp. spinaciae. Two newly identified races, designated race 5 (isolate CA1) and race 6 (isolate SP1), were detected in the United States. The isolate from the Netherlands also was distinct and designated race 7 (isolate JVN7). Some cultivars with resistance to races 1, 2, 3, and 4 were susceptible to race 5, whereas others were resistant, indicating that resistance to a given race may be governed by different genes (or alleles) depending on the source of resistance. A survey of races in California indicated that races 5 and 6 predominated. Although the majority of the cultivars examined were susceptible to race 6 based on the traditional qualitative cotyledon inoculation assay, significant quantitative differences in resistance to race 6 were observed using a true-leaf greenhouse screening procedure. Although more work is needed to confirm the results of the true-leaf assays, the quantitative resistance observed using this procedure appears to be race specific.

The Effect of Synthetic Surfactants on Disease Severity of White Rust on Spinach.

White rust, caused by Albugo occidentalis, is an economically important disease of spinach (Spinacia oleracea). Although cultural practices and partial host resistance are used for disease management, control strategies often rely on fungicides. An alternative approach, the use of ionic (cationic or anionic) and nonionic surfactants, was evaluated for its effect on white rust in greenhouse and field experiments. Surfactant treatments were compared with a water control and two commercial fungicides, azoxystrobin (Quadris) and 1,2,3-benzothiadiazole-7-carbothioic acid S-methyl ester (Actigard). Greenhouse plants were treated with a single surfactant application followed by inoculation with sporangia. Disease severity was rated on leaves 8 to 12 days after inoculation. Field tests were conducted in Arkansas and Texas and received three to five surfactant applications during the season. Disease severity was determined at the end of the growing season. In greenhouse and field tests, all surfactant treatments showed significant reductions in white rust severity compared with water controls. The surfactants Naiad and sodium dodecyl sulfate (SDS) were highly effective and comparable to fungicides in reducing white rust severity. In a laboratory assay, microscopic examination revealed that most of the surfactants at low concentrations caused rapid (<2 min) zoospore lysis.

Treatment of patients with refractory rheumatoid arthritis with extracorporeal protein A immunoadsorption columns: a pilot trial.

OBJECTIVE: To evaluate the safety and effectiveness of extracorporeal immunoadsorption with protein A (PROSORBA) columns in the treatment of refractory rheumatoid arthritis (RA). METHODS: Eleven patients with refractory RA were enrolled in an open prospective trial of 24 weeks' duration. Nine patients received 15 treatments over a 12-week period, 1 patient received 15 treatments over a 15-week period, and 1 patient received 12 treatments over a 9-week period. RESULTS: Using the composite criteria of Paulus, et al, 9 patients met the > or = 50% criteria when tested at Week 13, while 4 and 2 patients met the > or = 50% and > or = 20% criteria, respectively, when tested at Week 24. In addition, most of the clinical variables in the 9 responders at Week 13 had significantly improved (p < 0.05); 8 of these responders were able to continue for the entire 24-week study without a change in their arthritic medications, and 2 met the American College of Rheumatology criteria for clinical remission at Weeks 12, and 28 and have remained in remission for 6 and 5 months, respectively. Treatment associated side effects were of short duration and resolved without sequela. Four patients became symptomatically anemic during the treatments. CONCLUSION: Our preliminary study suggests that extracorporeal immunoadsorption therapy with protein A columns was well tolerated and may be effective in the treatment of RA. Further expanded and controlled trials are indicated to explore this new therapeutic modality.