RESUMEN
In August of 2011, the North Dakota State University Plant Diagnostic Lab received a hybrid corn (Zea mays) leaf sample from Burleigh County in south-central North Dakota (ND). The leaf had long, irregular, water-soaked lesions consistent with Goss's leaf blight of corn. Using a light microscope at 10× magnification, bacterial streaming was observed from the excised edge of leaf tissue. A bacterial suspension was created, streaked onto a semi-selective CNS medium (1), and incubated at 22°C. Dark yellow-orange colonies appeared on the medium after 5 days. Single colonies were subcultured onto additional CNS media. To verify the identity of the bacterial isolate, PCR amplification of the 16S ribosomal DNA from this isolate along with a known Clavibacter michiganensis spp. nebraskensis (Cmn) isolate collected in Indiana (4) was performed using the eubacterial universal primers 27f and 1525r (3). The 1,431-bp 16S rDNA region was obtained for each isolate and they were compared with each other and with those deposited in NCBI GenBank. Sequence alignment identified only one nucleotide difference between the ND isolate and the Indiana isolate. BLASTn search against the NCBI database showed the first 100 hits were described as C. michiganensis or unidentified Clavibacter sp. The ND isolate had a two-nucleotide difference with Cmn isolate NCPPB2581 (HE614873), and a three nucleotide difference was found with the C. michiganensis spp. michiganensis isolate NCPPB 382 (AM711867). To satisfy Koch's postulates, eight corn plants (Golden Cross Bantam) were grown in the greenhouse at 22 to 24°C. Four corn plants were inoculated at growth stage V4-V5 using a suspension of approximately 1 × 109 CFU/ml from cultures grown on CNS for 6 days. Wounds were created on the fifth leaf approximately 7 cm from the leaf tip using a tongue-seizing forceps outfitted with a rubber stopper composed of pins (2). Simultaneously, 1 ml of the bacterial suspension was delivered into the wounds through a hole on top of the rubber stopper. Four control plants were inoculated with sterile water in a similar fashion. No symptoms were observed on the control plants. After 6 days, long water-soaked symptoms were observed on leaves inoculated with the bacterial suspension. Using leaves with water-soaked lesions, the pathogen was re-isolated onto CNS media and subjected to PCR amplification, and the resulting amplicons were sequenced as before. The sequence of the amplicon from the re-isolation matched that of the original ND isolate. To our knowledge, this is the first account of Goss's leaf blight and wilt identified in ND. As the corn acreage and no-till production systems in the state have increased, the economic implications of this disease may become more significant. Recognition of symptoms and proper identification of this bacterial disease in the field should help reduce unnecessary foliar fungicide sprays. References: (1) D. C. Gross and A. K. Vidaver. Phytopathology 69:82, 1979. (2) W. A. Hagborg. Can. J. Bot. 48:1135, 1970. (3) X. Li and S. H. DeBoer. Can. J. Microbiol. 41:925, 1995. (4) G. Ruhl et al. Plant Dis. 93:841, 2009.
RESUMEN
Dollar spot disease is a major concern for golf courses nationwide, resulting in poor turf quality and significant damage to playing surfaces. To manage this disease effectively, fungicides need to be applied regularly. This management strategy represents a significant cost to turfgrass managers and may impact the economics of the industry in North Dakota. In the summer of 2011, small, circular, sunken brown patches of dead turf approximately 5 cm in diameter and resembling dollar spot disease were observed on a creeping bentgrass (Agrostis stolonifera L.) variety trial at the North Dakota State University Agricultural Experiment Station in Fargo, ND. Fresh individual leaf specimens with distinct lesions having straw colored centers and reddish brown margins were collected. Leaves were surface disinfected in a 0.05% sodium chloride solution for 60 s, rinsed three times in sterile distilled water, then placed onto potato dextrose agar (PDA). Three isolates were obtained from the disease infested leaves with similar morphology to that described for Sclerotinia homoeocarpa F.T. Bennett (1). Fungal colonies were initially colorless followed by the development of sparse white columnar aerial mycelia with cinnamon colored bases. Hyphae were 5 to 8 µm wide and thin walled with dense granular contents and septations at irregular intervals. Fourteen days after culturing, dark brown to black mycelial stroma developed. A single representative isolate was selected to conduct inoculations. Inoculum was produced by placing six 5-mm-diameter plugs of PDA with actively growing mycelium into an Erlenmeyer flask with 40 g of sterile millet seed. The inoculum was incubated for 14 days at ambient temperature (20 to 25°C). Three creeping bentgrass cultivars, Crenshaw, Declaration, and L-93, were inoculated (two pots per cultivar). Following inoculation, the pots were misted with water, sealed in separate plastic bags, and placed in the dark for 48 h. For the next 5 days, plants were placed for 8 h outside of bags on a bench with full spectrum fluorescent bulbs, followed by 16 h in plastic bags in the dark. Finally, pots were placed on a bench for 48 h. Signs and symptoms of S. homoeocarpa developed on all pots, whereas the controls remained asymptomatic. The same fungus was reisolated from grass leaves, satisfying Koch's postulates. To confirm the identity of the fungus, the internal transcribed spacer was amplified using the ITS4 and ITS5 primers (2). The amplicon was sequenced, generating a 549-bp sequence (Accession No. JQ735942) with 100% similarity to sequences of S. homoeocarpa in GenBank (Accession Nos. GQ924924.1, GQ924923.1, and EU123801.1). To our knowledge, this is the first confirmed report of dollar spot in North Dakota. References: (1) F. T. Bennett. Ann. Appl. Biol. 24:236, 1937. (2) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. Academic Press Inc., New York, 1990.
RESUMEN
Acreage of dry field pea (Pisum sativum) in North Dakota has increased approximately eightfold from the late 1990s to the late 2000s to over 200,000 ha annually. A coincidental increase in losses to root rots has also been observed. Root rot in dry field pea is commonly caused by a complex of pathogens which included Fusarium spp. and Rhizoctonia solani. R. solani isolates were obtained from roots sampled at the three- to five-node growth stage from North Dakota pea fields and from symptomatic samples received at the Plant Diagnostic Lab at North Dakota State University in 2008 and 2009. Using Bayesian inference and maximum likelihood analysis of the internal transcribed spacer (ITS) region of the ribosomal DNA (rDNA), 17 R. solani pea isolates were determined to belong to anastomosis group (AG)-4 homogenous group (HG)-II and two isolates to AG-5. Pathogenicity of select pea isolates was determined on field pea and two rotation hosts, soybean and dry bean. All isolates caused disease on all hosts; however, the median disease ratings were higher on green pea, dry bean, and soybean cultivars when inoculated with pea isolate AG-4 HG-II. Identification of R. solani AGs and subgroups on field pea and determination of relative pathogenicity on rotational hosts is important for effective resistance breeding and appropriate rotation strategies.
RESUMEN
Kidney transplantation in morbidly obese patients can be technically demanding. Furthermore, morbidly obese patients experience a high rate of wound infections and related complications, which mostly result from the longer length and extent of the incision. These complications can be avoided through minimally invasive surgery; however, conventional laparoscopic instruments are unsuitable for the safe performance of a kidney transplant in morbidly obese patients. Herein, we report the first minimally invasive, total robotic kidney transplant in a morbidly obese patient. A left, deceased donor kidney was transplanted into a 29-year-old woman with a body mass index (BMI) of 41 kg/m(2) who had been on hemodialysis for 5 years. The operation was performed intraabdominally using the DaVinci Robotic Surgical System with 4 trocars and a 7 cm midline incision. The operative time was 223 min, and the blood loss was less than 50 cc. The kidney had immediate graft function. No perioperative complications were observed, and the patient was discharged on postoperative day 5 with normal kidney function. Minimally invasive access and robotic technology facilitated the safe performance of a successful kidney transplant in a morbidly obese patient.