Identification of resistant germplasm containing novel resistance genes at or tightly linked to the <Emphasis Type="Italic">Pi2/9</Emphasis> locus conferring broad-spectrum resistance against rice blast

Background

The rice Pi2/9 locus harbors multiple resistance (R) genes each controlling broad-spectrum resistance against diverse isolates of Magnaporthe oryzae, a fungal pathogen causing devastating blast disease to rice. Identification of more resistance germplasm containing novel R genes at or tightly linked to the Pi2/9 locus would promote breeding of resistance rice cultivars.

Results

In this study, we aim to identify resistant germplasm containing novel R genes at or tightly linked to the Pi2/9 locus using a molecular marker, designated as Pi2/9-RH (Pi2/9 resistant haplotype), developed from the 5′ portion of the Pi2 sequence which was conserved only in the rice lines containing functional Pi2/9 alleles. DNA analysis using Pi2/9-RH identified 24 positive lines in 55 shortlisted landraces which showed resistance to 4 rice blast isolates. Analysis of partial sequences of the full-length cDNAs of Pi2/9 homologues resulted in the clustering of these 24 lines into 5 haplotypes each containing different Pi2/9 homologues which were designated as Pi2/9-A5, ?A15, ?A42, ?A53, and -A54. Interestingly, Pi2/9-A5 and Pi2/9-A54 are identical to Piz-t and Pi2, respectively. To validate the association of other three novel Pi2/9 homologues with the blast resistance, monogenic lines at BC₃F₃ generation were generated by marker assisted backcrossing (MABC). Resistance assessment of the derived monogenic lines in both the greenhouse and the field hotspot indicated that they all controlled broad-spectrum resistance against rice blast. Moreover, genetic analysis revealed that the blast resistance of these three monogenic lines was co-segregated with Pi2/9-RH, suggesting that the Pi2/9 locus or tightly linked loci could be responsible for the resistance.

Conclusion

The newly developed marker Pi2/9-RH could be used as a potentially diagnostic marker for the quick identification of resistant donors containing functional Pi2/9 alleles or unknown linked R genes. The three new monogenic lines containing the Pi2/9 introgression segment could be used as valuable materials for disease assessment and resistance donors in breeding program.

     

Accumulation of sorghum phytoalexins induced by Colletotrichum graminicola at the infection site

Microspectrophotometry was performed on intact, pigmented vesicle-like inclusions within living sorghum cells that were accumulating phytoalexins as a response to attempted fungal infection. The results indicate that the deoxyanthocyanidin phytoalexins are present in inclusions. Moreover, the phytoalexin concentration within a single inclusion, based on luteolinidin, was calculated to be 0·15 m. The amounts of luteolinidin and apigeninidin in cells involved in the phytoalexin response at individual infection sites were also determined. The data showed that luteolinidin accumulated to levels of 0·48–1·20 ng/cell whereas apigeninidin accumulated to levels of 0·24–0·91 ng/cell. The results of both analyses confirmed that at the infection site the deoxyanthocyanidins accumulate to levels in substantial excess of those required for inhibition of the fungus Colletotrichum graminicola.     

Some fluorine-containing pheromone analogues

Two analogues of the aphid alarm pheromone (E)-β-farnesene, a trifluorofarnesene and a difluoro-1-norfarnesene, were found to be highly active and were more readily detectable than the parent compound. For (Z)-hexadec-11-enal, a component of some lepidopteran sex attractant pheromones, replacement of the carbonyl oxygen with a difluoromethyl group to give a difluoroheptadecadiene resulted in loss of activity. A trifluoroacetoxyhexadecanolide was a more volatile analogue of the mosquito oviposition pheromone (?)-(5R,6S)-6-acetoxy-5-hexadecanolide and was highly active.     

XY male pseudohermaphroditism in a captive Arabian oryx (Oryx leucoryx).

A 2-yr-old Arabian oryx (Oryx leucoryx) was presented for evaluation of abnormal genitalia and infantile behavior. The oryx had a penis and a scrotum, but testes were not palpable within the scrotum or inguinal canal. The total serum testosterone for the individual was lower than in age-matched males of the same species. Surgical exploration showed markedly hypoplastic intra-abdominal gonads, which demonstrated both testicular and uterine tissue on histologic examination. After karyotype analysis, the individual was classified as an XY male pseudohermaphrodite. This condition resembles two human intersex syndromes: embryonic testicular regression syndrome and partial gonadal dysgenesis syndrome, which occur in familial lines.     

In Planta - Complementation of Clavibacter Michiganensis Subsp. sepedonicus Strains Deficient in Cellulase Production or HR Induction Restores Virulence

Clavibacter michiganensis subsp. sepedonicus, a Gram positive bacterium that causes bacterial ring rot of potato, was studied in eggplant, an alternate host, using strains that differed in phenotype. Two factors affecting virulence, the ability to induce a hypersensitive response (HR) and cellulase production, were studied. A plasmid-free isolate of C. michiganensis subsp. sepedonicus that causes HR on tobacco but is unable to produce cellulase multiplied efficiently in planta, but caused only weak symptoms. In contrast, a strain that is unable to induce HR on tobacco but produces cellulase was impaired in the ability to multiply in the host and caused no symptoms. When the two non-virulent strains were coinoculated into eggplants, typical disease symptoms developed. This enhancement was not due to formation of a new phenotype or significant increases in population density of either of the strains. Our results suggest that both cellulase production and the ability to induce HR are required for a successful infection process and disease induction by C. michiganensis subsp. sepedonicus. Our results additionally suggest that the ability to induce HR on non-host plants is required for multiplication in the host plant, whereas cellulase expression is necessary for induction of disease symptoms.     

Analysis of feline,canine and equine hemograms using the QBC VetAutoread

Blood samples form 120 consecutive clinical cases (40 cats, 40 dogs and 40 horses) were analyzed on the QBC VetAutoread analyzer and the results compared with those obtained by a Baker 9000 electronic resistance cell counter and a 100-cell manual differential leukocyte (WBC) count. Packed cell volume (PCV), hemoglobin (Hb) concentration, mean cell hemoglobin concentration (MCHC), and platelet, total WBC, granulocytes, and lymphocyte plus monocyte (L+M) counts were determined. Indistinct separation of red blood cell and granulocytes layers on the QBC VetAutoread was observed in samples from five cats (12.5%), two dogs (5%), and one horse. Significantly different (P=0.002) median values for the two methods were obtained for PCV, Hb concentration, MCHC and platelet count in cats; PCV, MCHC, WBC, count and granulocytes count in dogs; and PCV, Hb concentration, MCHC and WBC, granulocytes and platelet counts in horses. Results from the QBC VetAutoread should not be interpreted using reference ranges established using other equipment. Results were abnormal on a limited number of samples; however, when correlation coefficients were low, marked discrepancy existed between values within as well as outside of reference ranges. Spearman rank correlation coefficients were excellent (r=0.93) for PCV and Hb concentration in dogs, and Hb concentration and WBC count in horses. Correlation was good (r=0.80-0.92) for PCV and Hb concentration in cats, WBC count in dogs, and PCV, granulocytes count and platelet count in horses. For remaining parameters, correlation was fair to poor (r=0.79). Acceptable correlations (r>0.80) were achieved between the two test systems for all equine values except MCHC and L+M count, but only for PCV and HB concentration in feline and canine blood samples.     

Plasma Adrenocorticotropin Concentration in Healthy Horses and in Horses With Clinical Signs of Hyperadrenocorticism

Pituitary adenomas are commonly reported in older horses. The typical clinical signs associated with this condition, also known as equine Cushing's disease (ECD), are related to increased adrenocorticotropin (ACTH) production resulting in hyperadrenocorticism. The primary purpose of this study was to determine whether plasma ACTH concentrations differed between cushingoid and healthy horses. The second objective was to determine the effects of blood sample handling techniques on ACTH concentrations. A commercial human ACTH radioimmunoassay (RIA) was used to quantify equine plasma ACTH. Intra-assay and interassay variations, as well as dilutional parallelism were determined during the RIA validation. Plasma ACTH concentrations were evaluated in a group of healthy equids composed of 18 horses and 9 ponies, and in 22 equids with a clinical diagnosis of hyper adrenocorticism (11 horses and 11 ponies). The mean plasma ACTH concentrations in healthy horses and ponies, (18.68 ± 6.79 pg/mL (mean ± SD) and 8.35 ± 2.92 pg/mL, respectively), were significantly different (P = .009). The mean plasma ACTH concentration in horses and ponies with ECD, (199.18 ± 182.82 pglmL and 206.21 ± 319.56 pg/mL, respectively), were significantly higher than the mean ACTH concentration in the control animals (P < .001). Plasma ACTH concentrations appeared to be a sensitive and specific indicator of ECD in horses and ponies. ACTH concentrations measured in plasma samples kept at room temperature (19°C) as long as 3 hours after blood collection were not statistically different from those of samples kept at 1°C. J Vet Intern Med 1996;10:1–6. Copyright 1996 by the American College of Veterinary Internal Medicine .     

Monoclonal Gammopathy in a Dog With Chronic Pyoderma

A monoclonal gammopathy composed of immunoglobulin G, with concurrent light-chain proteinuria and generalized lymph node plasmacytosis, was associated with chronic pyoderma in a dog. A uniform population of plasma cells was observed cytologically and histologically in multiple lymph node specimens. A diagnosis of monoclonal gammopathy of unknown significance was eventually made by exclusion of other known causes of monoclonal gammopathy, resolution after antibiotic therapy, and no evidence of lymphoproliferative disease after 11 months of follow-up and subsequent necropsy. This report expands the diagnostic considerations for monoclonal gammopathies in the dog.     

Heinz Body Formation Associated With Ketoacidosis in Diabetic Cats

Oxidative damage plays an important role in the pathophysiology of diabetes and diabetic complications. Feline hemoglobin is uniquely susceptible to oxidative denaturation; therefore, Heinz body formation is a highly sensitive indicator of in vivo oxidative stress in this species. Heinz bodies also contribute to anemia. We investigated hematological and clinical biochemical changes in 30 cats with spontaneous diabetes mellitus (as compared to 15 healthy control cats) and evaluated the relationship of these changes to erythrocyte oxidative damage. Cats were categorized as ketoacidotic or nonketoacidotic based on their clinical presentation and the presence of urine ketones. Ketoacidotic cats had significantly ( P = .0009) more Heinz bodies (28.3% ± 9.1 %) than nonketotic diabetic cats (6.5% ± 1.60%) and healthy control cats (0.6% ± 0.2%). Percent Heinz bodies in diabetic cats directly correlated with plasma β-hydroxy-butyrate concentration ( r = .622; P = .0002), as well as with serum chloride concentration ( r = -0.576; P = 0.0009) and the number of monocytes ( r = .536; P = .0023). Percent Heinz bodies were negatively correlated with erythrocyte glutathione concentrations. Erythrocyte membrane lipid peroxidation was slightly but not significantly increased in diabetic cats. There were no significant associations between percent Heinz bodies and degree of anemia, hyperglycemia, or glycohemoglobin. These data indicate that ketones are associated with oxidative hemoglobin damage in cats, and suggest that ketone metabolism, ie by cytochrome P450 2E1, may be a potential source of in vivo oxygen radical generation in animals with ketosis.