Quantification of late blight resistance of potato using transgenic Phytophthora infestans expressing β-glucuronidase

A transgenic Phytophthora infestans strain that constitutively produces and secretes -glucuronidase (GUS) was used in detached leaf assays to quantify the levels of resistance to late blight in potato cultivars Surprise, Irene, Pimpernel, Alpha and Bintje. Four days after inoculation levels of GUS activity were determined in infected leaves. Significant differences between the various cultivars were observed. Discrimination between resistant and susceptible cultivars was possible based solely on levels of GUS activity. Regression analysis revealed a positive correlation between in planta GUS levels and field resistance expressed as Area under Disease Progress Curve (ADPC).     

Genetic dissection of bacterial speck disease resistance in tomato

The bacterial speck disease of tomato has been developed as a model system to elucidate the molecular basis of specificity in plant-bacterial interactions and to study signal transduction events involved in the expression of plant disease resistance. We have employed a mutagenic approach to define the steps involved in the expression of disease resistance to the bacterial pathogen, Pseudomonas syringae pv. tomato (Pst) Eleven disease susceptible mutants have been identified and characterized twith altered recognition of Pst strains that express the avirulence gene avrPto. Genetic analysis of these mutants has revealed that they fall into two complementation groups. Five of the mutants map at the Pto locus, while six map a new locus that we have termed Prf. Further characterization of these mutants has revealed that the mutants that map at Pto are still sensitive to the insecticide fenthion, while the prf mutants are altered in resistance and also are insensitive to fenthion. Genetic mapping has also determined that the Prf locus maps near Pto. We are currently employing a map-based cloning strategy to isolate the Prf locus.     

Phytophthora genome sequences uncover evolutionary origins and mechanisms of pathogenesis

Draft genome sequences have been determined for the soybean pathogen Phytophthora sojae and the sudden oak death pathogen Phytophthora ramorum. O?mycetes such as these Phytophthora species share the kingdom Stramenopila with photosynthetic algae such as diatoms, and the presence of many Phytophthora genes of probable phototroph origin supports a photosynthetic ancestry for the stramenopiles. Comparison of the two species' genomes reveals a rapid expansion and diversification of many protein families associated with plant infection such as hydrolases, ABC transporters, protein toxins, proteinase inhibitors, and, in particular, a superfamily of 700 proteins with similarity to known o?mycete avirulence genes.     

Photoperiod effects on2n pollen production,response to anther culture,and net photosynthesis of a diplandrous clone ofsolanum phureja

Flow-cytometric analysis of pollen samples obtained fromSolanum phureja, clone PP5, grown under 10, 14, and 18 hour photoperiods at the Southeastern Plant Environment Laboratories of North Carolina State University yielded two populations of pollen based on size parameters. These populations corresponded to pollen separation based on propidium iodide staining of DNA. Anther culture response from plants grown under 14 and 18 hour photoperiods for 8, 10, 12, and 14 weeks was greatest from 8 week old plants grown under a 14 hour photoperiod. Net photosynthesis was significantly higher for plants grown under a 10 hour photoperiod than for plants grown under either a 14, or 18 hour photoperiod. A significant positive correlation was found between net photosynthesis and tuber yield. Results suggest that vegetative growth is increased under short photoperiods, floral development is favored under long photoperiods, and androgenesis is greatest from young plants grown under an intermediate photoperiod.     

Thrombocytopenia and platelet activity in dogs experimentally infected with Rangelia vitalii

The aim of this study was to evaluate the platelet count, coagulation time and platelet activity in dogs experimentally infected with Rangelia vitalii during the acute phase of the disease. For this study, 12 young dogs (females) were used, separated in two groups. Group A (uninfected control) was composed by healthy dogs (n=5), and group B consisted of R. vitalii-infected animals (n=7). After being inoculated with R. vitalii-infected blood, animals were monitored by blood smear examinations, which showed intra-erythrocytic forms of the parasite five days post-inoculation (PI). Blood samples were collected on days 0, 10, 20 and 30 PI. The material collected was placed in tubes containing EDTA for quantification of platelets, citrate anticoagulant platelet aggregation, and measuring the clotting time. Right after blood collection on days 10 and 20 PI, dogs were anesthetized for collecting bone marrow samples. A significant reduction (P<0.01) of the number of platelets was observed in R. vitalii-infected blood, when compared with uninfected dogs on days 10 and 20 PI. Additionally, macro-platelets were observed only in infected dogs. Prothrombin time and activated partial thromboplastin time did not differ between infected and uninfected dogs. The megakaryocyte count increased (P<0.01) significantly in infected dogs when compared with uninfected ones on days 10 and 20 PI. Platelet aggregation decreased (P<0.01) significantly in infected dogs in comparison to the control on days 10 and 20 PI. Therefore, rangeliosis in dogs causes a severe thrombocytopenia during the acute phase of infection. This platelets reduction probably occurred due to splenic sequestration and/or immune-mediated thrombocytopenia.     

Potato Steroidal Glycoalkaloids: Biosynthesis and Genetic Manipulation

The potato steroidal glycoalkaloids (SGAs) are important components of plant resistance against pests and pathogens but can be toxic to humans at high levels. SGAs derive their toxicity from anticholinesterase activity affecting the central nervous system and the disruptive effects on cell membrane integrity affecting the digestive system and other organs. Accordingly, current safety regulations limit their content in the edible tuber to 20 mg per 100 g fresh weight. SGA composition and level are genetically determined, with unfavourable growth conditions and inappropriate postharvest management inducing the accumulation of SGAs at levels in the tubers of “safe” cultivars beyond the maximum level set by the industry. Hence, genetic alteration of potato to prevent toxic levels of SGAs in tubers is highly desirable. At the same time, maintaining high SGA levels in other plant organs will contribute to plant resistance against pathogen and pest attacks. To this end, SGA biosynthesis and degradation should be manipulated precisely to exploit tissue-specific expression rather than whole-plant suppression of SGA production, to produce potato cultivars with SGA content enriched in the foliage but diminished in the edible tubers. Only a few details are known about the SGA biosynthetic pathway, its genes and intermediates. Research on factors that regulate SGA biosynthesis and catabolism as well as searches for genetic markers linked to total and specific SGA levels have only recently been pursued. The present review summarizes current data on these issues to encourage further discussion on SGA manipulation for safer food products.     

Feeding ecology and population delineation of south‐eastern Atlantic and south‐western Indian Ocean humpback whales using stable isotope analysis

1. Population delineation is vital for effectively managing and protecting populations of all at‐risk species. Population boundaries of Southern Hemisphere humpback whales on their breeding and feeding grounds have not been fully resolved. A number of methods have been used to delineate breeding stocks of Southern Hemisphere humpbacks, but ecological characteristics determined via stable isotope analysis provide valuable information to contrast other data sources.
2. In this study, stable isotope analysis is used to investigate potential separation of humpback whale populations on Southern Hemisphere feeding grounds as evidenced by carbon and nitrogen isotope values in their skin as proxies of diet.
3. One hundred samples of whale skin obtained from biopsies in sampling localities off the coasts of Gabon, Mayotte (Mozambique Channel), and Madagascar were analysed for carbon (δ¹³C) and nitrogen (δ¹⁵N) stable isotope ratios. The results showed a statistically significant difference in the mean δ¹⁵N values for whales between the populations from Gabon and Madagascar (7.0 ± 0.1‰ and 7.6 ± 0.1‰), and Gabon and Mayotte (7.6 ± 0.1‰ and 7.2 ± 0.1‰), indicating that these breeding stocks are potentially visiting different areas of the feeding grounds outside of the breeding season.
4. The results from this study indicate that at least some breeding stocks may show fidelity to separate feeding areas and do not widely mix with individuals from other breeding stocks while feeding.