Increased tolerance to Phytophthora citrophthora in transgenic orange plants constitutively expressing a tomato pathogenesis related protein PR-5

Phytophthora citrophthora is the most widely spread oomycete plant pathogen over all the citrus growing areas and represents one of the major causes of crop losses. Constitutive over-expression of genes encoding proteins involved in plant defence mechanisms to disease is one of the strategies proposed to increase plant tolerance to oomycete and fungal pathogens. P23 (PR-5), a 23-kDa pathogenesis-related protein similar to osmotins, is induced in tomato (Lycopersicon esculentum Mill. cv. Rutgers) plants when they are infected with citrus exocortis viroid, and its antifungal activity has been demonstrated in in vitro assays. We have successfully produced transgenic orange (Citrus sinensis L. Obs. cv. Pineapple) plants bearing a chimeric gene construct consisting of the cauliflower mosaic virus 35S promoter and the coding region of the tomato pathogenesis-related PR-5. Nine regenerated transgenic lines constitutively expressed the PR protein. They were challenged with Phytophthora citrophthora using a detached bark assay. A significant reduction in lesion development was consistently observed in one transgenic line in comparison to the control plants. This same line achieved plant survival rates higher than control plants when transgenic trees were inoculated with oomycete cultures. These results provide evidence for the in vivo activity of the tomato PR-5 protein against Phytophthora citrophthora, and suggest that this may be employed as a strategy aimed at engineering Phytophthora disease resistance in citrus.     

Effect of temperature on life history of Cirrospilus vittatus (Hymenoptera: Eulophidae), an ectoparasitoid of Phyllocnistis citrella (Lepidoptera: Gracillariidae)

Cirrospilus vittatus is a generalist parasitoid detected on the invading Phyllocnistis citrella 1 yr after the introduction of this pest into Spain in 1993. In this study, the influence of temperature on parasitoid development, survival, and some selected life history parameters was determined. Development times shortened as the temperature increased from 15 to 30 degrees C, but increased between 30 and 35 degrees C. Larval development times varied the greatest over the range of temperatures, whereas egg development varied the least. The estimated upper and lower development thresholds were 38.2 and 8.2 degrees C, respectively, and the maximal developmental rate (8.75 d(-1)) occurred at 31.5 degrees C. The thermal constant was 275.1 +/- 4.6 degree-days. Cirrospilus vittatus appeared to be a synovigenic species. Mean fecundity at 25 degrees C was 39.17 eggs per female, and the oviposition rate fluctuated around five eggs per day. Superparasitism was quite common (42.8%), but fertility was high (85.6%). Immature C. vittatus survival was 58.1%. From these results, an intrinsic rate of increase of 0.126 females per female per day was estimated. Although under typical Mediterranean climatic conditions, development of C. vittatus could continue throughout the year, its reproductive fitness at 25 degrees C is lower than other leafminer parasitoids attacking P. citrella in Spain. These results could account for the progressive displacement observed in field populations occurring between C. vittatus and the predominant indigenous parasitoids of P. citrella, Cirrospilus sp. near lyncus.