Copper resistance genes from different xanthomonads and citrus epiphytic bacteria confer resistance to Xanthomonas citri subsp. citri

Genetic exchange is considered to be an important process in the selective adaptation of microorganisms to shifting and challenging environmental conditions. As a consequence of the copious use of copper bactericides, many species of plant pathogenic bacteria, including Xanthomonas citri subsp. citri (Xcc), have developed resistance to copper. This study assesses whether copper resistant (Cu^R) strains of other Xanthomonas species and citrus epiphytic bacteria pose a risk for the development of copper resistance in Xcc. Cu^R epiphytic bacteria were isolated on MGY agar from citrus leaves collected in two citrus groves treated with copper bactericides in Florida. Horizontal gene transfer of copper resistance genes was investigated within different Xanthomonas species and from citrus epiphytic bacteria to Xanthomonas. Cu^R epiphytic bacteria from citrus were screened for the presence of copper resistance genes homologous to copL, copA and copB genes from Xcc and characterized regarding tolerance to copper. Copper resistance determinants from a citrus epiphytic strain of Stenotrophomonas maltophilia (Stm) were cloned and expressed in Xcc and other Xanthomonas strains. Copper resistance genes in Xcc were determined to be present on a large (~300?kb) conjugative plasmid. Cu resistance was transferred via conjugation from two copper resistant citrus strains, Xcc and X. alfalfae subsp. citrumelonis (Xac), and two tomato pathogens, X. euvesicatoria (Xe) and X. perforans (Xp), to Xcc. PCR analysis revealed that two Cu^R strains from citrus, an epiphytic Xanthomonas ssp. and a strain of Stm, harboured homologs of the copper resistance genes found in Cu^R Xcc. The introduction of copLAB gene cluster from Stm into different xanthomonads conferred copper resistance to sensitive strains of Xcc, Xac, Xe and Xp. Based on these results there is a low, but significant, likelihood of horizontal gene transfer of copper resistance genes from other xanthomonads or epiphytic bacteria to Xcc in nature.     

Evaluation of uncalibrated preferential flow models against data for isoproturon movement to drains through a heavy clay soil

The uncalibrated predictive ability of four preferential flow models (CRACK‐NP, MACRO/MACRO_DB, PLM, SWAT) has been evaluated against point rates of drainflow and associated concentrations of isoproturon from a highly structured and heterogeneous clay soil in the south of England. Data were available for four plots for a number of storm events in each of three successive growing seasons. The mechanistic models CRACK‐NP and MACRO generally gave reasonable estimates of drainflow over the three seasons, but under‐estimated concentrations of isoproturon over a prolonged period in the first season and over‐estimated them in the two remaining seasons. CRACK‐NP simulated maximum concentrations of isoproturon over the first two events of each of the three seasons of 156, 527 and 24.4 µg litre^?1, respectively, and matched the observed data (465, 65.1 and 0.65 µg litre^?1) slightly better than MACRO (69.1, 566 and 58.5 µg litre^?1). Automatic selection of parameters from soils information within MACRO_DB reduced the emphasis on preferential flow relative to the stand‐alone version of MACRO. This gave a poor simulation of isoproturon breakthrough and simulated maximum concentrations were 0, 50.1 and 35.1 µg litre^?1, respectively. The capacity model PLM gave the best overall simulation of total drainflow for the first two events in each season, but over‐estimated concentrations of isoproturon (967, 808 and 51.3 µg litre^?1). The simple model SWAT represented total drainflow reasonably well and gave the best simulation of maximum isoproturon concentrations (140, 80.2 and 8.2 µg litre^?1). There was no clear advantage here in using the mechanistic models rather than the simpler models. None of the models tested was able to simulate consistently the data set, and uncalibrated modelling cannot be recommended for such artificially drained heavy clay soils. © 2001 Society of Chemical Industry     

The Validation of Pesticide Leaching Models

The validation of pesticide leaching models presents particular problems where the number of model predictions is far in excess of the observed data. Normally, however, there are more frequent field observations for other parameters (notably the site hydrology) than for pesticide concentrations in either water or soil. A five-stage validation procedure which takes advantage of the most frequently available observations and which tests each of the components of the model in a cumulative way, is thus advocated: Stage 1: Parameterisation of the model using only independently measured parameters. Stage 2: Hydrological validation: the validation of the predictions of water movement and water content of the soil. Stage 3: Solute movement validation: where field data are available for solutes other than pesticide, the model should first be validated for them, especially if they are more abundant than the pesticide observations. Conserved solutes such as chloride or bromide are preferred, although nitrate may be used for short periods. Stage 4: Pesticide fate in the soil: models should use parameters of pesticide fate derived from independent studies. Stage 5: Pesticide leaching: only in the last stage are the relatively small number of pesticide observations compared with the model predictions with respect to patterns and orders of magnitude of occurrence. With this scheme, the results of each stage are carried forward to the next, and confidence in the model is built with each stage. This is illustrated using the CRACK-P model and hydrological, nitrate and pesticide data from the Brimstone Farm Experiment Oxfordshire, UK.     

Conservation of marker synteny during evolution

Summary An aspect of cereal science that is becoming increasingly important is comparative genetics. Establishment of the relationship between genomes within polyploids, between species within tribes and between species within families will allow not only the integration of genetic maps but also the knowledge acquired of each of the species. Using a set of homoeologous probes, workers found the relationship between the three wheat genomes to be precisely collinear, after taking a few major translocation events into account. Transfer of the wheat map to rye led to the elucidation of similar relationships between the three wheat genomes and that of rye. Genome collinearity, however, extends even beyond tribes. In a comparison of the genomes of wheat, rice and maize, it was shown that despite the separation of these genomes for possibly 50 million years, gene order was still highly conserved. This collinearity between genomes can be exploited in a number of ways.     

Fungicide seed treatments to improve seedling emergence of perennial ryegrass Lolium perenne and the effect of different cultivars and soils

Seeds of perennial ryegrass Lolium perenne treated with four fungicides, benomyl, captan, iprodione and metalaxyl, used singly and in all combinations were sown in soil in pots. The soil was maintained at a low moisture content to predispose the seedlings to pre-emergence infection by soil-borne fungi. Thirteen of the fifteen fungicide treatments significantly increased seedling emergence and all but one contained benomyl and/or captan. In another pot experiment, a combination of benomyl and captan was tested against four similar treatments, carbendazim+captan, thiabendazole+captan, thiabendazole+metalaxyl and thiabendazole+thiram and also drazoxolon. Only benomyl+captan and drazoxolon did not increase seedling emergence significantly. A natural infection of the seedlings by powdery mildew Erysiphe graminis was reduced significantly by all treatments except thiabendazole+thiram and drazoxolon. Only car-bendazim+captan significantly increased the dry weight of seedlings per pot 98 days after sowing. Benomyl+captan seed treatment significantly increased seedling emergence in 9 of 16 soils collected from widespread sites. One perennial ryegrass cultivar, Parcour, was used in the above experiments and in a comparison with 12 other diploid cultivars its response to benomyl seed treatment was about mid-range.     

Use of pyrethroid analogues to identify key structural features for enhanced esterase resistance in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

It has been reported previously that the major resistance mechanism to pyrethroid insecticides by the cotton bollworm Helicoverpa armigera (Hübner) in Australia is a consequence of overproduction of esterase isoenzymes. This paper reports structure-activity relationships that support such a view, based on in vivo bioassays conducted with a range of pyrethroid structures containing a variety of acid and alcohol moieties and the correlation with in vitro esterase inhibition assays against the same structures, and identifies the critical regions of the molecule with regard to esterase inhibition, and hence resistance. The implications of this work in terms of possible resistance management are evaluated and discussed.     

The handling and fate of spermatophores in Neoentobdella diadema and N. apiocolpos (Monogenea: Capsalidae: Entobdellinae)

The elongated encased spermatophores of the capsalid (entobdelline) monogeneans Neoentobdella diadema (Monticelli, 1902) Kearn et Whittington, 2005 and N. apiocolpos (Euzet et Maillard, 1967) Kearn et Whittington, 2005 have been found attached by their proximal ends to the region of the vaginal opening, with the bulk of the spermatophore projecting from the vagina and therefore lying outside the body. In spite of previous reports, no spermatophores were found projecting from the common genital opening and if spermatophore exchange is as rapid as it is in the related entobdelline Entobdella soleae, then the chances of finding a spermatophore in this location are small. In N. diadema and N. apiocolpos it is likely that sperm enters the vagina through the open proximal end of an attached spermatophore, after which the empty spermatophore case is probably discarded. There is no evidence for a previous proposal that the whole spermatophore is engulfed by the vagina followed by digestion of the case to release the sperm. Three specimens of N. diadema were found each with two spermatophore cases projecting from the vagina and a specimen of N. apiocolpos carried three cases. Assuming that each parasite is able to donate or receive only one spermatophore at each mating, then the presence of one spermatophore does not prevent a further mating and acceptance of a fresh spermatophore. In spite of differences between the spermatophores of E. soleae and N. diadema/N. apiocolpos, the events of spermatophore exchange may be similar.