Superoxide dismutase polymorphism in field populations of the potato cyst nematodes Globodera rostochiensis and G. pallida

Superoxide dismutase (SoDase) polymorphism in reference populations of potato cyst nematodes (PCN) was studied by isoelectric focusing and compared with the banding pattern obtained from 26 English field populations of PCN. Qualitative and quantitative differences were found between the reference populations: a band detected only in the G. rostochiensis Ro4 and Ro5 reference populations was found in four field populations of varying pathotype composition and two field populations of G. rostochiensis Rol possessed extra bands that were not detected in the other field populations. Analysis of the band differences showed little correlation between pathotype and banding pattern in the field populations.     

Estimation of genomic variation between British populations of the potato cyst nematode Globodera pallida using RAPD-PCR

Random amplified polymorphic DNA (RAPD) has been used to investigate the interrelationships of 20 populations of Globodera pallida collected originally from field soils around the UK. RAPD analysis revealed a high level of relative genomic diversity within British G. pallida but there was no general correlation of genomic similarity with geographic distribution. Two populations of pathotype Pa1 were clearly divergent from the bulk of G. pallida and might represent a distinct introduction. Two other populations, from Scotland and Wales, were also dissimilar from each other and from the rest of the G. pallida populations.     

The management of potato cyst nematodes using resistant Solanaceae potato clones as trap crops

The concept of using a range of Solanaceae potato clones as trap crops for potato cyst nematode (PCN) management was investigated. A series of field trials were undertaken from 1999 to 2002 that evaluated 10 clones of either wild Solanum potato species, breeder’s hybrid lines or commercial cultivars. All had high resistance to all known PCN pathotypes (both Globodera rostochiensis and Globodera pallida) and the ability to stimulate high levels of PCN hatch. Investigations showed potential for the development of some clones as a means of reducing high PCN field population levels and for use by organic potato producers.     

Integrated control of potato cyst nematodes using small amounts of nematicide and potatoes with partial resistance

The control of potato cyst nematode (PCN) by less than approved amounts of nematicide combined with partially resistant potato clones was studied in a series of field experiments. On a site heavily infested with Globodera pallida only the most resistant clone (12380ac2) decreased the population density in untreated soil. With aldicarb at its full approved rate (3·36 kg ha^-1) numbers of PCN were decreased under all the genotypes, including the non-resistant Maris Piper. Aldicarb at 1·68 kg ha^-1significantly decreased populations on all clones except 12380ac2. Aldicarb at 0·84 kg ha^-1still significantly decreased population densities and multiplication rates of G. pallida on two clones with intermediate resistance (12243acl and 11233ab22). At two G. rostochiensis sites with light infestations nematode multiplication rates were greater and the control given by aldicarb and partially resistant genotypes of potato was not as great as that at the site with G. pallida. Tuber yields were not increased by the application of aldicarb at the G. rostochiensis infested sites. However, at the site heavily infested with G. pallida the yield of the most intolerant genotype (12380ac2) was increased seven-fold by the full rate of aldicarb (3·36 kg ha^-1) and four-fold by the quarter rate (0·84 kg ha^-1)-Yield of the most tolerant genotype (12243acl) was unaffected by the application of aldicarb.     

Glucose phosphate isomerase polymorphism in field populations of the potato cyst nematodes, Globodera rostochiensis and G. pallida

The isozymic variation of glucose phosphate isomerase (GPI) was compared in 20 field populations of potato cyst-nematode (PCN) and related to variation in reference pathotypes of PCN. Populations pathotyped as G. rostochiensis Ro 1 by conventional differential cultivar tests were found to be different from populations of G. pallida. Variation within populations pathoptyped as G. pallida was seen and three populations were found to be similar to the reference G. pallida Pal pathotype. This seemed to be confirmed for two populations using cysts formed on a cultivar resistant to the Pa 1 pathotype when an isoenzyme pattern characteristic of G. palida Pa2 and Pa3 was found. The feasibility of the use of isozyme identification of PCN pathotypes is discussed in the light of these results.     

Characterisation of a 34 kD protein from potato cyst nematodes, using monoclonal antibodies with potential for species diagnosis

Two monoclonal antibodies, which differentially recognise the two species of potato cyst nematodes (PCN), Globodera pallida and G. rostochiensis, are described. They have been shown to have potential for quantification of these two species, recognising proteins of the same molecular weight (34 kD) in both species. Further investigation showed these proteins to have isoelectric points at pH values of 5.7 in G. pallida and 5.9 in G. rostochiensis, in common with the proteins used by Fleming & Marks (1983) to differentiate the species of PCN. They are likely to be structurally very similar, with the same physiological function (and therefore similar concentrations) in the two species. In cross-reactivity tests with a wide range of soil nematode species, the antibodies reacted strongly only with species of the genus Globodera, and thereby confirmed their potential as the basis of a quantitative immunoassay likely to be useful in management of PCN populations.     

Effects of potnetial trap crops and planting date on soil infestation with potato cyst nematodes and root-knot nematodes

In 1997 and 1998 the stimulation of hatch of potato cyst nematodes (PCN) by a trap crop was studied at various times during the growing season in a container and a field experiment. Solanum nigrum‘90‐4750‐188’was used as the trap crop in both experiments and was sown on 1 May, 16 June or 1 August in two successive years on different plots. Neither experiment revealed much seasonal variation in hatchability of PCN juveniles under a trap crop. In the container experiment, the hatch of the Globodera pallida Pa3 population was equally and strongly stimulated (89%) at all sowing dates in both years, except for the 1 August sowing in 1998 (when the hatch was 77% under extremely wet soil conditions). In the control treatment with non‐hosts (flax followed by barley) the total spontaneous hatch was 50% over 2 yr. In the field experiment, the hatch of PCN, averaged over the four populations, was also equally stimulated (71%) at all sowing dates in both years. In the control treatment with non‐hosts (flax‐barley) the total spontaneous hatch was 36% over 2 yr. Total hatch under the trap crop over 2 yr varied between the four PCN populations from 63% to 80%. In 1998 and 1999, control of potato cyst nematodes (PCN) by the potential trap crops Solanum sisymbriifolium and S. nigrum‘90‐4750‐188’was studied in the field. Potato was also included as a trap crop. In the 1998 experiment, potato, S. sisymbriifolium and S. nigrum strongly stimulated the hatch of PCN compared with the non‐host white mustard (Sinapis alba). Roots of potato and white mustard were mainly found in the top 10 cm of soil, whereas roots of S. sisymbriifolium and S. nigrum were also abundant at depths of 10–20 cm and 20–30 cm. In the 1999 experiment, soil infestation with PCN decreased markedly with potato and S. sisymbriifolium as trap crops. In plots moderately to severely infested with 2‐yr old cysts (2–29 juveniles ml^?1 air dried soil), potato reduced soil infestation by 87% and S. sisymbriifolium by 77%. In plots moderately to severely infested with 1‐yr old cysts the reductions were 74% and 60%, respectively. The reduction was least on plots very severely infested with PCN (110–242 juveniles ml^?1 soil): 69% and 52% for potato and S. sisymbriifolium, respectively. Soil infestations of plots that were initially slightly to severely infested with the root‐knot nematode Meloidogyne hapla were greatly reduced under fallow and S. sisymbriifolium but increased under potato. From these and previous experiments it was concluded that, for several reasons, S. sisymbriifolium is a promising trap crop.     

Serological identification and quantification of potato cyst nematodes from clean cysts and processed soil samples

Two monoclonal antibodies which specifically recognise each of the two species of potato cyst nematodes (PCN) and do not cross react with other species of soil nematodes, were used successfully in an immunoassay to identify and quantify PCN species using clean cysts and mixed populations. These antibodies show reactivity only towards antigens prepared from live eggs and they recognise antigens which are easily released from the nematodes. The results presented in this paper show that serological identification and quantification of PCN, not only from clean cysts but also from processed soil samples is achievable. A simple procedure to recover nematodes from soil samples and then to release nematode antigens was devised. The use of these procedures and the immunoassay for quantification of PCN was validated in tests with soil samples from Northern Portugal. The flotation method proved to be as efficient as the Fen wick Can for cyst recovery from soil samples. A significant correlation was obtained between results from immunoassay estimates and the traditional method of cyst picking and egg counting. The amount of organic matter (OM) present in the soil samples affected the sensitivity of the immunoassay but quantification of nematodes extracted from soil samples was possible with soils containing up to 14% of OM. The challenge remains to optimise the extraction procedures and the immunoassay, in practical conditions with highly organic soils containing cysts of different sizes and ages.     

The interactions of potato cyst nematodes and Verticillium dahliae on early and maincrop potato cultivars

The effects of potato cyst nematodes (Globodera rostochiensis and G. pallida) on the growth of early and later maturing potato cultivars, with or without the H, resistance gene, were compared in pot experiments and the effects of introducing Verticillium dahliae into the system were studied. The influence of the nematodes on root function and structural integrity was inferred from their effects on calcium and water uptake. Early maturing cultivars were less tolerant of nematode attack than later maturing ones, and those with the H₁ resistance gene were more tolerant of G. rostochiensis than G. pallida. When V. dahliae was introduced, Verticillium wilt symptoms appeared much earlier on the early than the maincrop cultivars. Also, Maris Anchor (first early) died before Maris Peer (second early) and death of both when infected by nematodes and fungus was much earlier than uninfected control plants or plants infected with G. rostochiensis only. Maris Peer infected by both organisms snowed symptoms much more severe than when infected by either organism alone, whereas Maris Anchor was almost as severely affected by V. dahliae alone as both organisms together. Pentland Javelin (first early) developed Verticillium wilt symptoms several weeks later than other cultivars and the addition of nematodes failed to increase the severity of the symptoms. These differences in cultivar susceptibility to the combination of nematodes and fungus may be due to the nematodes causing different degrees of trauma in the roots of the three cultivars: measuring calcium uptake per unit water uptake showed that nematodes dramatically increase calcium uptake but that the proportional change is least in Pentland Javelin. If this is a reflection of less damage by nematodes to the endodermis of this cultivar, it may explain the greater resistance of its roots to penetration by V. dahliae.     

Effects of Zygorrhynchus moelleri on the growth of potato and reproduction of potato cyst nematodes

Laboratory, pot and field experiments investigated the effects of the fungus Zygorrhynchus moelleri on the growth of potato and on the reproduction of the potato cyst nematodes (PCN), Globodera pallida and G rostochiensis. Preliminary laboratory tests showed that Z. moelleri growth was favoured by temperatures and pH ranges commonly present in field soils. The fungus colonised potato roots in vitro and in compost or field soil. It also stimulated in vitro root growth of three potato cultivars. In pot experiments Z. moelleri stimulated potato growth, particularly in the presence of PCN attack. In field plots infested with a mixture of G pallida and G. rostochiensis, tuber yields were not increased after application of the fungus but, in G pallida‐infested plots, yields were significantly increased after drills were inoculated with Z. moelleri. The application of Z. moelleri had no apparent effects on nematode reproduction. Factors influencing the interactions between Z. moelleri, potato and potato cyst nematodes are discussed and the potential role of the fungus as a plant growth promoter in organic potato production considered.     

The relationship of potato yield with and without nematicide to density of potato cyst nematodes, lobodera rostochiensis and G. pallida

Trials relating response to nematicide to potato cyst nematode density were conducted initially in the West Midlands and later in other Regions. Thirteen trials, only three of which were within the intensive potato growing areas, conformed to a general pattern with yield losses being largely recouped by nematicide treatment. Five trials, four within the intensive areas, gave no correlation between potato yield and nematode density and an unpredictable response to nematicide. The control of the nematode appeared to be poorer in the latter trials but other factors affecting yield and nematode multiplication may have been involved. It is speculated the main factor might be interaction with fungal organisms e.g. Rhizoctonia or Verticillium. Evidence is presented to show that on several trials the nematicide has an effect other than by controlling potato cyst nematodes. Yield losses caused by potato cyst nematodes are more variable than previous work indicated, probably due to varietal, seasonal or environmental influences.     

Successful intercontinental transport of porcine embryos from Europe to South America

A total of 393 porcine embryos was transported in two trials each lasting more than 30h from Hannover, Federal Republic of Germany to Buenos Aires, Argentina. The embryos were shipped in phosphate buffered saline (PBS) medium supplemented with 10% lamb serum and packaged in 0.25ml straws placed in a modified thermos bottle at 36.5 degrees C. Upon arrival, 359 embryos (90.8%) were evaluated as morphologically intact and were transferred to 19 recipients. Twelve recipients remained pregnant. Three recipients aborted and nine recipients (47.4%) farrowed a mean number of 5.6 +/- 2.6 (x +/- SD) piglets after 115.1 +/- 1.8 d of gestation. The average birthweight was 1.1 +/- 0.2 kg. The percentage of embryos resulting in live piglets was 28.6% in farrowing recipients. These births represented the first piglets from embryos that had been stored for more than 30h in vitro and shipped from Europe to South America.     

Use of resistant and susceptible potato cultivars in the trap cropping of potato cyst nematodes, Globodera pallida and G. rostochiensis

The effects of planting date and growing period of potato cultivars on their efficiency as trap crops for potato cyst nematodes (PCN) were studied. Plots were planted with susceptible or resistant cultivars in April, June and August and these were grown for 5, 6 or 7 wk before removal of the plants by hand lifting. Crops planted in June provided the best overall reductions in PCN population density of up to 95%, with cv. Santé significantly more effective than the other cultivars. Population reductions from the August planting were only slightly less than from planting in June but the tuber yields obtained were much greater: Maris Piper and Maris Bard produced 16.4 and 21.4 t ha^-1 respectively, with 37% and 43% respectively, of a size useful for canning (i.e. between 20 and 40 mm diameter).     

Heavy metal tolerance and accumulation in metallicolous and non-metallicolous populations of Thlaspi caerulescens from continental Europe

In continental Europe, the heavy metal hyperaccumulator Thlaspi caerulescens occurs both on heavy-metal polluted soils (subsp. calaminare) and on soils with normal heavy metal content (subsp. caerulescens). In order to assess the extent and partitioning of variation in heavy metal tolerance and foliar mineral composition, twelve families from two populations of each subspecies were grown in pots in four soil treatments differing in heavy metal (Zn, Pb) and macronutrient concentrations. The two subspecies differed systematically in many respects. Subsp. calaminare had a higher survival at high levels of heavy metals and a higher tolerance index in all treatments. It also had three times lower foliar zinc and lead concentrations when grown at moderate levels of heavy metals. This, together with a negative correlation of foliar Pb concentration with growth in subsp. caerulescens, suggests that heavy metal accumulation per se is not a mechanism of tolerance in this species. Variation among families within populations accounted for a larger proportion of total variance in growth and mineral composition than variation between populations. Additionally, within population variation in heavy metal tolerance and accumulation was significantly lower in subsp. calaminare. This suggests that, adding to a background constitutive tolerance at the species level, natural selection has increased heavy metal tolerance in metallicolous populations of Thlaspi caerulescens.     

Skull morphology of bottlenose dolphins from different ocean populations with emphasis on South America

The bottlenose dolphin, genus Tursiops, is cosmopolitan occurring in tropical and temperate regions, with morphological variation between and within different oceans. Since the genus' taxonomy has been under discussion for a long time, this work aimed at analyzing the cranial variability of T. truncatus from different regions of the world. Geometric Morphometrics analyses were performed in 201 skulls of adult specimens, on dorsal, ventral, and lateral views, from the Eastern North Pacific, Eastern North Atlantic, Eastern South Atlantic, and Western South Atlantic oceans. The results indicate differences between individuals that inhabit the Atlantic and Pacific oceans. Within the Atlantic Ocean, there is an evident longitudinal differentiation of specimens from the eastern and western regions. A latitudinal separation was also observed, considering specimens from the North and South Atlantic Ocean. In the Western South Atlantic statistical differences were found between two morphological groups, identified as T. gephyreus (sensu Lahille, 1908) and T. truncatus, and the cross-validation presented 98% as minimum confidence for correct classification of these two groups. The present study provides strong morphological support to consider these two lineages as separate species.     

Modeling spatiotemporal dynamics of vole populations in Europe and America

The mathematical models proposed and studied in the present paper provide a unified framework to understand complex dynamical patterns in vole populations in Europe and North America. We have extended the well-known model provided by Hanski and Turchin by incorporating the diffusion term and spatial heterogeneity and performed several mathematical and numerical analyses to explore the dynamics in space and time of the model. These models successfully predicted the observed rodent dynamics in these regions. An attempt has been made to bridge the gap between the field and theoretical studies carried out by Turchin and Hanski (1997) [23] and Turchin and Ellner (2000) [24]. Simulation experiments, mainly two-dimensional parameter scans, show the importance of spatial heterogeneity in order to understand the poorly understood fluctuations in population densities of voles in Fennoscandia and Northern America. This study shed new light upon the dynamics of voles in these regions. The nonlinear analysis of vole data suggests that the dynamical shift is from stability to chaos. Diffusion driven model systems predict a new type of dynamics not yet observed in the field studies of vole populations carried out so far. This has been termed as chaotic in time and regular in space (CTRS). We observed CTRS dynamics in several simulation experiments. This directs us to expect that dynamics of this animal would be de-correlated in time and simultaneously mass extinctions might be possible at many spatial locations.     

Contrasting genetic structure in Plasmodium vivax populations from Asia and South America

Populations of Plasmodium falciparum show striking differences in linkage disequilibrium, population differentiation and diversity, but only fragmentary data exists on the genetic structure of Plasmodium vivax. We genotyped nine tandem repeat loci bearing 2-8 bp motifs from 345 P. vivax infections collected from three Asian countries and from five locations in Colombia. We observed 9-37 alleles per locus and high diversity (He=0.72-0.79, mean=0.75) in all countries. Numbers of multiple clone infections varied considerably: these were rare in Colombia and India, but > 60% of isolates carried multiple alleles in at least one locus in Thailand and Laos. However, only one or two of the nine loci show >1 allele in many samples, suggesting that mutation within infections may result in overestimation of true multiple carriage rates. Identical nine-locus genotypes were frequently found in Colombian populations, contributing to strong linkage disequilibrium. These identical genotypes were strongly clustered in time, consistent with epidemic transmission of clones and subsequent breakdown of allelic associations, suggesting high rates of inbreeding and low effective recombination rates in this country. In contrast, identical genotypes were rare and loci were randomly associated in all three Asian populations, consistent with higher rates of outcrossing and recombination. We observed low but significant differentiation between different Asian countries (standardized FST = 0.13-0.45). In comparison, we see greater differentiation between collection locations within Colombia (standardized FST = 0.4-0.7), and strong differentiation between continents (standardized FST = 0.48-0.79). The observed heterogeneity in multiple clone carriage rates, linkage disequilibrium and population differentiation are similar in some, but not all, respects to those observed in P. falciparum, and have important implications for the design of association mapping studies, and interpretation of P. vivax epidemiology.