Dynamics and management of the white potato cyst nematode Globodera pallida in commercial potato crops

Eight trials were conducted in commercial potato fields infested with the white potato cyst nematode (wPCN, Globodera pallida) and one in a field infested with the yellow PCN (yPCN, Globodera rostochiensis). Our aims were to produce data to validate and refine a computer‐based program (The Model) for the long‐term management of PCN, to determine nematicide effectiveness and to assess rates of PCN population decline between potato crops. Prior to planting, each farmer applied an overall nematicide treatment to his field, except for ten untreated plots that were widely spaced to encompass a range of PCN population densities. Each untreated plot was paired with a similar plot in the adjacent treated area and all plots were intensively sampled for PCN population densities at planting (P_i) and again at harvest (P_f) when tuber yields were determined. Four trials were re‐sampled 2–4 years later to determine PCN population decline rates. Regressions that form the basis of ‘The Model’ and described the relationship between P_i and tuber yield and PCN population density at harvest were fitted to the results from both the untreated and nematicide treated plots. These regressions also enabled us to estimate the yield potential at each site in the absence of PCN and showed that nematicide treatment generally did not increase yield potential and that both tuber yield and PCN multiplication decreased with increasing P_i. However, there were major differences between sites and cultivars. When untreated, the yield of cv. Maris Piper was hardly affected in a highly organic soil with P_i > 200 eggs g^?1 whereas the yield of partially resistant cv. Santé was decreased from a potential of c. 60 t ha^?1 to c. 20 t ha^?1 in a light silt with P_i = 20 egg g^?1 soil. Similarly, untreated wPCN multiplication rates at a low P_i ranged from 46‐fold to >100‐fold. Nematicide effectiveness was estimated from the regressions and, at several sites, yield was decreased despite nematicide treatment. Control of wPCN multiplication was even poorer. In only two of seven trials planted with susceptible cultivars was more than 50% control achieved – maximum populations in treated plots usually exceeded 250 eggs g^?1. Partially resistant Santé decreased the multiplication rate of wPCN in the two trials where it was planted. An alternative analysis using Genstat indicated that The Model tended to underestimate the maximum multiplication rate and overestimate the maximum population density. When four sites were re‐sampled 2–4 years after harvest the populations of wPCN had declined by between 15% and 33.5% per annum with a mean of 26% per annum. Modelling indicated that rotations longer than 8 years were required to control wPCN unless other effective control measures, such as growing a partially resistant cultivar, were used.     

Parasitism genes and host range disparities in biotrophic nematodes: the conundrum of polyphagy versus specialisation

This essay considers biotrophic cyst and root-knot nematodes in relation to their biology, host-parasite interactions and molecular genetics. These nematodes have to face the biological consequences of the physical constraints imposed by the soil environment in which they live while their hosts inhabit both above and below ground environments. The two groups of nematodes appear to have adopted radically different solutions to these problems with the result that one group is a host specialist and reproduces sexually while the other has an enormous host range and reproduces by mitotic parthenogenesis. We consider what is known about the modes of parasitism used by these nematodes and how it relates to their host range, including the surprising finding that parasitism genes in both nematode groups have been recruited from bacteria. The nuclear and mitochondrial genomes of these two nematode groups are very different and we consider how these findings relate to the biology of the organisms.     

Horizontal transmission, vertical inactivation, and stochastic loss of mariner-like transposable elements

Horizontal transmission has been well documented as a major mechanism for the dissemination of mariner-like elements (MLEs) among species. Less well understood are mechanisms that limit vertical transmission of MLEs resulting in the "spotty" or discontinuous distribution observed in closely related species. In this article we present evidence that the genome of the common ancestor of the melanogaster species subgroup of Drosophila contained an MLE related to the mellifera (honey bee) subfamily. Horizontal transmission, approximately 3-10 MYA, is strongly suggested by the observation that the sequence of the MLE in Drosophila erecta is 97% identical in nucleotide sequence with that of an MLE in the cat flea, Ctenocephalides felis. The D. erecta MLE has a spotty distribution among species in the melanogaster subgroup. The element has a high copy number in D. erecta and D. orena, a moderate copy number in D. teissieri and D. yakuba, and was apparently lost ("stochastic loss") in the lineage leading to D. melanogaster, D. simulans, D. mauritiana, and D. sechellia. In D. erecta, most copies are concentrated in the heterochromatin. Two copies from D. erecta, denoted De12 and De19, were cloned and sequenced, and they appear to be nonfunctional ("vertical inactivation"). It therefore appears that the predominant mode of MLE evolution is vertical inactivation and stochastic loss balanced against occasional reinvasion of lineages by horizontal transmission.      

Incorporation of axonally transported glycoproteins into axolemma during nerve regeneration

The insertion of axonally transported fucosyl glycoproteins into the axolemma of regenerating nerve sprouts was examined in rat sciatic motor axons at intervals after nerve crush. [(3)H]Fucose was injected into the lumbar ventral horns and the nerves were removed at intervals between 1 and 14 d after labeling. To follow the fate of the “pulse- labeled” glycoproteins, we examined the nerves by correlative radiometric and EM radioautographic approaches. The results showed, first, that rapidly transported [(3)H]fucosyl glycoproteins were inserted into the axolemma of regenerating sprouts as well as parent axons. At 1 d after delivery, in addition to the substantial mobile fraction of radioactivity still undergoing bidirectional transport within the axon, a fraction of label was already associated with the axolemma. Insertion of labeled glycoproteins into the sprout axolemma appeared to occur all along the length of the regenerating sprouts, not just in sprout terminals. Once inserted, labeled glycoproteins did not undergo extensive redistribution, nor did they appear in sprout regions that formed (as a result of continued outgrowth) after their insertion. The amount of radioactivity in the regenerating nerves decreased with time, in part as a result of removal of transported label by retrograde transport. By 7-14 d after labeling, radioautography showed that almost all the remaining radioactivity was associated with axolemma. The regenerating sprouts retained increased amounts of labeled glycoproteins; 7 or 14 d after labeling, the regenerating sprouts had over twice as much of radioactivity as comparable lengths of control nerves or parent axons. One role of fast axonal transport in nerve regeneration is the contribution to the regenerating sprout of glycoproteins inserted into the axolemma; these membrane elements are added both during longitudinal outgrowth and during lateral growth and maturation of the sprout.     

The metabolism of cyclohexanol by Nocardia globerula CL1

1. Nocardia globerula CL1, isolated by enrichment on cyclohexanol and grown with it as carbon source, oxidized it with a Q(o2) of 39mul/h per mg dry wt. and the overall consumption of 2.2mumol of oxygen/mol of substrate. Cyclohexanone, 2-hydroxycyclohexan-1-one dimer and cyclohexane-1,2-dione were oxidized with Q(o2) values similar to that for cyclohexanol whereas in-caprolactone and 6-hydroxycaproate were oxidized very slowly and adipate not all. 2. Disrupted cell suspensions could not be shown to catalyse the conversion of cyclohexanol into cyclohexanone. 3. A cyclohexanol-induced cyclohexanone oxygenase (specific activity 0.55mumol of NADPH oxidized/min per mg of protein) catalysed the consumption of 1mol of NADPH and 1mol of O(2) in the presence of 1mol of cyclohexanone. NADPH oxidation did not occur under anaerobic conditions. The only detected reaction product with 25000g supernatant was 6-hydroxycaproate. 4. Extracts of cyclohexanol-grown cells contained a lactone hydrolase (specific activity 15.6mumol hydrolysed/min per mg of protein), which converted in-caprolactone into 6-hydroxycaproate. 5. Incubation of 6-hydroxycaproate with 25000g supernatant in the presence of NAD(+) resulted in NAD(+) reduction under anaerobic conditions, oxygen consumption under aerobic conditions and the conversion of 6-hydroxycaproate into adipate. 6. Cyclohexanone oxygenase fractions devoid of in-caprolactone hydrolase catalysed the stoicheiometric formation of in-caprolactone from cyclohexanone in the presence of excess of NADPH. 7. The reaction sequence for the oxidation of cyclohexanone by N. globerula CL1 is: cyclohexanol --> cyclohexanone --> in-caprolactone --> 6-hydroxycaproate --> adipate. 8. It is suggested that the adipate may be further dissimilated by beta-oxidation.     

Nonrandom location of IS1 elements in the genomes of natural isolates of Escherichia coli

We have studied the spatial distribution of IS1 elements in the genomes of natural isolates comprising the ECOR reference collection of Escherichia coli. We find evidence for nonrandomness at three levels. Many pairs of IS1 elements are in much closer proximity (< 10 kb) than can be accounted for by chance. IS1 elements in close proximity were identified by long-range PCR amplification of the genomic sequence between them. Each amplified region was sequenced and its map location determined by database screening of DNA hybridization. Among the ECOR strains with at least two IS1 elements, 54% had one or more pairs of elements separated by < 10 kb. We propose that this type of clustering is a result of "local hopping," in which we assume that a significant proportion of tranposition events leads to the insertion of a daughter IS element in the vicinity of the parental element. A second level of nonrandomness is found in strains with a modest number of IS1 elements that are mapped through the use of inverse PCR to amplify flanking genomic sequences: in these strains, the insertion sites tend to be clustered over a smaller region of chromosome than would be expected by chance. A third level of nonrandomness is observed in the composite distribution of IS elements across strains: among 20 mapped IS1 elements, none were found in the region of 48-77 minutes, a significant gap. One region of the E. coli chromosome, at 98 min, had a cluster of IS1 elements in seven ECOR strains of diverse phylogenetic origin. We deduce from sequence analysis that this pattern of distribution is a result of initial insertion in the most recent common ancestor of these strains and therefore not a hot spot of insertion. Analysis using long- range PCR with primers for IS2 and IS3 also yielded pairs of elements in close proximity, suggesting that these elements may also occasionally transpose by local hopping.      

Enzymology of oxidation of tropic acid to phenylacetic acid in metabolism of atropine by Pseudomonas sp. strain AT3.

Pseudomonas sp. strain AT3 grew with dl-tropic acid, the aromatic component of the alkaloid atropine, as the sole source of carbon and energy. Tropic acid-grown cells rapidly oxidized the growth substrate, phenylacetaldehyde, and phenylacetic acid. Crude cell extracts, prepared from dl-tropic acid-grown cells, contained two NAD+-linked dehydrogenases which were separated by ion-exchange chromatography and shown to be specific for their respective substrates, dl-tropic acid and phenylacetaldehyde. Phenylacetaldehyde dehydrogenase was relatively unstable. The stable tropic acid dehydrogenase was purified to homogeneity by a combination of ion-exchange, molecular-sieve, and affinity chromatography. It had a pH optimum of 9.5 and was equally active with both enantiomers of tropic acid, and at this pH, phenylacetaldehyde was the only detectable product of tropic acid oxidation. The formation of phenylacetaldehyde from tropic acid requires, in addition to dehydrogenation, a decarboxylation step. By analogy with NAD+-specific isocitrate and malate dehydrogenases, phenylmalonic semialdehyde, a 3-oxoacid, would be expected to be the precursor of phenylacetaldehyde. Other workers have established that isocitrate and malate dehydrogenases catalyze the decarboxylation of enzyme-bound or added 3-oxoacid intermediates, a reaction that requires Mn2+ or Mg2+ ions. Studies with tropic acid dehydrogenase were hampered by lack of availability of phenylmalonic semialdehyde, but in the absence of added divalent metal ions, both enantiomers of tropic acid were completely oxidized and we have not, by a number of approaches, found any evidence for the transient accumulation of phenylmalonic semialdehyde.