Baubles, bangles, and biotypes: a critical review of the use and abuse of the biotype concept.

Pest species of insects are notoriously prone to escape the weapons deployed in management efforts against them. This is particularly true in herbivorous insects. When a previously successful tactic fails the insect population has apparently adapted to it and is often considered to be a new or distinct entity, and given the non-formal category 'biotype'. The entities falling under the umbrella term 'biotype' are not consistent either within or between biotypes, and their underlying genetic composition and origins, while generally unknown, are likely heterogeneous within and variable between biotypes. In some cases race or species may be more appropriate referents. Some examples of applications of the concept in the context of host plant resistance are discussed. It is argued here that the term 'biotype' and its applications are overly simplistic, confused, have not proved useful in current pest management, and lack predictive power for future management.

Flies in the ointment: a morphological and molecular comparison of Lucilia cuprina and Lucilia sericata (Diptera: Calliphoridae) in South Africa.

Complementary nuclear (28S rRNA) and mitochondrial (COI) genes were sequenced from blowflies that phenotypically resembled Lucilia cuprina (W.), Lucilia sericata (Meigen) or exhibited characters of both species. The aim was to test a long-held hypothesis that these species hybridize under natural conditions in South Africa (Ullyett, 1945). Blowflies were obtained predominantly from the Cape Town metropolitan area, but reference samples were acquired for L. sericata from Pretoria. Several L. cuprina-like flies were shown to possess a conflicting combination of nuclear and mitochondrial genes that has also been seen in Hawaiian specimens. Homoplasy, sampling of pseudogenes, hybridization and incomplete lineage sorting are discussed as possible hypotheses for the pattern and the latter is concluded to represent the most likely explanation.

Molecular systematics and evolution in an African cycad-weevil interaction: Amorphocerini (Coleoptera: Curculionidae: Molytinae) weevils on Encephalartos.

Weevils in the tribe Amorphocerini have been implicated in pollination of Encephalartos species in southern Africa. The services they render these plants and the unique attributes of the cycad-weevil interaction make them important from both conservation and evolutionary standpoints. Oberprieler [Oberprieler, R.G., 1996. Systematics and evolution of the tribe Amorphocerini, with a review of the cycad weevils of the world. Ph.D. dissertation, University of the Free State, Bloemfontein, South Africa], using morphological characters, proposed a tentative hypothesis of relationships among the Amorphocerini which is tested here using DNA sequence data. Sequences from one mitochondrial and three nuclear genes were used to infer phylogenetic relationships, levels of sequence divergence, evolution of host associations, and patterns of speciation in this tribe. The results are reasonably consistent with the morphological hypothesis of relationships and species concepts, though important differences are observed, particularly in relationships among a Porthetes hispidus Boheman species group, which is indicated to have experienced recent divergences. In general, low levels of sequence divergence among species within two of the three genera indicate a recent radiation of this tribe onto African cycads, thus while cycad-insect interactions have often been considered ancient this may not be the case for some extant interactions. A complex pattern of host shifts onto both closely related and more distantly related hosts is suggested.

Phylogenetic congruence of mealybugs and their primary endosymbionts.

Tight interactions between unrelated organisms such as is seen in plant-insect, host-parasite, or host-symbiont associations may lead to speciation of the smaller partners when their hosts speciate. Totally congruent phylogenies of interacting taxa have not been observed often but a number of studies have provided evidence that various hemipteran insect taxa and their primary bacterial endosymbionts share phylogenetic histories. Like other hemipterans, mealybugs (Pseudococcidae) harbour multiple intracellular bacterial symbionts, which are thought to be strictly vertically inherited, implying codivergence of hosts and symbionts. Here, robust estimates of phylogeny were generated from four fragments of three nuclear genes for mealybugs of the subfamily Pseudococcinae, and a substantial fragment of the 16S-23S rDNA of their P-endosymbionts. Phylogenetic congruence was highly significant, with 75% of nodes on the two trees identical, and significant correlation of branch lengths indicated coincident timing of cladogenesis. It is suggested that the low level of observed incongruence was influenced by uncertainty in phylogenetic estimation, but evolutionary outcomes other than congruence, including host shifts, could not be rejected.

Effects of short-term spontaneous mutation accumulation for life history traits in grape phylloxera, Daktulosphaira vitifoliae.

Mutation is the source of all genetic variation, but rate of input and effects of new mutations for phenotypic traits related to fitness and the role they play in the maintenance of genetic variation are still subject to controversy. These parameters are important in models of the evolution of sex and recombination, the persistence of asexual populations, and the extinction of small populations. Most estimates have come from a few model organisms. Here, mutation accumulation experiments were conducted with three clones of grape phylloxera, Daktulosphaira vitifoliae Fitch, a gall forming herbivore and pest of grapes, to estimate the rate of input and effects of spontaneous mutation on life history traits. This is perhaps the first such experiment using a non-model organism of economic importance. Significant heritable genetic variation accrued in one of three sets of lines for one of four traits measured, and deleterious effects of mutation were found for two of four traits in two of the three sets of lines. Estimates of the parameters by the Bateman-Mukai (BM) method were within the range found in previous studies but at the lower end for genomic mutation rate, U approximately 0.023 and mutational variance, V(M) approximately 0.0003, the upper end for average heterozygous effect, alpha, of approximately -0.11, and on the order of previous estimates for mutational heritability, hM approximately 0.007. Under a model of equal effects of mutations, maximum likelihood (ML) estimates of U were slightly higher, and of alpha lower, than the BM estimates. Support limits were too large to provide much confidence in the ML estimates, however, and models of mutational effects assuming a gamma distribution of effects under different values of the shape parameter, beta, could not be distinguished though likelihoods tended to be lower at lower values of beta (more leptokurtic). Rapid accumulation of deleterious mutations suggest that for many pest species, adaptive response under agricultural conditions may depend more on the standing variation derived from introductions than new mutation.

Locating the sources of an invasive pest, grape phylloxera, using a mitochondrial DNA gene genealogy.

Range expansions through human introductions have increased with global commerce and have led to the extinction of native species, alterations in community structure and pest status of the invasive species. Inferring the evolutionary history of invasive species can help to build a firmer footing for management tactics. This study used mitochondrial DNA (mtDNA) sequence comparisons of samples collected from the native and introduced ranges of a pest herbivore of cultivated grapes, grape phylloxera (Daktulosphaira vitifoliae Fitch, Phylloxeridae) to infer the sources and pattern of introductions into worldwide viticulture. Introductions into viticulture from its native North American range first occurred in the mid-19th century. The pattern of spread has suggested a focus of introduction into France, but independent introductions may have occurred elsewhere. The results show that the introduced population represents a limited subsample of the native genetic diversity. The data suggest that most grape phylloxera in viticulture, including all European, have originated in the northeastern USA where the grape species Vitis riparia dominates. There was evidence for independent introductions into South Africa and California. Most California haplotypes were most closely related to native grape phylloxera from the Atlantic Coast on V. vulpina. It is likely that subsequent spread from California into Australia, New Zealand and Peru has occurred.

Grapes, galls, and geography: the distribution of nuclear and mitochondrial DNA variation across host-plant species and regions in a specialist herbivore.

Studies of patterns of molecular variation in natural populations can provide important insights into a number of evolutionary problems. Among these, the question of whether geographic factors are more important than ecological factors in promoting population differentiation and ultimately speciation has been an important and contentious area in evolutionary biology. Systems involving herbivorous insects have played a leading role in this discussion. This study examined the distribution of molecular variation in a highly specialized gall-forming insect, grape phylloxera (Daktulosphaira vitifoliae Fitch), that is found on both sympatric and allopatric host-plant species of the genus Vitis. In addition, the relationship of insects in the introduced range in the United States to ancestral populations in the native range was examined. Evidence for differentiation along host-plant lines from both nuclear (RAPD) and mitochondrial (COI) DNA was confounded with the effect of geography. Differentiation was found where hosts were allopatric or parapatric, but no evidence was found for such differentiation on two hosts, V. vulpina and V. aestivalis, that are broadly sympatric. The question of population differentiation onto these sympatric hosts can be considered to be resolved--it has not occurred in spite of a long history of association. Evidence was equivocal, but suggestive of a period of divergence in allopatry prior to reestablishment of contact, for insects associated with another host plant species, V. cinerea, found in both sympatric and parapatric populations. A low level of diversity and placement of samples collected from the grape species V. riparia at the tip of a phylogenetic tree supports the hypothesis that this host has been recently colonized from populations from the Mississippi Valley. A polyphyletic origin for biotype B grape phylloxera was supported: Although most samples collected from vineyards in the introduced range in California had similar haplotypes, they were closely related to natives on V. vulpina from the Atlantic Coast-Piedmont region. All samples collected from vineyards in Oregon and Washington were closely related to natives on V. riparia in the northern United States.

Patterns of genetic variation in native grape phylloxera on two sympatric host species.

Random amplified polymorphic DNA (RAPD) markers were used to examine population genetic structure in populations of native grape phylloxera. This research asked: (i) do RAPD markers distinguish two groups corresponding to the two host plant species; and (ii) do RAPD markers distinguish groups according to spatial location, independent of host plant association? Forty-nine phylloxera clones were collected from five pairs of adjacent individuals of two sympatric grape species in five sites along a 145 km transect in Missouri, USA. A high level of polymorphism was observed, with some evidence for structuring between host plant species and no evidence for spatial structuring. An analysis of molecular variance (AMOVA) found that 6.52% of the variance in RAPD banding patterns was attributable to host species and 7.96% of the variance was attributable to spatial location. A cluster analysis did not result in two groups corresponding to the two hosts, or to five groups corresponding to the geographical sites sampled. A Mantel test showed a low correlation between genetic similarity and spatial location. Two of the 93 RAPD markers were nonrandomly associated between the hosts. It is suggested that there may be a small host-mediated effect on genetic variation but stochastic dispersal and a highly heterogeneous environment may be the primary influences on the observed polymorphism.

Performance of native grape phylloxera on host plants within and among terrestrial islands in Arizona, USA.

In Arizona, USA, the canyon grape, Vitis arizonica Englemann, and grape phylloxera (Daktulosphaira vitifoliae Fitch, Homoptera, Phylloxeridae) are distributed among mountain ranges that are surrounded by expanses of desert lacking Vitis habitat, thus forming a system of terrestrial islands. Both herbivore and host populations may have diverged genetically among mountain ranges under the influence of restricted gene flow and variable selection among sites. Herbivore adaptation to local hosts would be expected to ensue, with the potential to promote divergence, both in traits under selection and by further reducing the probability of interisland colonization. To test the hypothesis that phylloxera are adapted to local hosts, demographic components of fitness of field-collected native grape phylloxera were measured in the greenhouse on vines of V. arizonica that were categorized as either natal, neighboring, and or isolated hosts. There was no evidence for greater adaptation to natal or neighboring hosts but there were significant interactions between herbivore and host treatments in one experiment. There was genetic variation for gall formation among six clones tested. Though a failure to detect local adaptation could have resulted from low statistical power, benign experimental conditions, or inadequate genetic variation, the divergence of isolated grape populations is suggested to have been insufficient to promote local adaptation in grape phylloxera at the level of isolated mountain ranges. It is further suggested that, within populations, adaptation to individual host plants could be forestalled by selection for 'general purpose genotypes' through wind-borne displacement of colonizers into the unpredictable environment of a heterogeneous array of hosts. In addition, short-term extinction/colonization dynamics could promote gene flow such that time is insufficient for adaptive mutations or gene combination to arise.