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Dispersion of the Cf-4 disease resistance gene in Lycopersicon germplasm.

In the past, numerous Cf genes have been reported in tomato (Lycopersicon esculentum Mill.) that confer resistance against leaf mould (Cladosporium fulvum Cke.). We are interested in genetic variation at Cf loci. Therefore, previously uncharacterized Cf genes were further analysed. Recognition of the AVR4 elicitor, DNA gel blot analysis, PCR analysis and sequencing of part of the Cf-4 locus showed that a large proportion of the accessions tested harboured the Cf-4 resistance gene. We concluded that despite differences in nomenclature, all these accessions harbour the same Cf-4 locus, probably introgressed from the same donor. The origin of the Cf-4 locus and the reasons for discrepancies with earlier reports are discussed.

Blotting, Southern↗

Characterization and mapping of resistance to Oidium lycopersicum in two Lycopersicon hirsutum accessions: evidence for close linkage of two Ol-genes on chromosome 6 of tomato.

The cultivated tomato is susceptible to powdery mildew (Oidium lycopersicum). Several accessions of wild species are resistant. In this study we describe (i) the genetics and mapping of resistance to O. lycopersicum in G1.1290, one of the resistant accessions in Lycopersicon hirsutum, (ii) fine mapping of Ol-1 originated from L. hirsutum G1.1560, another resistant accession of L. hirsutum, and (iii) tests of allelism for resistance in G1.1290 and G1.1560. Initially, it is demonstrated that the resistance in G1.1290 to O. lycopersicum is controlled by an incompletely dominant gene, designated Ol-3. By using an advanced breeding line (ABL) containing introgression fragment(s) from G1.1290, Ol-3 was found to be associated with several RFLP and SCAR markers on chromosome 6. By using these markers, Ol-3 was mapped between markers TG25/SCAF10 and H9A11 on chromosome 6. Secondly, after testing some F3 lines and their progenies from the cross between L. esculentum cv Moneymaker and L. hirsutum G1.1560, we provided more evidence for the map position of Ol-1 to lie between SCAF10 and H9A11, indicating that Ol-1 and Ol-3 are in the same chromosome region. Thirdly, although allelism tests could not discriminate between Ol-1 and Ol-3, (indirect) evidence suggested that these two genes are not identical. They might instead represent functional genes of a cluster of Ol-homologues.

Alleles↗

Combined analyses of RAPDs, cpDNA and morphology demonstrate spontaneous hybridization in the plant genus Chaenomeles.

Evidence of spontaneous hybridization between two partially sympatric species of Chaenomeles, C. cathayensis and C. speciosa, has been obtained through analysis of offspring families from these two species, as well as from two presumed interspecific hybrid populations. A combination of different methods was applied. Analysis of diagnostic RAPD markers and of chloroplast DNA haplotypes supported the notion of spontaneous hybridization, and suggested that there has been symmetrical, rather than unidirectional, introgression between C. cathayensis and C. speciosa. RAPDs and morphological characters revealed concordant patterns of genetic relatedness among the studied offspring families. Some putative hybrid families had mainly intermediate characters, whereas others appeared to be later generation hybrids as they were genetically and phenotypically rather similar to families that appeared to represent pure species. The RAPD-based proportion of between-family variability was considerably higher in the putatively hybridogenous populations than in populations of the pure species. Within-family gene diversity estimates ranged from C. speciosa (max. Hj = 0.235) to C. cathayensis (min. Hj = 0.094) with the presumed hybrid families taking intermediate values.

Chloroplasts↗

Analysis of a contact zone in the Forficula auricularia L. (Dermaptera: Forficulidae) species complex in the Pyrenean Mountains.

The taxon Forficula auricularia L. (Dermaptera: Forficulidae) is a complex of two sibling species that differ in life history (number of clutches per year and imaginal diapause) and that have diverged at the molecular level. The study of a contact zone in the Pyrenean Mountains, using the PCR-RFLP method on two mitochondrial regions (the 16S rRNA and the Cytochrome Oxidase intergenic region), revealed the coexistence of the sibling species at intermediate altitude (1200 m) whereas at lower and higher altitudes only one species was found. An allozyme study, conducted simultaneously and based on four polymorphic loci (PGI1, AAT1, Est-P1 and Est-P2), showed no sign of nuclear introgression. The apparent lack of hybridization in the field is consistent with a postzygotic barrier observed in the laboratory (a nearly complete failure to produce F(1) hybrids). This contact zone is probably a sympatric zone between two genetically differentiated species.

Altitude↗

Characterization of a hybrid zone between two chromosomal races of the weta Hemideina thoracica following a geologically recent volcanic eruption.

Two chromosomal races (2n=17 and 2n=15; XO) of the weta Hemideina thoracica meet at the centre of a volcanic region in North Island, New Zealand. Five independent polymorphic genetic markers showed broadly coinciding, steep frequency clines from north to south across this zone beside the flooded crater, Lake Taupo. Three unlinked nuclear gene markers provide estimates of zone width that are at least twice the width of the chromosomal and mitochondrial clines, with cline centres displaced at least 2.5 km. The different zone widths and centres suggest that this hybrid zone is a semipermeable barrier reducing the introgression of the chromosomal markers more than genic markers. We estimate that this species of weta must have a dispersal rate of at least 100 m per generation using the time since the last Taupo eruption (1850 years ago), which covered an area of about 20 000 km2 with pyroclastic flow.

Animals↗

Allozyme analysis of the hybrid origin of Arisaema ehimense (Araceae).

Allozyme diversity was examined in the Japanese jack-in-the pulpit species, Arisaema serratum and A. tosaense, and their putative hybrid species, A. ehimense (all diploid). Arisaeme ehimense contains mostly alleles found in one or both of the putative parent species but few unique alleles, partially supporting the hypothesis that A. ehimense is of hybrid origin from the two species, although the possibility that A. ehimense arose via divergent speciation cannot be excluded. Because only limited information was gained from the survey of allozyme variation, additional taxon-specific markers from A. serratum and A. tosaense are required to test rigorously the hybrid origin of A. ehimense. A phenogram based on allele frequencies suggested that introgression occurs between A. serratum and A. tosaense where these species are sympatric.

Alleles↗

Chromosome races with Pliocene origins: evidence from mtDNA.

There are eight distinct chromosomal races of the New Zealand weta Hemideina thoracica. We used mtDNA sequence data to test the hypothesis that these races originated on islands during the early Pliocene (7--4 million years ago). Nine major mitochondrial lineages were identified from 65 cytochrome oxidase I sequences. Phylogenetic analysis of these lineages suggests that they arose at approximately the same time. The geographical distribution of some lineages coincides with areas that were islands during the Pliocene. Overall, hierarchical AMOVA analysis shows that chromosomal races and Pliocene islands describe only 28% and 24%, respectively, of the total current mtDNA variation. However, removing one widespread (A) and one putatively introgressed (F) lineage increases these estimates to 65% and 80%, respectively. Intraspecific sequence divergence was very high, reaching a maximum of 9.5% (uncorrected distance) and GC content was high compared to other insect mtDNA sequences. Average corrected distance among mtDNA lineages supports the Pliocene origins of this level of genetic diversity. In the southern part of the species range there is reduced mtDNA variation, probably related to local extinction of H. thoracica populations from recent volcanic activity and subsequent re-colonization from a leading edge. In contrast, in this southern part there are five chromosome races, suggesting that chromosome races here may be younger than those in the north.

Animals↗

Fecundity of transgenic wild-crop hybrids of Cucurbita pepo (Cucurbitaceae): implications for crop-to-wild gene flow.

Hybridization between crops and their weedy or wild relatives is an area of concern because the widespread use of genetically engineered crops may allow novel, beneficial transgenes to enter nearby populations. We compared fitness components of wild Cucurbita pepo from Arkansas, USA, with wild-crop hybrids derived from yellow squash (a cultivar of C. pepo with transgenic resistance to two viruses). Wild and hybrid progeny were grown in agricultural fields in Arkansas (1996-98) and Ohio (1996) in six similar experiments. Cross types (wild and hybrid) did not differ significantly in seedling survival, which exceeded 85% in all cases. In Ohio, where more detailed observations were made, hybrid plants produced 41% as many male flowers, 21% as many female flowers, and 28% as many seeds as wild plants. At all sites, flowering periods of the two cross types overlapped extensively. Putative virus symptoms were more common in wild plants than in hybrids. Lifetime fecundity varied considerably among sites and years. The average fecundity of hybrids ranged from 453 to 4497 seeds per plant and represented 15% - 53% of the numbers of seeds produced by wild plants in the same experiments. These results suggest that the F1 generation does not represent a strong barrier to the introgression of neutral or beneficial crop genes into free-living populations of C. pepo.

Crops, Agricultural↗

Reproductive processes in two oak (Quercus) contact zones with different levels of hybridization.

Patterns of reproductive isolation between two sympatric species of oaks, Quercus gambelii and Q. grisea, that exhibit strong ecological differentiation were examined. A full diallel cross using four trees of each species (i.e. all possible pollinations among eight trees) was performed. This design was repeated at two sites that represent different outcomes of sympatry: (1) a xeric mountain ridge where many hybrids are established (HZ); and (2) a mesic valley bottom where virtually no hybrids are established (MOCYN). By measuring fruit survival at several developmental stages, both the timing and strength of reproductive barriers within and between sites, species, cross types, and pollen dosage levels were examined. In three of four cases, heterospecific fruit set was significantly reduced compared to conspecific fruit set. This reduction occurred after the time of fertilization, but before the onset of embryo growth. Increasing the dose of pollen from an average of 9-194 grains/stigma did not affect this result. Thus, early postfertilization processes play a strong role in species fidelity in these oaks. Quercus gambelii experienced a five-fold decrease in conspecific fruit set at HZ relative to MOCYN. In contrast, heterospecific fruit set of Q. gambelii was the same at both sites. Poor Q. gambelii pollen performance is implicated as playing the major role in this result. One Q. gambelii individual at HZ was highly fecund, and had higher heterospecific than conspecific fruit set; slight introgression in this tree was detected uisng RAPD markers. The Environmental Emasculation Hypothesis that posits that environmental stress can increase the probability of hybrid formation by reducing the competitive ability of male gametes of one species is proposed.

Crosses, Genetic↗

Wolbachia infections in native and introduced populations of fire ants (Solenopsis spp.).

Wolbachia are cytoplasmically inherited bacteria that induce a variety of effects with fitness consequences on host arthropods, including cytoplasmic incompatibility, parthenogenesis, male-killing and feminization. We report here the presence of Wolbachia in native South American populations of the fire ant Solenopsis invicta, but the apparent absence of the bacteria in introduced populations of this pest species in the USA. The Wolbachia strains in native S. invicta are of two divergent types (A and B), and the frequency of infection varies dramatically between geographical regions and social forms of this host. Survey data reveal that Wolbachia also are found in other native fire ant species within the Solenopsis saevissima species complex from South America, including S. richteri. This latter species also has been introduced in the USA, where it lacks Wolbachia. Sequence data reveal complete phylogenetic concordance between mtDNA haplotype in S. invicta and Wolbachia infection type (A or B). In addition, the mtDNA and associated group A Wolbachia strain in S. invicta are more closely related to the mtDNA and Wolbachia strain found in S. richteri than they are to the mtDNA and associated group B Wolbachia in S. invicta. These data are consistent with historical introgression of S. richteri cytoplasmic elements into S. invicta populations, resulting in enhanced infection and mtDNA polymorphisms in S. invicta. Wolbachia may have significant fitness effects on these hosts (either directly or by cytoplasmic incompatibility) and therefore these microbes potentially could be used in biological control programmes to suppress introduced fire ant populations.

Animals↗

Patterns of DNA sequence variation in chromosomally recognized taxa of Anopheles gambiae: evidence from rDNA and single-copy loci.

Patterns of DNA sequence variation in the ribosomal DNA (rDNA) second internal transcribed spacer (ITS2) and five unlinked single-copy nuclear loci were examined for evidence of reproductive isolation among four chromosomally recognized taxa of Anopheles gambiae from West Africa: Savanna, Bamako, Mopti and Forest, as well as sibling species An. arabiensis and An. merus. Included among the single-copy loci were three sequence-tagged random amplified polymorphic DNA (RAPD) loci, two of which (R15 and R37) had been reported as discriminating between Mopti and other chromosomal forms. Each of the five single-copy sequences were highly polymorphic in most samples. However, the R15 and R37 loci had no diagnostic value, and therefore are not recommended as tools in recognition of field-collected An. gambiae chromosomal forms. Although pairwise comparisons between species generally revealed significant levels of differentiation at all five loci, variation was not partitioned by chromosomal form within An. gambiae at any single-copy locus examined. The few exceptions to these trends appear related to a location either inside or nearby chromosomal inversions. At the tryptophan oxygenase locus inside inversion 2Rb, variation was structured only by inversion orientation and not by taxonomic designation even between An. gambiae and An. arabiensis, providing the first molecular evidence that the 2Rb inversion was transferred between species by introgressive hybridization. By contrast, the rDNA showed fixed differences between species and a difference diagnostic for Mopti, consistent with effective, if not complete, reproductive isolation. The apparent disagreement between the data from this locus and multiple single-copy loci within An. gambiae may be explained by the much lower effective population size of rDNA, owing to concerted evolution, which confers increased sensitivity at much shorter divergence times. Taken together with the accompanying reports by della Torre et al. (2001), Favia et al. (2001) and Gentile et al. (2001), our data suggest that neutral molecular markers may not have the sensitivity required to detect isolation between these recently established taxa.

Animals↗

Assessment of rDNA IGS as a molecular marker in the Simulium damnosum complex.

For five cytospecies of the Simulium damnosum Theobald complex of blackflies (Diptera: Simuliidae) from West Africa, both ends of the intergenic spacer region (IGS) of the rDNA have been sequenced with the aim of developing specific molecular markers. No specific differences in these two regions were detected between Simulium sanctipauli V. & D., Simulium sirbanum V. & D., Simulium soubrense V. & D., Simulium squamosum Enderlein and Simulium yahense V. & D., except in the number of A subrepeats at the 5' end of the IGS (two in S. squamosum and four or five in the others) and in position 310 of the 3' end (a C in S. squamosum and a G in the others). However, genetic distances within and between species overlapped. These DNA sequences had no strong phylogenetic signal, and the trees obtained were mostly unresolved. Although most sequences from S. squamosum clustered together, a few of them were more similar to those in other cytospecies. These results could be explained either by hybridization with genetic introgression or by ancestral polymorphism and recent speciation.

Africa, Western↗

Genetic structure of an aphid studied using microsatellites: cyclic parthenogenesis, differentiated lineages and host specialization.

In a previous study, samples of the grain aphid Sitobion avenae (F.) were collected from wheat and adjacent cocksfoot hosts in a population thought to be primarily parthenogenetic, and DNA from individual aphids was analysed with a multilocus technique. Here we have applied single-locus microsatellites and a mitochondrial DNA marker to a subset of the same DNA extracts, and have made several additional inferences about important genetic and population processes in S. avenae. Microsatellite analysis indicated very high levels of genic and genotypic variation. S. avenae fell into three genotypic groups inferred to be almost noninterbreeding, while analysis of linkage and Hardy-Weinberg equilibria suggested high levels of sexual recombination within each genotypic group. Host specialization was evident: one lineage was found only on wheat, and one (bearing many alleles inferred to be introgressed from the blackberry-grass aphid S. fragariae (Walker)) was found only on cocksfoot. The third group of interrelated genotypes was found commonly on both hosts. Although most genotypes were found only once, some were much more numerous in the sample than expected from the frequency of the alleles they contained. This, and rapid temporal changes in genotypic composition of samples, indicates strong selective differences between genotypes and lineages. In the major genotypic group, the commonest genotypes were significantly more homozygous than were rare ones: thus these data may help to explain the frequent observation of homozygous excess in aphid allozymes. The genotype group showing S. avenae-like as well as S. fragariae-like alleles also carried S. fragariae-like mitochondrial DNA in at least 25/31 cases, indicating gender-asymmetrical hybridization.

Animals↗

Assessing hybridization in natural populations of Penstemon (Scrophulariaceae) using hypervariable intersimple sequence repeat (ISSR) bands.

Inferences regarding hybridization rely on genetic markers to differentiate parental taxa from one another. Intersimple sequence repeat (ISSR) markers are based on single-primer PCR reactions where the primer sequence is derived from di- and trinucleotide repeats. These markers have successfully been used to assay genetic variability among cultivated plants, but have not yet been tested in natural populations. We used genetic markers generated from eight ISSR primers to examine patterns of hybridization and purported examples of hybrid speciation in Penstemon (Scrophulariaceae) in a hybrid complex involving P. centranthifolius, P. grinnellii, P. spectabilis and P. clevelandii. This hybrid complex has previously been studied using three molecular data sets (allozymes, and restriction-site variation of nuclear rDNA and chloroplast DNA). These studies revealed patterns of introgression involving P. centranthifolius, but were unsuccessful in determining whether gene flow occurs among the other species, and support for hypotheses of diploid hybrid speciation was also lacking. In this study, we were able to fingerprint each DNA accession sampled with one to three ISSR primers and most accessions could be identified with a single primer. We found population- and species-specific markers for each taxon surveyed. Our results: (i) do not support the hybrid origin of P. spectabilis; (ii) do support the hypothesis that P. clevelandii is a diploid hybrid species derived from P. centranthifolius and P. spectabilis; and (iii) demonstrate that pollen-mediated gene flow via hummingbird vectors is prevalent in the hybrid complex.

California↗

Polymorphic chloroplast simple sequence repeat primers for systematic and population studies in the genus Hordeum

In this study we report the development of primers to amplify polymorphic chloroplast simple sequence repeats in the genus Hordeum, which includes cultivated barley (H. vulgare ssp. vulgare) and its wild progenitor H. vulgare ssp. spontaneum. Polymorphic products were amplified in a wide range of Hordeum spp. and intraspecific variation was detected in both cultivated and wild barley. A decrease in cytoplasmic diversity was observed between sspp. spontaneum and vulgare as well as between ssp. vulgare landraces and cultivars, which is characteristic of domestication processes in many corp species. We also observed possible evidence for reticulate evolution of H. brachyantherum polyploids, with apparent multiple cytoplasmic introgressions during successive polyploidization events.

Journal Article↗

Molecular variation in Leymus species and populations.

Icelandic populations of European lymegrass [Leymus arenarius (L.) Hochst.] were examined using amplified fragment length polymorphism (AFLP) and restriction fragment length polymorphism (RFLP) of the major ribosomal genes (18S-5.8S-26S rDNA), in comparison with Alaskan populations of its closely related species L. mollis (Trin.) Pilger. The AFLP profiles emerged as two distinct entities, clearly separating the two species, and based on species-specific bands it was simple to distinguish these two morphologically similar species. The rDNA-RFLPs also differentiated the species. Within species, the Icelandic L. arenarius was more homogeneous than the Alaskan L. mollis, and its variation was dispersed over geographically different populations, suggesting a common gene pool. The variation among the Alaskan L. mollis was more extensive and its interrupted pattern may be the result of gene introgression at subspecies level. Within a 40-year-old population of L. mollis established in Iceland from Alaskan material, the molecular profiles separated old and new genotypes. Both AFLP and rDNA revealed the new genotypes to be extremely similar. This rapid change in allele frequency is thought to be the result of adaptation to a new environment.

Alaska↗

Extensive intraspecific chloroplast DNA (cpDNA) variation in the alpine Draba aizoides L. (Brassicaceae): haplotype relationships and population structure.

Chloroplast DNA (cpDNA) sequence variation is currently the most widely used tool for the inference of phylogenetic relationships among plants at all taxonomic levels. Generally, noncoding regions tend to evolve faster than coding sequences and have recently been applied to the study of phylogenetic relationships among closely related taxa. An implicit assumption of many of these studies is that intraspecific cpDNA variation is either absent or low and therefore will not interfere with the reconstruction of interspecific relationships. A survey of cpDNA sequence variation in the common alpine plant species Draba aizoides L. was undertaken to assess levels of intraspecific cpDNA sequence variation. These levels were compared to levels of interspecific sequence divergence between D. aizoides and related alpine Draba species. Intraspecific cpDNA sequence divergence was extensive in D. aizoides, and intraspecific differences were often larger than interspecific differences. cpDNA haplotype relationships were explored using a maximum parsimony approach and minimum-spanning networks. Results from both methods were largely congruent but comparisons provided interesting insights into the presumed evolutionary history of cpDNA haplotypes. A combined effect of cpDNA introgression and complex lineage sorting was inferred to explain the pattern of cpDNA variation found in D. aizoides. Our results suggest that intraspecific cpDNA variation can be extensive and that intraspecific variation needs to be taken into account when inferring phylogenetic relationships among closely related taxa.

Brassicaceae↗

Chloroplast DNA variation in dactylis glomerata L. taxa endemic to the macaronesian islands

In the Dactylis glomerata infraspecific polyploid grass complex, restriction fragment length polymorphisms (RFLPs) of chloroplast DNA (cpDNA) were studied in diploid and tetraploid populations of several taxa endemic to Macaronesia (Madeira and the Canary islands) and in populations from the African and European continental areas closest to Macaronesia. Two chlorotypes, which differed by a single 290-bp length mutation, were observed in the Macaronesian and the continental populations. Chlorotype I, which is predominant in the whole D. glomerata complex, was found in the majority of continental populations. It was also observed in the most western Macaronesian islands, in the two diploid taxa endemic to the lowland scrub and the high elevation heath of Tenerife, respectively, and in tetraploids endemic to Madeira and La Palma. These island populations were growing under the influence of humid trade winds. Chlorotype II was found in the eastern part of the Archipelago (closer to Africa), which experienced subarid Mediterranean climate conditions, and in very few diploid and tetraploid Mediterranean populations growing at high elevation on the continent. This geographical and climatic distribution of chlorotype variation in Macaronesia is consistent with that reported previously for morphological, allozyme and phenolic variation in the same plant material. Chlorotype II was, however, also observed in tetraploid populations from La Gomera island and in one of the seven tetraploid populations analysed from Madeira, which all showed clearly subtropical characters for morphology, allozymes and phenolic compounds. This result suggests that cpDNA introgression has occurred more than once from the Mediterranean material into the subtropical one and may indicate that colonization between the mainland and islands, or among the islands, probably played a major role in the geographical pattern observed for that marker.

Journal Article↗