Detection of post-F1 fish hybrids in broodstock using molecular markers: Approaches for genetic management in aquaculture

Herein, we have developed molecular markers for nuclear genes to use in multiplex-PCR and PCR-RFLP, with the goal of characterising hybrid lines derived from crosses between pintado Pseudoplatystoma corruscans and cachara P. reticulatum. These markers, together with others described previously, were used to perform molecular identification analyses as genetic subsidies for Brazilian aquaculture. These analyses were performed due to the problems of high mortality in the offspring reported by the aquaculturist. From a total of 16 broodstock samples, 13 were genetically identified as hybrids; surprisingly, nine of these hybrids were found to be post-F1 lineages. These data show that the fertility of these animals can seriously affect the cultivated stocks, thus causing financial damage in this aquaculture system. The establishment of PCR-RFLP and multiplex-PCR as molecular techniques allows for both the correct management of these animals and the routine monitoring of production and trade of fish hybrids in aquaculture. Consequently, such tools will enable a sustainable development in the aquaculture industry. © 2012 Blackwell Publishing Ltd.

Genetic Identification of F1 and Post-F1 Serrasalmid Juvenile Hybrids in Brazilian Aquaculture

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cytogenetic characterization of F1, F2 and backcross hybrids of the Neotropical catfish species Pseudoplatystoma corruscans and Pseudoplatystoma reticulatum (Pimelodidae, Siluriformes)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Differences in the number of sensilla coeloconica and sensilla ampullacea of segment 10 of the antennae of Africanized and Caucasian bees and of their F1 hybrids

The numbers of sensilla coeloconica and sensilla ampullacea of segment 10 of the antennae of Caucasian and Africanized worker bees and of their hybrids were counted with the aid of a scanning electron microscope. Africanized bees have fewer sensilla than Caucasian bees and the continuous distribution obtained from the F1 values suggests polygenic inheritance for the control of this trait. There are also indications that the hybrid Brazilian Apis mellifera studied here are close to typical African bees (Apis mellifera scutellata) in terms of number of sensilla.

Detection of hybrids and genetic introgression in wild stocks of two catfish species (Siluriformes: Pimelodidae): The impact of hatcheries in Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Obtaining interspecific hybrids, and molecular analysis by microsatellite markers in grapevine

The objective of this work was to assess the potential of interspecific hybridization of Vitis labruscana and Muscadinia rotundifolia by using artificial cross-pollinations. Microsatellite markers were used to confirm interspecific hybridizations and the identity of the parental genotypes. In crosses in which M. rotundifolia was used as the female parent, no true hybrids were obtained. In the reciprocal crosses, 114 seedlings were identified as true V. labruscana x M. rotundifolia hybrids. Self pollination occurred in direct and in reciprocal crosses. The crossings between 'Bordo' x 'Carlos', 'Magnolia', 'Regale' and' Roanoke', and between' Isabel' x 'Bountiful', 'Carlos', 'Magnolia', 'Regale' and 'Roanoke' were confirmed. The 15 markers evaluated showed that two M. rotundifolia parental genotypes had the same fingerprint profile, indicating a like lyplanting error. The success of hybridization depends mainly on the species and on the cultivar used as the female parent. Microsatellite markers are efficient to confirm the paternity of interspecific F1 hybrids and to determine the correct identity of M. rotundifolia cultivars.

Expression of resistance in rice hybrids to Pyricularia grisea

Thirty-nine rice (Oryza sativa) hybrids and their restorers were assessed for vertical resistance to Pyricularia grisea in the rice blast nursery, and in artificial inoculation tests with two pathotypes, under controlled greenhouse conditions. The hybrids were developed from cytoplasmic genetic male sterile lines 046I and IR 58025A, derived from WA cytoplasm. In the rice blast nursery all hybrids showed susceptible reaction varying from 5 to 9. Compatible and incompatible leaf blast reactions of hybrids to two pathotypes, IC-1 and IB-45, were observed in inoculation tests. A majority of the hybrids were resistant when the restorer was resistant. However, seven of the 25 F1 hybrids exhibited susceptible reactions even when one of the parents was resistant to a pathotype. The partial resistance of 11 hybrids and their parents that showed compatible reactions to two pathotypes was analyzed. Differential interaction between isolates and genotypes was observed for partial resistance in relation to both disease severity and lesion number indicating the specific nature of partial resistance.

Evaluation of the courtship and of the hybrid male sterility among Drosophila buzzatii cluster species (Diptera, Drosophilidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Aplicação de marcadores moleculares no monitoramento genético de programas de hibridação interespecífica em pisciculturas brasileiras

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Produtividade em espécies de Drosophila do subgrupo saltans (grupo saltans, subgênero Sophophora): efeitos da infecção por Wolbachia em linhagens normais e introgredidas

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Synaptomenal complex analysis of four breeds of Bos taurus taurus x B. taurus indicus hybrids

The synaptonemal complex (SC) was analyzed in four F1 hybrids of Bos taurus taurus and B. taurus indicus including Gyr-Simmental (G-S), Nelore Simmental (N-S), Gyr-Holstein-Friesian (G-H) and Nelore-Piemontese (N-P). We analysed the frequency of various types of SC abnormalities and the frequency of cells with SC abnormalities. The results were compared with similar observations made on purebred animals. All the animals studied possessed 29 autosomal and one sex bivalent. The frequency of cells with abnormalities in the hybrids were 28.0% in the N-P, 29.1% in the G-S, 33.3% in the N-S and 40.0% in the G-H. The frequency of cells with abnormalities in the four hybrids was 31.5%; 57.9% of these abnormalities occurred in zygotene and 42.0% occurred in pachytene. The comparisons among the hybrids and among the hybrids and their parental breeds showed that the only significant difference was between Gyr and Gyr-Holstein-Friesian animals. Some aspects of the relationship between the frequency of cells with anomalies and the fertility of hybrids are discussed.

Spontaneous heterosis in larval life-history traits of hemiclonal frog hybrids

European water frog hybrids Rana esculenta (Rana ridibunda × Rana lessonae) reproduce hemiclonally, transmitting only their ridibunda genome to gametes. We compared fitness-related larval life-history traits of natural R. esculenta from Poland with those of the two sympatric parental species and of newly generated F1 hybrids. Compared with either parental species, F1 hybrid offspring had higher survival, higher early growth rates, a more advanced developmental stage by day 49, and earlier metamorphosis, but similar mass at metamorphosis. R. esculenta from natural lineages had trait values intermediate between those of F1 offspring and of the two parental species. The data support earlier observations on natural R. esculenta that had faster larval growth, earlier metamorphosis, and higher resistance to hypoxic conditions compared with either parental species. Observing larval heterosis in F1 hybrids in survival, growth rate, and time to metamorphosis, however, at an even higher degree than in hybrids from natural lineages, demonstrates that heterosis is spontaneous and results from hybridity per se rather than from subsequent interclonal selection; in natural lineages the effects of hybridity and of clonal history are confounded. This is compelling evidence for spontaneous heterosis in hybrid clonals. Results on hemiclonal fish hybrids (Poeciliopsis) showed no spontaneous heterosis; thus, our frog data are not applicable to all hybrid clonals. Our data do show, however, that heterosis is an important potential source for the extensively observed ecological success of hybrid clonals. We suggest that heterosis and interclonal selection together shape fitness of natural R. esculenta lineages.

Distribution and hybridization between Culex pipiens and Culex quinquefasciatus (Diptera: Culicidae) in Argentina

To better undesrtand the distribution of Culex pipiens and Cx. quinquefasciatus in Argentina, samples were collected from six localities situated in a North-South line from Castelli (Chaco Province) to Puerto Madryn (Chubut Province). Identification was based on the morphology of male genitalia. Only Cx. quinquefasciatus was found in Castelli and Esperanza, while in Rosario, 95.3% belonged to this species and 4.7% represented hybrid forms. Southern samples included only Cx. pipiens. With the purpose of verfying if Cx. pipiens and Cx. quinquefasciatus hybridize, different crosses between the two species were perfomed. All crosses produced viable egg rafts. Hatching ranged from 70 to 100%, except in one cross, female Cx. pipiens x male Cx. quinquefasciatus, where a high incompatibility was observed (11.1%hatch). The F1 hybrids obtained all crosses were fertile. The finding of hybrid forms in nature can be interpreted as evidence for subspecific status of Cx. pipiens and Cx. quinquefasciatus in Argentina.

Strong reproductive barriers in a narrow hybrid zone of West-Mediterranean green toads (Bufo viridis subgroup) with Plio-Pleistocene divergence

Background One key question in evolutionary biology deals with the mode and rate at which reproductive isolation accumulates during allopatric speciation. Little is known about secondary contacts of recently diverged anuran species. Here we conduct a multi-locus field study to investigate a contact zone between two lineages of green toads with an estimated divergence time of 2.7 My, and report results from preliminary experimental crosses. Results The Sicilian endemic Bufo siculus and the Italian mainland-origin B. balearicus form a narrow hybrid zone east of Mt. Etna. Despite bidirectional mtDNA introgression over a ca. 40 km North-South cline, no F1 hybrids could be found, and nuclear genomes display almost no admixture. Populations from each side of the contact zone showed depressed genetic diversity and very strong differentiation (FST = 0.52). Preliminary experimental crosses point to a slightly reduced fitness in F1 hybrids, a strong hybrid breakdown in backcrossed offspring (F1 x parental, with very few reaching metamorphosis) and a complete and early mortality in F2 (F1 x F1). Conclusion Genetic patterns at the contact zone are molded by drift and selection. Local effective sizes are reduced by the geography and history of the contact zone, B. balearicus populations being at the front wave of a recent expansion (late Pleistocene). Selection against hybrids likely results from intrinsic genomic causes (disruption of coadapted sets of genes in backcrosses and F2-hybrids), possibly reinforced by local adaptation (the ranges of the two taxa roughly coincide with the borders of semiarid and arid climates). The absence of F1 in the field might be due to premating isolation mechanisms. Our results, show that these lineages have evolved almost complete reproductive isolation after some 2.7 My of divergence, contrasting sharply with evidence from laboratory experiments that some anuran species may still produce viable F1 offspring after > 20 My of divergence.

Crocidura cossyrensis Contoli, 1989 (Mammalia, Soridiae): karyotype, biochemical genetics and hybridization experiments

Crocidura cossyrensis Contoli, 1989 (Mammalia, Soricidae): karyotype, biochemical genetics and hybridization experiments. - The shrew Crocidura cossyrensis Contoli, 1989 from Pantelleria (I), a Mediterranean island 100 km south of Sicily and 70 km west from Tunisia, was investigated in order to understand its origin and its relationship with C. russula from Tunisia, Morocco and Switzerland. With the exception of a single heterozygote centric fusion, C. cossyrensis had a karyotype identical with that of C russula from Tunisia (2N = 42, NF = 70 to 72), but it differed from C russula from Morocco and Switzerland (2N = 42, NF = 60). The former have 5-6 pairs of chromosomes with small arms that are acrocentric in the latter. Genetic comparisons with allozyme data revealed small genetic distance (0.04) between C cossyrensis and C russula from Tunisia. In contrast, this eastern clade (Tunisia and Pantelleria) is separated from the western clade (Switzerland and Morocco) by a genetic distance of 0.14. A hybridization experiment between shrews from Pantelleria and Switzerland lead rapidly to an F1 generation. From 12 F1 hybrids that were backcrossed, females reproduced normally, but none of the males did so. Concluding from the results, C. cossyrensis from Pantelleria and C. russula cf. agilis from Tunisia belong to the same taxon that may have reached the differentiation of a biological species within the C. russula group. More geographic samples are needed to determine the definitive taxonomic positions of these shrews.