Backcross Of F1 Hybrid Research Articles

First evidence for backcrossing of F1 hybrids in Acropora corals under sperm competition

Acropora is a species-rich genus of reef-building corals with highly diverse morphologies. Hybridization among intercrossing species potentially influences species diversity within Acropora. However, the mechanisms that allow hybridization/backcrossing remain unknown. Although we tested a limited number of species, we hypothesized that Acropora gametes in the Indo-Pacific may preferentially fertilize conspecific gametes despite their compatibility with heterospecific gametes, leading to infrequent hybridization between potentially intercrossing species. In this study, F1 hybrids of Acropora florida and A. intermedia showed specific fertilization trends. For example, sperm had the ability to backcross with the parental species even in the presence of sperm from the parental species. Also, eggs of the hybrids produced from A. florida eggs and A. intermedia sperm (“FLOint”) exhibited self-fertilization. Since a low ratio of hybridization between A. florida and A. intermedia is predicted, the population size of hybrids should be small. Therefore, self-fertilization would facilitate reproduction of the hybrid in nature, while remaining sperm could outcompete parental species sperm to backcross with eggs. Although we succeeded in breeding two colonies of hybrids, it is reasonable to speculate that hybrids show a high tendency to choose the most efficient sexual reproduction tactics.

Geographic patterns of postzygotic isolation between two closely related widespread dung fly species (Sepsis cynipseaandSepsis neocynipsea; Diptera: Sepsidae)

Identifying the contribution of pre‐ and postzygotic barriers to gene flow is a key goal of speciation research. The widespread dung fly species Sepsis cynipsea and Sepsis neocynipsea offer great potential for studying the speciation process over a range of opportunities for gene exchange within and across sister species (cross‐continental allopatry, continental parapatry and sympatry). We examined the role of postcopulatory isolating barriers by comparing female fecundity and egg‐to‐adult viability of F1 and F2 hybrids, as well as backcrosses of F1 hybrids with the parental species, via replicated crosses of sym‐, para‐ and allopatric populations. Egg‐to‐adult viability was strongly but not totally suppressed in hybrids, and offspring production approached nil in the F2 generation (hybrid breakdown), indicating yet unspecified intrinsic incompatibilities. Viable F1 hybrid offspring showed almost absolute male (the heterogametic sex) sterility while females remained largely fertile, in accordance with Haldane's rule. Hybridization between the two species in European areas of sympatry (Swiss Alps) indicated only minor reinforcement based on fecundity traits. Crossing geographically isolated European and North American S. neocynipsea showed similar albeit weaker isolating barriers that are most easily explained by random genetic drift. We conclude that in this system with a biogeographic continuum of reproductive barriers, speciation is mediated primarily by genetic drift following dispersal of flies over a wide (allopatric) geographic range, with some role of natural or sexual selection in incidental or direct reinforcement of incompatibility mechanisms in areas of European sympatry. S(ubs)pecies status of continental S. neocynipsea appears warranted.

Hybridization in a Commercial Production Field between Imidazolinone-Resistant Winter Wheat and Jointed Goatgrass (Aegilops cylindrica) Results in Pollen-Mediated Gene Flow ofImi1

Imidazolinone-resistant (IR) winter wheat allows selective control of jointed goatgrass with the herbicide imazamox. However, the spontaneous hybridization between jointed goatgrass and IR winter wheat threatens the value of the IR technology. The objectives of this study were to determine if F1hybrids collected in a commercial production field under IR winter wheat–fallow rotation in Oregon and their first-backcross progeny (BC1) carried theImi1gene and were resistant to imazamox, and to analyze the parentage of F1and BC1plants. The average seed set of the F1spikes was 3.3%, and the average germination of BC1seed was 52%. All F1and BC1plants tested carriedImi1. Jointed goatgrass plant mortality was 100% when treated with imazamox at 0.053 kg ai ha−1, compared to 0% for IR winter wheat and BC1progeny. All F1plants had jointed goatgrass as the maternal parent; whereas, most BC1plants (85.7%) were produced with IR winter wheat as the paternal backcross parent. Although the backcrossing of F1hybrids with jointed goatgrass is very low, it demonstrates the potential for introgression ofImi1from IR winter wheat into jointed goatgrass under natural field conditions.

Cytogenetic and breeding studies with Brassica

A summary of studies made with Brassica napocampestris (B. nc.), 2n=58, obtained by colchicine treatment of the F1 hybrid between B. napus and B. campestris is given. Cytological studies of the primary hybrids reveal that the single genome of B. oleracea in B. napus—“c1”– may be utilized for elucidating the basic genomic number of the genus Brassica. Based on the mode of chromosome pairing at MI in the primary hybrids, the conclusion was drawn of a basic genomic number of 6. From the backcrosses of these F1 hybrids to B. campestris types, the same conclusion was indirectly arrived at. The backcross experiments with the F1 hybrids indicated the possibility of extracting not only a whole genome “a1” from B. napus but also, the supposed basic genome. For the hexaploid plants (2n=58), meiotic studies revealed that besides many bivalents there were some multivalents. Anaphase movements were slightly disturbed by bridges, presumed to result from inversions. On the whole, secondary balance accompanied by chromosomal alterations, may be responsible for the present serial polyploidy in the tribe Brassiceae. Aneuploid plants were not encountered in the hexaploid populations used. However, by continuous backcrosses of F1 hybrids (2n=29) to the hexaploids, plants with chromosome numbers close to 58 were obtained which showed fairly good seed fertility. It is suggested that selection against aneuploidy occurs at the gametic stage and this would account for the relatively low pollen fertilities found in the hexaploid B. nc. types.

Hybridization and Introgression in a Managed, Native Population of Yellowstone Cutthroat Trout: Genetic Detection and Management Implications

Since the mid-1920s, the Idaho Department of Fish and Game has cultured Yellowstone cutthroat trout Oncorhynchus clarki bouvieri at Henrys Lake to offset declines in natural production and for use in stocking programs throughout Idaho. Since the mid-1970s, they have also produced F1 hybrids: female Yellowstone cutthroat trout × male rainbow trout O. mykiss. The ability of fishery managers, when selecting broodstock, to visually distinguish returning cutthroat trout from F1 hybrids is, therefore, crucial to avoid accidental introduction of rainbow trout genes into the hatchery-supplemented cutthroat trout population. To evaluate this ability, fish identified by staff as putative cutthroat trout or hybrids (an array of phenotypic characters are used), were sampled during two spawning seasons. Phenotypically identified fish were genetically tested using species-specific restriction fragment length polymorphisms (RFLPs) of nuclear and mitochondrial DNA gene loci and diagnostic allozyme loci. Current levels of rainbow trout introgression in the cutthroat trout population at Henrys Lake were also investigated by analyzing samples collected from the lake and several of its tributaries. Results indicated that staff's phenotypic identifications were highly accurate in distinguishing cutthroat trout from F1 hybrids when selecting broodstock (no F1 hybrids were detected among 80 samples identified as pure). However, backcrosses of F1 hybrids were identified in random collections of adults from the lake as well as fry from Henrys Lake tributaries, indicating introgression. Present levels of rainbow trout introgression are most likely the product of past rainbow trout introductions and limited, intermittent spawning of hatchery-produced F1 hybrids with wild Yellowstone cutthroat, rather than the accidental crossing of F1 hybrids with cutthroat trout at the hatchery. Current levels of introgression are inadvertently maintained by (1) the inability of managers to phenotypically identify and exclude as broodstock individuals with low levels of rainbow trout introgression and (2) the limited, intermittent reproductive success of straying, hatchery-produced F1 hybrids.

Integration of the feline radiation hybrid and linkage maps.

The recent development of genome mapping resources for the domestic cat provides a unique opportunity to study comparative medicine in this companion animal which can inform and benefit both veterinary and human biomedical concerns. We describe here the integration and order comparison of the feline radiation hybrid (RH) map with the feline interspecies backcross (ISB) genetic linkage map, constructed by a backcross of F1 hybrids between domestic cat (Felis catus) and the Asian leopard cat (Prionailurus bengalensis). Of 253 microsatellite loci mapped in the ISB, 176 equivalently spaced markers were ordered among a framework of 424 Type I coding markers in the RH map. The integration of the RH and ISB maps resolves the orientation of multiple linkage groups and singleton loci from the ISB genetic map. This integrated map provides the foundation for gene mapping assessments in the domestic cat and in related species of the Felidae family.

Selection of Anopheles dirus for refractoriness and susceptibility to Plasmodium yoelii nigeriensis.

Two lines of the Oriental malaria vector mosquito Anopheles dirus species A (Diptera: Culicidae), one fully refractory and one fully susceptible to Plasmodium yoelii nigeriensis (an African rodent malaria parasite), were established after 17 generations of mass selection, followed by single female selection for one or two generations. Prior to selection, the stock colony of An. dirus was 17% refractory. Both lines of An. dirus produced abundant ookinetes that started to invade the midgut within 24h post-infection, as seen in histological sections. In most of the refractory mosquitoes, oocysts stopped development <12 h post-invasion, indicating a rapid defence mechanism. Dead P. y. nigeriensis parasites were apparently localized as small melanized spots (2-5 microm) seen in wet preparations of mosquito midguts dissected 5-7 days post infective bloodmeal. In some refractory An. dirus females, apart from the spots, a small number of totally encapsulated oocysts (c. 10 microm) were also present. These larger melanized parasites predominated in a few females: they appeared 2-3 days post-infection as a secondary delayed defence mechanism. The progeny of reciprocal matings between susceptible and refractory lines had approximately 50% susceptibility. Backcrosses of F1 hybrids with susceptible or refractory lines increased or decreased the susceptibility of backcross progeny accordingly. Overall, these results suggest polygenic control of susceptibility to P. y. nigeriensis infection. The refractory line of An. dirus showed normal susceptibility to natural infections of the human malarias P. falciparum and P. vivax from local patients.

Trypanosoma congolense: Inheritance of susceptibility to infection in inbred strains of mice

Inbred strains of mice have shown marked differences in susceptibility to infection with Trypanosoma congolense, as judged by survival and levels of parasitemia. The underlying genetic basis of the susceptibility was examined with F1 hybrids and backcrosses derived from mouse strains of high and low susceptibility. The influence of H-2 haplotype on susceptibility was studied using H-2 congenic resistant strains of mice. F1 hybrids between the most susceptible strain (A/J) and the least susceptible strain (C57B l/6) showed similar survival to that of the C57B l/6 parent. This was reflected in a similar undulating pattern of parasitemia, although the level of parasitemia was consistently higher in the F1 hybrids than in the C57B l/6. Backcrosses of the F1 hybrids with C57B l/6 also had a similar pattern of parasitemia although there was a greater scatter in survival times so that a few animals survived longer than either of the parental strains. Backcrosses of F1 hybrids with A/J showed a range of survival times; approximately 25% of these animals died during the period when the A/J mice died, approximately 25% had a similar survival to that of C57B l/6, while the remaining animals showed an intermediate duration of survival. All these backcrosses had a high initial peak of parasitemia; in about 70% of the mice the early parasitemia showed a distinct undulating pattern. F1 hybrids of A/J and C57B l/6 with C3H/He mice, which are known to be of intermediate susceptibility, were also examined. The degree of dominance for low susceptibility was much less pronounced in these hybrid combinations than in the A/J × C57B l/6 hybrids. The H-2 congenic resistant strains, all of which were on a C57B l/10 genetic background, showed a similar pattern of parasitemia and survival. However, although the majority of all these strains survived for more than 100 days, there was a significant difference in survival between the C57B l/10 mice and the H-2 congenic resistant strains. It was concluded that susceptibility of mice to T. congolense infection is likely to be under complex genetic control and that, at least in C57B l/mice, H-2 haplotype has little influence on susceptibility.