Enzyme patterns in two species of Xenopus and their hybrids

Differences in the isozyme patterns of Xenopus laevis and Xenopus mulleri have been utilized to examine the expression of alleles of both species in hybrid animals. Mitochondrial MDH and tetrazolium oxidase phenotypes were examined during the development of non-hybrid embryos of each species and of reciprocal hybrids. Early stages of the hybrids resemble the enzyme phenotype of the maternal parent. Appearance of paternal enzyme takes place just prior to the active feeding tadpole stage for both mitochondrial MDH and oxidase. The maternal effect disappears shortly thereafter in early feeding tadpoles, at which point reciprocal hybrids have identical isozyme patterns. There is no evidence for a predominance of one species over the other. Examination of feeding tadpoles and adult toads indicates that both laevis and mulleri expression is stable. The appearance of paternal mitochondrial MDH does not correspond to the time when other mitochondrial components begin to increase in Xenopus. Multiple bands of MDH in both species and of oxidase in laevis are probably not due to the aggregation of subunits produced by different alleles at the same locus. There is no evidence for the formation of “hybrid” molecules consisting of subunits of both species.     

Expression of the lactate dehydrogenase genes in Xenopus species hybrids

The lactate dehydrogenase isozyme patterns of six Xenopus species-hybrid combinations have been investigated. The genes from both parental species were equally expressed in all of them. “Hybrid” isozymes originating from aggregation of subunits from both parents could be visualized and demonstrated random aggregation of enzyme subunits. Reciprocal crosses displayed the same pattern demonstrating also random aggregation of subunits.     

Cytogenetics of semi-fertile triploid and aneuploid intergeneric vine cacti hybrids

Crosses between the diploid Hylocereus polyrhizus, as the female parent, and the tetraploid Selenicereus megalanthus, as the male parent, yielded triploid and aneuploid hybrids. The fruits of these hybrids combined the attractive appearance of Hylocereus fruits with the delicious taste of S. megalanthus fruits. The aim of this work was to assess the fertility and breeding potential of the triploid and aneuploid hybrids with a view to developing an improved vine cactus crop. Pollen mother cells at metaphase I revealed univalents, bivalents, trivalents, and occasionally quadrivalents. Chromosome distribution at anaphase I revealed different classes of chromosome segregation as well as lagging chromosomes. At metaphase II, parallel and tripolar spindles were observed. The occurrence of triads was frequent, whereas dyads were rarely observed. Pollen stainability varied among the clones studied ranging from 9.8% to 18.6%. The diameters of the stained pollen grains varied widely, probably as a result of the number of chromosomes. Despite the allotriploid origin of our hybrids, functional female and male gametes were produced in considerable proportions, most likely as a result of balanced chromosome segregation. The triploid and aneuploid clones studied yielded viable seeds whose number per fruit was strongly dependent on the pollen donor.     

Developmental characteristics of two closely related species of Allonemobius and their hybrids

Summary Two species of ground crickets, Allonemobius fasciatus (from Corvallis, 44.3 N) and A. socius (from Gainesville, 30.3 N), and their hybrids were compared for nymphal development, wing form, and embryonic diapause. A. fasciatus matured more rapidly at a short photoperiod than at a long photoperiod and also than A. socius which showed no developmental response to photoperiod. The proportion of long-winged forms varied with nymphal photoperiod greatly in A. fasciatus but slightly in A. socius. A. fasciatus laid only diapause eggs, while A. socius responded to a long parental photoperiod and a high incubation temperature by averting diapause. A. fasciatus showed temperature-dependent shift of the diapause stage butA. socius entered diapause only at the early stage. These differences are related to their voltinism and distribution.Reciprocal crosses between the two species produced viable hybrids. In development rate, female hybrids were intermediate between their parents while male hybrids were closer to their mothers, suggesting that the development rate is controlled primarily by the X chromosome. Overdominance occurred in wing form; reciprocal crosses produced either higher or lower proportions of long-winged forms than their parents. Diapause of F1 hybrid embryos was determined only by the female parent and the paternal effect appeared in F2 embryos.     

A developmental analysis of the histone patterns of two species of Xenopus and their hybrids

The histone patterns in Xenopus laevis and X. borealis have been examined at various developmental stages using acid-urea polyacrylamide electrophoresis. Qualitative differences between the two species have been demonstrated in the H1 fraction. These differences are not affected by alkaline phosphatase digestion. Quantitative changes during development in both H1 and H4 fractions have also been observed. In addition, histones from hybrids of the two species have been examined. H1 histones characteristic of both species are present by the neurula stage in laevis ♀ × borealis ♂ hybrids. Quantitative changes observed in borealis development are mimicked in laevis ♀ × borealis ♂ hybrid development.     

Elimination and duplication of particular Hordeum vulgare chromosomes in aneuploid interspecific Hordeum hybrids

Summary Seeds formed in crosses Hordeum lechleri (6x) x H. vulgare (2x and 4x), H. arizonicum (6x) x H. v. (2x), H. parodii (6x) x H. v. (2x), and H. tetraploidum (4x) x H. v. (2x) produced plants at high or rather high frequencies through embryo rescue. Giemsa C-banding patterns were used to analyze chromosomal constitutions and chromosomal locations on the methaphase plate. Among 100 plants obtained from H. vulgare (2x) crosses, 32 plants were aneuploid with 2n=29 (1), 28 (3), 27 (13), 26 (5), 25 (4), 24 (4), or 22 (2); 50 were euploid (12 analyzed), and 18 were polyhaploid (5 analyzed). Four plants had two sectors differing in chromosome number. Two of four hybrids with H. vulgare (4x) were euploid and two were aneuploid. Parental genomes were concentrically arranged with that of H. vulgare always found closest to the metaphase centre. Many plants showed a certain level of intraplant variation in chromosome numbers. Except for one H. vulgare (4x) hybrids, this variation was restricted to peripherally located non-H. vulgare genomes. This may reflect a less firm attachment of the chromosomes from these genomes to the spindle. Interplant variation in chromosome numbers was due to the permanent elimination or, far less common, duplication of the centrally located H. vulgare chromosomes in all 34 aneuploids, and in a few also to loss/gain of non-H, vulgare chromosomes. This selective elimination of chromosomes of the centrally located genome contrasts conditions found in diploid interspecific hybrids, which eliminate the peripherally located genome. The difference is attributed to changed genomic ratios. Derivatives of various H. vulgare lines were differently distributed among euploid hybrids, aneuploids, and polyhaploids. Chromosomal constitutions of hypoploid hybrids revealed a preferential elimination of H. vulgare chromosomes 1, 5, 6, and 7, but did not support the idea that H. vulgare chromosomes should be lost in a specific order. H. vulgare SAT-chromosomes 6 and 7 showed nucleolar dominance. Aneuploidy is ascribed to the same chromosome elimination mechanism that produces haploids in cross-combinations with H. vulgare (2x). The findings have implications for the utilization of interspecific Hordeum hybrids.     

Performance of aneuploid backcross hybrids between the crop Brassica napus and its wild relative B. rapa

• Crossings between the diploid wild Brassica rapa (AA , 2n = 20) and the tetraploid cultivar B. napus (AACC , 2n = 38) can readily be made. Backcrosses to the wild B. rapa (BC ₁) produce aneuploids with variable chromosome numbers between 20 and 29. How does survival and performance relate to DNA content of plants?
• Growth of the BC ₁ plants was measured in the lab. One plant in the F₁ self‐pollinated spontaneously and produced abundant F₂ seeds that were also examined. The number of C‐chromosomes was estimated from DNA values obtained with flow cytometry.
• Average DNA value of the BC ₁ was similar to that of the parents, which shows that C‐chromosomes do not reduce success of pollen or embryos. The average DNA value in the F₂ was 13% higher than in the F₁, suggesting that extra C‐chromosomes facilitated gamete success and/or embryo survival. Under both optimal and drought stress conditions growth and survival of BC ₁ hybrids was similar to that of B. rapa . No significant correlations existed between growth or survival and DNA value.
• Aneuploid plants were not inferior under the conditions of the growth room and may persist in nature. We discuss other factors, such as herbivory, that could prevent hybrid establishment in the field.

     

Mitochondrial ribosomal proteins in Xenopus laevis/X. mulleri interspecific hybrids.

The large subunits of mitochondrial ribosomes were isolated from two related frog species, Xenopus laevis and X. mulleri, and their proteins were compared by two-dimensional polyacrylamide gel electrophoresis. Three of the proteins observed in X. laevis are absent from X. mulleri, and four of the proteins observed in X. mulleri are absent from X. laevis. More than these seven such species-specific proteins may occur.Reciprocal crosses between frogs of the two species gave two groups of F1 hybrids. Nuclear genes in these hybrids derive equally from both species, while mitochondrial DNA (and therefore mitochondrial rRNA) derived exclusively from the maternal species. Electrophoretic analyses of the large subunit proteins of these F1 animals revealed that four of the species-specific proteins are present only when their corresponding species was the mother. While this result is consistent with the coding of these four proteins by mitochondrial DNA, it does not provide evidence against nuclear coding of these proteins. A fifth protein is absent from both F1 hybrids. A sixth is present in both F1 hybrids, and a seventh is present only when its corresponding species was the father. We conclude that at least these latter two mitochondrial ribosomal proteins are encoded by nuclear genes.     

Developmental stability in hybrids between the sibling species pair,Drosophila melanogaster and Drosophila simulans

Drosophila melanogaster and its sibling species D. simulans were hybridized in the laboratory to test the hypothesis that developmental homeostasis in hybrids between two species having no prior gene flow would be significantly reduced. Developmental stability was assessed by measuring fluctuating asymmetry for three bilateral traits: sternopleural chaetae, wing length, and fronto-orbital plus frontal chaetae. Male F₁ hybrids showed no decrease in developmental stability compared to males of parental species. Female hybrids showed significant fluctuating asymmetry compared to other flies. The results are discussed with respect to ideas about coadaptation and gene flow based upon previous studies of hybrid developmental stability.     

Developmental basis of pronephric defects in Xenopus body plan phenotypes.

We have used monoclonal antibodies that recognize the pronephric tubules or pronephric duct to explore the induction of the embryonic kidney in developing Xenopus embryos. Morphogenesis of the pronephros was examined in UV-ventralized and lithium-dorsalized embryos. We find that the pronephric tubules are present in all but the strongest UV-induced phenotypes, but absent from relatively moderate lithium phenotypes. Interestingly the pronephric duct, which develops from the ventroposterior portion of the pronephric anlage, is missing from more of the mild UV phenotypes than are pronephric tubules. The loss of the capacity to form pronephroi in UV-ventralized embryos is caused by the loss of tissues capable of inducing the pronephric mesoderm, as marginal zone explants from ventralized embryos are still competent to respond to pronephric-inductive signals. Explant recombination experiments indicate that the tissue responsible for both the loss of pronephroi in UV-ventralized embryos and the induction of pronephroi during normal development is the anterior somites. The absence of pronephroi in relatively mild lithium phenotypes has a developmental basis different from that of the UV phenotype, as explants from lithium-treated embryos are effective inducers of pronephroi in recombinants with competent mesoderm, even though they themselves do not form pronephroi in isolation. Together these data indicate that dorsal tissues, especially the anterior somites, are responsible for the establishment of the intermediate mesoderm and the induction of the embryonic kidneys and that even mild dorsalization destroys the capacity to form cells competent to receive this signal.     

Developmental Capacity of Aneuploid Xenopus Species Hybrids

Diploid as well as triploid Xenopus interspecific hybrids generate aneuploid eggs because of the presence, at meiosis, of univalent chromosomes which are presumably distributed at random. Zygotes obtained from such eggs, fertilized by either normal or UV-irradiated sperm, were analysed for their developmental capacities. All monosomics die in the course of embryogenesis, whereby optimum capacities correspond closely with those observed in monosomic mammalian embryos, especially in mice. In contrast, hyperdiploid Xenopus are relatively viable: although many die exhibiting the'haploid syndrome'or various other abnormalities, 8% of them reach metamorphosis, and 1–2% become adults. Of the latter, the karyotype was established in 13 individuals. Among them, 8–16 supernumerary chromosomes were found to be present.     

Developmental expression of fibrillarin and U3 snRNA in Xenopus laevis.

Fibrillarin is one of the protein components that together with U3 snRNA constitute the U3 snRNP, a small nuclear ribonucleoprotein particle involved in ribosomal RNA processing in eucaryotic cells. Using an antifibrillarin antiserum for protein detection and a fibrillarin cDNA and a synthetic oligonucleotide complementary to U3 snRNA as hybridization probes, the expression of these two components has been studied during Xenopus development. Fibrillarin mRNA is accumulated early in oogenesis, like many other messengers, and translated during oocyte growth. Fibrillarin protein is thus progressively accumulated throughout oogenesis to be assembled with U3 snRNA and used for ribosome production in the amplified nucleoli. After fertilization, the amount of U3 snRNA decreases while the maternally accumulated fibrillarin mRNA is maintained and utilized to produce more protein. After the mid-blastula transition, stored fibrillarin is assembled with newly synthesized U3 snRNA and becomes localized in the prenucleolar bodies and reforming nucleoli.