Inversion polymorphism in populations of Drosophila sulphurigaster albostrigata and Drosophila nasuta albomicans from Phuket,Thailand

Two species of the Drosophila nasuta subgroup of the Drosophila immigrans group, D. sulphurigaster albostrigata and D. nasuta albomicans were investigated in this study. Collections of both species were made from Phuket, Thailand. Both species have similar salivary chromosomes, with four autosomal arms and one sex chromosome arm, and both are highly polymorphic for paracentric inversions. D. s. albostrigata accounted for the majority of the isolines collected and exhibited the greater number of inversions. One inversion, C₁, was common to both species, indicating common ancestry.A non-random distribution of inversions was observed on the proximal end of chromosome II in both D. s. albostrigata and D. n. albomicans. An inter-collection comparison revealed that both rigid and flexible chromosomal polymorphism were operating in the two species, with a seasonal variation noted for one inversion in D. s. albostrigata. A non-random association of two inversions was observed in D. n. albomicans.Based on a comparison of the indices of crossing over, both D. s. albostrigata and D. n. albomicans were found to be more heterozygous than in previous studies, with D. n. albomicans appearing to have evolved further than D. s. albostrigata.Based on a thesis submitted for the degree of Ph. D. at the University of Queensland.     

Chromosomal polymorphism in species of the Drosophila nasuta complex in East and South East Asia

This paper is presented as a review of ten years collection of the D. nasuta complex from the Asian region by Dr. Wharton B. Mather. It incorporates the analysis of chromosomal polymorphisms with regard to their geographical distribution and then uses the polymorphisms to establish phylogenetic relationships between D. sulfurigaster albostrigata, D. albomicans and D. kohkoa.     

Inversion polymorphism in species of the Drosophila nasuta subgroup from Thailand

Thailand populations of three species of the D. nasuta complex have been analysed for the presence of paracentric inversions. D. albomicans and D. sulfurigaster albostrigata were collected from Phuket, Chiang Mai and the River Kwai, whilst D. kohkoa was found only in Phuket and the River Kwai. Chromosomal polymorphism was studied in respect to geographical distribution. The Phuket populations of all three species proved to be highly polymorphic by comparison with the River Kwai and Chiang Mai populations. The heterozygosity frequencies of inversions were calculated and the variations interpreted as a result of adaptation to local ecogeographical conditions. Shared polymorphisms revealed that D. kohkoa and D. s. albostrigata are more closely related to D. albomicans than they are to each other.Based on a thesis submitted for the degree of Ph.D. in the University of Queensland.     

Inversion polymorphism in Southeast Asian populations of the Drosophila nasuta subgroup

Heterozygosity for chromosomal arrangements was investigated in four species of the nasuta complex. D. sulfurigaster albostrigata, D. albomicans and D. kohkoa are highly polymorphic whereas D. pulaua is monomorphic. Inversions were identified with the aid of photographic chromosome maps. The geographic distribution and frequencies of inversions detected and their possible phylogenies were discussed. Most inversions in D.s. albostrigata and D. albomicans occur on chromosome IIL. In D. kohkoa there is marked preponderance of inversions on chromosome III. Non-random association of certain inversions was noted. Variation in frequencies of inversions is interpreted as a result of adaptation to local ecogeographic conditions. Shared extant polymorphisms indicate phylogenetic relationships between species.The contents of this paper were incorporated in a Ph.D. thesis accepted by the University of Queensland in 1978. The author's present address is: Unit Genetik, Universiti Kebangsaan Malaysia, Jalan Pantai Baru, Kuala Lumpur, Malaysia.     

Linkage disequilibrium in laboratory populations of Drosophila nasuta

Summary Natural populations of Drosophila nasuta are polymorphic for many gene arrangements. Two non-overlapping inversions of the third chromosome, III-2 and III-35, are most common and display extreme linkage disequilibrium. Six randomly mating laboratory stocks, each founded by one gravid female heterozygous in coupling for both III-2 and III-35, were observed after 32 generations. Significant linkage disequilibrium was observed in all stocks. Recombinants were found in only two stocks. The absence of effective recombination in some stocks and its presence in others might be due to different genotypic backgrounds. We suggest that natural selection, influencing recombination rates in several ways, and intrachromosomal epistasis between the two inversions were the main factors for the maintenance of linkage disequilibrium in D. nasuta.     

Hybridization,transgressive segregation and evolution of new genetic systems in<Emphasis Type="Italic">Drosophila</Emphasis>

Introgressive hybridization facilitates incorporation of genes from one species into the gene pool of another. Studies on long-term effects of introgressive hybridization in animal systems are sparse.Drosophila nasuta (2n = 8) andD. albomicans (2n = 6)—a pair of allopatric, morphologically almost identical, cross-fertile members of thenasuta subgroup of theimmigrans species group-constitute an excellent system to analyse the impact of hybridization followed by transgressive segregation of parental characters in the hybrid progeny. Hybrid populations ofD. nasuta andD. albomicans maintained for over 500 generations in the laboratory constitute new recombinant hybrid genomes, here termed cytoraces. The impact of hybridization, followed by introgression and transgressive segregation, on chromosomal constitution and karyotypes, some fitness parameters, isozymes, components of mating behaviour and mating preference reveals a complex pattern of interracial divergence among parental species and cytoraces. This assemblage of characters in different combinations in a laboratory hybrid zone allows us to study the emergence of new genetic systems. Here, we summarize results from our ongoing studies comparing these hybrid cytoraces with the parental species, and discuss the implications of these findings for our understanding of the evolution of new genetic systems. This paper is dedicated to the memory of our teacher, Prof. N. B. Krishnamurthy.     

The chromosomes of two Drosophila races: D. nasuta nasuta and D. nasuta albomicans III. Localization of nucleolar organizer regions

In the two Drosophila races D. n. nasuta and D. n. albomicans the nucleolar-organizer regions (NORs) have been localized on metaphase chromosomes by a combined acid treatment and silver-staining technique. In both Drosophila races one NOR is present on the heterochromatic Y chromosomes and another on the microchromosomes. Such a NOR distribution has not yet been reported in Drosophila. It is suggested that this distribution has evolved from an original type with NORs on the X and Y chromosomes.     

Male accessory gland secretory protein polymorphism in natural populations of <Emphasis Type="Italic">Drosophila nasuta nasuta</Emphasis> and <Emphasis Type="Italic">Drosophila sulfurigaster neonasuta</Emphasis>

Male accessory gland secretory protein polymorphism was analysed in natural populations of Drosophila nasuta nasuta and D. sulfurigaster neonasuta for the first time, using SDS-PAGE to score polymorphism of these proteins in 2788 individuals of D. n. nasuta and 2232 individuals of D. s. neonasuta from 12 different populations from southern India. A total of 25 and 18 variant protein phenotypes were identified in D. n. nasuta and D. s. neonasuta, respectively. Protein fractions of group III were more polymorphic than those from groups I and II. The results show that accessory gland secretory proteins show high levels of polymorphism, irrespective of species or habitat. Moreover, we have used the variation in the accessory gland proteins to assess the extent of divergence between the species and to infer their population structure. The study suggests that though both D. n. nasuta and D. s. neonasuta belong to the same subgroup, they differ in population structure, as far as accessory gland protein polymorphism is concerned.     

Evidence for no sexual isolation between Drosophila albomicans and D. nasuta

Sexual isolation, the reduced tendency to mate, is one of the reproductive barriers that prevent gene flow between different species. Various species‐specific signals during courtship contribute to sexual isolation between species. Drosophila albomicans and D. nasuta are closely related species of the nasuta subgroup within the Drosophila immigrans group and are distributed in allopatry. We analyzed mating behavior and courtship as well as cuticular hydrocarbon profiles within and between species. Here, we report that these two species randomly mated with each other. We did not observe any sexual isolation between species or between strains within species by multiple‐choice tests. Significant difference in the courtship index was detected between these two species, but males and females of both species showed no discrimination against heterospecific partners. Significant quantitative variations in cuticular hydrocarbons between these two species were also found, but the cuticular hydrocarbons appear to play a negligible role in both courtship and sexual isolation between these two species. In contrast to the evident postzygotic isolation, the lack of sexual isolation between these two species suggests that the evolution of premating isolation may lag behind that of the intergenomic incompatibility, which might be driven by intragenomic conflicts.     

The chromosomes of two Drosophila races: D. nasuta nasuta and D. nasuta albomicana

Interracial hybridization between Drosophila nasuta nasuta (2n=8) and D. n. albomicana (2n=6) has resulted in the evolution of two new karyotypic strains, called Cytoraces I and II. Males and females of Cytorace I have 2n=7 and 2n=6 respectively. The reconstituted karyotype is totally new in its composition, the chromosomes being drawn from both the parental races. The individuals of Cytorace II have 2n=6. Even though the chromosomes of the parental races are duly represented in the F₁, there is selective retention/elimination of certain chromosomes in the succeeding generations during which repatterning of the karyotype has taken place. Dynamics of each one of the parental chromosomes are presented and its implications re discussed.We dedicate this paper to the memory of the founder of our Department, the late Prof. M.R. Rajasekarasetty on the occasion of the Silver Jubilee of our Department     

Mitochondrial DNA evolution in theDrosophila nasuta subgroup of species

Summary TheDrosophila nasuta group consists of about 12 closely related species distributed throughout the Indo-Pacific region. They are of great interest because of their evolutionary idiosyncrasies including little morphological differentiation, the ability to intercross in the laboratory often producing fertile offspring, and substantial chromosomal evolution. Studies of metric traits, reproductive isolation, and chromosomal and enzyme polymorphisms have failed to resolve the phylogeny of the species. We report the results of a survey of the mitochondrial DNA (mtDNA) restriction patterns of the species. The phylogeny obtained is consistent with other available information and suggests thatD. albomicans may represent the ancestral lineage of the group. The amount of polymorphism in local populations (=1.0% per site) is within the typical range observed in other animals, includingDrosophila. The degree of differentiation between species is, however, low: the origin of the group is tentatively dated about 6–8 million years ago. This study confirms the usefulness of mtDNA restriction patterns for ascertaining the phylogeny of closely related species.     

Comparative ecophysiology ofLarix kaempferi (Lamb.) Carr. andAbies veitchii Lindl. III. Analysis of factors determining the differences in root depth

A new method for analyzing growth is proposed to elucidate factors responsible for the differences in root depth ofLarix kaempferi andAbies veitchii seedlings. Root depth (D) divides into three factors;

Glue proteins inDrosophila nasuta

Larval glue protein fractions ofDrosophila nasuta nasuta were analyzed by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. Seven major and at least four minor glue protein fractions were recognized. Six of the major fractions are glycosylated. They migrate as three prominent doublets (>100, 43, and 30/28 kd). The synthesis of traceable amounts of these major fractions begins already during the second as well as during the early stages of the third larval instar. The 43-kd and the 30/28-kd fractions are coded by X-chromosomal genes. They are probably clustered within the huge puff of division 10, which is the most prominent X-chromosomal puff in the polytene chromosomes of the third larval instar. Complex posttranslational modification of all but one major glue protein fraction (14 kd) leads to the formation of about 15 different protein fractions in the final glue product. The amount of glue protein produced byD. n. nasuta larvae (in relation to the total saliva proteins) is nearly twice the amount produced byD. melanogaster larvae (ca. 55 and 32%, respectively). This work was supported by the University Grants Commission, New Delhi, India, the Deutscher Akademischer Austauschdienst, FR Germany (to S.R.R.), and the Deutsche Forschungsgemeinschaft (Ka 309/9-1).     

Drosophila virilis histone gene clusters lacking H1 coding segments

Summary Approximately 30–40% ofDrosophila virilis DNA complementary to clonedDrosophila histone genes is reduced to 3.4-kilobase-pair (kbp) segments by Bgl I or Bgl II digestion. The core histone genes of a 3.4-kbp Bgl II segment cloned in the plasmid pDv3/3.4 have the same order as theD. melanogaster core histone genes in the plasmid cDm500: . Nonetheless, pDv3/3.4 and cDm500 have different histone gene configurations: In pDv3/3.4, the region between the H2B and H3 genes contains 0.35 kbp and cannot encode histone H1; in cDm500, the region contains 2.0 kbp and encodes histone H1. The lack of an H1 gene between the H2B and H3 genes in 30–40% ofD. virilis histone gene clusters suggests that changes in histone gene arrays have occurred during the evolution ofDrosophila. The ancestors of modernDrosophila may have possessed multiple varieties of histone gene clusters, which were subsequently lost differentially in thevirilis andmelanogaster lineages. Alternatively, they may have possessed a single variety, which was rearranged during evolution. The H1 genes ofD. virilis andD. melanogaster did not cross-hybridize in vitro under conditions that maintain stable duplexes between DNAs that are 75% homologous. Consequently,D. virilis H1 genes could not be visualized by hybridization to an H1-specific probe and thus remain unidentified. Our observations suggest that the coding segments in the H1 genes ofD. virilis andD. melanogaster are >25% divergent. Our estimate of sequence divergence in the H1 genes ofD. virilis andD. melanogaster seems high until one considers that the coding sequences of cloned H1 genes from the closely related speciesD. melanogaster andD. simulans are 5% divergent.     

Introgressive hybridization and evolution of a novel protein phenotype: Glue protein profiles in thenasuta-albomicans complex ofDrosophila

Glue proteins are tissue-specific proteins synthesized by larval salivary gland cells ofDrosophila. InDrosophila nasuta nasuta andD. n. albomicans of thenasuta subgroup, the genes that encode the major glue protein fractions are X-linked. In the present study, these X-linked markers have been employed to trace the pattern of introgression ofD. n. nasuta andD. n. albomicans genomes with respect to the major glue protein fractions in their interracial hybrids, called cytoraces. These cytoraces have inherited the chromosomes of both parents and have been maintained in the laboratory for over 400–550 generations. The analysis has revealed that cytoraces withD. n. albomicans X chromosome show eitherD. n. nasuta pattern or a completely novel pattern of glue protein fractions. Further, quantitative analysis also shows lack of correlation between the chromosomal pattern of inheritance and overall quantity of the major glue protein fractions in the cytoraces. Thus, in cytoraces the parental chromosomes are not just differentially represented but there is evidence for introgression even at the gene level.     

The chromosomes of two Drosophila races: Drosophila nasuta nasuta and Drosophila nasuta albomicana V. Introgression and the evolution of new karyotypes

Reciprocal crosses were made between an Indian strain of D. n. nasuta (2n=8) and the Thailand strain of D. n. albomicana (2n=6). Hybrids were fertile. They were inbred for over four years. Later, the karyotypes of the hybrid populations were analysed. In the hybrid progeny of the cross between D. n. nasuta females and D. n. albomicana males, there were six types of kaotypes. Of these, only two types had a diploid content of chromosomes. They were males with 2n=7 and females with 2n= 8 , while others were aneuploids. This hybrid population is designated as Cytorace III. On the other hand, hybrid progeny of the reciprocal cross had 2n-8 in both males and females; and there was no karyotypic variation. This hybrid population is named as Cytorace IV. The composition of these new karyotypes of Cytorace III and IV have been presented and compared with those of Cytorace I and II reported by Ramachandra and Ranganath (1986).     

Cultivation of the sapropelic ciliate Plagiopyla nasuta Stein and isolation of the endosymbiont Methanobacterium formicicum

The sapropelic ciliate Plagiopyla nasuta was isolated and cultured in monoculture. Optimal conditions for growth were: 15–20°C, pH about 7, and about 2% of oxygen in the headspace. Cultures of P. nasuta produced methane. Epifluorescence microscopy revealed the presence of methanogenic bacteria as endosymbionts. An endosymbiont of the ciliate was isolated and identified as Methanobacterium formicicum. In the ciliate cell these methanogens were found to be closely associated with microbody-like organelles. No mitochondria could be detected.     

The chromosomes of two Drosophila races: D. nasuta nasuta and D. nasuta albomicana

The microchromosomes of the totally cross fertile Drosophila races, D. nasuta nasuta and D. nasuta albomicana have been studied in nietaphase and polytene nuclei. In metaphase the microchromosome of D. n. albomicana is nearly five times longer than the homologous chromosome in D. n. nasuta. As shown by C-banding these length differences are mainly due to a massive addition of heterochromatin to the D. n. albomicana chromosome. In polytene nuclei these striking heterochromatin differences between the microchromosomes of the two Drosophila races cannot be observed. Analysis of the polytene banding pattern shows that the microchromosomes of both races differ by an inversion and by a duplication, present only in D. n, albomicana. The location and orientation of the duplicated regions in D. n. albomicana leads to a specific loop like chromosome configuration. On the basis of these differences within the Drosophila races studied it is assumed that the karyotype of D. n. albomicana is a more recent evolutionary product.     

The inheritance of mating songs in two cryptic,sibling lacewing species (Neuroptera: Chrysopidae: Chrysoperla)

Speciation often involves incremental responses to natural selection and results in large scale genomic changes, but it may also occur abruptly and with little genetic imprint, as seen in some complexes of cryptic species. Recent attention has focused on sexual selection in rapid speciation, because it can disrupt premating signals that mediate reproductive isolation. Some models require that environmental adaptation assist sexual selection during speciation, while others show that populations can diverge through mate choice alone. We propose that speciation involving environmental adaptation in premating signals is likely to have a polygenic basis, while speciation due to arbitrary changes in premating signals could be accompanied by changes at just a few loci. The sibling species Chrysoperla plorabunda and Chrysoperla johnsoni belong to a large complex of cryptic species of green lacewings, and meet all criteria for speciation via sexual selection. We perform a genetic analysis of line crosses between them, focusing on their substrate-borne premating songs. Measurements of seven song features and their principal components are compared among parentals, F₁ and F₂ hybrids, and backcrosses. The distributions of phenotypes are consistent with a model of more than one, but not many, genes. Sex linkage and/or maternal effects are negligible. C. plorabunda is dominant for most traits affecting mate choice. Bayesian analyses of segregation variance show significant additive and epistatic effects on line means. A Bayesian Castle–Wright estimate suggests that relatively few effective factors are responsible for variation in volley period ( ), number of volleys per song ( ), and PC-1 ( ). Our results are consistent with simple genetic architecture of songs, supporting a role for major genes in premating isolation and strengthening the notion that mate choice alone, without significant environmental adaptation, has been responsible for generating new lacewing species.     

Analysis of cytoplasmic membrane from wild type and mutant Paracoccus denitrificans containing excess nitrate reductase protein and cytochrome b

Cytoplasmic membranes were isolated from wild type and mutants strain M-1 of Paracoccus denitrificans grown with low aeration to promote synthesis of nitrate reductase protein and cytochrome b. The presence of 10-100-fold excess of nitrate reductase in the wild type or the corresponding enzymically inactive protein in the mutant did not significantly affect respiratory oxidase activities with NADH, succinate or TMPD-ascorbate as electron donor. A cytochrome b-nitrate reductase complex was resolved by isoelectric focussing of Triton X-100 solubilized membranes from the wild type grown with azide and from the mutant, whereas the enzyme complex from nitrate-grown wild type was not resolved from cytochrome c. Preparations from azideinduced wild type or from the mutant could be a suitable source of the cytochrome b associated with nitrate reductase for more detailed studies.Non standard abbreviations IEF isoelectric focussing - TMPD N, N, N, N-tetramethylphenylenediamine - SDS-PAGE Sodium dodecyl sulphate polyacrylamide gel electrophoresis