Balbiani rings in Drosophila: the Balbiani ring 1 is a common characteristic in several montium species.

Drosophila bicornuta, D. jambulina, D. biauraria, D. triauraria and D. quadraria, belonging to the montium subgroup of the melanogaster species, have a well-formed Balbiani ring (BR) in their salivary gland chromosomes. The BR has many similarities to the BR1 of D. auraria and D. serrata, two other montium species, indicating that the BR1 is a common characteristic in several species of the montium subgroup. It is suggested that the BR1 structure and its possible function(s), in contrast to the BR2, may have been selected during evolution.     

Isolation, characterization, and localization of beta-tubulin genomic clones of three Drosophila montium subgroup species.

Genomic libraries were constructed from three Drosophila species, namely Drosophila auraria, Drosophila serrata, and Drosophila kikkawai, belonging to the Drosophila montium subgroup of the Drosophila melanogaster species group. Clones containing beta-tubulin specific sequences were isolated, characterized by restriction endonuclease digestions and Southern hybridizations, and mapped by in situ hybridization on the polytene chromosomes of the species studied. The distribution of the beta-tubulin loci was found to be similar in D. montium species and D. melanogaster.     

The prominent role of ectopic pairing in the toroidal conformation of Drosophila polytene chromosome bands.

An examination of the polytene chromosomes of some Drosophila species from the montium subgroup, namely, D. auraria, D. serrata, D. jambulina and D. bicornuta, revealed an intimate relationship between ectopic pairing and the toroidal conformation of the polytene chromosome bands. The possible significance of these events is discussed.     

Phylogenetic biogeography and classification of the Drosophila montium species group (Diptera: Drosophilidae)

The Drosophila montium group is the largest clade of the subgenus Sophophora consisting of 94 palaeotropical species, whose phylogenetic relationships remain unclear. Here, I used a recent tree inferred from three nuclear genes and one mitochondrial gene for almost half of the species to reconstruct the historical biogeography of the group and propose a comprehensive classification for the totality of its species. The group originated in South-East Asia nearly 20 million years ago (mya), and dispersed to Africa in the Late Miocene. A second northward expansion into East Asia took place in the Pliocene. Based on morphological (male abdominal pigmentation and genitalia) and chorological traits congruent with the molecular tree, I divide the montium group into seven subgroups: parvula, montium, punjabiensis, serrata, kikkawai, seguyi and orosa. The polyphyletic status of some of the previously defined complexes (auraria, jambulina, serrata, kikkawai and nikananu) is also resolved.     

Reproductive isolation in the saltans group of Drosophila. I. The saltans subgroup

In order to study isolation within the salians subgroup, pair mating crosses involving its 7 species were performed. The results of testing different interspecific combinations varied from complete isolation at the insemination level to the production of fertile hybrids. Some isolation indices are proposed: Ii (isolation index at the insemination level), Fi (isolation index at the fertilization level), Ti (total isolation index), and Ai (average isolation index). A schema of the biological relationships on the basis of the isolation degree is also presented for the 7 species of the subgroup.Part of this study was included in the thesis submitted for the PH.D. degree at the University of São Paulo and was performed during the tenure of a fellowship supported by the Fundação de Amparo à Pesquisa do Estado de São Paulo.     

Mitochondrial DNA evolution in the Montium-species subgroup of Drosophila

Mitochondrial DNA (mtDNA) restriction-site maps for six species (10 strains) of the Drosophila montium subgroup were established. A total of 50 restriction sites were mapped, corresponding to 1.67% of the mtDNA genome. On the basis of differences in the restriction sites, nucleotide divergence (delta) was calculated for each pair of species (strains), and phylogenetic trees were constructed by using distance- matrix and parsimony methods. Comparison of the resultant phylogenetic trees shows that the sibling species D. auraria and D. quadraria are closely related. At the other extreme, considerable divergence was observed between the two strains of D. serrata and between D. serrata and D. birchii, a finding that contrasts with their grouping within the same species complex. Nevertheless, our data indicate that these six oriental montium species are rather closely related.      

Molecular phylogeny of the Drosophila auraria species complex and allied species of Japan based on nuclear and mitochondrial DNA sequences

We studied the phylogenetic relationships among the six species of the Drosophila auraria species complex and three species of the montium species subgroup endemic to Japan. To do this, we examined the DNA sequences of the nuclear ITS1 and the mitochondrial COII of these species. Intraspecific variation in ITS1 was observed for most of the species studied. However, interspecific variation was small and of the same magnitude as intraspecific variation among D. auraria, D. triauraria, and D. quadraria. In addition, the amount of intraspecific variation between COII sequences from D. auraria and D. triauraria was almost the same as that of the interspecific variation between these species. Phylogenetic trees based on ITS1 or COII data reveal that, with the exception of D. pectinifera, these species can be separated into two taxonomic groups. D. rufa formed a taxonomic lineage with two Japanese endemic species, D. asahinai and D. lacteicornis. Physiological and behavioral differences between D. auraria and D. triauraria have been reported, but our results differed from those previously reported. Such differences may be better explained by hybridization between D. auraria and D. triauraria than by similarity due to recent common ancestry. Both the ITS1- and COII-based phylogenetic trees placed D. pectinifera outside the auraria complex. However, D. pectinifera sequences were more similar to those of the auraria complex than those of D. punjabiensis, another species thought to be closely related to members of the auraria complex.     

Evolution of intrinsic reproductive isolation among four North American populations of Rhagoletis pomonella (Diptera: Tephritidae)

Across its range in North America, four geographically separated, ecologically and genetically diverged populations of hawthorn (Crataegus)‐infesting Rhagoletis pomonella (Diptera: Tephritidae) flies inhabit the Eje Volcánico Trans Mexicano (EVTM), the Sierra Madre Oriental (SMO), the Chiapas Highlands (CHIS) and the USA. Here, we tested whether these four populations are reproductively isolated by any intrinsic, nonhost‐related, pre‐ or postmating barriers to gene flow. Crossing experiments suggested that a low level of host‐independent prezygotic isolation may exist between hawthorn flies from EVTM and the three other populations, but only with respect to a slight reduction in copulation duration in EVTM matings. Some evidence for postmating isolation was found, again primarily involving EVTM crossed to SMO, CHIS and US flies. Certain crosses produced no (SMO male × EVTM female) or few (EVTM male × CHIS female; CHIS male × SMO female) F1 hybrid offspring. F2 crosses were generally fertile, except for US male × CHIS female matings. Inherent reproductive isolation therefore appears to be quantitative rather than absolute between populations, as the possibility for gene flow exists through at least some combinations of mating among EVTM, SMO, CHIS and US flies. Our results are consistent with a recently advanced hypothesis that episodic introgression from Mexico into the USA has played a role in providing genetic variation, facilitating sympatric host race formation and the adaptive radiation of the R. pomonella sibling species' complex in the USA. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 100 , 213–223.     

Effects of Courtship Song in Interspecific Crosses Among the Species of the Drosophila auraria Complex (Diptera: Drosophilidae)

The courtship behavior and the effects of courtship song in inter- and intraspecific crosses were studied in the four sympatric species of the Drosophila auraria complex: D. auraria, D. biauraria, D. subauraria, and D. triauraria. Orientation, tapping, and vibration (the repertoires of male courtship) were observed in both inter- and intraspecific crosses, suggesting that signals from heterospecific females were enough to elicit such male behaviors. The crossability tests with wingless or winged heterospecific males (tests for wing effects) revealed that winged heterospecific males copulated less than wingless ones in all four species but not all the pairwise cases. Since the crossability tests with aristaless females (deaf) or normal females showed essentially the same results as the tests for wing effects, we concluded that the sound produced by wing vibration plays an important role and that the wing movement itself is less important. These findings suggest that courtship songs are of great importance in mate discrimination and the sexual isolation between the species of this complex.     

Hybridization studies in the planthopper genus Ribautodelphax (Homoptera,Delphacidae)

Seven known Ribautodelphax species and four recently discovered taxa were crossed. Forty-three percent of the interspecific crosses were successful. Interspecific crosses generally resulted in low numbers of hybrids. In some combinations prolonged larval development of the hybrid males was observed. Backcrosses showed that both male and female hybrids were fertile. Reproductive isolation was mainly maintained by premating barriers. Insemination tests revealed that prolonged confinement of males and females lowered mating thresholds. The taxonomic status of the known species and new taxa was shown to be in need of revision. No indications for a hybrid origin of the pseudogamous triploid Ribautodelphax females were found.     

Fusarium subglutinans f. sp. Pini represents a distinct mating population in the gibberella fujikuroi species complex

Fusarium strains in the Gibberella fujikuroi species complex cause diseases on a variety of economically important plants. One of these diseases, pitch canker of Pinus spp., is caused by strains identified as Fusarium subglutinans f. sp. pini. Fertile crosses were detected between F. subglutinans f. sp. pini strains from South Africa, California, and Florida. F. subglutinans f. sp. pini strains were not cross-fertile with the standard tester strains of six of the seven other mating populations of G. fujikuroi. Sporadic perithecia with ascospores were obtained in two crosses with the mating population B tester strains. These perithecia were homothallic, and the ascospores derived from these perithecia were vegetatively compatible with the mating population B tester strain parent. We concluded that fertile F. subglutinans f. sp. pini isolates represent a new mating population (mating population H) of G. fujikuroi and that they belong to a unique biological species in a distinct taxon.     

Sexual reproduction in the soybean sudden death syndrome pathogen Fusarium tucumaniae

We investigated the sexual reproductive mode of the two most important etiological agents of soybean sudden death syndrome, Fusarium tucumaniae and Fusarium virguliforme. F. tucumaniae sexual crosses often were highly fertile, making it possible to assign mating type and assess female fertility in 24 South American isolates. These crosses produced red perithecia and oblong-elliptical ascospores, as is typical for sexual members of the F. solani species complex. Genotyping of progeny from three F. tucumaniae crosses confirmed that sexual recombination had occurred. In contrast, pairings among 17 U.S. F. virguliforme isolates never produced perithecia. Inter-species crosses between F. tucumaniae and F. virguliforme, in which infertile perithecia were induced only in one of the two F. tucumaniae mating types, suggest that all U.S. F. virguliforme isolates are of a single mating type. We conclude that the F. tucumaniae life cycle in S. America includes a sexual reproductive mode, and thus this species has greater potential for rapid evolution than the F. virguliforme population in the U.S., which may be exclusively asexual.     

Genetic architecture of the cryptic species complex of Acanthocyclops vernalis (Crustacea: Copepoda). II. Crossbreeding experiments, cytogenetics, and a model of chromosomal evolution

Collectively, populations of Acanthocyclops vernalis, a species complex of freshwater copepods, are remarkably similar as to morphology and DNA content, despite variability in chromosome number. Reproductive isolation had been reported among some populations, but with each new investigation the species boundaries and factors that may influence them appeared less clear. To clarify the pattern of biological species within this group of populations, we adopted a comprehensive approach and examined patterns of reproductive isolation in populations for which morphology, chromosome number, DNA content, and 18S rDNA sequences are known. In this study we established nine isofemale lines from four sites in Wisconsin and performed 266 crosses. Crosses within and among these lines were used to relate the degree of reproductive isolation to chromosome differences and to construct a model to explain the origin and maintenance of chromosome number variability. Different gametic and somatic chromosome numbers were observed among specimens within some isofemale lines. In a few cases, gametes with different haploid numbers were produced by a single female. Matings within isofemale lines always produced at least some reproductively successful replicate crosses (produced viable, fertile offspring). Crosses between lines from the same site showed reduced success relative to within-line crosses. Crosses between populations from distant sites showed limited genetic compatibility, producing viable, fertile F1 offspring but infertile F2 adults. One cross between lines with different chromosome numbers (one with 2n = 8 and one with 2n = 10) produced fertile viable offspring, which reproduced for at least 60 generations. These hybrids had either eight or nine chromosomes in the third generation of inbreeding, and eight chromosomes after 20 generations. These hybrids also had reduced nuclear DNA contents at the third generation, a level that persisted through the 20th generation. Successful backcrosses between some hybrids and their parental lines further demonstrated the potential for genetic compatibility among forms with different chromosome numbers. We propose a model in which alterations due to Robertsonian fusions, translocations, and/or loss of chromosomal fragments generate heritable variation, only some of which leads to reproductive isolation. Hence, some of the criteria traditionally used to recognize species boundaries in animals (morphology, DNA content, chromosome number) may not apply to this species complex.     

The Balbiani ring and the polytene chromosomes of Drosophila bicornuta.

A study of the BR1 and of the most prominent puffs during larval development and after in vitro ecdysterone treatment, as well as of the banding pattern and inverted tandem chromosomal duplications of the salivary gland chromosomes of Drosophila bicornuta, is presented in this report. These data are compared and discussed with those of D. auraria and D. serrata, two other montium species.     

Developmental constraints and convergent evolution in Drosophila sex comb formation

SUMMARY The most complex and diverse secondary sexual character in Drosophila is the sex comb (SC), an arrangement of modified bristles on the forelegs of a subclade of male fruit flies. We examined SC formation in six representative nonmodel fruit fly species, in an effort to understand how the variation in comb patterning arises. We first compared SC development in two species with relatively small combs, Drosophila takahashii , where the SCs remain approximately transverse, and Drosophila biarmipes , where two rows of SC teeth rotate and move in an anterior direction relative to other bristle landmarks. We then analyzed comb ontogeny in species with prominent extended SCs parallel to the proximodistal axis, including Drosophila ficusphila and species of the montium subgroup. Our study allowed us to identify two general methods of generating longitudinal combs on the tarsus, and we showed that a montium subgroup species ( Drosophila nikananu ) with a comb convergently similar in size, orientation and position to the model organism Drosophila melanogaster , forms its SC through a different developmental mechanism. We also found that the protein product of the leg patterning gene, dachshund (dac) , is strongly reduced in the SC in all species, but not in other bristles. Our results suggest that an apparent constraint on SC position in the adult may be attributable to at least two different lineage-specific developmental processes, although external forces could also play a role.     

Horizontal transmission versus vertical inheritance of P elements in Drosophila and Scaptomyza: has the M-type subfamily spread from East Asia?

Polymerase chain reaction (PCR) screening for P elements was carried out in 77 species with a primer set highly specific for the M-type subfamily. In the course of this search M-type elements were detected in 29 species: In the melanogaster (subgroups montium and rhopaloa ) and obscura species groups of Drosophila (25 out of 71 species examined), and in the genus Scaptomyza (four out of six species). M-type elements are present in all species of the montium subgroup investigated so far (21), but occur only sporadically in other groups. Within the montium subgroup 20 species possess only incomplete copies, only one species ( D. lacteicornis ) harbours apparently full-sized elements. In contrast, outside the montium subgroup almost all species with M-type elements carry full-sized copies suggesting transpositional activity, at least in the recent past. The interior section of the full-sized M-type element of D. lacteicornis was partially sequenced (936 bp). In addition, the complete sequences of four internally deleted M-type elements of D. lacteicornis, D. rufa, D. quadraria , and D. triauraria were determined. Sequence comparisons (including sequence data from previous investigations) revealed striking discrepancies between P element phylogeny and the cladogenesis of their host species. Among several possible pathways for interspecific transfers of M-type elements, we favour the hypothesis assuming the invasion of Scaptomyza as well as the obscura group species of Drosophila via independent transmission routes originating from Asian species of the montium subgroup of Drosophila . The logical geographic scenario for these events would be East-Asia.     

用RAPD标记探讨金色果蝇复合种内的遗传分化

以随机扩增多态性DNA（RAPD）技术分析了金以果蝇复合种的5个姊妹种（Drosophile auraria、D．biauraria、D.triauraria、D．quadraria和D．subauraria）共12个地理种群的遗传变异。在40种10碱基随机引物中有30种引物对每个种群都可获得令人满意的扩增结果。在161个RAPD标记中有129个呈多态性。以遗传距离为尺度用UPGMA法构建了该复合种的聚类关系图。聚类分析结果结果表明：D.subauraria处于聚类关系图的基部,为该复合种中最早分化的种类;其后才是D.biauraria的分化,而D.auraria和D.quadraria的分化相对较晚。此结果不支持以往认为D.quadraria为金色果蝇复合种祖先种的观点。RAPD分子标记为金色果蝇复合种的系统演化的研究提供了新的、更有价值的信息。     

Neurospora species in bakeries

S. YASSIN AND A. WHEALS. 1992. Neurospora species may occur as spoilage organisms in bakeries and bakery products. A survey of two bakeries over a 4 month period produced 315 individually isolated strains which were characterized with respect to six features: conidial colour, protoperitheciality, mating type and percentage dark-coloured (fertile) ascospores in crosses with type specimens of Neurospora crassa, N. sitophila and N. intermedia. We concluded that (i) N: crassa of both mating types was by far the most abundant but N. sitophila and N. intermedia were also represented; (ii) conidial colour was not a good species criterion; (iii) the percentage of fertile ascospores produced in crosses with several type specimens provided a reliable and simple species indicator; (iv) identification of individuals based on these six characteristics suggested that a large number of genetically different organisms were present in the collection; and (v) bakery infection was not due to endemic contamination in the bakery or in raw materials but from repeated external infection, perhaps on contaminated returned goods.     

CROSSING RELATIONSHIPS AMONG DIPLOID SPECIES OF THE SOLANUM NIGRUM COMPLEX IN NORTH AMERICA

Crosses were made in all combinations of the six diploid species of the Solarium nigrum complex that occur in North America. Some interspecific pollinations failed to yield viable seed; successful crosses gave rise to moderately to highly sterile F₁ hybrids. Results of interspecific crosses suggested phylogenetic relationships that were not completely in accord with those suggested by morphology. Interspecific crosses also gave varied results. All interpopulational crosses within S. interius and S. sarachoides produced fully fertile hybrids. In contrast, hybrids within S. americanum and S. douglasii varied from fully fertile to almost completely sterile. Populations of S. pseudogracile could be divided into two groups which are geographically separated but not morphologically differentiated. Fully fertile hybrids resulted from crosses within a group, whereas crosses between groups gave hybrids with reduced fertility. Four crossing groups were observed within 5. nodiflorum; three of the parental groups are sympatric and are morphologically differentiated. Although hybrid sterility in interspecific crosses is sometimes used to support delimitation of species, the presence of sterility in intraspecific crosses suggests that such an interpretation is unwarranted for the S. nigrum complex. Hybrid sterility, therefore, is not considered to have special taxonomic significance in this complex.     

江苏省稻瘟病菌有性态的研究*

用标准菌株对１９９７～１９９９年在江苏省吴江市,宜兴市、通州市、高邮市和赣榆县采集的３２５个猪瘟病菌单孢分离菌株的可育性和交配型进行了测定,结果表明江苏省稻瘟病菌菌株的育性较低,可交配率为２２．７７％,可育率仅为７．０８％。不同年份、不同地区采集的猪瘟病菌茵株的性亲和力和交配型有较大的差异,三年的交配率分别为２６．６１％、８．２６％和３３．６４％;温州地区和赣榆地区菌株的交配率相对较高,分别为２６．１５％和２５．４２％,宜兴地区菌株的交配率较低,只有１５．３８％。江苏省稻瘟病菌菌株的交配型在不同年份亦出现很大差别,１９９７年２９个可交配菌株中有２１个菌株表现为ＭＡＴ１－２ 交配型,而１９９９年３６个可交配茵株均为ＭＡＴ１－１交配型。用江苏省稻瘟病菌的可育菌株进行互交,２５个组合中只有６个组合能产生子囊壳和子囊,但均不产生子囊孢子,提示江苏省稻瘟病菌在田间产生健康有性后代的几率不大。对杂交后代的遗传学分析表明,菌株的交配型是受单基因控制的。