Postmating Reproductive isolation between strains of Drosophila willistoni

Speciation can occur through the presence of reproductive isolation barriers that impede mating, restrict cross-fertilization, or render inviable/sterile hybrid progeny. The D. willistoni subgroup is ideally suited for studies of speciation, with examples of both allopatry and sympatry, a range of isolation barriers, and the availability of one species complete genome sequence to facilitate genetic studies of divergence. D. w. willistoni has the largest geographic distribution among members of the Drosophila willistoni subgroup, spanning from Argentina to the southern United States, including the Caribbean islands. A subspecies of D. w. willistoni, D. w. quechua, is geographically separated by the Andes mountain range and has evolved unidirectional sterility, in that only male offspring of D. w. quechua females × D. w. willistoni males are sterile. Whether D. w. willistoni flies residing east of the Andes belong to one or more D. willistoni subspecies remains unresolved. Here we perform fecundity assays and show that F1 hybrid males produced from crosses between different strains found in Central America, North America, and northern Caribbean islands are reproductively isolated from South American and southern Caribbean island strains as a result of unidirectional hybrid male sterility. Our results show the existence of a reproductive isolation barrier between the northern and southern strains and suggest a subdivision of the previously identified D. willistoni willistoni species into 2 new subspecies.     

Drosophila subpulchrella, a new species of the Drosophila suzukii species subgroup from Japan and China (Diptera: Drosophilidae)

Drosophila (Sophophora) subpulchrella Takamori and Watabe, sp. nov., of the D. suzukii subgroup in the D. melanogaster species group, is described from Japan and southern China, and compared with its sibling species, D. pulchrella Tan et al. distributed in the Yun‐Gui Highland, south‐western China. The results of cross‐experiments show a complete pre‐mating isolation between D. subpulchrella and D. pulchrella.     

Rapid evolution of courtship song pattern in Drosophila willistoni sibling species

Courtship behaviours may provide a more reliable means of identifying reproductively isolated taxa than traits such as morphology or many genetic markers. Here we describe the courtship songs of the Drosophila willistoni sibling species group, which consists of several species and subspecies. We find that song pattern is species-specific, despite significant differences among strains within species. D. paulistorum has the most variable song pattern, which reflects this species' traditional subdivision into semispecies. All the other species could be unambiguously identified by song. The major differences among these species was in the interpulse interval, as has been found in other studies of fly song. However, the interpulse intervals of the species studied here were often multimodal. This was partly due to the presence of multiple song types within the courtship repertoire, but it also reflected changes in interpulse interval within a song type by some males. Unusually, some species had distinctively patterned variation in interpulse interval. Song must have evolved rapidly within the species complex, probably due to sexual selection.     

Divergence in wing morphology among sibling species of the Drosophila buzzatii cluster

The Drosophila buzzatii species cluster consists of the sibling species D. buzzatii, D. koepferae, D. serido, D. borborema, D. seriema, D. antonietae and D. gouveai, all of which breed exclusively in decaying cactus tissue and, except for D. buzzatii (a colonizing subcosmopolitan species), are endemic to South America. Using a morphometric approach and multivariate analysis of 17 wing parameters, we investigated the degree of divergence in wing morphology among the sibling species of this cluster. Significant differences were obtained among the species and discriminant analysis showed that wing morphology was sufficiently different to allow the correct classification of 98.6% of the 70 individuals analysed. The phenetic relationships among the species inferred from UPGMA cluster analysis based on squared Mahalanobis distances (D²) were generally compatible with previously published phylogenetic relationships. These results suggest that wing morphology within D. buzzatii cluster is of phylogenetic importance.     

果蝇Drosophila nasuta亚群系统发育问题的探讨

果蝇Ｄｒｏｓｏｐｈｉｌａｎａｓｕｔａ亚群由在１４个处于不同物种分化阶段的种,亚种和分类元组成,这个亚群的物种有许多进化上的独特之处,使得它在物种分化研究方面倍受关注,然而,在形态学,生殖隔离,染色体和同工酶多态,线粒体ＤＮＡＲＦＬＰ,求偶歌特征惟及线粒体和核基因序列分析等方面的研究都未能清楚地阐明这一亚群的系统进化关系,本文综合分析了关于这一亚群的进化遗传学的研究结果,并提出了有待进一步的一些问题     

Mutagenesis of the catalytic subunit of rabbit muscle protein phosphatase-1

We have generated site-directed mutants of the catalytic subunit of rabbit muscle ppase-1. Since it is known that ppase-1 and ppase-2A are highly susceptible to inactivation by sulfhydryl reagents, we have mutagenized the six cysteine residues conserved between these two enzymes to serines. The six mutants were purified to near homogeneity by affinity chromatography on inhibitor-2-Sepharose and characterized. All six exhibited enzymatic activity. These results indicate that the catalytic mechanism of ppase-1 is different from that of the protein tyrosine phosphatases which involve a cysteinyl phosphate intermediate.     

Determinants of the nucleolar targeting of protein phosphatase-1

The ubiquitously expressed protein Ser/Thr phosphatase-1 isoforms PP1alpha, PP1beta and PP1gamma1 are dynamically targeted to distinct, but overlapping cellular compartments by associated proteins. Within the nucleus of HeLa cells, EGFP-tagged PP1gamma1 and PP1beta were predominantly targeted to the nucleoli, while PP1alpha showed a more diffuse distribution. Using PP1 chimaeras and point mutants we show here that a single N-terminal residue, i.e., Gln20 for PP1alpha, Arg19 for PP1beta and Arg20 for PP1gamma1 accounts for their distinct subnuclear distribution. Our data also suggest that the N-terminus of PP1beta and PP1gamma1 harbours an interaction site for one or more nucleolar interactors.     

Identification of the interaction sites of Inhibitor-3 for protein phosphatase-1

Inhibitor-3 is a potent inhibitor of protein phosphatase-1, with an IC₅₀ in the nanomolar range for the inhibition of the dephosphorylation of phosphorylase a. Human Inhibitor-3 possesses a putative protein phosphatase-1 binding motif, ³⁹KKVEW⁴³. We provide direct evidence that this sequence is involved in PP1 interaction by examining the effects of site-directed mutations of Inhibitor-3 on its ability to inhibit protein phosphatase-1. A second interaction site whose deletion led to loss of inhibitory potency was identified between residues 65 and 77. The existence of two interaction sites is consistent with the high inhibitory potency of Inhibitor-3, and with current models for other inhibitor and targeting proteins that interact with protein phosphatase-1 with high affinity.     

Molecular, morphological and behavioural data reveal the presence of a cryptic species in the widely studied Drosophila serrata species complex

The Drosophila serrata species complex from Australia and New Guinea has been widely used in evolutionary studies of speciation and climatic adaptation. It is believed to consist of D. serrata, D. birchii and D. dominicana, although knowledge of the latter is limited. Here we present evidence for a previously undescribed cryptic member of the D. serrata species complex. This new cryptic species is widespread in far north Queensland, Australia and is likely to have been previously mistaken for D. serrata. It shows complete reproductive isolation when crossed with both D. serrata and D. birchii. The cryptic species can be easily distinguished from D. serrata and D. birchii using either microsatellite loci or visual techniques. Although it occurs sympatrically with both D. serrata and D. birchii, it differs from these species in development time, viability, wing size and wing morphology. Its discovery explains patterns of recently described mitochondrial DNA divergence within D. serrata, and may also help to clarify some ambiguities evident in early evolutionary literature on reproductive incompatibility within the D. serrata species complex.     

Diagnosis of sibling species of Drosophila involved in the colonization of North America by D. subobscura

To determine the effects of the recent colonization of the west coast of North America by the Palaearctic species Drosophila subobscura on the dynamics of the Drosophila populations, the sibling species D. athabasca and D. azteca must be classified unambiguously. We have characterized these two species using three molecular techniques: allozymes, mtDNA and RAPDs. All three techniques allow the classification of any individual as belonging to either species. The study of five localities in northern California and southern Oregon show that the area of overlap is larger than previously described.     

Interspecific and intraspecific comparisons of the period locus in the Drosophila willistoni sibling species

The period (per) locus has received much attention in molecular evolution studies because it is one of the best studied "behavioral genes" and because it offers insight into the evolution of repetitive sequences. We studied most of the coding region of per in Drosophila willistoni and confirmed previously observed patterns of conservation and divergence among distantly related species. Five regions are so highly diverged that they cannot be aligned, whereas a region encompassing the PAS domain is very conserved. Structural and nucleotide polymorphism patterns in the willistoni group are not the same as those observed in previously studied species. We sequenced the region homologous to the highly polymorphic threonine-glycine repeat of D. melanogaster in multiple strains of D. willistoni, as well as in other members of willistoni group, and found an unusual amount of conservation in this region. However, the next nonconserved region downstream in the sequence is quite variable and polymorphic for the number of repeated glycines. The glycine codon usage is significantly different in this glycine repeat as compared to other parts of the gene. We were able to plot the directionality of change in the glycine repeat region onto a phylogeny and find that the addition of glycines is the general trend with the diversification of the willistoni group.      

Naturally occurring quantitative variants of acid phosphatase-1 in Drosophila melanogaster

We have examined 111 wild Drosophila melanogaster lines for cis-acting quantitative variants of the Acph-1 gene, which codes for acid phosphatase-1 (ACPH). Three variants with obvious, reproducible phenotypes were isolated. All variants acted equally on all tissues and developmental stages examined. No recombinants were detected between one quantitative variant and the site determining the electrophoretic mobility of Acph-1 among 3885 flies examined. Several enzymatic properties of the variant enzymes were tested, including the K _m values for two substrates, inhibition by three different inhibitors, and thermal stability; the variant enzymes behaved identically to the wild-type enzyme in all cases. Immunological titration experiments showed that the variant enzymes had the same enzyme activity per molecule of ACPH as the wild-type enzyme. These results suggest that the quantitative variants we have identified are altered in the regulatory portion of Acph-1 so as to produce altered numbers of normal ACPH molecules.This work was supported by NIH Grant 21548. MAJ was supported by NIH Predoctoral Training Grant GM07413.     

萼花臂尾轮虫种复合体内三个姐妹种生活史特征的比较研究

应用生命表统计学方法,在13℃、18℃、23℃和28℃下对采自芜湖市莲塘湖和荷花塘水体中的萼花臂尾轮虫三个姐妹种HE1、HE3和LE9的生活史特征进行了比较研究。结果表明,三个姐妹种生命表参数间的差异因温度的不同而异。13℃下,姐妹种LE9的生命期望、平均寿命和世代时间都显著长于姐妹种HE3,LE9的净生殖率和种群内禀增长率均最大,三个姐妹种所产后代中的混交雌体百分率无显著的差异。18℃下,姐妹种LE9的生命期望和平均寿命显著长于姐妹种HE3,姐妹种LE9的世代时间显著长于姐妹种HE1和HE3,姐妹种 HE3的种群内禀增长率显著高于姐妹种LE9,姐妹种HE3所产后代中的混交雌体百分率最高,三个姐妹种的净生殖率无显著的差异。23℃下,姐妹种HE1的生命期望、平均寿命和世代时间均最长,姐妹种HE1所产后代中的混交雌体百分率显著低于HE3,三个姐妹种的净生殖率和种群内禀增长率均无显著的差异。28℃下,三个姐妹种的生命期望、平均寿命、世代时间和种群内禀增长率均无显著的差异;姐妹种HE1的净生殖率最低;姐妹种HE3所产后代中的混交雌体百分率最高。三个姐妹种的生命表参数对升高的温度的响应也不相同。温度、姐妹种及两者间的交互作用对轮虫的世代时间、平均寿命、出生时的生命期望和轮虫所产后代中的混交雌体百分率均有显著的影响;温度和姐妹种对轮虫的净生殖率和种群内禀增长率均有极显著影响,但两者的交互作用对其无显著性影响。     

萼花臂尾轮虫(Brachionus calyciflorus)种复合体内三个姐妹种生活史特征比较

采自芜湖市莲塘湖和荷花塘水体中的萼花臂尾轮虫3个姐妹种HE1、HE3和LE9在13℃、18℃、23℃和28℃下的生命表参数间的差异表现为13℃和18℃下姐妹种LE9的生命期望和平均寿命都显著长于姐妹种HE3,23℃下姐妹种HE1的生命期望和平均寿命均最短;13℃下姐妹种LE9的世代时间显著长于姐妹种HE3,18℃下和23℃姐妹种LE9和HE1的世代时间分别最长和最短;13℃下姐妹种LE9的净生殖率最高,28℃下姐妹种HE1的净生殖率最低;13℃下姐妹种LE9的种群内禀增长率最高,18℃下姐妹种HE3的种群内禀增长率显著高于姐妹种LE9;18℃和28℃下姐妹种HE3所产后代中的混交雌体百分率最高,23℃下姐妹种HE1所产后代中的混交雌体百分率显著低于HE3.3个姐妹种的生命表参数对升高的温度的响应也不相同.温度、姐妹种及两者间的交互作用对轮虫的世代时间、平均寿命、出生时的生命期望和轮虫所产后代中的混交雌体百分率均有显著的影响;温度和姐妹种对轮虫的净生殖率和种群内禀增长率均有极显著影响,但两者的交互作用对其无显著性影响.     

Ⅰ型蛋白磷酸酶研究进展

Ⅰ型蛋白磷酸酶(PP1)属丝/苏氨酸磷酸酶的一种,在生物体中广泛存在,参与调节多种重要的生理功能,包括转录、翻译、代谢、细胞生长及分化等.PP1分子结构表面的3个凹槽及β 12-β 13 Loop环结构,它在底物与抑制剂的结合方面起决定作用.近期研究发现,Loop环结构除了是抑制剂的结合部位之外,对整个酶分子的结构和性质都起重要作用.功能研究也证明PP1还参与HIV-1转录过程的调节,并且与老年性痴呆等多种疾病密切相关.主要对PP1的组织分布、分子结构、酶学特性、催化机制以及生物学功能等方面进行了相应的综述.     

Molecular evolution of the histone 3 multigene family in the Drosophila melanogaster species subgroup

Molecular evolution of the histone multigene family was studied by cloning and sequencing regions of the histone 3 gene in the Drosophila melanogaster species subgroup. Analysis of the nucleotide substitution pattern showed that in the coding region synonymous changes occurred more frequently to A or T in contrast to the GC-rich base composition, while in the 3' region the nucleotide substitutions were most likely in equilibrium. These results suggested that the base composition at the third codon position of the H3 gene, i.e., codon usage, has been changing to A or T in the Drosophila melanogaster species subgroup.     

Evolution of abdominal pigmentation differences across species in the Drosophila dunni subgroup

The Drosophila dunni subgroup displays a nearly perfect latitudinal cline in abdominal pigmentation that likely resulted from selective forces acting in the habitat of each species during speciation. Here we characterize the nature of this clinal variation by developing a quantitative measure to assess variation in abdominal pigmentation within and between the D. dunni subgroup species. Using discriminant analysis, we confirm the existence of a cline and find that our quantitative measure of pigmentation distinguishes each of the species with singular efficacy. We then combine our quantitative phenotypic analysis of pigmentation with the phylogeny of the D. dunni subgroup species and map the species relationships into the three-dimensional morphological space defined by our pigmentation measures. In this manner, we can visualize how the species have traversed the morphological pigmentation space during the course of speciation. Our analysis reveals that natural selection has caused overall intensity of pigmentation among the northernmost species of the cline to converge. Along with this convergence in phenotype has been a relaxation in expression of sexual dimorphism in these species, indicating a possible shift in the relative intensity of natural and sexual selection. Our analysis indicates an accelerated rate of change in pigmentation for the darkest species in addition to this species evolving a novel abdominal pigmentation trait.     

Life cycle differences among Brazilian sandflies of the Lutzomyia longipalpis sibling species complex

The developmental cycles of five Brazilian populations of the Lutzomyia longipalpis Lutz & Neiva species complex (Diptera: Psychodidae) were compared under laboratory conditions. Three of the populations were derived from insects collected in allopatric sites at Natal (Rio Grande do Norte State), Jacobina (Bahia State) and Lapinha Cave (Minas Gerais State). The other two originated from Sobral (Ceará State), where the males of two sympatric species can be distinguished by the presence of one (1S) or two (2S) pairs of abdominal spots. The results of the present study clearly show that all three populations whose males produce C₁₆ pheromones and use pulse-type copulation songs (Jacobina, Lapinha Cave and Sobral 1S) are more easily adapted to the colonization conditions used in our laboratory, producing larger egg batches, with higher survival and an overall faster developmental cycle. This contrasts with populations producing C₂₀ male pheromones and using burst-type copulation songs (Natal and Sobral 2S) that produce smaller egg batches, have higher oviposition mortality and a slower rate of development under identical laboratory conditions. In conclusion, these phenological differences are a further indication of the differentiation of the siblings within the Lu. longipalpis species complex.