Evolutionary Potential of Parthenogenesis-Bisexual Lineages within Triploid Apomictic Thelytoky in Cacopsylla ledi (Flor, 1861) (Hemiptera, Psylloidea) in Fennoscandia.

A widely accepted hypothesis is that parthenogenesis is an evolutionary dead end since it is selectively advantageous in the short term only but results in lowered diversification rates. Triploid apomictic parthenogenesis might represent an exception, as in favorable environments, triploid females are able to produce rare males and diploid females. The aim of the present study was to analyze the modes of reproduction and their evolutionary implications in the parthenogenetic psyllid Cacopsylla ledi (Flor, 1861) from Fennoscandia. The cytogenetic assessment of ploidy levels and the analysis of the COI haplotype revealed two geographically separated bisexual lineages implying genuine bisexual populations. The southern lineage occurring south of latitude 65° N in Finland showed a COI haplotype different from that of parthenogenetic triploids in the same population but identical to the haplotype of specimens in a genuine bisexual population in the Czech Republic. This allows us to suggest that bisexuals in southern Fennoscandia represent the original bisexual C. ledi. By contrast, in the northern bisexual lineage north of latitude 65° N, rare males and diploid females carried the same haplotype as triploids in the same population, having been produced by the triploids. In the Kola Peninsula, a genuine bisexual population of presumably rare male/diploid female origin was discovered. As this population is geographically isolated from populations of the ancestral bisexual C. ledi, it can develop into a new bisexual species through peripatric speciation during evolution. Our findings demonstrate that apomictic triploid parthenogenesis is not necessarily an evolutionary dead end but is able to lead to the emergence of a new bisexual species of parthenogenetic origin.

The Incidence of Wolbachia Bacterial Endosymbiont in Bisexual and Parthenogenetic Populations of the Psyllid Genus Cacopsylla (Hemiptera, Psylloidea).

Wolbachia is one of the most common intracellular bacteria; it infects a wide variety of insects, other arthropods, and some nematodes. Wolbachia is ordinarily transmitted vertically from mother to offspring and can manipulate physiology and reproduction of their hosts in different ways, e.g., induce feminization, male killing, and parthenogenesis. Despite the great interest in Wolbachia, many aspects of its biology remain unclear and its incidence across many insect orders, including Hemiptera, is still poorly understood. In this report, we present data on Wolbachia infection in five jumping plant-lice species (Hemiptera, Psylloidea) of the genus Cacopsylla Ossiannilsson, 1970 with different reproductive strategies and test the hypothesis that Wolbachia mediates parthenogenetic and bisexual patterns observed in some Cacopsylla species. We show that the five species studied are infected with a single Wolbachia strain, belonging to the supergroup B. This strain has also been found in different insect orders (Lepidoptera, Hemiptera, Plecoptera, Orthoptera, Hymenoptera, Diptera) and even in acariform mites (Trombidiformes), suggesting extensive horizontal transmission of Wolbachia between representatives of these taxa. Our survey did not reveal significant differences in infection frequency between parthenogenetic and bisexual populations or between males and females within bisexual populations. However, infection rate varied notably in different Cacopsylla species or within distinct populations of the same species. Overall, we demonstrate that Wolbachia infects a high proportion of Cacopsylla individuals and populations, suggesting the essential role of this bacterium in their biology.

Description of two new species of the genus Cacopsylla Ossiannilsson, 1970 (Hemiptera, Psylloidea) from northern Fennoscandia recognized by morphology, cytogenetic characters and COI barcode sequence.

Based on chromosomal, molecular and morphological analyses, two new Cacopsylla Ossiannilsson, 1970 species are described, C. lapponica S. Nokkala & Ch. Nokkala, sp. nov. and C. borealis S. Nokkala et Ch. Nokkala, sp. nov. (Hemiptera, Psylloidea). C. lapponica is a rare bisexual alpine species living on Vaccinium uliginosum Linnaeus, 1753 above tree line on northern hills, where it forms sympatric populations with C. myrtilli W. Wagner, 1947. So far, the species has been found in northern Finland, Utsjoki and Kilpisjärvi, and in northern Sweden, Abisko. The chromosome number in males is 2n = 12+X(0), characteristic of psyllids. The species is easily distinguished from C. myrtilli by its conspicuously smaller size mainly due to difference in wing size. Additional morphological differences are found in the length of antennae, female genital plates and male parameres. C. borealis, in turn, is a relatively common apomictic parthenogenetic species with 5n = 60 + XXXXX living on the same host plant, Ledum palustre Linnaeus, 1753, as C. ledi (Flor, 1861) and occasionally forming sympatric populations with it. No males have been recorded in C. borealis. Its distribution range reaches at least from northern Fennoscandia to Lake Baikal in the East. C. borealis can be distinguished from C. ledi by differences in the length and width of antennae, dark brown markings on the wing and female terminal structures. For molecular analysis, a 638 bp fragment of the mitochondrial COI gene was sequenced. C. lapponica differs from the cohabitating C. myrtilli by 20 fixed nucleotide substitutions (uncor rected p-distance 3.13 %), while C. borealis differs from C. ledi by 21 fixed nucleotide substitutions (uncorrected p-distance 3.29 %). Molecular phylogeny construction (ML and BI) reveals two highly divergent clades, one comprising two bisexual species, C. lapponica and C. fraudatrix Labina & Kuznetsova, 2012, and the other clade comprising the parthenogenetic species C. borealis, C. myrtilli, and C. ledi. Within this clade, C. borealis is more closely associated with C. myrtilli than with C. ledi.

Characteristics of parthenogenesis in Cacopsylla ledi (Flor, 1861) (Hemiptera, Sternorryncha, Psylloidea): cytological and molecular approaches.

Characteristics of parthenogenesis in Cacopsylla ledi (Flor, 1861) were analyzed using cytological and molecular approaches. In all three populations studied from Finland, i.e. Turku, Kustavi and Siikajoki, males were present at a low frequency but were absent from a population from Vorkuta, Russia. In a follow-up study conducted in the Turku population during 2010-2016, the initial frequency of males was ca. 10 % and showed no intraseasonal variation, but then dramatically decreased down to approximately 1-2 % level in seasons 2015-2016. Male meiosis was chiasmate with some traces of chromosomal fragmentation and subsequent fusions. In most females, metaphase in mature eggs included 39 univalent chromosomes which indicated apomictic triploidy. Only a small fraction of females was diploid with 13 chiasmate bivalents. The frequency of diploid females approximately equaled that of males. COI barcode analyses showed that triploid females (N = 57) and diploids (7 females and 5 males) displayed different haplotypes, demonstrating that triploid females reproduced via obligate parthenogenesis. The rarity of diploids, along with the lack of males' preference towards diploid females, suggested that most likely diploids were produced by rare triploid females which shared the same haplotype with the diploids (not found in the present analysis). Minimum haplotype diversity was detected in the Turku population, but it was much higher in Vorkuta with some indication for the mixed origin of the population. We suggest that functional diploids produced in a parthenogenetic population can give rise either to a new parthenogenetic lineage or even to a new bisexual species.

Meiosis in rare males in parthenogenetic Cacopsylla myrtilli (Wagner, 1947) (Hemiptera, Psyllidae) populations from northern Europe.

For studying meiosis in males, large samples of Cacopsylla myrtilli (Wagner, 1947) (Hemiptera, Psyllidae) were collected in Norway, Sweden, Finland and northwest Russia. In addition to all-female populations, males were present in 10 out of 47 populations; still, all populations were highly female-biased, the proportion of males varying from 0.1% to 9.1%. These males are thus rare or so-called spanandric males. Males in northern Norway, Finland and northwest Russia showed normal chiasmate meiosis, while complete absence of chiasmata due to asynapsis was found in males collected in Norway and northern Sweden. In asynaptic meiosis, all univalent chromosomes divided during the first meiotic division resulting in incomplete second meiotic division and formation of diploid sperms. Hence, males in these populations are nonfunctional and do not contribute to the genetic constitution of the population, but appear in every generation as reversals from apomictic parthenogenesis and the mode of parthenogenesis is of obligatory type.

Cytogenetics of the true bug infraorder Cimicomorpha (Hemiptera, Heteroptera): a review.

The Cimicomorpha is one of the largest and highly diversified infraorders of the Heteroptera. This group is also highly diversified cytogenetically and demonstrates a number of unusual cytogenetic characters such as holokinetic chromosomes; m-chromosomes; multiple sex chromosome systems; post-reduction of sex chromosomes in meiosis; variation in the presence/absence of chiasmata in spermatogenesis; different types of achiasmate meiosis. We present here a review of essential cytogenetic characters of the Cimicomorpha and outline the chief objectives and goals of future investigations in the field.

The distribution and population sex ratio of Cacopsylla myrtilli (W. Wagner, 1947) (Hemiptera: Psylloidea).

In Cacopsylla myrtilli (W. Wagner, 1947) bisexual populations, all-female populations and populations heavily biased towards females have been described. In the present paper all the available data on the distribution and population sex ratio of C. myrtilli are summarized. New data obtained by the authors are also presented. First records for Russia are described from Siberia and the southern part of the Far East.

Dm nxf1/sbr gene affects the formation of meiotic spindle in female Drosophila melanogaster.

The small bristles (sbr) gene of Drosophila melanogaster belongs to the family of nuclear export factor (NXF) genes that participate in mRNA nuclear export. During meiosis, females of Drosophila melanogaster that carry various combinations of mutant alleles of the Dm nxf1/sbr gene exhibit disruption of the division spindle and misalignment of chromosomes at the metaphase plate. Meiosis of sbr ( 5 ) /+ females is characterized by the formation of tripolar spindles during the first cell division. According to the sequencing results, the sbr ( 5 ) (l(1)K4) lethal allele is a deletion of 492 nucleotides. In SBR(5) protein, 57 of the 146 amino acids that have been lost by deletion belong to the NTF2-like domain.

First evidence of polyploidy in Psylloidea (Homoptera, Sternorrhyncha): a parthenogenetic population of Cacopsylla myrtilli (W. Wagner, 1947) from northeast Finland is apomictic and triploid.

This paper reports results of the first cytogenetic study of parthenogenetic psyllids, carried out on an asexual population of the holarctic species Cacopsylla myrtilli W. Wagner from northeast Finland. Preparations of mature eggs extracted from females revealed 39 univalent chromosomes in prophase and metaphase cells. Hence, female meiosis is of apomictic type and replaced by a modified mitosis. The karyotype consists of 3n = 39 (36 + XXX). Clearly, the population is triploid, the haploid number being n = 12 + X as characteristic of the genus Cacopsylla as a whole. As typical for Psylloidea, the chromosomes are holokinetic, only slightly varying in size and without any visible markers, rendering impossible the precise identification of triplets of homologous chromosomes in the triploid complement. The distribution of bisexual and parthenogenetic populations of C. myrtilli throughout the world is briefly given, and a possible origin of the triploid parthenogenetic population is discussed.

Cytogenetic characterization of the Trinidad endemic, Arachnocoris trinitatus Bergroth: the first data for the tribe Arachnocorini (Heteroptera: Cimicomorpha: Nabidae).

As an extension of the ongoing cytogenetic studies of the bug family Nabidae (Heteroptera: Cimicomorpha), the first evidence for the tribe Arachnocorini (the subfamily Nabinae), with reference to the Trinidad endemic, Arachnocoris trinitatus Bergroth, is provided. This is an attempt to gain a better insight into the evolution, systematics and within-family relationships of the family Nabidae. The studies were conducted using a number of cytogenetic techniques. The male karyotype (chromosome number and size; sex chromosome system; NOR location; C-heterochromatin amount, distribution and characterization in terms of the presence of AT-rich and GC-rich DNA), and male meiosis with particular emphasis on the behavior of the sex chromosomes in metaphase II are described. Also investigated are the male and female internal reproductive organs with special reference to the number of follicles in a testis and the number of ovarioles in an ovary. A. trinitatus was found to display a number of characters differentiating it from all hitherto studied nabid species placed in the tribe Nabini of the subfamily Nabinae, and in the tribe Prostemmatini of the subfamily Prostemmatinae. Among these characters are chromosome number 2n = 12 (10 + XY), the lowest within the family, nucleolus organizer regions (NORs) situated on the autosomes rather than on the sex chromosomes as is the case in other nabid species, and testes composed of 3 follicles but not of 7 as in other nabids. All the data obtained suggest many transformations during the evolution ofA. trinitatus.

First evidence of sex chromosome pre-reduction in male meiosis in the Miridae bugs (Heteroptera).

The karyotype and male meiosis of Macrolophus costalis Fieber (Insecta, Heteroptera, Miridae) were studied using C-banding, AgNOR-banding and DNA sequence specific fluorochrome staining. The chromosome formula of the species is 2n = 28(24+X1X2X3Y). Male meiotic prophase is characterized by a prominent condensation stage. At this stage, two sex chromosomes, "X" and Y are positively heteropycnotic and always appeared together, while in autosomal bivalents homologous chromosomes were aligned side by side along their entire length, that is, meiosis is achiasmatic. At metaphase I, "X" and Y form a pseudobivalent and orient to the opposite poles. At early anaphase I, the "X" chromosome disintegrates into three separate small chromosomes, X1, X2, and X3. Hence both the autosomes and sex chromosomes segregate reductionally in the first anaphase, and separate equationally in the second anaphase. This is the first evidence of sex chromosome pre-reduction in the family Miridae. Data on C-heterochromatin distribution and its composition in the chromosomes of this species are discussed.

Automictic and apomictic parthenogenesis in psocids (Insecta: Psocoptera).

Karyotypes and meiosis patterns in three obligatory thelytokous Psocoptera species have been studied for the first time. Females of Aaroniella badonneli (Danks) display 9 chiasmatic bivalents in oocyte metaphase I (2n = 18), hence meiosis is of the automictic type. Females of Ectopsocus meridionalis Ribaga and Valenzuela sp. display 3n = 27, and 27 univalent chromosomes are present in oocyte metaphase I. Thus, meiosis in these species is of the apomictic type.

Role of Jun N-terminal Kinase (JNK) signaling in the wound healing and regeneration of a Drosophila melanogaster wing imaginal disc.

When a fragment of a Drosophila imaginal disc is cultured in growth permissive conditions, it either regenerates the missing structures or duplicates the pattern present in the fragment. This kind of pattern regulation is known to be epimorphic, i.e. the new pattern is generated by proliferation in a specialized tissue called the blastema. Pattern regulation is accompanied by the healing of the cut surfaces restoring the continuous epithelia. Wound healing has been considered to be the inductive signal to commence regenerative cell divisions. Although the general outlines of the proliferation dynamics in a regenerating imaginal disc blastema have been well studied, little is known about the mechanisms driving cells into the regenerative cell cycles. In this study, we have investigated the role of Jun N-terminal Kinase (JNK) signaling in the wound healing and regeneration of a Drosophila wing imaginal disc. By utilizing in vivo and in vitro culturing of incised and fragmented discs, we have been able to visualize the dynamics in cellular architecture and gene expression involved in the healing and regeneration process. Our results directly show that homotypic wound healing is not a prerequisite for regenerative cell divisions. We also show that JNK signaling participates in imaginal disc wound healing and is regulated by the physical dynamics of the process, as well as in recruiting cells into the regenerative cell cycles. A model describing the determination of blastema size is discussed.

Dynamics of decapentaplegic expression during regeneration of the Drosophila melanogaster wing imaginal disc.

Regeneration of an imaginal disc involves highly ordered proliferation and pattern regulation of the newly formed tissue. Although the general principles of imaginal disc regeneration have been extensively studied, knowledge of the underlying molecular mechanisms is far from complete. Results from other model organisms suggest that regeneration is the result of local recapitulation of the normal patterning genes. To analyze the dynamics of one major Drosophila patterning gene, decapentaplegic (dpp), in wing imaginal disc regeneration, a vital GFP reporter together with iontophoretic cell labeling were used. Our observations reveal that the restoration of compartment-border-specific dpp expression is a common event in imaginal disc regeneration. However, we did not find evidence of an upregulation of dpp expression during the regeneration process.

Holocentric chromosomes of psocids (Insecta, Psocoptera) analysed by C-banding, silver impregnation and sequence specific fluorochromes CMA3 and DAPI.

The pattern of nucleolus attachment and C-heterochromatin distribution and molecular composition in the karyotypes of psocid species Psococerastis gibbosa (2n = 16+X), Blaste conspurcata (2n = 16+X) and Amphipsocus japonicus (2n = 14+neo-XY) were studied by C-banding, silver impregnation and sequence specific fluorochromes CMA3 and DAPI. Every species was found to have a single nucleolus in male meiosis. In P. gibbosa the nucleolus is attached to an autosomal bivalent; in B. conspurcata to the X-chromosome; in A. japonicus to the neo-XY bivalent. The species show a rather small amount of constitutive heterochromatin, C-blocks demonstrating telomeric localization with rare exceptions. P. gibbosa is characterized by a polymorphism for C-blocks occurrence and distribution. In the autosomes of this species, C-heterochromatin consists of AT-rich DNA except for the nucleolus organizing region, which is also GC-rich; the X-chromosome shows both AT- and GC-rich clusters. In A. japonicus and B. conspurcata, C-heterochromatin of the autosomes and sex chromosomes consists of both GC-rich and AT-rich DNA clusters, which are largely co-localized.

The origin of the achiasmatic XY sex chromosome system in Cacopsylla peregrina (Frst.) (Psylloidea, Homoptera).

The status of an extra univalent, if it is a B chromosome or an achiasmatic Y chromosome, associating with the X chromosome in male meiosis of Cacopsylla peregrina (Frst.) (Homoptera, Psylloidea) was analysed. One extra univalent was present in all males collected from three geographically well separated populations, it was mitotically stable, and showed precise segregation from the X chromosome. These findings led us to propose that the univalent represents in fact a Y chromosome. The behaviour of the X and Y chromosomes during meiotic prophase suggested that their regular segregation was based on an achiasmatic segregation mechanism characterised by a 'touch and go' pairing of segregating chromosomes at metaphase I. To explain the formation of the achiasmatic Y within an insect group with X0 sex chromosome system, it was suggested that the Y chromosome has evolved from a mitotically stable B chromosome that was first integrated into an achiasmatic segregation system with the X chromosome, and has later become fixed in the karyotype as a Y chromosome.

C-heterochromatin and extra (B) chromosome distribution in six species of the Nabis (Heteroptera, Nabidae) with the modal male karyotype 2n = 16 + XY.

The basic male karyotype of the six Nabis species (Heteroptera, Nabidae) is confirmed as being 2n=16+XY. The chromosomes are holokinetic while male meiosis is achiasmatic. The sex chromosomes undergo postreduction and in second metaphase show distance pairing, registered in all nabid species examined so far. Using C-banding technique for the first time in the family Nabidae, the heterochromatin was revealed on chromosomes of six species. The species showed different amount and distribution of C-heterochromatin. Only in Nabis (Dolichonabis) limbatus did the C-bands distribution make possible the identification of every chromosome pair in the karyotype. In other species, C-bands were found in some of the autosomes and the X, localized either interstitially or at telomeres. Only the Y usually showed relative stability ofthe C-banding pattern. In four of six species, extra (B) chromosomes were observed and their behaviour in meiosis described.

A new approach to the auchenorrhyncha (Hemiptera, Insecta) cytogenetics: chromosomes of the meadow spittlebug Philaenus spumarius (L.) examined using various chromosome banding techniques.

The karyotype of the meadow spittlebug Philaenus spumarius (L.) was studied using conventional chromosome staining, C- and AgNOR- banding, and fluorescent CMA3- and DAPI- techniques. This is the first report on differential staining of the holocentric chromosomes of Auchenorrhyncha. The karyotype of Ph. spumarius includes 2n = 22 + XX/X0. The autosomal pair 1 is large and carries a gap in every homologue. After silver staining, NORs were revealed in both this chromosome pair and a middle-sized pair, most likely 6 or 7. In spermatocyte meiosis, the majority of bivalents formed one chiasma each. The bivalent 1 showed from 1 to 4 chiasmata, the value of 1 or 2 being prevalent. Two further bivalents also showed two chiasmata in some cells. After C-banding, terminal and interstitial dot-type C-heterochromatic blocks were revealed in the chromosomes. In 4 of 11 studied males, the autosomal pair 1 was polymorphic for an extra segment attached to one of the homologues. The segment consisted of both heterochromatic and euchromatic portions. No defined signals were observed in any chromosome treated with DAPI. After CMA3- staining, bright fluorescent signals were obtained in the NOR-bearing chromosomes, suggesting GC-rich DNA bound to the NORs.