Atypically Shaped Setae in Gall Mites (Acariformes, Eriophyoidea) and Mitogenomics of the Genus Leipothrix Keifer (Eriophyidae).

The setae in Eriophyoidea are filiform, slightly bent and thickened near the base. Confocal microscopy indicates that their proximal and distal parts differ in light reflection and autofluorescence. Approximately 50 genera have atypically shaped setae: bifurcated, angled or swollen. These modifications are known in the basal part of prosomal setae u', ft', ft″, d, v, bv, ve, sc and caudal setae h2. We assessed the distribution of atypically shaped setae in Eriophyoidea and showed that they are scattered in different phylogenetic lineages. We hypothesized that the ancestral setae of eriophyoid mites were bifurcated before later simplifying into filiform setae. We also proposed that hypo-furcating setae are a synapomorphy that unites Eriophyoidea with Nematalycidae. We analyzed four new mitochondrial genomes of Leipothrix, the largest genus with bifurcated d, and showed that it is monophyletic and has a unique mitochondrial gene order with translocated trnK. We exclude Cereusacarus juniperensisn. comb. Xue and Yin, 2020 from Leipothrix and transfer five Epitrimerus spp. to Leipothrix: L. aegopodii (Liro 1941) n. comb., L. femoralis (Liro 1941) n. comb., L. geranii (Liro 1941) n. comb., L. ranunculi (Liro 1941) n. comb., and L. triquetra (Meyer 1990) n. comb.

Deuterogyny and the Association of Two Vagrant Eriophyoid Mites (Acariformes, Eriophyoidea) with the Host-plant Generative Organs of Two Broad-leaved Trees in North-West Russia.

Phytoparasitic mites of the superfamily Eriophyoidea Nalepa live and feed on mature leaf surfaces, between leaf bud scales, and (though less commonly) on flowers or fruits. In this study, we focused on the seasonal associations of two eriophyoid species, Shevtchenkella serrata (Nalepa 1892) with the Norway maple tree (Acer platanoides L.), and Brevulacus reticulatus Manson 1984 with the common oak (Quercus robur L.). These species have complex life cycles with two morphologically different, seasonal female forms, the protogyne and deutogyne. In B. reticulatus, both forms retain all the major generic characteristics but in S. serrata only the protogynes conform to the diagnosis of Shevtchenkella, whereas the deutogynes have the typical traits of Anthocoptes. We confirmed the conspecificity of the protogynes and deutogynes of both eriophyoid species by sequencing a barcode fragment of the Cox1 gene from which we obtained four pairwise identical sequences: ON920305/ON920306 (S. serrata) and ON920307/ON920308 (B. reticulatus). In addition, taxonomical studies on Shevtchenkella and Brevulacus resulted in new synonymies and combinations: (1) Oxypleurites obtusus Roivainen 1947 is considered a deutogyne of S. serrata and treated as a junior synonym of S. serrata; (2) two rhyncaphytoptine species from North America are transferred from the genus Rhyncaphytoptus to Brevulacus: B. albus (Keifer 1959) comb. nov. and B. atlanticus (Keifer 1959) comb. nov.; and (3) one species, B. salicinus Soika et al. 2017, is excluded from Brevulacus and transferred to Rhyncaphytoptus: Rhyncaphytoptus salicinus (Soika et al. 2017) comb. nov. Apart from distinct morphological deuterogyny in S. serrata and B. reticulatus, we observed the persistent association of S. serrata with the generative organs of the maple tree, A. platanoides, leading to transmission to the next host generation via the seed-containing winged fruits (samaras) and subsequent colonization of seedlings. In B. reticulatus, similar synchronization with host-plant dispersal was not detected; however, in mid-summer, temporary colonization of immature acorns and feeding was observed. Additional studies conducted in various ecosystems and including different ecological groups of plants, especially anemochorous plants, are needed to estimate the frequency of the association of eriophyoids with plant generative organs, seeds and seedlings to better understand what role in mite ecology such associations may play.

Molecular Aspects of Gall Formation Induced by Mites and Insects.

Recent publications on gall formation induced on the leaves of dicotyledonous flowering plants by eriophyoid mites (Eriophyoidea) and representatives of four insect orders (Diptera, Hemiptera, Hymenoptera, Lepidoptera) are analyzed. Cellular and molecular level data on the stimuli that induce and sustain the development of both mite and insect galls, the expression of host plant genes during gallogenesis, and the effects of these galling arthropods on photosynthesis are considered. A hypothesis is proposed for the relationship between the size of galls and the volume of secretions injected by a parasite. Multistep, varying patterns of plant gene expression and accompanying histo-morphological changes in the transformed gall tissues are apparent. The main obstacle to better elucidating the nature of the induction of gallogenesis is the impossibility of collecting a sufficient amount of saliva for analysis, which is especially important in the case of microscopic eriophyoids. The use of modern omics technologies at the organismal level has revealed a spectrum of genetic mechanisms of gall formation at the molecular level but has not yet answered the questions regarding the nature of gall-inducing agents and the features of events occurring in plant cells at the very beginning of gall growth.

Integrative Taxonomy of the Gall Mite Nothopoda todeica n. sp. (Eriophyidae) from the Disjunct Afro-Australasian Fern Todea barbara: Morphology, Phylogeny, and Mitogenomics.

Eriophyoidea is a group of phytoparasitic mites with poorly resolved phylogeny. Previous studies inferred Eriophyidae s.l. as the largest molecular clade of Eriophyoidea, and Nothopodinae as the basal divergence of Eriophyidae s.l. We investigate the morphology and molecular phylogeny of Nothopoda todeican. sp. (Nothopodinae, Nothopodini), associated with a disjunct Afro-Australasian fern Todea barbara (Osmundaceae) from South Africa. Our analyses (1) determine new erroneous sequences (KF782375, KF782475, KF782586) wrongly assigned to Nothopodinae instead of Phyllocoptinae, (2) confirm the basal position of Nothopodinae in Eriophyoidea s.l., (3) question the monophyly of the Colopodacini and Nothopodini tribes, and (4) show the nested position of African fern-associated Nothopoda within a clade dominated by Asian nothopodines from angiosperms, which implies (a) a secondary association of nothopodines with ferns and (b) no relation between geography (continents) and the phylogenetic relationships of Nothopodinae species. Finally, we obtained a first complete mitochondrial genome for Nothopodinae and revealed a new gene order in the mitogenome of N. todeican. sp., notably deviating from those in other investigated eriophyoids. Our results contribute to resolving the phylogeny of Eriophyoidea and provide an example of an integrative study of a new taxon belonging to an economically important group of acariform mites.

Where Eriophyoidea (Acariformes) Belong in the Tree of Life.

Over the past century and a half, the taxonomic placement of Eriophyoidea has been in flux. For much of this period, this group has been treated as a subtaxon within Trombidiformes. However, the vast majority of recent phylogenetic analyses, including almost all phylogenomic analyses, place this group outside Trombidiformes. The few studies that still place Eriophyoidea within Trombidiformes are likely to be biased by incomplete taxon/gene sampling, long branch attraction, the omission of RNA secondary structure in sequence alignment, and the inclusion of hypervariable expansion-contraction rRNA regions. Based on the agreement among a number of independent analyses that use a range of different datasets (morphology; multiple genes; mitochondrial/whole genomes), Eriophyoidea are almost certain to be closely related to Nematalycidae, a family of vermiform mites within Endeostigmata, a basal acariform grade. Much of the morphological evidence in support of this relationship was apparent after the discovery of Nematalycidae in the middle of the 20th century. However, this evidence has largely been disregarded until very recently, perhaps because of overconfidence in the placement of Eriophyoidea within Trombidiformes. Here, we briefly review and identify a number of biases, both molecular- and morphology-based, that can lead to erroneous reconstructions of the position of Eriophyoidea in the tree of life.

Anatomy of the miniature four-legged mite Achaetocoptes quercifolii (Arachnida: Acariformes: Eriophyoidea).

Miniaturization is one of the important trends in the evolution of terrestrial arthropods. In order to study adaptations to microscopic sizes, the anatomy of the smallest insects was previously studied, but not the anatomy of the smallest mites. Some of the smallest mites are Eriophyidae. In this study we describe for the first time the anatomy of the mite Achaetocoptes quercifolii, which is about 115 µm long. For this purpose, we used light, scanning, and transmission electron microscopy and performed 3D reconstructions. The anatomy of A. quercifolii is compared with the anatomy of larger representatives of Eriophyoidea. Despite the small size of the studied species, there is no considerable simplification of its anatomy compared to larger four-legged mites. A. quercifolii has a number of miniaturization effects similar to those found in microinsects: a strong increase in the relative volume of the reproductive system, an increase in the relative volume of the brain, reduction in the number and size of cells of the nervous system. As in some larger four-legged mites, A. quercifolii undergoes midgut lysis at the stage of egg production. On the other hand, in A. quercifolii a greater number of opisthosomal muscles are preserved than in larger gall-forming four-legged mites.

First rhyncaphytoptine mite (Eriophyoidea, Diptilomiopidae) parasitizing American hazelnut (Corylus americana): molecular identification, confocal microscopy, and phylogenetic position.

The plant genus Corylus is an economically important crop, valued especially for its nuts. Numerous pathogens and harmful phytophagous arthropods are known to damage hazelnuts. We report on a new eriophyoid mite, Rhyncaphytoptus corylivagrans n. sp., and the first record of Coptophylla lamimani both collected from leaves of American hazelnut (Corylus americana) in North Carolina, USA. Including our new data, the complex of eriophyoids from Corylus comprises 15 species from three families: Phytoptidae (2 spp.), Eriophyidae (11 spp.), and Diptilomiopidae (2 spp.). We obtained sequences of three genes (Cox1, D1-D5 28S, and ITS1-5.8S-ITS2), applied BLAST and tree-based approaches for identification of R. corylivagrans n. sp., and performed the first molecular phylogenetic analysis focused on Rhyncaphytoptinae. Among the three genes, Cox1 showed better power when used for BLAST searches. Combined molecular phylogenetic analyses inferred R. corylivagrans n. sp. as sister to R. betulae, determined several moderately supported host-specific lineages of rhyncaphytoptines, and indicated a close relationship of the new species with members of the genus Rhinotergum. In two Rhinotergum spp. from Rosaceae, confocal microscopy revealed a new structure, the needle-like anterior process of the prodorsal shield, which is absent in R. corylivagrans n. sp. Additionally, the elements of the anal secretory apparatus presumably associated with silk-production and hypothesized as a synapomorphy of Eriophyoidea, were detected in the new species, providing the first documented report of this structure in Diptilomiopidae. Our study contributes to knowledge on the biodiversity of phytoparasites associated with hazelnuts and calls for future comparative phylogenetics of Diptilomiopidae.

Vertical transmission and seasonal dimorphism of eriophyoid mites (Acariformes, Eriophyoidea) parasitic on the Norway maple: a case study.

Eriophyoid mites are highly host-specific, microscopic phytoparasites that primarily disperse to new hosts passively via wind. This seems paradoxical, as the likelihood of landing on an appropriate host species needed to survive appears low. Here we investigate two eriophyoids found on the Norway maple Acer platanoides: Aceria platanoidea and Shevtchenkella serrata. For 14 months, we observed mite phenotypical changes and micro-habitat distribution on host plants and their propagules. Both mite species hibernate on twigs or samaras fallen on the ground, and, in the spring, feed on buds or seedlings, respectively. This apparently novel association with plant seeds indicates that the mites can exploit the host dispersal mechanism and colonize the next generation of hosts (vertical transmission). Our seasonal and DNA sequence data also indicate that S. serrata has two distinct morphotypes that partially overlap seasonally. This work can provide new insights into the dispersal routes of eriophyoid mites and transmission patterns of plant pathogens vectored by these mites, with implications for better pest mite species control.

Symbiotic bacteria of the gall-inducing mite Fragariocoptes setiger (Eriophyoidea) and phylogenomic resolution of the eriophyoid position among Acari.

Eriophyoid mites represent a hyperdiverse, phytophagous lineage with an unclear phylogenetic position. These mites have succeeded in colonizing nearly every seed plant species, and this evolutionary success was in part due to the mites' ability to induce galls in plants. A gall is a unique niche that provides the inducer of this modification with vital resources. The exact mechanism of gall formation is still not understood, even as to whether it is endogenic (mites directly cause galls) or exogenic (symbiotic microorganisms are involved). Here we (i) investigate the phylogenetic affinities of eriophyoids and (ii) use comparative metagenomics to test the hypothesis that the endosymbionts of eriophyoid mites are involved in gall formation. Our phylogenomic analysis robustly inferred eriophyoids as closely related to Nematalycidae, a group of deep-soil mites belonging to Endeostigmata. Our comparative metagenomics, fluorescence in situ hybridization, and electron microscopy experiments identified two candidate endosymbiotic bacteria shared across samples, however, it is unlikely that they are gall inducers (morphotype1: novel Wolbachia, morphotype2: possibly Agrobacterium tumefaciens). We also detected an array of plant pathogens associated with galls that may be vectored by the mites, and we determined a mite pathogenic virus (Betabaculovirus) that could be tested for using in biocontrol of agricultural pest mites.

Phylogenetic Position of a New Trisetacus Mite Species (Nalepellidae) Destroying Seeds of North American Junipers and New Hypotheses on Basal Divergence of Eriophyoidea.

Eriophyoid mites of the genus Trisetacus Keifer are widespread parasites of conifers. A new oligophagous species, T. indelis n. sp., was discovered severely damaging seeds of North American junipers (Juniperus osteosperma, J. occidentalis, and J. californica) in the western USA. It has two codon deletions in the mitochondrial gene Cox1 rarely detected in Eriophyoidea and includes distinct morphological dimorphism of females. A phylogenetic analysis based on amino acid alignment of translated Cox1 sequences using a large set of out-groups (a) determined that two North American congeners, T. batonrougei and T. neoquadrisetus, were the closest known relatives of T. indelis n. sp., and (b) indicated that Old and New World seed-inhabiting Trisetacus from junipers do not form a distinct clade, suggesting a possible independent transition to living in seeds of junipers in America and Eurasia by Trisetacus spp. Our analysis produced a new topology consistent with a scenario assuming gradual reduction of prodorsal shield setation in Eriophyoidea and an ancient switch from gymnosperms to other hosts. Additionally, our analysis did not support monophyly of Trisetacus; recovered a new host-specific, moderately supported clade comprising Trisetacus and Nalepellinae (Nalepella + Setoptus) associated with Pinaceae; and questioned the monophyly of Trisetacus associated with Cupressaceae.

Molecular phylogeny of the phytoparasitic mite family Phytoptidae (Acariformes: Eriophyoidea) identified the female genitalic anatomy as a major macroevolutionary factor and revealed multiple origins of gall induction.

Phytoptidae s.str. is a lineage of eriophyoid mites associated with angiosperms. Based on representative taxon sampling and four gene markers (COI, HSP70, 18S, and 28S), we inferred the molecular phylogeny of this group and performed comparative analyses of cuticle-lined female internal genitalia. Although basal relationships were unclear, several well supported clades were recovered. These clades were supported by geography, host associations, and female genital anatomy, but contradicted the current morphology-based systematics. The monophyly of each of five conventional supraspecific groupings (Fragariocoptes, Phytoptus, Phytoptinae, Sierraphytoptinae, and Sierraphytoptini) is rejected based on a series of statistical tests. Additionally, four morphological characters (the absence of tibial solenidion φ and opisthosomal seta c1, presence of telosomal pseudotagma, and 'morphotype') were found to be homoplasies that cannot be used to confidently delimit supraspecific lineages of phytoptids. However, our molecular topology was highly congruent with female genital characters. Eight molecular clades were unambiguously supported by the shapes and topography of the spermathecal apparatus and genital apodemes. This suggests that the female genital anatomy could be an important factor affecting cladogenesis in Phytoptidae, a conclusion contrasting with the general expectation that host characteristics should be a major macroevolutionary force influencing the evolution of host-specific symbionts. Indeed, despite the high host-specificity, there were no apparent cophylogenetic patterns. Furthermore, we show that gall-inducing ability evolved multiple times in phytoptids. Because gall formation creates nearly instantaneous niche partitioning and the potential loss or reduction of gene flow, we hypothesize that it could be an important evolutionary factor affecting speciation within different host-associated clades of phytoptid mites.

Molecular phylogenetic analyses reveal a deep dichotomy in the conifer-inhabiting genus Trisetacus (Eriophyoidea: Nalepellidae), with the two lineages differing in their female genital morphology and host associations.

We analyzed the phylogenetic relationships of the genus Trisetacus using two genes [cytochrome c oxidase subunit I (COI) and D1-D2 region of 28S rDNA (D1-D2 28S)], a representive taxon sampling (nearly 40% of known diversity), and a large set of close and distant outgroups. Our analyses suggest the presence of a dichotomy between Trisetacus associated with Cupressaceae and Pinaceae. The following smaller molecular clades were found: Pin-1 (bud mites, twig sheath mites, bark gall mites, and endoparasitic mites from pinaceans), Pin-2 (needle sheath mites from pines), Pin-2a (putative Nearctic group of needle sheath mites), Pin-2b (putative Palearctic group of needle sheath mites), Cup-1 and 2 (bud, cone, seed mites and mites living under bark scales from cupressaceans). The monophyly of the recently proposed subgenus Brevithecus nested within clade Cup-2 was confirmed. Ancestral character reconstruction analyses recovered: (1) Pinaceae as the ancestral hosts of Nalepellidae and Trisetacus, (2) repetitive reductions of the spermathecal tube independently occurred in two lineages of Trisetacus from Cupressaceae, and (3) several mite habitats on host (galls, cones, twig sheaths, seeds, inside leaves, and under scales) are evolutionarily derived states, whereas living in buds or needle sheaths are ancestral states for Trisetacus clades Cup and Pin. Using confocal microscopy, we identified six basic types of the female internal genitalia of Trisetacus based on shapes of the spermatheca and spermathecal tube. These genitalic types are strongly correlated with lineages recovered by molecular phylogenetic analyses, suggesting that the female genital morphology is both evolutionarily conserved and is a factor influencing macroevolutionary patterns in this group of mites.

Comprehensive phylogeny of acariform mites (Acariformes) provides insights on the origin of the four-legged mites (Eriophyoidea), a long branch.

Eriophyoid, or four-legged mites, represent a large and ancient radiation of exclusively phytophagous organisms known from the Triassic (230 Mya). Hypothesizing phylogenetic relatedness of Eriophyoidea among mites is a major challenge due to the absence of unambiguous morphological synapomorphies, resulting in ten published hypotheses placing eriophyoids in various places in the acariform tree of life. Here we test the evolutionary relationships of eriophyoids using six genes and a representative taxonomic sampling of acariform mites. The total evidence analysis places eriophyoids as the sister group of the deep soil-dwelling, vermiform family Nematalycidae (Endeostigmata). This arrangement was supported by the rDNA and CO1 partitions. In contrast, the nuclear protein partition (genes EF1-α, SRP54, HSP70) suggests that Eriophyoidea is sister to a lineage including Tydeidae, Ereynetidae, and Eupodidae (Eupodina: Trombidiformes). On both of these alternative topologies, eriophyoids appear as a long branch, probably involving the loss of basal diversity in early evolution. We analyze this result by using phylogenetically explicit hypothesis testing, investigating the phylogenetic signal from individual genes and rDNA stem and loop regions, and removing long branches and rogue taxa. Regardless of the two alternative placements, (i) the cheliceral morphology of eriophyoids, one of the traits deemed phylogenetically important, was likely derived directly from the plesiomorphic acariform chelicerae rather than from the modified chelicerae of some trombidiform lineages with a reduced fixed digit; and (ii) two potential synapomorphies of Eriophyoidea+Raphignathina (Trombidiformes) related to the reduction of genital papillae and to the terminal position of PS segment can be dismissed as result of convergent evolution. Our analyses substantially narrow the remaining available hypotheses on eriophyoid relationships and provide insights on the early evolution of acariform mites.

Suboral fork: a newly discerned gnathosomal structure from the proboscis of eriophyoid mites (Acari, Eriophyoidea).

The infracapitulum of eriophyoid mites comprises a cone-like basal infracapitulum, containing a pharynx, and a distal infracapitulum, forming a proboscis ensheathing a bunch of stylets. A well-developed basal labral section was observed in all studied specimens. A newly discerned structure, the suboral fork, situated in the ventral part of the proboscis was discovered. It is larger in diptilomiopids and Nalepella and notably smaller in eriophyids and phytoptids. This structure presumably determines the site of piercing and functions in a similar way to the pressure foot of a sewing machine which controls the movements of a needle. In diptilomiopids the suboral fork might have an additional function: it is a stopper which prevents the proboscis from further penetrating into plant tissues. It is possible that the suboral fork is homologous with the labium of early derivative acariform mites. The proboscis might be a fusion product of the infracapitular lateral lips, malapophyses and the labium. The proboscis serves as a feeding structure in eriophyoids; two ways of sucking plant cell sap, depending on shapes of proboscis and labrum, are hypothesized. Further work is needed to draw conclusions on homologies and the function of all gnathosomal structures in eriophyoids.

New species and records of phytoptid mites (Acari: Eriophyoidea: Phytoptidae) on sedges (Cyperaceae) from the Russian Far East.

Two new phytoptine species, Oziella virgata n. sp. and O. ovalis n. sp., were collected from the Russian Far East on sedges, Carex appendiculata (Trautv. & C.A. Mey) Kükenthal and Kobresia myosuroides (Villars) Fiori, respectively, and are described herein using conventional light microscopy (LM) and confocal laser scanning microscopy (CLSM). In addition, Oziella cf. rigida (Roivainen 1950) was recorded from Carex scita var. riishirensis (Franch.) Kük. in the Kamchatka Peninsula whilst Novophytoptus rostratae Roivainen 1947 was found on Carex saxatilis L. and C. appendiculata in the Sakha Republic (Yakutia) of Russia and Carex soczavaeana Gorodkov on the Kamchatka Peninsula. Oziella virgata n. sp. has a unique rod-like seta u' on the medial-lower surface of tarsi I & II, a character not previously described in eriophyoid mites. Additionally, the position of setae 3a in nymphs and females differs: these are located on the same annulus as tubercles of setae c2 in nymphs whereas in females, those are situated notably ahead of tubercles c2 (closer to coxae II). Males of O. virgata n. sp. possess a well-developed genital coverflap, resembling that of Mackiella reclinata Chetverikov & Craemer, 2014, Pentasetacus araucariae (Schliesske, 1985) and Loboquintus subsquamatus Chetverikov & Petanovic, 2013. The original slidemounted specimens of the new Oziella species described herein were inappropriate for LM study. However, CLSM microscopy images obtained prior to remounting were sharp enough for diagnostic purposes indicating that this is a useful method for studying poor quality specimens which may otherwise be difficult to remount or, in some cases, are very rare.

Generic delimitation between Fragariocoptes and Sierraphytoptus (Acari: Eriophyoidea: Phytoptidae) and a supplementary description of Fragariocoptes gansuensis with remarks on searching for mummified eriophyoid mites in herbaria under UV light.

Generic concepts of Fragariocoptes Roivainen, 1951 and Sierraphytoptus Keifer, 1939 are discussed and the correct delimitation between these two genera is given. A supplementary description of Fragariocoptes gansuensis Wei, Chen & Luo, 2005 is included based on fresh specimens from Astrakhan, Russia and dried mummies found in old herbaria collected in 1919 from southern European Russia of the cinquefoil, Potentilla bifurca L. (Rosaceae) with pathological stem proliferation. The male of this species is described for the first time. The cuticle of eriophyoid mummies emitted a faint glow under UV light wavelength equal to 365 nm of a common UV Light-Emitting diode (LED) lamp showing that this characteristic could be useful for quickly detecting eriophyoids in old herbaria which would otherwise be almost indistinguishable against the background under the regular white light source of a stereomicroscope. This was only possible for plant material stored in appropriate conditions enabling the autofluorescent signal of the dried mite cuticle to remain strong enough for observation.

Description of a new early-derivative mite, Pentasetacus plicatus n. sp. (Acariformes, Eriophyoidea), and remarks on the systematic position of pentasetacines.

A new vagrant early-derivative eriophyoid mite species Pentasetacus plicatus n. sp. (GeneBank accession number KT070291) is described from Araucaria araucana (Molina) K.Koch from Chile and Peru with the aid of conventional and confocal laser scanning microscopy. Observations on the gnathosoma of the new species indicate that the oral stylet is bent in the middle and twice shorter than the infracapitular and cheliceral stylets. A trough-like subcapitulum originated from the palpcoxae, and swollen basal parts of chelicerae bearing putative cheliceral seta, were observed under CLSM in specimens with disjointed mouth parts. Males with an exposed gonopore under an uncovered genital coverflap and remnants of soft genital organs were registered. Contrary to other eriophyoids, the plates of the longitudinal bridge in P. plicatus n. sp. and other pentasetacines are not fused so that the genital slit continues posterior to the spermathecal apparatus. The previously established taxon Pentasetacidae Shevchenko, Bagnyuk, Sukhareva, 1991 is proposed for the eriophyoids bearing five prodorsal shield setae (Loboquintus and Pentasetacus); a refined diagnosis of Pentasetacidae is given.

A study of embryonic development in eriophyoid mites (Acariformes, Eriophyoidea) with the use of the fluorochrome DAPI and confocal microscopy.

The embryonic development of four eriophyoid mite species, Cecidophyopsis ribis, Phytoptus avellanae, Oziella liroi and Loboquintus subsquamatus, has been studied with the use of fluorochrome DAPI and confocal microscopy. The first three nuclear divisions occur on the egg periphery (the groups of 2, 4, and 6 nuclei have been recorded), while the biggest part of yolk remains undivided. After four or five nuclear divisions all nuclei are situated only in one sector of the embryo, while other sectors contain only yolk suggesting possible meroblastic cleavage. Later, the formation of superficial blastoderm takes place. A few large yolk cells are situated inside the embryo. Germ band formation initiates as funnel-like cell invagination and leads to formation of a typical stage with four paired prosomal buds (chelicerae, palps, legs I and II). Each palp contains two lobes (anterior and posterior), the adult subcapitulum is presumably a fusion product of the anterior pair of the lobes. Neither rudiments of legs III and IV, traces of opisthosomal segments nor remnants of the prelarval exuvium under the egg shell were detected. Overall, the pattern of embryonic development in eriophyoids re-emphasizes the peculiarity of this ancient group of miniaturized phytoparasitic animals, and invites researches to pursue a deeper investigation of various fundamental aspects of this aberrant group of Acari. Further studies using various fluorescent dyes and transmission electron microscopy are needed to visualize plasma membranes and clarify the pattern of early cleavage of eriophyoids.

Hidden diversity of endoparasitic eriophyoid mites: two new Novophytoptus Roivainen, 1947 (Acari: Eriophyoidea: Phytoptidae) species from the parenchymatous tissues of rushes (Juncaceae).

The monogeneric subfamily Novophytoptinae is a separate lineage of phytoptids restricted to endoparasitism on herbaceous monocots of the order Poales. Novophytoptines live under the epidermis of their hosts where they feed on parenchymatous cells and reproduce therein. It is unknown yet how novophytoptines penetrate the plant epidermis, but preliminary observations indicate that they might be able to penetrate through circular holes which they cut in the epidermis using their modified gnathosoma. Two new species, Novophytoptus luzulis n. sp. from Luzula pilosa L. and Novophytoptus maritimus n. sp. from Juncus maritimus Lam., are described and illustrated. Two small pores, presumably representing external openings of spermathecal tubes, were found in the postero-medial genital cuticle (sensu Chetverikov 2014b) at the level between the posterior margin of the genital coverflap and the genital rim, in both new species. This is the first documented report of such structures in slide-mounted eriophyoid mites. CLSM and DIC microscopy-based observations showed that novophytoptines possess a peculiar spermathecal apparatus, including greatly expanded sack-shaped spermathecae and thick, bent spermathecal tubes directed anteriad, and a semicircular anterior genital apodeme perpendicular to the long body axis. Similarity in the structure of the spermathecal apparatus among novophytoptines, phytoptines and sierraphytoptines (all Phytoptidae from angiosperms) apparently supports their assignment to a common group. Additional examples of endoparasitism among Eriophyoidea are listed. The hypothesis of a primary endoparasitic life style in the eriophyoid basal stalk and a secondary shift to free living forms on exposed surfaces of plants is briefly discussed. Research on grass-associated endoparasitic mites is important because they may include new vectors of pathogens. SketchUp Free Software is recommended as one of the most simple and promising 3D drawing tools for modeling the internal genitalia and other complex anatomical structures of microarthropods (especially eriophyoids) based on digital data obtained using various microscopic techniques such as CLSM.