A New Aculodes Species (Prostigmata: Eriophyidae) Described from an Invasive Weed by Morphological, Morphometric and DNA Barcode Analyses.

A new species of eriophyoid mite, Aculodes marcelli sp. nov., was discovered on cheatgrass, Anisantha tectorum (L.) Nevski (syn. Bromus tectorum L.), an annual grass that is native to Eurasia and Northern Africa. This grass was introduced to North America near the end of the 19th century and now is widespread and associated with the observed increases in the size, frequency, and intensity of wildfires in western N. America. In this paper, A. marcelli sp. nov., is morphologically described and illustrated. Compared with other Aculodes spp., it differs based on morphology and the sequence of the mitochondrial cytochrome oxidase gene, subunit I (MT-CO1). Results of morphometric analysis showed clear differentiation between A. marcelli sp. nov., and the most similar congener, A. altamurgiensis from Taeniatherum caput-medusae. Analysis of MT-CO1 sequence divergence revealed significant levels of genetic variation (17.7%) and supported the results from the morphometric analysis; therefore, it is determined that they are two different species. Aculodes marcelli sp. nov., is a new candidate agent for classical biological control of A. tectorum.

Integrative Taxonomy and Synonymization of Aculus mosoniensis (Acari: Eriophyidae), a Potential Biological Control Agent for Tree of Heaven (Ailanthus altissima).

The taxonomy of Aculus mosoniensis appears to be an unresolved question and its clarification is required, owing to the potential relevance of this mite species as a biological control agent of the tree of heaven. This paper is aimed at giving accurate details on a previously and shortly announced synonymization with Aculops taihangensis, using a morphological and molecular approach. A fusiform morph of A. mosoniensis was distinguished from a vermiform morph and this latter was recognized as deutogyne, which was herein documented. Phylogenetic relationships between Chinese Ac. taihangensis and all A. mosoniensis mites collected in twenty localities in Europe were examined through the analysis of the mitochondrial cytochrome c subunit I (CO1) protein and the nuclear ribosomal internal transcribed spacer 1 region (ITS1). CO1 sequences of Ac. taihangensis from the Shandong province in China and those from mites collected in Austria and Slovenia were 100% identical; the ITS1 sequence of an Ac. taihangensis paratype matched for 99.8% with those obtained from protogynes and deutogynes of A. mosoniensis collected in Italy. All these data supported the announced synonymization of A. mosoniensis with Ac. taihangensis. Aculusmosoniensis was found genetically variable, with five CO1 haplotypes in Europe (becoming eight along with those of Ac. taihangensis) clustering in two highly supported maternal lineages and eight ITS1 haplotypes (becoming nine along with those of Ac. taihangensis) distributed in four supported clades. No overlap between intra- and interspecies distances was observed for both markers and all studied A. mosoniensis populations clustered in one monophyletic mitochondrial clade, suggesting that only one single species might occur in Europe. However, more mite clades may be related to more tree of heaven biotypes with potential ecological differences, which might have potential effects on the biological control and should be investigated.

Field Assessment of the Host Range of Aculus mosoniensis (Acari: Eriophyidae), a Biological Control Agent of the Tree of Heaven (Ailanthus altissima).

Tree of heaven (Ailanthus altissima) is a fast-growing deciduous tree native to China, considered a serious invasive species worldwide, with several socio-economic and ecological impacts attributed to it. Chemical and mechanical methods have limited efficacy in its management, and biological controls may offer a suitable and sustainable option. Aculus mosoniensis (Ripka) is an eriophyid mite that has been recorded to attack tree of heaven in 13 European countries. This study aims to explore the host range of this mite by exposing 13 plant species, selected either for their phylogenetic and ecological similarity to the target weed or their economic importance. Shortly after inoculation with the mite, we recorded a quick decrease in mite number on all nontarget species and no sign of mite reproduction. Whereas, after just one month, the population of mites on tree of heaven numbered in the thousands, irrespective of the starting population, and included both adults and juveniles. Significantly, we observed evidence of damage due to the mite only on target plants. Due to the specificity, strong impact on the target, and the ability to increase its population to high levels in a relatively short amount of time, we find A. mosoniensis to be a very promising candidate for the biological control of tree of heaven.

Eriophyid Mites in Classical Biological Control of Weeds: Progress and Challenges.

A classical biological control agent is an exotic host-specific natural enemy, which is intentionally introduced to obtain long-term control of an alien invasive species. Among the arthropods considered for this role, eriophyid mites are likely to possess the main attributes required: host specificity, efficacy, and long-lasting effects. However, so far, only a few species have been approved for release. Due to their microscopic size and the general lack of knowledge regarding their biology and behavior, working with eriophyids is particularly challenging. Furthermore, mites disperse in wind, and little is known about biotic and abiotic constraints to their population growth. All these aspects pose challenges that, if not properly dealt with, can make it particularly difficult to evaluate eriophyids as prospective biological control agents and jeopardize the general success of control programs. We identified some of the critical aspects of working with eriophyids in classical biological control of weeds and focused on how they have been or may be addressed. In particular, we analyzed the importance of accurate mite identification, the difficulties faced in the evaluation of their host specificity, risk assessment of nontarget species, their impact on the weed, and the final steps of mite release and post-release monitoring.

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.

A new species of Aculus mite (Acari: Eriophyidae), a potential biocontrol agent for Australian swamp stonecrop, Crassula helmsii (Crassulaceae).

A new, gall-forming eriophyoid mite species is described from Australia. Aculus crassulae sp. nov. was found causing significant leaf deformation in Crassula helmsii (Kirk) Cockayne (Crassulaceae), a semi-aquatic, succulent plant. Native to Australia and New Zealand, this plant is now a highly invasive weed in the United Kingdom and Western Europe. The host specificity of the new mite species, and damage caused to the host plant, infer its potential to be a valuable biological control agent in countries where Australian swamp stonecrop is threatening native flora. The species description provided here, which also includes a revised diagnosis for the genus Aculus, incorporates line drawings and scanning electron micrographs (SEM). This is supplemented by a partial mitochondrial gene sequence of cytochrome c oxidase subunit I (mtCOI) and the sequence was compared with Aculus amygdali Xue Hong and Aculus ichnocarpi (Ghosh Chakrabarati) available in the NCBI database. Pairwise comparison of mtCOI sequences between A. crassulae sp. nov. and two congeneric species revealed 22.6% and 23.1% genetic divergence, respectively.

Phenetic and phylogenetic relationships among Aceria spp. (Acari: Eriophyoidea) inhabiting species within the family Brassicaceae in Serbia.

We examined morphological and genetic differences among Aceria spp. inhabiting six Brassicaceae species in Serbia. Five of them have been already mentioned in the literature as original or alternate hosts of Aceria spp. (Berteroa incana (L.) DC., Capsella bursa-pastoris (L.) Medik., Cardamine hirsuta L., Lepidium draba L. and Sisymbrium orientale L.), whereas Aurinia petraea (Ard.) Schur was registered here for the first time as a host for Aceria sp. Results of morphometric analyses indicated clear differentiation of Aceria spp. from B. incana, L. draba, A. petraea and S. orientale, whereas the other two entities were less diverse and clustered together. Molecular analyses indicated that the average mean divergence over all sequence pairs was 18.3% (11.8-25.9%) and disclosed that the observed mtCOI distances between the six host-associated Aceria entities are large enough to represent differences between species. The Aceria sp. hosted by B. incana was morphologically and genetically most distinctive, whereas discrepancy between phenetic and phylogenetic relationships was found in the position of Aceria sp. hosted by L. draba when compared with those inhabiting C. hirsuta, S. orientale and C. bursa-pastoris.

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.

Cryptic speciation within Phytoptus avellanae s.l. (Eriophyoidea: Phytoptidae) revealed by molecular data and observations on molting Tegonotus-like nymphs.

Hazelnut big bud mite, Phytoptus avellanae Nalepa, is one of the most harmful pests of Corylus spp. (Corylaceae) worldwide. Herein, we show that this species represents a complex of two cryptic species: one that lives and reproduces in buds causing their enlargement ('big buds') and drying, whereas the other is a vagrant living on leaves, under bud scales and in catkins, based on phylogenetic analyzes of mitochondrial cytochrome c oxidase subunit I (COI) DNA and the nuclear D2 region of 28S rDNA sequences. A molecular assessment based on mtCOI DNA and nuclear D2 28S rDNA revealed consistent differences of 16.8 and 3.5% between the two species, respectively. Molecular analysis also revealed that atypical flattened nymphs (Tegonotus-like nymphs sensu Keifer in Mites Injurious to Economic Plants, University of California Press, Berkeley, pp 327-562, 1975) with differently annulated opisthosoma, which appear in the life cycle of P. avellanae s.l., belong to the 'vagrant' lineage, i.e. vagrant cryptic species. Light microscopy images of Tegonotus-like nymphs molting into males and females are presented for the first time. Our results suggest that the name P. avellanae comprise two species. Big bud mite should keep the name P. avellanae, and the vagrant cryptic species should be re-named after a proper morphological description is made.

Eriocaenus (Acari: Trombidiformes: Eriophyoidea), a new genus from Equisetum spp. (Equisetaceae): morphological and molecular delimitation of two morphologically similar species.

Surveys conducted on horsetails, Equisetum spp. (Equisetaceae), in Serbia led to the discovery of a new eriophyoid mite genus while searching for a classical biological control agent against these weeds in New Zealand. Eriocaenus gen. n. is described based on the type species Aceria equiseti Farkas, 1960 (transferred to Eriophyes by Farkas 1965; herein reassigned to the new genus) and Eriocaenus ramosissimi n. sp., a new species discovered on Equisetum ramosissimum Desf. in Serbia. Eriocaenus equiseti (Farkas, 1960), previously only known from Hungary, was found in Serbia for the first time on Equisetum arvense L. and Equisetum telmateia Ehrh., and is redescribed. Species descriptions include line drawings as well as phase contrast (PCLM), differential interference contrast (DIC) and scanning electron (SEM) micrographs. The differential diagnosis between the two Eriocaenus species is supplemented by molecular differentiation of 28S rDNA sequences including D2 fragments for both mites.

Geometric morphometric study of geographic and host-related variability in Aceria spp. (Acari: Eriophyoidea) inhabiting Cirsium spp. (Asteraceae).

The russet mite, Aceria anthocoptes (Nalepa), is the only eriophyoid that has been recorded on Cirsium arvense (L.) Scop. It has been noted in several European countries and recently in the USA. In this study we explored the geographic and host-related variability of Aceria spp. inhabiting different Cirsium spp. We applied landmark-based geometric morphometric methods to study morphological variability of three body regions (ventral, coxigenital and prodorsal) of 13 Aceria spp. populations inhabiting five Cirsium spp. in Serbia (Europe) and four Cirsium spp. in Colorado (North America). Analyses of size and shape variation revealed statistically significant differences between Aceria spp. living on European native and North American native Cirsium spp., as well as between A. anthocoptes s.s. inhabiting European C. arvense and North American C. arvense. The coxigenital region was the most informative when considering inter-population shape differences. European Aceria spp. dwelling on Cirsium spp., including A. anthocoptes s.s. from C. arvense, are characterized by higher inter-population size and shape variability than their North American counterparts. This finding supports a Eurasian origin of A. anthocoptes, presumed to consist of a complex of cryptic taxa probably coevolved with host plants in the native environment. Morphological similarity among Aceria spp. inhabiting North American native Cirsium spp. may indicate that speciation of A. anthocoptes started relatively soon after the host shift to plants different from C. arvense in the invaded region.

Description of a new relict eriophyoid mite, Loboquintus subsquamatus n. gen. & n. sp. (Eriophyoidea, Phytoptidae, Pentasetacini) based on confocal microscopy, SEM, COI barcoding and novel CLSM anatomy of internal genitalia.

A new pentasetacine mite Loboquintus subsquamatus n. gen. & n. sp. was found living under scale-like leaves of 2-3 years old twigs of Cupressus sempervirens in Montenegro. This mite species possesses a number of morphological features (uncommon teardrop-shaped body, traits of prosoma, atypical primitive anatomy of the genital apparatus and morphological traits of immatures) which clearly distinguish it from all other known eriophyoids. Adults of L. subsquamatus have seta vi situated on the anterior margin of a uniquely elongate lingua-like thin frontal lobe, three pits on the posterior prodorsal shield margin, a remarkable tube-like structure in the basal part of gnathosoma, a complicated three-layered epigynium, spermathecae directed antero-laterad, short spermathecal tubes and setae eu suppressed in males and possibly expressed in females. External genitalia of males and females of L. subsquamatus are fundamentally similar. Hypothesized remnants of coxisterna III or IV (forming a postgenital plate) are remarkably distinct in males. Two new morphometrical variables are proposed to supplement the CLSM protocol for description of internal genitalia of eriophyoids proposed by Chetverikov et al. (Zootaxa 3560:41-60, 2012b): (a) the length of ventral projection of the transvers genital apodeme and (b) the length of the posterior (=postspermathecal) part of the longitudinal bridge which in L. subsquamatus is remarkably long, whereas in many other eriophyoids it is reduced.

Plant-eriophyoid mite interactions: cellular biochemistry and metabolic responses induced in mite-injured plants. Part I.

This review is a comprehensive study of recent advances related to cytological, biochemical and physiological changes induced in plants in response to eriophyoid mite attack. It has been shown that responses of host plants to eriophyoids are variable. Most of the variability is due to individual eriophyoid mite-plant interactions. Usually, the direction and intensity of changes in eriophyoid-infested plant organs depend on mite genotype, density, or the feeding period, and are strongly differentiated relative to host plant species, cultivar, age and location. Although the mechanisms of changes elicited by eriophyoid mites within plants are not fully understood, in many cases the qualitative and quantitative biochemical status of mite-infested plants are known to affect the performance of consecutive herbivorous arthropods. In future, elucidation of the pathways from eriophyoid mite damage to plant gene activation will be necessary to clarify plant responses and to explain variation in plant tissue damage at the feeding and adjacent sites.

Phenotypic variability in five Aceria spp. (Acari: Prostigmata: Eriophyoidea) inhabiting Cirsium species (Asteraceae) in Serbia.

From about 250 Cirsium spp., only two Aceria spp. (Acari: Eriophyoidea) have been described, Aceria anthocoptes (Nal.) and Aceria cirsii Pet. B. & Shi. Host specificity, which generally characterizes eriophyoid mites, potentially leads to speciation, so we may expect more than two Aceria spp. and/or other infraspecific taxa. Furthermore, studies on host-related variability in the morphology of Aceria mites are generally lacking. The purpose of this study was to investigate quantitative morphological traits of five Aceria populations inhabiting five Cirsium spp. in Serbia. MANOVA analysis revealed significant differences in 23 commonly used morphological traits as well as four additional traits related to the prodorsal shield design. In addition, the most important qualitative traits using scanning electron microphotographs were studied in order to clarify phenotypic differences among five Aceria spp. Discriminant analysis identified eight traits that significantly differentiate five populations. UPGMA cluster analysis of the squared Mahalanobis distances indicates that A. cirsii was morphologically the most divergent, while A. anthocoptes populations from Cirsium arvense and Aceria sp. from Cirsium heterophyllum were isolated from the branch clustering Aceria spp. populations from Cirsium eriophorum and Cirsium creticum. Analysis of qualitative traits using SEM confirmed results obtained from the analysis of morphometric features.

Plant-eriophyoid mite interactions: specific and unspecific morphological alterations. Part II.

The paper presents recent advances related to both specific and unspecific morphological alterations of plant organs caused by eriophyoid mites. Based on old and new case studies, the diversity of plant malformations, such as galls, non-distortive feeding effects and complex symptoms induced by eriophyoids and/or pathogens vectored by them, is analysed and summarised.

Morphological variation in different populations of Aceria anthocoptes (Acari: Eriophyoidea) associated with the Canada thistle, Cirsium arvense, in Serbia.

The russet mite, Aceria anthocoptes (Nal.), is the only eriophyid that has been recorded on Canada thistle, Cirsium arvense (L.) Scop. It has been noted in several European countries and recently in the USA. With its apparent host specificity and because of the damage it causes to its host plant, A. anthocoptes is being studied as a potential candidate for classical biological control. The aim of the present study was to examine quantitative morphological traits in four populations of A. anthocoptes living on two infraspecific host plant taxa (C. arvense var. arvense and C. arvense var. vestitum) in two geographically separate areas of Serbia in order to test the hypothesis of absence of the possible host plant impact on mite morphology. MANOVA analysis revealed significant differences between populations from different localities in Serbia. Populations of A. anthocoptes inhabiting two thistle varieties in the vicinity of Belgrade differed significantly from mites inhabiting the same two host varieties in the vicinity of the town of Ivanjica. Canonical discriminant analysis showed that the trait which best discriminates the populations of A. anthocoptes is the number of dorsal annuli. It was not possible to ascribe morphological differences to the impact of the host plant.

Anatomical injury induced by the eriophyid mite Aceria anthocoptes on the leaves of Cirsium arvense.

Anatomical injury of the leaves of the invasive species, Cirsium arvense (L.) Scop., caused by the eriophyid mite Aceria anthocoptes (Nal.), which is the only eriophyid mite that has been recorded on C. arvense worldwide, is described. The injury induced by the mite feeding on the leaves of C. arvense results in visible russeting and bronzing of the leaves. Other conspicuous deformations are folding and distortion of the leaf blade and curling of leaf edge, as well as gradual drying of leaves. The anatomical injury of the mature leaves of field-collected plants was limited to the epidermis of the lower leaf surface. However, on young leaves of experimentally infested plants, rust mite injuries extend to epidermal cells on both leaf surfaces and to those of deeper mesophyll layers. On these leaves, lesions on the lower leaf surface even affected the phloem of the vascular bundles. Leaf damage induced by A. anthocoptes is discussed with regard to the mite's potential as a biological control agent of C. arvense.