Rapid diagnosis of the invasive wax scale,Ceroplastes rusci Linnaeus (Hemiptera: Coccoidea: Coccidae) using nested PCR

The fig wax scale, Ceroplastes rusci (Linnaeus) (Hemiptera: Coccoidea: Coccidae), is an invasive fruit pest of Afrotropical origin and potentially could become a serious threat to commercial fruit crops in China. C. rusci is difficult to identify owing to the shortage of easily distinguishable morphological characters. A rapid, accurate and reliable method to identify C. rusci in quarantine work is needed to detect further spread. In the present study, we describe a nested PCR method for the molecular identification of C.rusci. The nested PCR primers were designed based on variations in the barcode region of COI sequences between C. rusci and five other Ceroplastes species. A 200‐bp fragment was successfully amplified from 96 C. rusci individuals of seven geographical populations in China and Vietnam, and 13 individuals of two populations in Italy (the type country for C. rusci). These provided diagnostic bands that were not observed in any of five other Ceroplastes species widely distributed in China, namely, C. ceriferus (Fabricius), C. floridensis Comstock, C. japonicus Green, C. pseudoceriferus Green and C. rubens Maskell. Sensitivity tests revealed that diagnostic bands were generated even with a DNA template concentration of ~1.5 × 10^?5 ng/μl, and with average DNA template concentrations for adult females, single first‐instar nymphs and eggs of 14.7, 6.3 and 3.0 ng/μl, respectively. Our study demonstrates that the molecular diagnosis of C. rusci using nested PCR is rapid and accurate and shows potential in plant quarantine programmes.     

外来入侵新害虫——无花果蜡蚧

无花果蜡蚧Ceroplastes rusci (Linnaeus)原产于非洲,是危害经济林和园林树种的重要害虫。2007年被列入《中华人民共和国进境植物检疫性有害生物名录》中。2012年4月下旬,该虫于广东省茂名市和四川省攀枝花市的榕树上被首次发现,为我国的新入侵害虫。本文介绍了其形态特征、地理分布、寄主植物、生物学和经济重要性等。     

A new species of Coccus (Hemiptera,Coccoidea,?Coccidae) from China

A new species of soft scale, Coccus multisetus Wang & Feng, sp. n. is described and illustrated from Yunnan, China. A key to adult females of all Coccusknown from China is provided.     

中国蜡蚧科一新记录属（半翅目：蚧总科）

记述采自中国云南的1新记录属:类白蜡蚧属Ericeroides Danzig,1990及1新记录种:越南类白蜡蚧E.zaitzevi Danzig,1990。该新记录属近似于白蜡蚧属Ericerus Guérin-Méneville,二属均有成群气门刺及爪齿;主要区别是:类白蜡蚧属Ericeroides无背面管状腺及肛前孔,而白蜡蚧属Ericerus则有。文中对雌成虫形态特征进行了详细描述和绘图。     

我国发现一新入侵蜡蚧:三列鬃软蚧(半翅目,蚧总科,蜡蚧科)

本文记述采自中国惠州的1新记录属鬃软蚧属Trijuba De Lotto,该属目前只有1种,区别于蜡蚧科其他属的特征是雌成虫背面具有3列长刚毛。新记录种三列鬃软蚧Trijuba oculata(Brain)寄生于猴耳环Pithecellobium clypearia的枝干和叶柄上。     

The hinged back plate mechanism in glassy wax tests of New Zealand male soft scale insects (Hemiptera: Coccoidea: Coccidae)

Male scale insects (Hemiptera: Coccoidea) undergo a metamorphosis of the neometabola type, from scale-like nymph through prepupa and pupa to winged adult. The nymphal instar before prepupa secretes a waxy protective covering that remains in place throughout metamorphosis and these covers are characteristic of each family of scale insects. Most scale insect families (e.g. mealybugs, eriococcids, diaspidids) have rather loosely woven male covers, but male nymphs in the family Coccidae (soft scales) construct more rigid, glassy wax tests, which need a special mechanism for adult emergence. In the New Zealand male soft scales, a suture across the posterior quarter of the test enables the back plate to flex at a pair of hinges, to be raised up off the substrate, and so allow egress. The waxy back plate hinges are secreted by groups of tubular ducts on the abdominal dorsum of 2nd-instar males, during construction of the test. Scanning electron micrographs (SEMs) show the detail and diversity of hinge types. The wax tests of most New Zealand Coccidae, both female and male, are apparently unique in that they are constructed in rows of hexagonal plates, separated by sutures, however in the male test, the sutures are all fused except for the back plate suture. The two species in the endemic New Zealand genus Pounamococcus have male tests more like those of species in the Australian genus Austrolecanium.     

A cladistic analysis of wax scales (Hemiptera: Coccoidea: Coccidae: Ceroplastinae)

Seven genera of wax scales (Ceroplastinae) have been accepted as valid by some or all coecidologists: Ceroplastes Gray, Ceroplastidia Cockerell, Cerostegia De Lotto, Gascardia Targioni Tozzetti, Paracerostegia Tang, Vinsonia Signoret and Waxiella De Lotto. The status of most these genera is controversial. The present study reviews the taxonomic history of wax scales and presents a new classification based on cladistic analysis of morphological data from the adult females of eighty-three species. The analysis indicates that the wax scales form a monophyletic group, that Waxiella is monophyletic but that Ceroplastidia, Gascardia and Vinsonia are not, and fails to support the generic status of Cerostegia and Paracerostegia. Apart from the type species of Vinsonia , which was placed as a sister to the rest of wax scales, the only other species assigned to Vinsonia and most species of all other genera fall within Ceroplastes in the cladogram. Recognition of any of these genera renders Ceroplastes paraphyletic or polyphyletic. Thus, cladistically, the wax scales should be classified into one genus, Ceroplastes.     

DNA barcoding of six Ceroplastes species (Hemiptera: Coccoidea: Coccidae) from China

Ceroplastes Gray (wax scales) is one of the genera of Coccidae, most species of which are considered to be serious economic pests. However, identification of Ceroplastes species is always difficult owing to the shortage of easily distinguishable morphological characters. Mitochondrial cytochrome c oxidase I (COI) sequences (or DNA barcodes) and the D2 expansion segments of the large subunit ribosomal RNA gene 28S were used for accurate identification of six Ceroplastes species (C. floridensis Comstock, C. japonicus Green, C. ceriferus (Fabricius), C. pseudoceriferus Green, C. rubens Maskell and C. kunmingensis Tang et Xie) from 20 different locations in China. For COI data, low G·C content was found in all species, averaging about 20.4%. Sequence divergences (K2P) between congeneric species averaged 12.19%, while intra‐specific divergences averaged 0.42%. All 112 samples fell into six reciprocally monophyletic clades in the COI neighbour‐joining (NJ) tree. The NJ tree inferred from 28S showed almost same results, but samples of two closely related species, C. ceriferus and C. pseudoceriferus, were clustered together. This research indicates that the standard barcode region of COI can efficiently identify similar Ceroplastes species. This study provides an example of the usefulness of barcoding for Ceroplastes identification.     

Preliminary study of wing interference patterns (WIPs) in some species of soft scale (Hemiptera,Sternorrhyncha, Coccoidea,Coccidae)

The fore wings of scale insect males possess reduced venation compared with other insects and the homologies of remaining veins are controversial. The hind wings are reduced to hamulohalterae. When adult males are prepared using the standard methods adopted to females and nymphs, i.e. using KOH to clear the specimens, the wings become damaged or deformed, an so these structures are not usually described or illustrated in publications. The present study used dry males belonging to seven species of the family Coccidae to check the presence of stable, structural colour patterns of the wings. The visibility of the wing interference patterns (WIP), discovered in Hymenoptera and Diptera species, is affected by the way the insects display their wings against various backgrounds with different light properties. This frequently occurring taxonomically specific pattern is caused by uneven membrane thickness and hair placement, and also is stabilized and reinforced by microstructures of the wing, such as membrane corrugations and the shape of cells. The semitransparent scale insect’s fore wings possess WIPs and they are taxonomically specific. It is very possible that WIPs will be an additional and helpful trait for the identification of species, which in case of males specimens is quite difficult, because recent coccidology is based almost entirely on the morphology of adult females.     

中国克里蜡蚧属二新记录种（半翅目：蚧总科：蜡蚧科）

记述采自海南的克里蜡蚧属Kilifia 2新记录种:Kilifia deltoides De Lotto, 1965和K. americana Ben-Dov, 1979。对雌成虫形态特征进行了详细描述和绘图,提供了克里蜡蚧属中国已知种的检索表。研究标本保存在西北农林科技大学昆虫博物馆。     

An annotated checklist of the scale insects of Iran (Hemiptera,Sternorrhyncha, Coccoidea) with new records and distribution data

A list of scale insects (Hemiptera: Sternorrhyncha: Coccoidea) of Iran is present based mainly on the literature records since 1902. In total, 13 families and 275 species have been recorded and these are listed along with their locality data and host plants. The families are as follows: Asterolecaniidae, Cerococcidae, Coccidae, Diaspididae, Eriococcidae, Kermesidae, Margarodidae, Monophlebidae, Ortheziidae, Phoenicococcidae, Pseudococcidae, Putoidae and Rhizoecidae. The following ten species are recorded for the first time from Iran: Diaspidiotus lenticularis (Lindinger), Diaspidiotus wuenni (Lindinger), Fiorinia proboscidaria Green, Koroneaspis lonicerae Borchsenius, Eriococcus cingulatus Kiritchenko, Eriococcus pamiricus (Bazarov), Eriococcus reynei Schmutterer, Eriococcus sanguinairensis Goux and Eriococcus saxidesertus (Borchsenius) and Porphyrophora victoriae Jashenko.     

A molecular phylogenetic study of armoured scale insects (Hemiptera: Diaspididae)

Abstract.  Armoured scale insects are economically important parasites of woody plants and grasses. They are promising subjects for the evolutionary study of physiology (no complete gut), genetics (chimerism, paternal genome elimination, frequent parthenogenesis) and coevolution (with host plants, parasitoids, Septobasidium fungi, endosymbiotic bacteria). Little phylogenetic work has been accomplished with armoured scales, and uncertainty surrounds their classification. Here, we report the phylogenetic results of Bayesian and parsimony analyses of 705 base pairs of Elongation Factor 1α and 660 base pairs of 28S from eighty-nine species of armoured scale insects, representing forty-seven genera and five tribes in the subfamilies Diaspidinae and Aspidiotinae, together with two outgroups. 28S was aligned based on a secondary structural model. Our results broadly corroborate the major features of the existing classification, although we do not find perfect monophyly of any of the traditionally recognized subfamilies or tribes. The subfamily Aspidiotinae is paraphyletic with respect to the subfamily Diaspidinae. Diaspidinae consists of two main clades that only roughly correspond to the tribes Lepidosaphidini and Diaspidini. Diaspidini is nearly monophyletic, except that it includes a single aspidiotine species. Other members of the tribe Aspidiotini form a clade, except that the clade includes a single species of Leucaspidini and excludes Maskellia and Pseudaonidia . Our results weakly support the hypothesis that the most recent common ancestor of the Diaspididae had adult females that were permanently enclosed within the derm of the second instar (the pupillarial habit) and had diploid adult males that eliminated their paternal genomes during spermatogenesis (late paternal genome elimination).     

Gonoid thelytoky in soft scale insects (Coccidae: Homoptera)

Cytological analysis of the thelytokous soft-scale insects Coccus hesperidum L. (2n=14) and Saissetia coffeae (Walker) (2n=16) revealed that while in both species the chromosomes did not pair during prophase I, meiosis consisted of two divisions, the chromosome number was reduced, and diploidy was restored by the fusion of the female pronucleus with the polar nucleus II. The difficulty of trying to classify this type of thelytoky as either automictic or apomictic led to the proposal that a new criterion and new terms be used to classify thelytoky (and parthenogenesis). The new criterion is whether the number of chromosome elements present in the first (or only) metaphase of oogenesis is the same as that present in the oogonia (gonoid thelytoky) or different from it (agonoid thelytoky). The new criterion is superior to the existing criteria because it is unambiguous, and because it groups together forms with a similar tendency towards heterozygosity (or homozygosity). The possible evolution of the forms analyzed as well as the two other thelytokous forms of each species described by Thomsen (1927) are discussed. Another soft-scale insect, Physokermes hemicryphus Dalam, consisted of a diploid (2n=18) and a triploid (3n=27) form, in both of which the chromosomes also did not pair. Each of the three species contained a strain in which only a single nucleolus was present per cell. In C. hesperidum some strains with two nucleoli differed in the size of the nucleoli.     

Description of a new soft scale insect of the genus Pulvinaria Targioni Tozzetti (Hemiptera,Coccoidea, Coccidae) from Bogota,Colombia

A new soft scale (Hemiptera: Coccoidea: Coccidae) species, Pulvinaria caballeroramosae Tanaka & Kondo, sp. n., is described from specimens collected on twigs of Ficus soatensis Dugand (Moraceae) in Bogota, Colombia. The new species resembles Pulvinaria drymiswinteri Kondo & Gullan, described from Chile on Drimys winteri J.R. Forst. & G. Forst. (Winteraceae), but differs in the distribution of preopercular pores on the dorsum, the presence of dorsal tubular ducts, dorsal microducts, and reticulation on the anal plates; and in its feeding habits, i.e., Pulvinaria caballeroramosae feeds on the twigs whereas Pulvinaria drymiswinteri feeds on the leaves of its host. A key to the Colombian species of Pulvinaria Targioni Tozzetti is provided.     

A taxonomic revision of Limacoccus Bondar with a cladistic analysis of its relationships with other scale insects (Hemiptera: Coccoidea)

The scale insect genus Limacoccus and its four palm pest species are revised and all instars with sexual dimorphism are described and illustrated. A key to Limacoccus species is provided. The unique life-history of this genus with construction of a protective tunnel by the second-instar female is described. A cladistic analysis of Limacoccus and eleven representative species of Diaspididae, Halimococcidae, Phoenicococcidae and Beesoniidae, was undertaken using fifty-four traditional and new characters from first and second instars and adult females. Character polarity was determined by outgroup comparison with Margarodidae, Pseudococcidae, Eriococcidae and Asterolecaniidae as outgroups. Nine most parsimonious trees were found. Analysis of the eleven terminal taxa of the strict consensus tree using Phoenicococcidae, Halimococcidae and Diaspididae as outgroups produced one parsimonious tree. As result of this analysis, Limacoccus is believed to be a monophyletic group of characterized by seven autapomorphies and is assigned to the family Beesoniidae fam.rev. The redefined family is divided into two tribes: Beesoniini stat.nov. with genera Beesonia and Mangalorea and Limacocciini stat.nov. with the genus Limacoccus. The Beesoniidae are here regarded as a sister group of Phoenicococcidae.     

Phylogeny of ensign scale insects (Hemiptera: Coccoidea: Ortheziidae) based on the morphology of Recent and fossil females

The Ortheziidae (ensign scale insects) is a morphologically well‐defined family. The morphology and occurrence in the fossil record suggests a probable early origin of the family in scale insect evolution. The present phylogenetic analysis – based on 69 morphological characters of female ortheziids, using 39 exemplar Recent species – provides the first analytical assessment of relationships among living genera of the family, as well as the relationships of eight fossil species, based on complete, well‐preserved specimens in amber. Monophyly of the subfamilies Newsteadiinae, Ortheziinae and Ortheziolinae is supported, but Nipponortheziinae is found to be paraphyletic by inclusion of the Ortheziolinae. Thus, the subfamily Ortheziolinae is reduced in rank to tribe Ortheziolini stat.n. , which now includes Matileortheziola Kozár & Foldi, Ortheziolacoccus Kozár, Ortheziolamameti Kozár and Ortheziola?ulc. Consequently, the tribes Matileortheziolini, Ortheziolacoccini and Ortheziolamametini are synonymized ( syn.n. ) here under Ortheziolini. Five new species and one new genus of fossil ensign scales are described from three amber deposits: Burmorthezia gen.n. with type species Burmorthezia kotejai sp.n. and also B. insolita sp.n ., both in mid‐Cretaceous Burmese amber (98 Ma) and Arctorthezia baltica sp.n. in Eocene Baltic amber (c. 43 Ma) based on second‐instar nymphs; Mixorthezia kozari sp.n . and M. dominicana sp.n . in Miocene Dominican amber (c. 17 Ma) based on adult females. Fossil placements are unambiguous, with Burmorthezia forming a stem to crown‐group (Recent and Tertiary) Ortheziidae. A summary of described fossil ortheziids is provided.     

A preliminary phylogeny of the scale insects (Hemiptera: Sternorrhyncha: Coccoidea) based on nuclear small-subunit ribosomal DNA

Scale insects (Hemiptera: Sternorrhyncha: Coccoidea) are a speciose and morphologically specialized group of plant-feeding bugs in which evolutionary relationships and thus higher classification are controversial. Sequences derived from nuclear small-subunit ribosomal DNA were used to generate a preliminary molecular phylogeny for the Coccoidea based on 39 species representing 14 putative families. Monophyly of the archaeococcoids (comprising Ortheziidae, Margarodidae sensu lato, and Phenacoleachia) was equivocal, whereas monophyly of the neococcoids was supported. Putoidae, represented by Puto yuccae, was found to be outside the remainder of the neococcoid clade. These data are consistent with a single origin (in the ancestor of the neococcoid clade) of a chromosome system involving paternal genome elimination in males. Pseudococcidae (mealybugs) appear to be sister to the rest of the neococcoids and there are indications that Coccidae (soft scales) and Kerriidae (lac scales) are sister taxa. The Eriococcidae (felt scales) was not recovered as a monophyletic group and the eriococcid genus Eriococcus sensu lato was polyphyletic.     

Cellular immune response of brown soft scale Coccus hesperidum L. (Hemiptera: Coccidae) to eggs of Metaphycus luteolus Timberlake (Hymenoptera: Encyrtidae)

The parasitic wasp, Metaphycus luteolus Timberlake, is an endoparasitoid of various soft scale insects including brown soft scale, Coccus hesperidum L. Development of this parasitoid in scale hosts is hindered by encapsulation. In the present study, using transmission and scanning electron microscopy, we show that hemocytes are responsible for encapsulation, which is mediated by the direct deposition of cells and melanin on the surface of M. luteolus eggs. By 12 h post-oviposition, scale hemocytes, presumably granulocytes, aggregate, spread and directly lyse on the surface of parasitoid eggs. This process continues for at least 1 day and results in the gradual formation of a capsule. Two to three days post-oviposition, a melanized capsule is well formed and signs of chemical deposition are evident by examination of the outer surface of the capsule. These results demonstrate that soft scale insects are fully capable of melanotic encapsulation of foreign material mediated by hemocytes.     

Aschersonia basicystis on scale insects (Homoptera: Coccidae) in Venezuela

The fungus Aschersonia basicystison scale insects (Homoptera: Coccidae) is described and illustrated on the basis of the examination of Venezuelan collections, using transmitted light and differential interference contrast optical microscopy.     

热带平刺粉蚧的检疫鉴定

【目的】明确热带平刺粉蚧Rastrococcus tropicasiaticus Williams的鉴别特征,建立平刺粉蚧属种类检索表。【方法】随机抽样、形态学观察和测量。【结果】从进口越南龙眼鲜果中截获热带平刺粉蚧,头胸部腹面多格腺以单排排列,体背面大五格腺少是其识别的主要形态学特征;编制了平刺粉蚧属16种检索表。【结论】热带平刺粉蚧为我国口岸首次检出,研究结果为进口水果中该类粉蚧的检疫鉴定提供了重要的参考资料。