DNA barcoding of stoneflies (Plecoptera) in a general genetics course

DNA Barcoding is a powerful molecular biology tool for the identification of species, analysis of gene flow from within and between populations and evaluating species concepts. The process can be incorporated into a college genetics or bioinformatics core curriculum. Here we demonstrate how, by simply using Plecoptera, we actively engaged students in DNA extraction, amplification and DNA Barcoding. As a result, 38 new DNA sequences were accepted by an international DNA database, with potentially three of them being new species to the database. More importantly, through multiple assessment measurements, students showed a high degree of learning took place and that all student learning objectives were met.     

The stoneflies (Plecoptera) of Belarus: Fauna and biotopic distribution

A total of 29 species of stoneflies (Plecoptera) from 7 families were found in Belarus: Perlodidae—6, Perlidae—1, Chloroperlidae—3, Taeniopterygidae—2, Nemouridae—11, Capniidae—2, and Leuctridae—4 species. The fauna is fairly diverse and includes a number of rare species in Belarus and other European countries: Diura bicaudata, Marthamea vitripennis, Isoptena serricornis, Xanthoperla apicalis, Brachyptera risi, Protonemura intricata, Nemoura cambrica, Nemoura marginata, and Capnia bifrons.     

Biometric studies of some stoneflies and a mayfly (Plecoptera and Ephemeroptera)

Aspects of the nymphal/adult developmental change were investigated in biometric studies of several species of Plecoptera: Nemouridae near Schlitz, Hesse, Germany. Preliminary information on the mayfly, Baetis vernus Curtis, is also provided. Nemourid nymphs pass through 3 wing bearing stages before reaching adulthood. Instars can be identified by their characteristic shapes, as expressed by the wing length/head width (WL/HW) ratio. Size does not allow instar discrimination, mainly due to sexual size differences. HW is ca 10% larger in last instar female than in male nemourid nymphs; exuviae shed at the moult to adult represent about 14% of nymphal ash free dry weight (AFDW). Biomass lost with exuviae during the many larval moults should be accounted for in estimates of production. Freshly emerged nemourid females are about 6% larger and 30% heavier than males. The HW/AFDW relationship is the same in both sexes. Through terrestrial feeding during adult life, males double their weight on average. Mature females are up to three times heavier than freshly emerged ones. They invest about 30% of their final AFDW in reproduction.Shape of last instar nymphal Baetis was expressed as the ratio wing length/mesonotum length. It is size-dependent, a characteristic, instar-specific shape may not occur in this mayfly. Nymphal and subimaginal exuviae together represent about 14% of last instar nymphal dry weight. Females of Baetis are about 55% heavier than males. Unlike in Plecoptera, the size/weight (ML/AFDW) relationship differs between sexes.     

Post-emergence migration of stoneflies (Plecoptera) into the nearby forest

Abstract. 1. The post-emergence lateral migration of both sexes of eight stonefly species was examined in a dystrophic, fourth-order forest river in eastern Finland.
2. For this purpose, 7351 stonefly adults were collected with eighteen trunk funnels positioned in rows of six at distances of approximately 1, 15 and 60 m from the river. A further 1880 adults were also caught from the vegetation of the bank zone by sweep netting and with slit traps.
3. The species could be grouped into two types with respect to migration distance: Isoperla difformis, I.grammatica and Leuctra fusca tended to stay in the bank area, whereas Nemoura flexuosa, N.avicularis, Amphinemura borealis, L.hippopus and N.cinerea tended to disperse into the forest, so that the majority were found some distance away from the shore.
4. The males of the leuctrids, N.flexuosa and N.cinerea migrated farther than the females.
5. The sex ratio was significantly biased in all species except N.flexuosa. The isoperlids, leuctrids and A.borealis showed a significant predominance of females in the trunk funnel catches, but males were significantly dominant in N.avicularis and N.cinerea. The material caught by other methods reversed the ratio for I.difformis and N.avicularis. Comparison of the sex ratios of the species with other reports revealed marked variation and deviation from unity, much of which could be attributed to bias introduced by the sampling methods.
6. Lateral migration seems to be the first phase in the colonization cycle, although the latter as such was not studied here. Adults of Euholognatha species migrate farther than those of Systellognatha, a difference of which may be due to their ability to feed as adults.     

Global diversity of stoneflies (Plecoptera; Insecta) in freshwater

Plecoptera, or stoneflies, is a small order of hemimetabolous insects: according to our data, more than 3,497 species have been described so far in the world. The total number of species has enormously increased in the last 30 years (2,000 species estimated in 1976) and, if the trend continues, then it will nearly double in the near future. The order is divided into the suborders Arctoperlaria and Antarctoperlaria, and includes 16 families: 12 arctoperlarian and 4 antarctoperlarian. The Arctoperlaria account for a total number of 3,179 species, and Antarctoperlaria, only 318 species. The total number of genera is 286. We give in this article the estimated number of species for each family. The fauna and diversity of stonefly in North America (650 species reported) and Europe (426 species) are best known. Nevertheless, in the last 25 years, a mean of 2.6 Plecoptera species per year were described in Europe. Stonefly-faunas of Australia (191 species, Tasmania included) and New Zealand (104 species) are relatively well-known, while our knowledge of the Plecoptera of Central and South America (95 and 378 species respectively) is poor and still not representative of the real diversity. Africa has a reduced stonefly fauna (126 species). Asian stonefly diversity (approximately 1,527 species) is much greater than that of Europe or North America despite the fact that, except for Japan and Asiatic Russia that have been well studied, our knowledge of the remaining Asiatic areas is extremely poor. Even though our data indicate the Holarctic Region as the diversity hot-spot for the order, the analysis of the specific diversity divided by family suggests also an important role of tropical stoneflies. Guest editors: E.V. Balian, C. Lévêque, H. Segers & K. Martens Freshwater Animal Diversity Assessment     

Resource division by perlid stoneflies (Plecoptera) in a lake outlet ecosystem

In a stream draining a productive lake, two abundant (250/m²) stoneflies Calineuria californica and Hesperoperla pacifica differed in population size structure, habitat use and food although overlap in resource use was extensive. Life cycles of two (Calineuria) and three years (Hesperoperla) were indicated. A multivariate analysis of habitat use demonstrated size dependent habitat selection in both species and between species habitat differences. Feeding habits differed although only the largest Hesperoperla had a food refuge. Diets of individual nymphs represented an interaction of species, size and habitat.     

Implications of imaginal-size variation over the flight period in stoneflies (Plecoptera)

In the present study, the decrease in the mean size of specimens over the species flight period in Plecoptera (12 species, 5 families) was analysed, revealing a significant decrease in the adult mean size, for both males and female, in almost all the species without an extended flight-period pattern. Some of the data show that, in addition to temperature, other regulatory factors must govern these variations. Data support the explanation previously proposed for other insect groups that the individuals with appropriate development emerge at the beginning of the flight period or slightly afterwards, when environmental conditions are suitable, whereas individuals with suboptimal size emerge later trying to increase in size but under pressure to emerge during a specific period of the year. Nevertheless, our data suggest that maximum size does not necessarily coincide with the peak flight period (maximum reproductive possibility).     

Molecular phylogeographic analysis of East Asian cryptoperlan stoneflies (Insecta: Plecoptera, Peltoperlidae)

Cryptoperlan stoneflies inhabit the headwaters or upper stream areas of rivers. A molecular phylogeographic study of cryptoperlans in the Japanese archipelago and on Taiwan Island has been conducted. Altogether the mtDNA 16S rRNA region of 71 individuals from 61 populations, the mtDNA COI region of 76 individuals from 41 populations, and the nDNA Histone 3 region of 56 individuals from 52 populations were sequenced and analyzed. The respective ML, NJ, MP and Bayesian dendrograms were proposed from the sequencing data for the 16S rRNA region (362-bp), the COI region (540-bp), and the Histone 3 region (322-bp), estimated using Yoraperla uenoi as an outgroup. Based upon those data and the resulting dendrograms, it has become clear that the cryptoperlan stoneflies of the Japanese archipelago and those of Taiwan Island comprise two major clades. The first of these two major clades consists of a number of OTUs [operational taxonomic units: Cryptoperla japonica (Honshu, Shikoku and Kyushu Islands) + C. ishigakiensis (Ishigaki-jima Island) + Cryptoperla spp. (Okinawa-jima and Taiwan Islands)]. The other clade consists of the species Cryptoperla kawasawai inhabiting only Shikoku Island. Of particular note, C. kawasawai was observed to be significantly genetically differentiated from all other cryptoperlans examined. Yet, despite the fact that the specimens of C. japonica were taken from a very broad range of populations, their genetic diversity was relatively low, similar to that of C. kawasawai, which inhabits only a limited region within Shikoku Island. Furthermore, even the species C. kawasawai was revealed to be composed of two significantly genetically differentiated subclades. It is considered that this genetic structure among cryptoperlans largely reflects the geological history from the middle to upper Miocene Epoch (i.e., Tortonian stage) of the Japanese archipelago and Taiwan Island.     

New stoneflies (Insecta: Perlida = Plecoptera) from Eocene Rovno amber,Ukraine

A new genus, Rovnocapnia gen. nov., with new species R. atra sp. nov. and R. ambita sp. nov., from the family Capniidae, and a new genus and species Palaeoleuctra acuta gen. sp. nov., from the family Leuctridae, of the suborder Nemourina (Euholognatha), are described from Late Eocene Rovno amber. Familial assignment of one additional specimen (suborder Perlina, infraorder Perlomorpha (Systelognatha)), represented by a nymphal skin, is obscure due to its incomplete preservation.     

Contribution to the knowledge of the adult feeding of Nemouroidea stoneflies (Insecta: Plecoptera)

The feeding habits of 17 species of Nemouroidea stoneflies (belonging to the families Nemouridae, Leuctridae and Capniidae) from the north‐western Iberian Peninsula are described and quantified by means of gut content analyses. As a whole, fungi, followed by pollen (both from angiosperm and, in a lesser degree, Pinaceae), leaf fragments (CPOM), detritus and, more punctually, lichens and animal matter are the most common items found in their guts. Nevertheless, variations among species were observed. For a species with a flight period that extends throughout the year, Protonemura globosa, variations in the diet were noticed among seasons, with pollen being more important during the spring and summer period. Together with a few previous studies, the present one reaffirms that feeding is an important aspect of the adult life in males and females of Nemouroidea.     

A new genus of African Lasiocampidae (Lepidoptera)

The species Gastroplakaeis idakum Bethune-Baker, 1913, G. balia Tams, 1929, and G. elongata Hering, 1941 are transferred from the genus Gastroplakaeis Möschler, 1887 into the newly erected genus Weirdonia gen. n. (the type species is Gastroplakaeis idakum Bethune-Baker, 1913), and corresponding new combinations are established. New species, Weirdonia baccara sp. n. and W. hypocrita sp. n. (type locality: Democratic Republic of the Congo), W. murena sp. n. (type locality: Northern Nigeria), and W. helicea sp. n. (type locality: Gabon) are described. A new synonymy is established: Gastroplakaeis balia Tams, 1929 = Gastroplakaeis elongata Hering, 1941 syn. n. Diagnoses and distribution maps are given for all the species.     

中国诺衤责属两新种记述（衤责翅目：卷衤责科）

记述中国卷襀科Leuctridae诺襀属Rhopalopsole 2新种:须毛诺襀Rhopalopsole intonsa sp. nov.和短剑诺襀Rhopalopsole siculiformis sp. nov.。须毛诺襀与诺襀属其它种类的区别在于:第9背板后缘有1 横长方形骨化斑;第10背板中骨片下方的1对横向三角形骨片具有浓密的刷毛,侧面观能清晰看到1 簇刷毛;肛下叶基部有1对小叶状膜片,肛下叶末端钝圆且膜质;尾须端有小刺。短剑诺襀区别于诺襀属其它种类的特征是:第9背板中部有1个"T"形弱骨化区域,肛下叶末端膜质,背面观形似1柄短剑状;尾须端无小刺。模式标本保存于扬州大学昆虫标本室。     

Dispersal,feeding and parasitism of adult stoneflies (Plecoptera) at a New Zealand forest stream

The temporal occurrence and dispersal of stoneflies were investigated at a forest stream in the South Island, New Zealand. Although 12 species were taken in Malaise traps, only Spaniocerca zelandica and Cristaperla fimbria (Notonemouridae) were abundant. Adults of S. zelandica were present throughout the year and a wide range of different-sized nymphs, including very small individuals, was also found in all months. In contrast, the emergence period of C. fimbria was limited to about four months in summer and as nymphs are mainly hyporheic, few were taken in Surber samples. Malaise trapping on a 16 m long transect away from the stream and with more distantly located traps in forest and grassland resulted in a very high proportion of males and females of both species being caught within 1 m of the stream edge. Captured females of both species included immature, gravid, and spent individuals. Males and females of both species had almost identical diets dominated by sooty mould fungi, fungal spores, pollen and fine particulate organic matter. High proportions of them were also infected by the encysted larvae of a gordian worm (Nematomorpha) and may be important vectors in its life cycle.     

Structure of the ovaries in larvae and mature females of euholognathan stoneflies (Plecoptera)

The morphology of ovaries, oviducts and egg capsules in four species of euholognathan stoneflies was investigated. The characteristic features found were as follows: (i) numerous, long ovarioles, that open individually to the extensively folded, lateral oviducts; (ii) a thin, morphologically undifferentiated chorion; (iii) a thick gelatinous layer (extrachorion) which acts as an adhesive layer fixing the eggs to the substrate. Additionally, in the larval ovariole of Leuctra sp. the terminal filament anlage and clusters of germ cells have been found. These observations are in agreement with the classification of stonefly ovaries as primary (true) panoistic.     

中国华襀属一新种记述（襀翅目：襀科）

记述中国襀科 Perlidae 华襀属 Chinoperla 1 新种:勐满华襀 Chinoperla mengmanensis sp. nov.。本新种与其他华襀属种类的区别在于:第9背板前缘中部骨片短小且强骨化,呈倒V形;半背片细长,紧挨第 9 背板中部骨片;阳茎部分骨化,阳茎囊未完全翻出,但是明显呈三叶,背外侧两叶完全覆盖小棘刺,中叶呈球形囊泡,无棘刺。模式标本保存于扬州大学昆虫标本室。     

A new genus for a rare African vespertilionid bat: insights from South Sudan

A new genus is proposed for the strikingly patterned African vespertilionid “Glauconycteris” superba Hayman, 1939 on the basis of cranial and external morphological comparisons. A review of the attributes of a newly collected specimen from South Sudan (a new country record) and other museum specimens of “Glauconycteris” superba suggests that “Glauconycteris” superba is markedly distinct ecomorphologically from other species classified in Glauconycteris and is likely the sister taxon to Glauconycteris sensu stricto. The recent capture of this rarely collected but widespread bat highlights the need for continued research in tropical sub-Saharan Africa and in particular, for more work in western South Sudan, which has received very little scientific attention. New country records for Glauconycteris cf. poensis (South Sudan) and Glauconycteris curryae (Gabon) are also reported.     

Diet and predation by three leaf-associated stoneflies (Plecoptera) in an Arkansas mountain stream

SUMMARY. 1. Life cycles, food habits, mouthpart morphologies, prey preferences, and predator-prey size relationships were investigated for the three most common stoneflies in leaf packs in the Little Missouri River, Arkansas: Clioperla clio (Newman), Isoperla namata Prison, and Perlesta spp. Each species was univoltine and had a fast seasonal cycle, with extended egg or nymphal diapause, rapid nymphal growth and emergence in spring to early summer.
2. Foregut analysis showed predictable dietary shifts by C. clio and Perlesta spp., from detritus and diatoms in early instars to invertebrate prey in later instars. Isoperla namata fed facultatively on insects, detritus and diatoms, with no conspicuous ontogenetic shifts. The mouthparts of all three species strongly reflected their diet.
3. Electivity analysis indicated opportunistic feeding by the three species, with feeding rarely differing significantly from random.
4. Sizes of stonefly predators and their ingested prey were highly correlated ( P <0.01). However, rather than shifting toward larger prey sizes, growing predators expanded their size thresholds and continued to include numerous small prey items in their diet. Prey choice was governed most directly by prey availability, because stoneflies selected the most abundant groups, irrespective of size or taxon.
5. The likelihood of competition among the three stoneflies for prey was minimized by timing of their life cycles, differential food use and probably the temporary nature of leaf pack habitats.     

Killickia (Lamiaceae): a new genus from KwaZulu-Natal, South Africa

The genus Killickia is described to accommodate the South African endemic species formerly placed in Micromeria sect. Hesperothymus . Morphological data, as well as results from unpublished phylogenetic studies support its separation from the genera Micromeria and Clinopodium . A new species Killickia lutea Bräuchler is described and three new combinations are made. Killickia is characterised as comprising solitary- or few-flowered cymes, a campanulate to subcampanulate (obconical) calyx with similar teeth, a corolla tube with two pubescent ridges and nutlets with scattered minute hairs. A thickened marginal vein in the leaves as typical for Micromeria is absent. As currently understood all species are restricted to the Drakensberg mountains and KwaZulu-Natal Midlands in eastern South Africa. A key to the species, brief notes on anatomy and ecology are provided.  © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society , 2008, 157 , 575–586.