Biological control of Tipula paludosa (Diptera: Nematocera) using entomopathogenic nematodes (Steinernema spp.) and Bacillus thuringiensis subsp. israelensis

Tipula paludosa (Diptera: Nematocera) is the major insect pest in grassland in Northwest Europe and has been accidentally introduced to North America. Oviposition occurs during late August and first instars hatch from September until mid-October. Laboratory and field trials were conducted to assess the control potential of entomopathogenic nematodes (EPN) (Steinernema carpocapsae and S. feltiae) and Bacillus thuringiensis subsp. israelensis (Bti) against T. paludosa and to investigate whether synergistic effects can be exploited by simultaneous application of nematodes and Bti. Results indicate that the early instars of the insect are most susceptible to nematodes and Bti. In the field the neonates prevail when temperatures tend to drop below 10 °C. S. carpocapsae, reaching >80% control, is more effective against young stages of T. paludosa than S. feltiae (<50%), but the potential of S. carpocapsae might be limited by temperatures below 12 °C. Mortality of T. paludosa caused by Bti was not affected by temperature even at 4 °C but the lethal time increased with decreasing temperatures. Synergistic effects of Bti and EPN against T. paludosa were observed in 3 out of 10 combinations in laboratory assays but not in a field trial. The potential of S. carpocapsae was demonstrated in field trials against early instars in October reaching an efficacy of >80% with 0.5 million nematodes m⁻² at soil temperatures ranging between 3 and 18 °C. Results with Bti were strongly influenced by the larval stage and concentration. Against early instars in autumn between 74 and 83% control was achieved with 13 kg ha⁻¹ Bti of 5,700 International Toxic Units (ITUs) and 20 kg ha⁻¹ of 3,000 ITUs. Applications in spring against third and fourth instars achieved between 0 and 32% reduction. The results indicate that application of Bti and nematodes will only be successful and economically feasible during the early instars and that the success of S. carpocapsae is dependent on temperatures >12 °C. Synergistic effects between S. carpocapsae and Bti require more detailed investigations in the field to determine maximal effect.     

Die Quellflur-Gesellschaft des Cratoneuro-Arabidetum bellidifoliae (Koch 1928) in der subalpinen Stufe Graubündens

Summary Two subassociations of the Cratoneuro-Arabidetum bellidifoliae from the subalpine belt in Graubünden (Switzerland) are described for the first time. The association occurs along springs with running, base-rich water and is usually bordering the equally basiphilous Caricetum davallianae. The association also occurs in the Pyrenees. Synsystematically it is related to a Cortusa Matthioli—Crepis paludosa association, occurring as an endemic community in the Lower Engadin.The two subassociations are cratoneuretosum commutati and cratoneuretosum falcati, characterized by the dominance of Cratoneurum commutatum and C. falcatum, respectively.

Communication Nr. 219.     

青藏高原不同生境下湿生扁蕾(Gentianopsis paludosa)个体大小依赖的繁殖分配

植物对资源的投资和分配是生态学中的重要问题,它反映了植物应对环境变化时的生活史策略。选择青藏高原东缘同一海拔下的嵩草草甸(Kobresia sp.meadow)、金露梅灌丛(Potentilla fruticosa shrub meadow)以及草甸-灌丛交错带3种生境类型,并以3种生境下的湿生扁蕾(Gentianopsis paludosa)为对象,研究了其繁殖分配特征。结果发现:(1)在种群水平上,在生境从草甸经交错带到灌丛的变化中,湿生扁蕾个体大小和繁殖分配比例逐渐增加;3个种群湿生扁蕾的总花数目没有显著差异,但草甸生境湿生扁蕾的蕾期花数目显著高于灌丛生境,而果期花数目则显著低于灌丛生境;(2)在个体水平上,湿生扁蕾的繁殖绝对投入与个体大小显著正相关,且各种群植株都存在繁殖所需的个体大小阈值,而繁殖阈值在生境从草甸经交错带到灌丛的过渡中逐渐减小;湿生扁蕾的繁殖相对投入与个体大小负相关,但相关系数随着生境从草甸经交错带到灌丛的过渡中逐渐减小;各种群花数目与湿生扁蕾植株个体大小显著正相关。研究表明,湿生扁蕾的繁殖投资存在大小依赖效应,但生境差异会对其繁殖投资和生活史策略造成显著影响,而这种影响主要是由不同生境下自然条件的不同造成的。同时,资源分配也与湿生扁蕾的遗传特性和延迟自交的繁育系统特征有关。湿生扁蕾这种不同生境下个体大小依赖的繁殖投资差异是湿生扁蕾与其生境长期适应和进化(生境选择)的结果。     

15. Three Rumanian Mallomonas species (Synurophyceae), with special distribution patterns

Mallomonas is the largest genus of the Synurophyceae (chrysophytes) and includes more than 120 species. Most of them are cosmopolitan, living in freshwater habitats. The Rumanian taxa occur usually in spring or early summer at water temperatures ranging between 4 ° and 10 °C. They are most frequent in neutral or slightly acid, clear-watered, aquatic habitats of various types. However, Mallomonas paludosa Fott appears mostly in brown watered peatbog pools of low mineral contents, having a particular geographical area, with its southernmost limit in the Rumanian Eastern Carpathians. Its distribution, like that of other boreal elements, is strictly connected with the occurrence of genuine ombrogenic Sphagnum-bogs, which have their southern limit also in Romania. The other two species dealt with — M. cyathellata Wujek & Asmund and M. portae-ferreae Péterfi & Asmund —, occurring in circumneutral or moderately alkaline waters of eutrophic character, are apparently warm water forms, mostly distributed in tropical, subtropical or mediterranean zones. The Rumanian localities (in the Danube Delta and at Eşelniţa near the Danube, up stream Drobeta Turnu-Severin) might be the northern limits of M. portae-ferreae in Eastern Europe. The Danube Delta is the only locality of M. cyathellata in Europe, and its northernmost occurrence as well. The occurrence of both species in Rumania is presumably due to migratory waterfowl.     

Effect of eight benthic diatoms as feed on the growth of red abalone (Haliotis rufescens) postlarvae

The growth rate of abalone post larvae of Haliotis rufescens fed ad libitum with a benthic monoalgal diatom culture maintained as monocultures on a semi-commercial scale, was evaluated and correlated with the biochemical composition of the diatoms. The cell size (7.0 × 4.0 μm to 21.0 × 7.5 μm), protein percentage (7.42% to 13.66%), and ash content (49.03% to 59.61%) were different among diatom strains; lipid percentage, nitrogen free extract, and energy content (Kcal g⁻¹) were similar among diatom strains. The values of essential and non-essential amino and fatty acids composition differed among diatom strains. Differences in the abalone shell length and orthogonal analyses revealed postlarval growth was dependent on the quality of the food source. Postlarvae abalone displaying the longest shell lengths were fed Nitzschia thermalis var. minor and Amphiprora paludosa var. hyalina (1,712.0 ± 61 μm and 1,709 ± 67 μm, respectively), followed by Navicula incerta (1,413.3 ± 43 μm). The fatty acid content of benthic diatoms and abalone growth rate were not correlated.     

The mitotic apparatus during unequal microspore division observed by a confocal laser scanning microscope

Summary The structure of the mitotic apparatus during the microspore division ofTradescantia paludosa, which has a distinctively unequal division of large vegetative and small generative cells, was studied using -tubulin immunofluorescence methods and confocal laser scanning microscopy. Mitotic apparatuses began to develop asynchronously during early prophase at the vegetative pole (VP) and during prometaphase at the generative pole (GP). Both, however, reached completion together at the same time during metaphase. At the VP from prophase to prometaphase, microtubules (MTs) did not converge on the pole, and there was a circular area containing only a few MTs. The prophase spindles on the VP side were in the form of domes or cones that lacked the top. In the metaphase, however, the MTs concentrated at the pole to form a representative cone-shaped half-spindle. At the GP from prometaphase to metaphase, the MTs did not concentrate, and a circular area existed that lacked MTs. The half-spindles formed truncated cones. When the phragmoplast developed and curved around the generative nucleus during the telophase. it first grew toward the long axis of the ellipsoidal-shaped microspore; and after it arrived at the inner membrane of the microspore, it again curved past the generative nucleus toward the short axis. In conclusion, it was found that the mitotic apparatus ofT. paludosa microspores with its asynchronous growth and asymmetrical spindle structure and with its three dimensional growth of phragmoplasts had a peculiar developmental manner related to unequal division.     

Allopolyploid speciation of <Emphasis Type="Italic">Calypogeia sphagnicola</Emphasis> (Jungermanniopsida,Calypogeiaceae) based on isozyme and DNA markers

Calypogeia sphagnicola is one of nine species of the genus Calypogeia known in Europe. Occurrence of the species is closely connected with peat bogs. Nowadays, two forms of this species are distinguished—C. sphagnicola f. sphagnicola and C. sphagnicola f. paludosa. The results of the present study, based on two classes of markers—isozymes and sequences of chloroplast genom (trnH-psbA, rpoC1)—unanimously support the genetic differentiation within the taxon and show that the present-day forms represent genetically distinct species. Phylogenetic analysis resolved two lineages that correspond with the present-day forms with high bootstrap support, which differ in ploidy level: C. sphagnicola f. sphagnicola is haploid, whereas C. sphagnicola f. paludosa is a diploid form. Allopolyploid origin of the diploid form was revealed by the isozyme pattern. Nei’s genetic distance between the two present-day forms of C. sphagnicola was 0.472. The forms in Poland have an allopatric pattern of geographic distribution: C. sphagnicola f. sphagnicola occurs exclusively in the lowlands of the northern part of the country on raised peat bogs, whereas C. sphagnicola f. paludosa is found only in the mountains of southern Poland, mainly in the subalpine zone, where it grows on Sphagnum-Polytrichum hummocks on the upper part of north-facing slopes. Plants regarded in this study as C. sphagnicola f. sphagnicola morphologically correspond to the syntype specimen of C. sphagnicola.     

N-terminal polyhedrin sequences and occludedBaculovirus evolution

Summary A phylogenetic tree for occluded baculoviruses was constructed based on the N-terminal amino acid sequence of occlusion body proteins from six baculoviruses including three lepidopteran nuclear polyhedrosis viruses (NPVs), [two unicapsid (Bombyx mori andOrgyia pseudotsugata) and one multicapsid (Orgyia pseudotsugata)]; one granulosis virus (Pieris brassicae); and NPVs from a hymenopteran (Neodiprion sertifer) and a dipteran (Tipula paludosa). Amino acid sequence data for theB. mori NPV were from a report by Sere-bryani et al. (1977) and that for theO. pseudotsugata NPVs were reported previously by us (Rohrmann et al. 1979). The other N-terminal amino acid sequences are presented in this paper. The phylogenetic relationships determined based on the molecular evolution of polyhedrin were also investigated by antigenic comparisons of the proteins using a solid phase radioimmune assay. The results indicate that the lepidopteran NPVs are the most closely related of the above group of viruses and are related to these viruses in the following order:N. sertifer NPV,P. brassicae granulosis virus, andT. paludosa NPV. These data, in conjunction withBaculovirus distribution and evidence concerning insect phylogeny, suggest that theBaculovirus have an ancient association with insects and may have evolved along with them.     

Differentiation between larvae of Tipula paludosa and Tipula oleracea (Diptera: Tipulidae) using isoelectric focusing, and their occurrence in grassland in northern Britain

Thin layer agarose isoelectric focusing (IEF) was used to separate proteins from the larvae of Tipula oleracea Linnaeus and Tipula paludosa Meigen (Diptera, Nematocera). Silver staining revealed protein banding patterns which consistently distinguished these two species at any stage in their life cycles. Within the pH range 5–6 Tipula paludosa contained one major protein band and T. oleracea two bands, with the pI value of the T. paludosa protein being slightly higher than those of T. oleracea. The IEF method is particularly useful for the larval stages of these species because they are often visually inseparable. Leather jackets obtained during a survey of Northern Ireland and north-eastern and western Scotland were tested to determine the distribution and frequency of these species occurring in grassland. In each area surveyed T. paludosa predominated with only one field in Northern Ireland and north-eastern Scotland found to contain T. oleracea. In western Scotland T. oleracea occurred in five fields each year of the survey, and was locally abundant on the Isle of Bute in the 1990/91 survey making up 15% of the total larvae tested.     

The effect of calcium and pH on Florida apple snail, Pomacea paludosa (Gastropoda: Ampullariidae), shell growth and crush weight

Pomacea (Ampullariidae) snails, commonly referred to as apple snails, serve as prey for many freshwater-dependent predators, and some species are highly invasive. Identifying limits to apple snail distribution and abundance are pertinent to understanding their ecology. Calcium (Ca²⁺) availability and pH generally influences freshwater snail populations, yet scant data exist for Pomacea snails. We measured 6-week change in shell length (ΔSL) in P. paludosa in two laboratory experiments with varying Ca²⁺ and pH levels. ΔSL was significantly higher in ≥28 mg Ca²⁺/l compared with treatments ≤14 mg/l. Snails from populations living in low Ca²⁺/pH waters did not appear genetically predisposed at growing faster in these conditions. Smallest ΔSL was in snails treated with 3.6 mg Ca²⁺/l and pH < 6.5 water; these snails had signs of shell erosion. Shell crush weights (CWs) were lowest for snails grown in the lowest Ca²⁺/pH treatment. Smaller shells and lower CWs have implications for predation vulnerability and reproductive success. Our results are consistent with reports associating relatively low snail densities with relatively low Ca²⁺/pH waters, and they are consistent with the geographic distribution of P. paludosa as related to the underlying water chemistry as influenced by geology.     

Microsatellites for eastern Australian Banksia species

We developed eight primer pairs for Banksia microsatellite markers (five using DNA from Banksia oblongifolia and three from Banksia robur) in order to study the processes of speciation within hybridizing B. oblongifolia, B. robur and Banksia paludosa complex. We genotyped four populations of B. oblongifolia and B. robur, and three of B. paludosa. Numbers of alleles ranged from 1 to 13 across the three species and observed average heterozygosities ranged from 0.000 to 0.833. At least four loci completely discriminated B. robur from B. oblongifolia and three discriminated B. paludosa from B. oblongifolia. Seven of these primers amplified DNA from at least two of three other local species.     

Induced parthenogenesis in mandarin for haploid production: induction procedures and genetic analysis of plantlets

This study focused on haploid induction in mandarin through in situ gynogenesis by pollination with irradiated pollen of ‘Meyer’ lemon. Pollination was carried out for three genotypes of mandarin with four levels of gamma-ray-irradiated pollen (150, 300, 600, and 900 Gy). The resulting seeds were characterised by a small size. Embryos were rescued in vitro and the ploidy level of the plantlets was determined by flow cytometry analysis. Haploid, diploid, triploid plantlets were obtained. The haploid parthenogenetic origin was confirmed using microsatellite marker analysis and chromosome count. Diploid and triploid plants were the result of crosses between mandarin and lemon. The induction of gynogenetic haploids of ‘Fortune’ (Citrus clementina Hort ex Tan. × Citrus tangerina Hort ex Tan.) and ‘Ellendale’ (Citrus reticulata Blanco × Citrus sinensis L. Osb) is reported here for the first time.     

Two types of matter economy for the wintering of evergreen shrubs in regions of heavy snowfall

Plant adaptation to an environment subject to heavy snowfalls was investigated in four species of evergreen shrubs growing in a Fagus crenata forest in an area of Honshu on the Sea of Japan. These shrubs stored carbohydrates in some organs before the snowy season and were covered with snow for 4–5 months. Aucuba japonica var. borealis, Camellia rusticana, and Ilex crenata var. paludosa maintained a reserve of carbohydrates during the snowy season. In Daphniphyllum macropodum var. humile, the reserve of carbohydrates decreased during winter. The respiration rates in the first three species decreased from autumn to winter, whereas the decrease in D. macropodum was slight. It was found that the first three species could use reserve carbohydrates for the growth of new shoots after the thaw, whereas in the last species the growth of new shoots depends on high photosynthetic activity in late spring. Our findings suggest some types of matter economy in evergreen shrubs for wintering in an environment of heavy snow.     

中国禾本科植物锈菌补遗

本文对中国禾本科植物锈菌增补了五种,其中两个新种:1、单穗拂子茅夏孢锈Uredo calamagrostidis-emodensis 2、多花剪股颖夏孢锈U. agrostidis-myrianthae;三个中国新记录:3、青篱竹柄锈Puccinia arundinariae 4、阿切尔单胞锈Uromyces archerianus 5、龙爪茅单胞锈U. dactyloctenii。每个种都有描述及附图。标本保藏在中国科学院微生物研究所真菌标本室。     

Systematics and distribution of the New Zealand onychoteuthid fauna (Cephalopoda: Oegopsida), including a new species, Notonykia nesisi sp. nov.

Onychoteuthid squids are among the most common cephalopods found in New Zealand waters, and comprise a major portion of the regional diets of teuthophagous marine mammals. Although several recent publications have addressed aspects of various species’ biology and reproduction, the systematics of the group remains poorly understood. Herein the ontogenetic and adult morphologies of regionally occurring known species of the genera Moroteuthis, Onychoteuthis, and Notonykia are redescribed, and Notonykia nesisi sp. nov. is described for the first time. Ontogenetic and sexually dimorphic variation in characters and character states associated with body proportions, and beak, radula, tentacular hook, palatine tooth and gladius morphologies are also described and compared between local onychoteuthid taxa for the first time.     

Occurrence of “mammalian” lignans in plant and water sources

Enterolignans, also called “mammalian” lignans because they are formed in the intestine of mammals after ingestion of plant lignans, were identified for the first time in extracts of four tree species, i.e., in knot heartwood of the hardwood species Fagus sylvatica and in knot or stem heartwood of the softwood species Araucaria angustifolia, Picea smithiana, and Abies cilicia. They were also identified for the first time in grain extracts of cultivated plants, i.e., in 15 cereal species, in 3 nut species, and in sesame and linseeds. Furthermore, some plant lignans and enterolignans were identified in extracts of water from different sources, i.e., in sewage treatment plant influent and effluent and in humic water, and for the first time also in tap and seawater. They were present also in water processed through a water purification system (ultrapure water). As enterolignans seem to be abundant in the aquatic environment, the occurrence of enterolignans in plant sources is most likely due to uptake by the roots from the surrounding water. This uptake was also shown experimentally by treating wheat (Triticum aestivum ssp. vulgare) seeds with purified lignan-free water spiked with enterolactone (EL) during germination and growth. Both the remaining seeds and seedlings contained high EL levels, especially the roots. They also contained metabolites of EL, i.e., 7-hydroxy-EL and 7-oxo-EL.     

“Meganthropus” cranial fossils from Java

There are now twelve significant hominid cranial fossils from the Lower and Middle Pleistocene of Java, all but two being from the Sangiran site. Most of this material is well-known in the literature, but three skulls, possibly representing “Meganthropus” are here described in detail for the first time. Most scholars have assigned them all toHomo erectus, while others have suggested that they represent as many as four different hominoid taxa. The author argues that they represent two possible species of hominids. “Meganthropus” I, II, and III are more massive than any of the knownH. erectus specimens. They are also relatively higher vaulted, apparently smaller brained, and have unusually thick lower occipital planes. “Meganthropus” may represent a species that separated fromH. erectus upon its arrival to Java.     

Sulphated flavonoid glycosides from leaves of Atriplex hortensis

Two flavonoid sulphates, i.e. quercetin 3-O-sulphate-7-O-α-arabinopyranoside and kaempferol 3-O-sulphate-7-O-α-arabinopyranoside, were isolated from leaves of Atriplex hortensis L. The structures of these compounds were established by UV, ¹H and ¹³C NMR, 2D NMR and MS spectra. The compounds were isolated for the first time from plant material.     

Somatic embryogenesis in Jatropha curcas Linn., an important biofuel plant

Jatropha curcas L. is one potential source of non-edible biofuel-producing energy crop. Its importance also lies in its medicinal properties. The species is primarily propagated through heterozygous seeds, and thus the seed oil content varies from 4 to 40%. Moreover, due to its perennial nature, seed setting requires 2 to 3 years time. The seed viability and rate of germination are low, and quality seed screening is another laborious task; thus, seed propagation alone cannot provide quality planting material for sustainable use. Somatic embryogenesis, a powerful tool of plant biotechnology for faster and quality plant production has been successfully applied to regenerate plants in Jatropha curcas for the first time. Embryogenic calli were obtained from leaf explants on MS basal medium supplemented with only 9.3 μM Kn. Induction of globular somatic embryos from 58% of the cultures was achieved on MS medium with different concentrations of 2.3–4.6 μM Kn and 0.5–4.9 μM IBA; 2.3 μM Kn and 1.0 μM IBA proved to be the most effective combination for somatic embryo induction in Jatropha curcas. Addition of 13.6 μM adenine sulphate stimulated the process of development of somatic embryos. Mature somatic embryos were converted to plantlets on half strength MS basal medium with 90% survival rate in the field condition. The whole process required 12–16 weeks of culture for completion of all steps of plant regeneration. This protocol of somatic embryogenesis in Jatropha curcas may be an ideal system for future transgenic research.     

Notes on the morphology of the early stage of the rare bramid genus Eumegistus

The morphology of the early stage of Eumegistus was described from three specimens [E. brevorti: 23.0 mm in standard length (SL) juvenile; E. illustris: 5.8 mm SL postflexion larva, and 40.0 mm SL juvenile] recently rediscovered in museum collections. Larval and juvenile pigmentation patterns were reported for the first time for this genus. The 5.8 mm SL postflexion larva of E. illustris had pigmentation on the head and anterior half of the body, through to the middle of the dorsal fin base. In larvae and juveniles of both species, the outer side of the pelvic fin was pigmented. The two juveniles possessed several spines on the lachrymal and protruding rays in the middle of the caudal fin. Although it is known previously that the notochord flexion occurs at 5.0–6.0 mm SL in E. brevorti, the reexamined 5.0 mm SL specimen had the notochord completely flexed. Furthermore, we could not confirm whether the previously studied 4.0 mm SL specimen was E. brevorti because it was badly damaged.