Accelerated nrDNA evolution and profound AT bias in the medicinal fungus Cordyceps sinensis

Cordyceps sinensis is a reputed medicinal fungus growing parasitically on buried larvae of ghost moths in Asian high-altitude grassland ecosystems. We have analysed the intraspecific ITS nrDNA (ITS1, 5.8S gene, ITS2) variation among 71 sequences of C. sinensis available in EMBL/Genbank. The ITS sequences, submitted to Bayesian ML analyses, were distributed into five groups, referred to as A–E. Nine of the sequences (groups D and E) grouped with distantly related hypocrealean/clavicipitalean taxa and are interpreted as sequences erroneously accessioned under wrong taxon names. The remaining 62 sequences constituted three highly supported clades (groups A–C), that may represent cryptic (phylogenetic) species currently ascribed to C. sinensis. A remarkably high sequence divergence occurred in the 5.8S gene between the three groups. Sequences of groups B and C showed accelerated substitution rates and high AT nucleotide bias. We hypothesize that the accelerated evolution and AT bias have been caused by a shift in life historical attributes or ecology. We also suggest that the recorded differences in medicinal effects among C. sinensis populations may be attributed to the existence of genetically differentiated chemotypes in this morphotaxon.     

Genetic diversity and structure of Cordyceps sinensis populations from extensive geographical regions in China as revealed by inter-simple sequence repeat markers

Cordyceps sinensis is one of the most valuable medicinal caterpillar fungi native to China. However, its productivity is extremely limited and the species is becoming endangered. The genetic diversity of eighteen C. sinensis populations across its major distributing regions in China was evaluated by inter-simple sequence repeat (ISSR) markers. A total of 141 markers were produced in 180 individuals from the 18 populations, of which 99.3% were polymorphic. The low average of Shannon (0.104) and Nei index (0.07) of the 18 populations indicates that there are little genetic variations within populations. For all 18 populations, estimates of total gene diversity (H_T), gene diversity within populations (H_S), coefficient of genetic differentiation (G_ST), and gene flow (Nm) were 0.170, 0.071, 0.583, and 0.357, respectively. This pattern suggests that the genetic diversity of C. sinensis is low and most of the ISSR variations are found among populations with little gene exchange. The 18 populations are divided into five groups based on the genetic distance and the grouping pattern matches with the geographic distribution along the latitudinal gradient. The five groups show obvious difference in the G_ST and Nm values. Therefore, the genetic diversification of C. sinensis populations may be determined by geographic isolation and the combined effects of life history characters and the interaction with host insect species. The information illustrated by this study is useful for selecting in situ conservation sites of C. sinensis.     

Juvenile Chinese mitten crabs (<Emphasis Type="Italic">Eriocheir sinensis</Emphasis>) in the Thames estuary: distribution,movement and possible interactions with the native crab <Emphasis Type="Italic">Carcinus maenas</Emphasis>

The Chinese mitten crab, Eriocheir sinensis is a successful invader. Whilst non-breeding adult E. sinensis have been associated with the destruction of riverbanks, little is known about the ecology of this species in its invaded areas. This is especially true of the juveniles which are a key migratory stage. Intertidal surveys along the Thames estuary indicated an increase in the population since the 1990s. Juvenile E. sinensis were abundant in the sampled upper tidal region of the Thames, refuging under boulders in the intertidal at low tide. Seasonal differences in sampled populations were observed, with a significantly lower abundance of crabs found during winter compared to summer. Mark-recaptures indicated movement in the intertidal occurring during high tide, with an influx of new crabs evident after a single tidal cycle. Endogenous rhythms were also apparent, with peaks in activity occurring corresponding with night-time high tide conditions. E. sinensis juveniles can successfully exclude similar sized native Carcinus maenas from shelters in the laboratory, regardless of which species originally inhabited the shelter. This may have implications for native estuarine Carcinus populations, which can rely on such intertidal shelters for refuges.     

Isolation and Characterization of cDNAs Encoding Ribosome Inactivating Protein from Dianthus sinensis L.

To isolate a ribosome inactivating protein (RIP) gene, six plant species were surveyed for antiviral activity. Crude proteins extracted from these plants were tested for the antiviral activity against tobacco mosaic virus (TMV) in Nicotiana glutinosa. All the plants, Spinacia oleracea, Amaranthus lividus, Dianthus superbus, Dianthus sinensis and Celosia cristata, with an exception of Oenanthe stolonifera, presented 70–90% inhibition of viral infectivity. In an attempt to search for the RIP gene from D. sinensis, partial cDNA was obtained by polymerase chain reaction (PCR) of the poly(A)⁺ RNA from D. sinensis leaves. DNA gel blot analysis showed that D. sinensis has multi-copy RIP genes. The expression of RIP gene was investigated in the flower, leaf, root and stem of D. sinensis, and was found to be most abundant in the leaf. Using the partial cDNA as a probe, seven full-length cDNAs were isolated from a library prepared from D. sinensis leaves. They were divided into three groups on the basis of their nucleotide sequence homology. The three representative clones, cDsRIP1, cDsRIP2 and cDsRIP3 were completely sequenced. They all had an open reading frame of 882 bp. The cDsRIP2 showed 79% homology with dianthin 30 and saporin genes; 59% with PAP and Mirabilis antiviral protein MAP genes. From the analysis of deduced amino acid sequences, it was predicted that D. sinensis RIP cDNAs might have a putative signal peptide of 23 amino acid residues at their N-terminus. When the cDNA was expressed in E. coli, the bacteria was unable to grow upon IPTG induction, suggesting that expression of the gene renders toxicity to E. coli cells.     

Adaptive evolution of osmoregulatory-related genes provides insight into salinity adaptation in Chinese mitten crab, <Emphasis Type="Italic">Eriocheir sinensis</Emphasis>

Osmoregulation is an important mechanism by which euryhaline crustaceans regulate osmotic and ionic concentrations. The Chinese mitten crab (Eriocheir sinensis) is a strong osmoregulating animal model among crustacean species, as it can maintain its hemolymph composition and survives well in either seawater or freshwater. Osmoregulation by E. sinensis during physiological adaptation has been studied extensively. However, the genetic basis of osmoregulation in E. sinensis for acclimating to changing salinities remains unclear. The current study investigated five genes involved in E. sinensis osmoregulation and compared them with a representative marine crab Portunus trituberculatus to test whether adaptive evolution has occurred changing salinity conditions. The results showed that carbonic anhydrase (CA), cytochrome P450 4C (CYP4C), glutamate dehydrogenase (GDH), and the Na⁺/H⁺ exchanger (NHE) have undergone positive selection (i.e., directional selection) in E. sinensis. Thus, the positive selection in CA and NHE suggests that E. sinensis has enhanced capacity for maintaining systemic acid-base balance and ion regulation. GDH and CYP4C also demonstrated positive selection in E. sinensis, suggesting that E. sinensis might have acquired an enhanced capacity to metabolize glutamate and synthesize ecdysteroids in response to a change in osmotic concentration. The present study provides new insight into the molecular genetic basis of salinity adaption in E. sinensis.     

Circadian rhythms of melatonin in haemolymph and optic lobes of Chinese mitten crab (Eriocheir sinensis) and Chinese grass shrimp (Palaemonetes sinensis)

This study is the first to examine the circadian rhythms of melatonin in Eriocheir sinensis and Palaemonetes sinensis, two economically important crustaceans. We collected haemolymph and optic lobes from both species every 4 h over a whole day cycle. Melatonin content was measured with high-performance liquid chromatography. E. sinensis haemolymph exhibited significant (p < 0.05) peaks in melatonin at 16:00 (0.180 ± 0.020 μg·mL^?1) and 24:00 (0.244 ± 0.055 μg·mL^?1), while eyestalks had significant peaks at 16:00 (72.377 ± 18.100 μg·eyestalk^?1) and 24:00 (98.756 ± 30.271 μg·eyestalk^?1). In P. sinensis, melatonin peaked significantly only at 16:00 in optic lobes (12.493 ± 1.475 μg·eyestalk^?1) (p < 0.05); no significant peaks were present in haemolymph. Thus, both E. sinensis and P. sinensis exhibit species-specific melatonin rhythms. Time of day should therefore be considered when examining the physiological status of both crustaceans, given the potential influence of fluctuating daily melatonin concentrations.     

Secondary Production and Its Trophic Basis of Two Mayfly Species in a Subtropical Stream of China

During June 2003 to June 2004, an investigation on life cycle, production and trophic basis of two species of mayfly in a second‐order river of Hanjiang River Basin, Hubei, China was conducted. The results showed Epeorus sinensis Umler and Caenis nigropunctata Wu both developed two generations a year. The mean annual production and P/B ratio of E. sinensis were 9.154 g m^–2 a^–1 dry weight and 16.0, and those of C. nigropunctata were 1.554 g m^–2 a^–1 and 9.6, respectively. For E. sinensis , the proportions contributing to secondary production of the main food types were: amorphous detritus 33.46%, fungi 10.8%, vascular plant detritus 1.8%, diatoms 53.9%; for C. nigropunctata , the proportions were 70.8%, 6.90%, 3.5% and 18.8%, respectively. Compared with those species reported in North America and Europe, although land use mode and local climate were greatly different in China, life history and trophic basis of the mayflies seemed roughly similar, yet secondary production appeared to be much higher. (© 2006 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Characterization of microsatellite loci for the Chinese mitten crab,Eriocheir sinensis

The Chinese mitten crab, Eriocheir sinensis is an important human food source in Asia and causes considerable ecological and economical damage as a recent invader of North America and Europe. Here we report the isolation and characterization of 12 highly polymorphic microsatellite loci for E. sinensis. The number of observed alleles per locus ranged from 7 to 40 and the observed heterozygosity from 0.50 to 0.95. These markers should prove a useful tool to investigate the colonization process and its evolutionary implications.     

Early herbaceous succession along a topographical gradient on forest clear-felling sites in mountainous terrain,central Japan

Early successional patterns of herbaceous communities in forest clear-felling sites were investigated along a topographical gradient, which included ridge, slope and valley habitat types, in warm-temperate evergreen forest regions of central Japan for 5 years. Three dominant species with wind-dispersed seeds played a major role in the succession: an annual,Crassocephalum crepidioides, a biennial,Erigeron canadensis, and a perennial,Miscanthus sinensis. Pioneer herbs that have a seed-bank strategy, which are common in old field succession, were not found in the mountainous sites. The persistence of the annual or biennial dominants during the very early stages of secondary succession was different in the three topographical habitat types. In the ridge habitat,M. sinensis dominated from the first year state, andC. crepidioides andE. canadensis were less prominent.Crassocephalum crepidioides andE. canadensis became dominant as one moved down the slope. In the valley habitat,C. crepidioides dominated in the first year stage, was succeeded byE. canadensis in the second year, and thenM. sinensis gradually replaced it in later years. As all three wind-dispersed dominants simultaneously invaded in all the habitat types after clear-felling, the different successional patterns along the topographical gradient might have resulted from differences in the establishment ability and the growth rate of the three dominants depending on the three habitat types.     

Biomass yield in a genetically diverse Miscanthus sinensis germplasm panel evaluated at five locations revealed individuals with exceptional potential

To breed improved biomass cultivars of Miscanthus ×giganteus, it will be necessary to select the highest‐yielding and best‐adapted genotypes of its parental species, Miscanthus sinensis and Miscanthus sacchariflorus. We phenotyped a diverse clonally propagated panel of 569 M. sinensis and nine natural diploid M. ×giganteus at one subtropical (Zhuji, China) and five temperate locations (Sapporo, Japan; Leamington, Ontario, Canada; Fort Collins, CO; Urbana, IL; and Chuncheon, Korea) for dry biomass yield and 14 yield‐component traits, in trials grown for 3 years. Notably, dry biomass yield of four Miscanthus accessions exceeded 80 Mg/ha in Zhuji, China, approaching the highest observed for any land plant. Additionally, six M. sinensis in Sapporo, Japan and one in Leamington, Canada also yielded more than the triploid M. ×giganteus ‘1993‐1780’ control, with values exceeding 20 Mg/ha. Diploid M. ×giganteus was the best‐yielding group at the northern sites. Genotype‐by‐environment interactions were modest among the five northern trial sites but large between Zhuji, and the northern sites. M. sinensis accessions typically yielded best at trial sites with latitudes similar to collection sites, although broad adaptation was observed for accessions from southern Japan. Genotypic heritabilities for third year yields ranged from 0.71 to 0.88 within locations. Compressed circumference was the best predictor of yield. These results establish a baseline of data for initiating selection to improve biomass yield of M. sinensis and M. ×giganteus in a diverse set of relevant geographies.     

Distribution of overwintering mosquitoes (Diptera: Culicidae) in grassy fields in the Republic of Korea, 2007–2008

Overwintering mosquitoes in a grassy field were surveyed at 13 different localities from December 2007 to February 2008 in the Republic of Korea (ROK). Mosquitoes were collected by putting a white colored tent on the grass, without the base, and heating the inside of the tent. A total of 784 female adults comprising seven species were collected during the study period, including 395 (45.6%) Anopheles pullus, 338 (43.2%) An. sinensis, 21 (2.7%) An. belenrae, 2 (0.3%) An. kleini, 43 (5.5%) Culex tritaeniorhynchus, 20 (2.5%) Cx. inatomii, and 1 (0.3%) Cx. pipiens complex. The mosquitoes showed different overwintering distributions by temperature zone and locality. Predominant species, the malaria vectors An. pullus and An. sinensis, showed different overwintering distributions. Anopheles pullus had a 90.9% collection rate except for Youngju and an average – 0.6°C temperature between December and February. Anopheles pullus showed >50% collection rate in most regions maintained below an average of ?1.4°C, and the rate of An. sinensis showed more than 50% collection rate in all regions maintained at an average above ?1.1°C. Other species showed regional characteristics; An. belenrae was distributed in the northwest part of the ROK, and An. kleini was collected rarely at one locality in the middle part of ROK. Culex tritaeniorhynchus, a Japanese encephalitis virus vector, was distributed in the southern and costal region and maintained at an above average of ?1.0°C. Culex inatomii was collected on the coastal region of the ROK.     

Effects of several establishment modes of Miscanthus × giganteus and Miscanthus sinensis on yields and yield trends

Miscanthus is a C4 perennial grass originating from East Asia, the yields of which progressively increase in the first years of growth. Several species for bioenergy have been studied since the mid‐1980s in Europe, in particular (Miscanthus × giganteus [M. × giganteus]), due to its high yields. M. × giganteus is mainly cultivated in France and established from rhizomes. Our study aimed to assess, in field conditions, alternative establishment methods combined with an alternative species, Miscanthus sinensis (M. sinensis). We set up a multi‐environment experimental network. On each trial, we tested two treatments with M. × giganteus, established from rhizomes (G_r‐sd) and from plantlets obtained from rhizomes (G_p‐sd), and two treatments with M. sinensis seedlings transplanted in single (S_p‐sd) and double density (S_p‐dd). ANOVA was performed to compare establishment and regrowth rates across treatments, as well as yields across treatments and site‐years. A logistic model was used to describe yield trends and to compare the maximum yield reached and the rate of yield increase of both species. Results showed that miscanthus establishment from plantlets resulted in higher establishment (between 87% and 92%) and regrowth (between 91% and 94%) rates compared to establishment from rhizomes. Treatments with M. × giganteus obtained higher average yields across site‐years than those with M. sinensis, but more variable yields across site‐years. We showed a strong species effect on yields, yield components (shoot weight, shoot density and shoot number per plant) and light interception (through leaf area index). Lastly, to use M. sinensis established from transplanted plantlets as an alternative to M. × giganteus, research would be required on the breeding of M. sinensis sterile seeds to avoid risks of invasiveness.     

Arbuscular mycorrhizal status of wild plants in saline-alkaline soils of the Yellow River Delta

A survey was made of the arbuscular mycorrhizal (AM) status of five dominant wild plants Tamarix chinensis, Phragmites communis, Suaeda glauca, Aeluropus littoralis var. sinensis and Cirsium setosum in saline-alkaline soils of the Yellow River Delta that show low plant diversity. All of the species were colonized and showed typical AM structures (arbuscules, vesicles). The colonization percentage ranged from 0.2% to 9.5%, where C. setosum was the highest. The species richness of AMF at the different sites ranged from 2.00 to 2.40 per 50 ml soil, with an average of 2.16. Species diversity ranged from 1.99 to 2.22 per 50 ml soil, with an average of 2.13. Spore density ranged from 3 to 30 per 50 ml soil, with an average of 12. Glomus was the dominant genus, with a frequency and relative abundance of 88.1% and 68.4%, respectively. G. caledonium, with a frequency and relative abundance of 15.0% and 4.6%, respectively, was the dominant species. Differences were also observed in the distribution of AMF in different soil layers. Although there were still AM fungal spores in the layer 40 cm below the surface, most spores were found at a depth of 0–40 cm.     

基于MaxEnt模型的中国沙棘潜在适宜分布区分析

中国沙棘主要分布于我国华北、西北、西南等地森林—草原过渡地带,是我国北方地区退耕还林、生态修复等工程的重要造林树种,对维持干旱、半干旱地区的生态环境稳定具有重要意义。探讨限制中国沙棘分布的主导气候因子,模拟其潜在适宜分布区,以期为中国沙棘在林业生态工程和生态经济林建设中的合理种植和推广提供理论依据。基于中国沙棘自然分布的328个地理样点,利用最大熵(MaxEnt)模型对中国沙棘的潜在分布区的主导气候因子进行分析,并预测中国沙棘的潜在分布范围。结果表明,基于气候变量的MaxEnt模型训练集和测试集受试者工作特征曲线下面积(AUC值)分别为0.962±0.001和0.949±0.001,均大于0.9,表明MaxEnt模型对中国沙棘潜在分布区的预测具有极高的准确度,可信度好。基于环境变量贡献率和刀切法的结果表明年降雨情况、生长季的水热状况、最干季降雨和最冷月最低温等是限制中国沙棘分布的主要气候因素,其中年降雨是限制中国沙棘分布的主导气候因子。通过模拟得到现代中国沙棘潜在地理分布的总适生区面积为165.1万km~2;其中高适生区和中适生区面积共93.3万km~2,主要集中分布于河北西部、北部,...     

Sperm ultrastructure in honeycomb (foam) oysters (Mollusca,Bivalvia, Gryphaeidae,Pycnodontinae): comparison with other Ostreoidea and taxonomic implications

Sperm ultrastructural features of the honeycomb (foam) oysters Hyotissa hyotis, H. sinensis, and H. mcgintyi (Gryphaeidae) are described and compared with other Ostreoidea and more generally with other pteriomorphian Bivalvia. Spermatozoa of H. sinensis and H. mcgintyi (the type species of Parahyotissa Harry 1985) exhibit (1) a broad, low‐conical acrosomal vesicle; (2) subacrosomal material (very electron‐dense granular material and an almost electron‐lucent axial rod); (3) a spheroidal nucleus with a wide anterior invagination (filled with subacrosomal components); (4) a midpiece composed of four spherical mitochondria surrounding a pair of centrioles (rootlet associated with proximal centriole); and (5) a flagellum. Sperm of Hyotissa hyotis (type species of Hyotissa Stenzel 1971) differ markedly from those of H. sinensis and H. mcgintyi, in having (1) a conical acrosomal vesicle showing coarse granular texture anteriorly; (2) a very electron‐dense axial rod; (3) a barrel‐shaped nucleus with a long, narrow anterior invagination (filled with both subacrosomal components) and a basal invagination partly housing the proximal centriole; and (4) five midpiece mitochondria and no proximal centriolar rootlet. Results indicate that H. sinensis should be relocated to another genus, possibly a revised genus Parahyotissa, and also show that the sperm of H. sinensis and H. mcgintyi show many similarities to those of the Ostreidae, with the exception that the ‘axial rod’ component of the subacrosomal material is less electron‐dense than the surrounding substance (more dense in Ostreidae, as in H. hyotis). No family defining sperm features of the Gryphaeidae can be identified.     

桂西北矿区3种草本植物对铅镉胁迫的生理响应和综合评价

为探讨矿区优势植物粽叶芦(Thysanolaena latifolia)、类芦(Neyraudia reynaudian)和芒草(Miscanthus sinensis)的抗重金属污染能力及生态恢复潜力,对生长在铅镉污染土壤中3种植物的生理指标和重金属含量进行了测定.结果表明,同一铅、镉污染水平下3种植物的过氧化物酶活...