由12S rDNA序列探讨中国狼蛛科主要类群的分子系统发育关系

将自测的我国蜘蛛目狼蛛科4属6个种和从互联网GenBank中检索到相关物种的线粒体基因组12SrDNA的序列进行同源性比较,计算核苷酸使用频率。然后据此进行分子分析,利用2个外群(漏斗蛛科的机敏漏斗蛛Agelena difficilis和缘漏斗蛛Agelena limbat)和2种建树方法(邻近法Neighbour Joining,NJ和最大简约法Maximum Parsimony,MP)分析我国狼蛛科内的亲缘关系。获得平均为291.6 bp的序列中,A T平均含量为78.13%,而G C含量只要21.87%,颠换取代(tranversion)的速度多数大于或接近转换取代(transition)的速度,其中161个核苷酸位点存在变异。研究结果表明:在蜘蛛目狼蛛科有差异的161 bp中,属内种间仅为1.08%,狼蛛科属间为6.85%~14.80%。所构建的分子系统树表明:科内的属和属内的种均优先聚在一起;狼蛛科现行分类系统中各亚科的演化关系顺序为:马蛛亚科→狼蛛亚科→豹蛛亚科;狼蛛科各属的演化关系顺序为:水狼蛛属→马蛛属(或水狼蛛属和马蛛属)→獾蛛属→狼蛛属→豹蛛属;水狼蛛属为最早分出的一支或者水狼蛛属和马蛛属...     

蛛形纲动物含能值及其季节变动规律研究

研究干旱地区蛛形纲动物的含能值变动规律。菜纲蝎目和蜘蛛目动物含能值之间的差异不显著,平均能值为２１．３２Ｊ／ｍｇ无灰干重。但是蝎目钳蝎科（２２．１３Ｊ／ｍｇ无灰干重）、蜘蛛目的圆蛛科（２２．８２Ｊ／ｍｇ无灰干重）、巨眼蛛科（２２．６５Ｊ／ｍｇ无灰干重）、跳蛛科（２３．０８Ｊ／ｍｇ无灰干重）和狼蛛科（２１．７６Ｊ／ｍｇ无灰干重）的含能值显著地高于蟹蛛科（１９．０４Ｊ／ｍｇ无灰干重）和盗蛛科（１９．６     

中国狼蛛科娲蛛属(娲蛛亚科)分类地位的探讨

采用DNA测序方法,获得了中国狼蛛科Lycosidae4亚科6属26种mtDNA-16S rRNA基因的部分序列,比较来自北美狼蛛科豹蛛属2种豹蛛的同一基因序列,并选取漏斗蛛科1种蜘蛛作为外群,采用Bayesian方法和最大简约法(MP)构建分子系统树.两种建树方法均支持娲蛛属和豹蛛属形成一大的单系;这一结果与现行狼蛛科传统分类体系中娲蛛属的分类地位有差别.据此,作者认为:娲蛛属和豹蛛属可以归为同一个分类亚单位.狼蛛科6属间的分子系统关系为(Rirata(Hippasa(Trochsa Arctosa(Pardosa Wadicosa)))).     

基于线粒体16S rRNA基因对中国狼蛛科(Lycosidae)系统发育研究

将自测的中国狼蛛科Lycosidae4亚科6属26种和从GenBank中检索到的北美2种豹蛛的mtD-NA-16S rRNA序列进行比较;以漏斗蛛科1种蜘蛛作为外群,对碱基序列的组成和遗传距离进行了分析,采用Bayesian方法和最大简约法(MP)构建分子系统树。研究结果表明:16SrRNA基因的部分序列为340bp到360bp,A T含量平均为75%,存在较强的A T含量偏向性;序列共有157个碱基存在变异,其中79个简约信息位点。狼蛛科各属间的遗传距离介于0.026￣0.200之间。2种建树方法均表明:科内的属及属内的种优先聚在一起;水狼蛛属相对马蛛属是狼蛛科中较为原始的类群,分化较早;獾蛛属作为1个单系群与熊蛛属合为1个并系,属于狼蛛亚科。狼蛛科6属间的分子系统关系为(Pirata(Hippasa(Trochsa Arctosa(Pardosa Wadicosa))))。     

江西水浆自然保护区蜘蛛多样性研究

本文报道了江西水浆自然保护区蜘蛛14科48属111种,并利用Margalef物种丰富度指数、Simpson物种多样性指数、Shannon-Wiencr指数及Pielou均匀度指数对园蛛科进行了多样性分析.保护区内蜘蛛分布具有水平和垂直地带性特点.在14科中,物种丰富度指数高低依次为:园蛛科、跳蛛科、狼蛛科等.该保护区有17种汀西新记录种和25种未确定种蜘蛛.     

须脚目动物系统学研究进展

须脚目是一类体型微小、形态特殊、生活于土壤或洞穴中的蛛形动物,目前全世界仅记录109种,隶属于2科8属,包括2个化石记录种。由于对该类群系统学研究比较欠缺,其系统发育地位仍不明确。基于形态学的支序系统学分析大部分支持须脚目与四肺类(蜘蛛目+无鞭目+鞭蝎目+裂盾目)聚为一支,但也有研究支持须脚目与真螨类关系较近;分子系统学方面的研究较少,其分析结果也存在分歧。目前中国须脚目动物仅有零星的报道,研究匮乏。现简要报道最近在广西发现的1种类须脚蝎属动物,分析须脚目已知科、属及其分布,比较世界各地区已知须脚目的物种数,列举须脚目研究学者及其研究区域,从经典分类学和系统发育两个方面对须脚目系统学研究的最新进展进行综述,同时给出了须脚目各科、属的中文学名翻译,以期推动中国须脚目动物系统学的开展。     

农田蜘蛛群落结构及其多样性研究

本文对南宁地区农田自然蜘蛛群落和旱地狼蛛亚群落结构及其多样性进行了研究。 1.农田区蜘蛛群落有14科、41属、80种。主要成分是食虫瘤胸蛛、拟环纹狼蛛、拟水狼蛛、奇异獾蛛和豹蛛属种类,个体数以食虫瘤胸蛛最多。旱地狼蛛亚群落主要成分是奇异獾蛛和豹蛛属种类。 2.稻田区蜘蛛群落多样性指数以田埂杂草群落最高,早稻田群落最低。多样性、均匀度和种类丰富度时间格局是早、晚稻皆随时间序列增加。平均多样性、均匀度和个体数,晚稻均高于早稻。狼蛛更群落多样性和均匀度以草地最高。 3.应用群落比例相似性和聚类分析表明,拟环纹狼蛛与拟水狼蛛和奇异獾蛛与豹蛛属种类是组成当地农田区的水田和旱地生境具有代表性的两个狼蛛亚群落主要成分。 本文试图揭示农田生态系统中的稻田蜘蛛,在不受化学农药干扰下群落结构及其多样性变化,并以狼蛛亚群落为代表,比较农田区内不同生境的群落结构及多样性与生境生态特征的关系,为害虫防治和动物资源的利用提供参考。     

中国豹蛛属新种记述(蜘蛛目:狼蛛科)

狼蛛科Lycosidae是蜘蛛目中一个大科,豹蛛属Pardosa,又是本科的一个大属。由于狼蛛是农田害虫的重要捕食性天敌,以前的分类基础又比较薄弱,因而笔者认为有必要进行系统的整理。经初步研究,笔者鉴定狼蛛112种,其中豹蛛52种,本文限制篇幅,只报道豹蛛属的9个新种。     

钳蝎系统分类研究概况

传统观点认为蝎类是蛛形纲中最原始的类群,但是最近的形态分析和分子系统发生研究表明蝎类与单胫类(Haplocnemata=Solifugae Pseudoscorpiones)构成姐妹群,并与盲蛛类一起在蛛形纲形成一个独立的分支.钳蝎科是蝎目中种类最多分布最广的一个科,目前已知82属600余种.钳蝎一直被认为是蝎目中最原始的类群,但是随着Pseudochactidae的发现和建立,其原始的地位也被取代.钳蝎科中亚科的划分一直存在很大的争议,至今尚没有一个亚科划分意见被广泛接受.同样,钳蝎科中属种的界定也存在问题,一些建立较早的属亟待修订.本文对钳蝎的系统发生和分类进行了综述,并对国内外文献中已记载的中国钳蝎种类进行了汇总和记述,即东亚钳蝎Mesobuthus martensii(Karsch,1879),条斑钳蝎蒙古亚种M.eupeus mongolicus(Birula,1911)、凶恶亚种M.e.thersites(C.L.Koch,1839),高加索钳蝎间型亚种M.caucasicus intermedius(Birula,1897)、普氏亚种M.c.przewaklskii(Birula,1897),细尖狼蝎Lychas mucronatus(Fabricius,1798),斑等蝎Isometrus maculates(DeGeer,1778),凹直钳蝎Orthochirus scrobiculosus(Grube,1873).     

中国狼蛛科的系统发育

运用分支分析法,采用支序分析软件Phylip程序,以盗蛛科和褛网蛛科为外群,从形态、生态等方面选取了18个特征对中国狼蛛11个属(脉狼蛛属未在其列)进行支序分析。分析结果表明,马蛛亚科(包括马蛛属和水狼蛛属)最为原始,潮浪蛛亚科次之;熊蛛属和獾蛛属构成的分支更次,狼蛛亚科和豹蛛亚科构成姊妹群最为进化。所以潮浪蛛属、熊蛛属和獾蛛属应分别从狼蛛亚科中独立出来,皆自成亚科,分别名潮浪蛛亚科和獾蛛亚科。     

海南岛2种蝎子地理分布及生境分析

通过调查细尖狼蝎Lychas mucronatus和澳链尾蝎Liocheles australasiae在海南岛的分布,获得了植被和土壤类型,部分土壤湿度,以及分布地的海拔高度等数据。对数据进行分析,可以看出,细尖狼蝎在全岛大部分地区均有分布,澳链尾蝎集中分布于岛的东南部;两者基本分布于低海拔地区;澳链尾蝎可以生存的环境空间较广阔,而澳链尾蝎对环境条件的要求要高一些,需要在一定植被类型、土壤类型和湿度以及一定的海拔范围内才能生存。     

Adaptive evolution of insect selective excitatory β-type sodium channel neurotoxins from scorpion venom

Insect selective excitatory β-type sodium channel neurotoxins from scorpion venom (β-NaScTxs) are composed of about 70–76 amino acid residues and share a common scaffold stabilized by four unique disulfide bonds. The phylogenetic analysis of these toxins was hindered by limited sequence data. In our recent study, two new insect selective excitatory β-NaScTxs, LmIT and ImIT, were isolated from Lychas mucronatus and Isometrus maculatus, respectively. With the sequences previously reported, we examined the adaptive molecular evolution of insect selective excitatory β-NaScTxs by estimating the nonsynonymous-to-synonymous rate ratio (ω = d_N/d_S). The results revealed 12 positively selected sites in the genes of insect selective excitatory β-NaScTxs. Moreover, these positively selected sites match well with the sites important for interacting with sodium channels, as demonstrated in previous mutagenesis study. These results reveal that adaptive evolution after gene duplication is one of the most important genetic mechanisms of scorpion neurotoxin diversification.     

中国等蝎属一新种(蝎目,钳蝎科)

描述了采于中国西藏等蝎属1新种——西藏等蝎,Isometrus(Reddyanus) tibetanus sp.nov.。为了比较,根据原始模式标本和保存在法国自然博物馆的系列标本,重新描述了原始描述于印度和尚分布于尼泊尔的阿萨姆等蝎Isometrus(Reddyanus) assamensis Oates,1888。西藏等蝎,新种Isometrus(Reddyanus) tibetanus sp.nov.(图1~13,27~31,33)正模♂,中国西藏车苏(Chesu),1970年9月,Lingberg采(现保存在河北大学博物馆)。体长42·70mm。根据一般形态,新种近似于原始描述产于印度和尚分布于尼泊尔的阿萨姆等蝎Isometrus(Reddyanus) assamensis Oates,1888(图14~26,32,33),但如下特征不同于后者:1)身体颜色更浅,呈浅黄色到浅红黄色,具不明显或荒废的斑点,而阿萨姆等蝎I.(R.)assamensis相反,其身体和附肢具褐色斑点,且背甲和中体背板主要为浅褐色,2)后体节的脊和背脊的后顶颗粒较弱,3)背甲和中体背板的颗粒稀少或荒废。词源:新种的种名以模式标本的产地而拟定。     

Isolation and characterization of the insecticidal,two-domain toxin LaIT3 from the Liocheles australasiae scorpion venom

ABSTRACT

A novel insecticidal peptide (LaIT3) was isolated from the Liocheles australasiae venom. The primary structure of LaIT3 was determined by a combination of Edman degradation and MS/MS de novo sequencing analysis. Discrimination between Leu and Ile in MS/MS analysis was achieved based on the difference in side chain fragmentation assisted by chemical derivatization. LaIT3 was determined to be an 84-residue peptide with three intrachain disulfide bonds. The sequence similarity search revealed that LaIT3 belongs to the scorpine-like peptides consisting of two structural domains: an N-terminal α-helical domain and a C-terminal cystine-stabilized domain. As observed for most of the scorpine-like peptides, LaIT3 showed significant antibacterial activity against Escherichia coli, which is likely to be caused by its membrane-disrupting property.     

Reproductive biology of the viviparous scorpion,Liocheles australasiae (Fabricius) (Arachnida,Scorpiones, Scorpionidae). I. Absence of males in two natural populations

Summary

Sex of the viviparous scorpion, Liocheles australasiae, collected in two localities in Asia at various seasons during the years 1977 to 1983, was examined on the basis of morphology of gonads and germ cells. A total of 569 specimens consisted of 408 pregnant and 7 interpregnant adult females, 126 immature or nymphal females, and 28 young nymphs in which the sex was not distinguishable. Distinct males were never collected in either locality at any season. No spermatozoa were found in any region of the female reproductive systems. Judging from these results, males do not occur in these populations, and it is presumed that these populations reproduce by thelytokous parthenogenesis.     

SdPI, the first functionally characterized Kunitz-type trypsin inhibitor from scorpion venom

Background

Kunitz-type venom peptides have been isolated from a wide variety of venomous animals. They usually have protease inhibitory activity or potassium channel blocking activity, which by virtue of the effects on predator animals are essential for the survival of venomous animals. However, no Kunitz-type peptides from scorpion venom have been functionally characterized.

Principal Findings

A new Kunitz-type venom peptide gene precursor, SdPI, was cloned and characterized from a venom gland cDNA library of the scorpion Lychas mucronatus. It codes for a signal peptide of 21 residues and a mature peptide of 59 residues. The mature SdPI peptide possesses a unique cysteine framework reticulated by three disulfide bridges, different from all reported Kunitz-type proteins. The recombinant SdPI peptide was functionally expressed. It showed trypsin inhibitory activity with high potency (K_i = 1.6×10⁻⁷ M) and thermostability.

Conclusions

The results illustrated that SdPI is a potent and stable serine protease inhibitor. Further mutagenesis and molecular dynamics simulation revealed that SdPI possesses a serine protease inhibitory active site similar to other Kunitz-type venom peptides. To our knowledge, SdPI is the first functionally characterized Kunitz-type trypsin inhibitor derived from scorpion venom, and it represents a new class of Kunitz-type venom peptides.     

LC/MS/MS identification of 20-hydroxyecdysone in a scorpion (Liocheles australasiae) and its binding affinity to in vitro-translated molting hormone receptors

Recent advances in mass spectrometry (MS) technology have facilitated the detection and quantification of minor components in organisms and the environment. In this study, we successfully identified 20-hydroxyecdysone (20E) in first instar nymphs (7 days after hatching) of the scorpion Liocheles australasiae, using tandem mass spectrometry combined with high-performance liquid chromatography (LC/MS/MS). This substance was not found in adults after the fifth stage. Other possible molting hormone candidates such as makisterone A (MaA) and ponasterone A (PoA), both of which are reported to be the molting hormones of a few arthropod species, were not detected in this scorpion. The ligand-receptor binding of 20E and its analogs was quantitatively evaluated against the in vitro-translated molting hormone receptor, the heterodimer of ecdysone receptor (EcR) and the retinoid X receptor (RXR) of L. australasiae (LaEcR/LaRXR). The concentrations of ecdysone (E), MaA, 20E, and PoA that are required to inhibit 50% of [(3)H]PoA binding to the LaEcR/LaRXR complex were determined to be 1.9, 0.69, 0.05, and 0.017 μM, respectively. The activity profiles of these 4 ecdysteroids are consistent with those obtained for the molting hormone receptors of several insects. The binding of a non-steroidal E agonist, tebufenozide, to EcR was not observed even at high concentrations, indicating that the structure of the ligand-binding pocket of LaEcR is not favorable for interaction with tebufenozide.     

ALCAPs induce mitochondrial apoptosis and activate DNA damage response by generating ROS and inhibiting topoisomerase I enzyme activity in K562 leukemia cell line

The solution structure of an insecticidal toxin LaIT1, a 36-residue peptide with a unique amino-acid sequence and two disulfide bonds, isolated from the venom of the scorpion Liocheles australasiae was determined by heteronuclear NMR spectroscopy. Structural similarity search showed that LaIT1 exhibits an inhibitory cystine knot (ICK)-like fold, which usually contains three or more disulfide bonds. Mutational analysis has revealed that two Arg residues of LaIT1, Arg¹³ and Arg¹⁵, play significant roles in insecticidal activity.