中国无尾两栖类新记录种——朝鲜侧褶蛙北大核心CSCD

2021年9月在沈阳市蒲河湿地公园(41°41′54″N,122°52′19″E,海拔35 m)采集到3号无尾两栖类动物,经形态特征比较确认为侧褶蛙属(Pelophylax)金线侧褶蛙种组(P.plancyi species complex)的物种。基于线粒体12S rRNA和16S rRNA基因联合构建的侧褶蛙属部分物种最大似然系统发育树显示,采集到的侧褶蛙标本与朝鲜侧褶蛙(P.chosenicus)聚为一支,且具有较高的支持率(0.95)。综合形态和系统发育比较,确定采集到的标本为无尾目(Anura)蛙科(Ranidae)侧褶蛙属的朝鲜侧褶蛙,系中国两栖动物分布新记录种。     

金线侧褶蛙和泽陆蛙的两性异形与生育力

比较金线侧褶蛙和泽陆蛙的两性异形和生育力,首次通过种间比较来验证生育力选择理论是否能解释两栖动物两性异形的进化。金线侧褶蛙和泽陆蛙均为雌体大于雄体的两性异形类型,其两性异形程度存在显著差异,金线侧褶蛙两性异形程度指数为0 .30 ,而泽陆蛙仅为0 .0 8。然而,两者的怀卵量无显著差异。因此,结果与生育力选择理论预测不一致。此种不一致性可能由下述原因导致:选择压力作用于雄体而非雌体、能量分配策略和死亡率的种间差异、以及系统发育历史的种间差异。金线侧褶蛙和泽陆蛙雄体的身体状态指数显著高于雌体。金线侧褶蛙的头,眼和四肢都显著大于泽陆蛙;两者雌体的前后肢长度显著大于雄体。两性金线侧褶蛙的前后肢随体长呈同速生长,而泽陆蛙的前后肢随体长呈异速生长     

A cryptic invasion within an invasion and widespread introgression in the European water frog complex: consequences of uncontrolled commercial trade and weak international legislation

In Western Europe, many pond owners introduce amphibians for ornamental purposes. Although indigenous amphibians are legally protected in most European countries, retailers are circumventing national and international legislation by selling exotic nonprotected sibling species. We investigated to what extent non‐native species of the European water frog complex (genus Pelophylax) have become established in Belgium, using morphological, mitochondrial and nuclear genetic markers. A survey of 87 sampling sites showed the presence of non‐native water frogs at 47 locations, mostly Marsh frogs (Pelophylax ridibundus). Surprisingly, at least 19% of all these locations also harboured individuals with mitochondrial haplotypes characteristic of Anatolian water frogs (Pelophylax cf. bedriagae). Nuclear genotyping indicated widespread hybridization and introgression between P. ridibundus and P. cf. bedriagae. In addition, water frogs of Turkish origin obtained through a licensed retailer, also contained P. ridibundus and P. cf. bedriagae, with identical haplotypes to the wild Belgian populations. Although P. ridibundus might have invaded Belgium by natural range expansion from neighbouring countries, our results suggest that its invasion was at least partly enhanced by commercial trade, with origins as far as the Middle East. Also the invasion and rapid spread of Anatolian lineages, masked by their high morphological similarity to P. ridibundus, is likely the result of unregulated commercial trade. We expect that Anatolian frogs will further invade the exotic as well as the native range of P. ridibundus and other Pelophylax species elsewhere in Western and Central Europe, with risks of large‐scale hybridization and introgression.     

黑斑侧褶蛙的两性异形和雌性繁殖特征

测定了黑斑侧褶蛙成体的体长、体重、头长、头宽、眼径、鼓膜径、前肢长、后肢长等形态指标以及雌体的怀卵数量。黑斑侧褶蛙雌体的体长和体重显著大于雄体。其它局部形态特征指标与体长呈正相关,协方差分析表明,雄体的鼓膜径大于雌体,其余形态指标不存在明显的两性差异。黑斑侧褶蛙雌体怀卵数量与体长和体重皆成正相关关系,表明黑斑侧褶蛙通过增加个体大小增加繁殖输出。     

黑斑蛙消化系统蛋白酶的活力

用福林 酚试剂法对黑斑蛙 (Rananigromaculata)消化系统蛋白酶的活力进行了分析。结果表明 ,黑斑蛙食道、胃、前肠、后肠、直肠和胰脏蛋白酶的最适pH值分别为 1 5、1 5、7 4、7 4、7 4和 9 6 ,最适温度分别为 55、55、50、50、50和 50℃。在各自最适pH值和最适温度条件下 ,各部位蛋白酶活力由高到低的顺序为 :胰脏 >食道 >胃 >前肠 >后肠 >直肠。文中对黑斑蛙蛋白酶的特性进行了讨论 ,并对蛙的人工养殖提出了几点建议     

When a clonal genome finds its way back to a sexual species: evidence from ongoing but rare introgression in the hybridogenetic water frog complex

Besides several exceptions, asexual metazoans are usually viewed as ephemeral sinks for genomes, which become ‘frozen’ in clonal lineages after their emergence from ancestral sexual species. Here, we investigated whether and at what rate the asexuals are able to introgress their genomes back into the parental sexual population, thus more or less importantly affecting the gene pools of sexual species. We focused on hybridogenetic hybrids of western Palaearctic water frogs (Pelophylax esculentus), which originate through hybridization between P. ridibundus and P. lessonae, but transmit only clonal ridibundus genome into their gametes. Although usually mating with P. lessonae, P. esculentus may upon mating with P. ridibundus or another hybrid produce sexually reproducing P. ridibundus offspring with the introgressed ex‐clonal genome. We compared the rate of nuclear amplified fragment length polymorphism (AFLP) and mitochondrial introgression in two types of populations, that is, those where P. ridibundus occurs in isolation and those where it lives with the hybridogens. Although significant differentiation (Φpt) between sexual and clonal ridibundus genomes suggested limited gene flow between sexuals and hybridogens, a non‐negligible (~5%) proportion of P. ridibundus bore introgressed mtDNA and AFLP markers. Whereas transfer of mtDNA was exclusively unidirectional, introgression of nuclear markers was bidirectional. The proportion of introgressed P. ridibundus was highest in syntopic populations with P. esculentus, proving an ongoing and site‐specific interspecific genetic transfer mediated by hybridogenetic hybrids. It turns out that asexual hybrids are not just a sink for genes of sexual species, but may significantly influence the genetic architecture of their sexual counterparts.     

Expressions and localizations of Bax/Bcl-2 proteins during metamorphosis of Pelophylax ridibundus

Bcl-2 and Bax proteins are expressed in cells of the tails of Pelophylax ridibundus larvae. We investigated the levels of these proteins in tails undergoing apoptosis. Apoptotic cells were observed in the epidermis, muscle and notochord of tails of different lengths. The apoptotic cells in epidermis exhibited the typical features of apoptosis. Amorphous masses and irregularities in striated muscle tissue undergoing apoptosis and apoptotic remnants in the notochord also were observed. In general, Bax staining in the epidermis, subepidermal fibroblast layer, muscle and notochord cells increased, while Bcl-2 staining decreased as the tail regressed. Our results suggest that during tail regression due to metamorphosis, Bcl-2 and Bax proteins play key roles in the apoptosis of tail epidermis, subepidermal fibroblast layer, muscle and notochord cells.     

A new PCR-RFLP-based method for an easier systematic affiliation of European water frogs

We describe a non‐invasive, PCR‐RFLP‐based method that allows reliable determination of the European water frog species Pelophylax lessonae and Pelophylax ridibundus and the hybrid form Pelophylax esculentus. Maximum‐likelihood analysis of ITS2 sequences revealed two robust monophyletic clades corresponding to water frogs of the P. lessonae and P. ridibundus groups. Three restriction enzymes (KpnI, HaeII, and SmaI) were used to digest three conserved ITS2 domains. Taxonomic identification was unambiguous; the three restriction enzymes gave the same results. A French reference sample was identified using allozyme electrophoresis. Our PCR‐RFLP method confirmed circa 83% of identification of the allozyme method. We conclude that the difference between identifications was caused by introgression.     

两种蛙Sox基因的PCR-SSCP分析

采用PCR技术,以参考人SRI,基因HMG-box的保守区序列而设计一对特异引物,扩增了黑斑蛙和金线蛙的Sox基因。结果两种蛙的雌雄个体均扩增出217bp的基因片段,与人对照相同。对扩增产物进行SSCP分析显示,两种蛙雌雄个体间的单链迁移率相同,两种蛙之间差异较小,而与人有较大差异。测序表明,两种蛙的Sox序列之间以及与各类物种的Sox基因都有非常高的相似性,充分显示出Sox基因在系统进化上的高度保守性。     

黑斑蛙肾的组织结构

应用组织学和组织化学方法研究了黑斑蛙(Rana nigromaculata)肾的显微结构。结果表明,黑斑蛙的肾除具有肾单位和集合小管外,还见有淋巴样组织分散于肾实质中。在肾腹侧发现有与真骨鱼类斯坦尼斯小体相似的结构,其中聚集有较多的肥大细胞。说明黑斑蛙的肾具有多种生理功能。