Population genetic diversity and regional differentiation of Chinese forest frog s (Rana chensinensis) in Heilongjiang Province

The Random Amplified Polymorphism DNA (RAPD) markers were used to stu dy the intra-specific diversity and regional differentiation of the Chinese fore st frogs (Rana chensinensis), which were sampled from the fields of 8 regions in Heilongjiang Province. Totally 78 polymorphic DNA loci were amplified by 10 RAP D primers. By genetic distance analysis and phylogenetic tree reconstruction wit h the Neighbor-Joining (NJ) method the results showed that the populations of Ch inese forest frogs distributed in 8 regions existed great differentiation (Avera ge Fst=0.347, SD=0.235) while there was the paradox between geographic distances and genetic distances. Based on geographic and geological data, a hypothesis wa s posed that it is very possible that the hilly lands in the downstream of the S onghua River and the Heilong River were the center of the origin of the Chinese forest frogs in Heilongjiang Province. And mainly through the Songhua River syst em, the Chinese forest frogs dispersed into the Songneng Plain from the Sanjiang Plain whereas the Fangzheng region became a sub-center for the western dispersi on.

中国林蛙Rana chensinensis系统发育及物种形成和分化的研究

运用分支分类法,通过形态、染色体和同工酶性状分析寻找出姐妹群,建立中国林蛙各亚种系统发育分支图。并根据地理分布、古地理和古气候特点,认为中国林蛙起源于西伯利亚勒拿河以东。讨论了中国林蛙的分化及与其近缘种的亲缘关系。根据形态推测起源中心时,应考虑环境变化因素,当分布区环境不变或呈单向性变化时,保留祖征最多的亚种所在地就是该物种的起源中心。

中国林蛙(Rana chensinensis)消化道嗜银细胞的研究

本文应用 Grimelius 法观察了中国林蛙消化道嗜银细胞的形态及分布密度．结果显示：嗜银细胞见于消化道全长,其各部密度分布大体呈一系列高低变动的“M”形曲线,有两个分布密度高峰,分别是胃体和回肠,其中胃体分布密度最高；而在食管,十二指肠和直肠分布密度较低,其中食管处分布密度最低．嗜银细胞主要呈锥体形和梭形,多位于上皮细胞之间,腺泡上皮之间或上皮细胞基部,还有的位于固有层中．根据其形态我们认为,中国林蛙消化道嗜银细胞具有内、外分泌两种功能．

中国林蛙RANA CHENSINENSIS四亚种性状主成分分析

本文对中国林蛙指名亚种、兰州亚种、康定亚种和长白山亚种22项在外形上有差异的性状进行了主成份分析。结果表明体长、足长、蹠部腹面痣粒是否明显、鼓膜大小。第四趾两侧蹼缘缺刻深度、眼径、前臂及手长和头宽八项性状在中国林蛙亚种分类上有较重要的意义。作出了主成分分析坐标图,讨论了二维坐标图与形态特征并同和演化顺序的关系。

Distribution of Methyl Mercury in Rana chensinensis and Environmental Media in Gold-mining Areas of Upper Reaches of Songhua River,China

The distribution characteristics of methyl mercury in Rana chensinensis and water,sediment and soil in gold-mining areas of the upper reaches of the Songhua River,China were studied by field sampling and laboratory testing.The results show that the methyl mercury contents in water,sediment and soil in gold-mining areas are much higher than those in the control site,indicating that gold-mining activities intensify the methyl mercury pollution in the study area.Methyl mercury contents are in a descending order of sediment > soil > water in the environment,and in a descending order of brain > viscera > muscle > skin in Rana chensinensis.There are significant correlation between methyl mercury contents in Rana chensinensis and those in water and sediment.In particular,the methyl mercury content in the skin of Rana chensinensis is positively correlated with those in water and sediment in spring.Therefore,skin is one of main intake pathways for methyl mercury due to its high permeability.

中国林蛙Rana chensinensis四亚种聚类分析

采用Q模式聚类方法对中国林蛙四亚种进行了聚类分析。文中试提出一种权值公式对数量特征进行加权处理,结果表明康定亚种和兰州亚种最相似,指名亚种次之,长白山亚种与前三者差异最大。由聚类分析得出的表相图与分支系统学总结出的系统发育分支图吻合,即表征关系反映了系统发育关系。

Histological Architecture of the Nasal Region in Rana chensinensis

The nasal region of Rana chensinensis is divided into the nasal capsules and nasal cavities. In this study, we investigated the adult histological structure of the nasal capsules and nasal cavities in the frog R. chensinensis under the microscope. We found that an eminentia olfactoria is present in this frog and the presence of the eminentia olfactoria may be connected to a terrestrial life style. The double staining method using alcian blue and alizarin red showed that the septomaxilla, the most important bone associated with the olfactory capsules, is an intramembranous bone in R. chensinensis. The opening of the nasolacrimal duct showed a close proximity to the apertura nasalis externa. The presence of the nasolacrimal duct in the olfactory region may be an adaptation to a terrestrial environment. The function of the vomeronasal and olfactory organs is discussed in the paper.

Differentiations of 5-HT and GAS cells in the digestive canals of Rana chensinensis tadpoles

In the current study, 5-nydroxytryptamine （5-HT） and gastrin （GAS） cells in the digestive canals of Rana chensinensis tadpoles at different developmental stages were investigated by immunohistochemistry. Results showed that the 5-HT cells were only detected in the duodenum before metamorphosis began, and were extensively distributed in the stomach, duodenum, small intestine, and rectum thereafter, with the highest counts found in the duodenum and rectum when metamorphosis was completed. The GAS cells were only distributed in the stomach and duodenum, and only rarely detected in the duodenum before metamorphosis began, but increased in the stomach during metamorphosis and showed zonal distribution in the gastric mucosa when metamorphosis was completed. Metamorphosis is a critical period for amphibians, during which structural and functional physiological adaptations are required to transition from aquatic to terrestrial environments. During metamorphosis, the differentiations of 5-HT cells in the gastrointestinal canals of tadpoles could facilitate mucus secretion regulation, improve digestive canal lubrication, and help water- shortage food digestion in terrestrial environments. Conversely, GAS cell differentiations during metamorphosis might contribute to the digestive and absorptive function transition from herbivore to omnivore.

Agro-climatic Division of Rana chensinensis in Fushun

Based on data collected by 3 basic weather stations in Fushun City and 47 automatic weather stations in the whole region,we selected indexes suitable for agro-climatic division of Rana chensinensis,and carried out a study on agro-climatic division of Rana chensinensis in this region.Annual active accumulated temperature ≥10 ℃ and precipitation during April and September are used as basic calculation factors,and we use climatic aridity to make climatic division of Rana chensinensis in Fushun City.According to relevant indexes,we divide Fushun City into 3 regions,namely most appropriate,appropriate and general regions.Finally,countermeasures and suggestions are put forward for providing reference for agricultural production.

A New Brown Frog of the Genus Rana(Anura,Ranidae)from North China,with a Taxonomic Revision of the R.chensinensis Species Group

The Rana chensinensis species group is widely distributed throughout North China.However,its taxonomy and composition remain controversial.In recent field investigations of the Taihang Mountains,a series of Rana specimens were collected,which were once identified as R.chensinensis.However,these samples showed significant differences from R.chensinensis of the type locality(Shaanxi Province in the Qinling Mountains)in both morphology and genetics.In this paper,based on analyses of seventeen geographic populations from the Taihang and Qinling Mountains,we describe a new species(namely R.taihangensis sp.nov.)in the R.chensinensis species group.A phylogenetic analysis of the R.chensinensis species group based on mitochondrial genes—COI,16S rRNA and Cytb—revealed the monophyly of the cryptic species,which formed the sister taxon to R.kukunoris.Morphological comparisons indicated that the cryptic species can be distinguished from its congeners by a combination of characteristics.Additionally,the distribution patterns of the Rana species in North China were clarified.The populations of the southwestern Taihang Mountains,Xiaoqinling Mountains,and Funiu Mountains in Henan Province remain R.chensinensis,whereas the populations recorded as R.chensinensis in Beijing City,Hebei Province,and the southeastern Taihang Mountains of Henan Province should be revised as R.taihangensis sp.nov.

Transcriptome Sequencing and de novo Analysis for Oviductus Ranae of Rana chensinensis Using Illumina RNA-Seq Technology

Oviductus Ranae is the dried oviduct of female Rana tem-poraria chensinensis （David）, distributed mainly in North- eastern China. Oviductus Ranae is one of the best-known and highly valued oriental foods and medicines. Traditional Chinese medicine holds that Oviductus Ranae can nourish yin, moisten lung and replenish the kidney essence. Meanwhile, activities of Oviductus Ranae such as anti-aging, anti-lipemic, anti-oxidation and anti-fatigue have also been demonstrated by modern phar-macological studies. Previous studies have shown that Oviductus Ranae is mainly composed of proteins, which are up to 50% or more.

Mercury pollution in Rana Chensinensis in Weisha River reach, in the upstream region of Songhua River

Twice a year continuous samples of Rana Chensinensis and sediments have been collected near gold mine in the upstream region of Songhua River from 2000 to 2002, for analyzing the total mercury (THg), methylmercury (MeHg) concentration and correlation in sediments, muscles, livers of the frog. The study indicates that THg and MeHgconcentrations in polluted samples near the gold mine are higher than those in unpolluted ones. THg and MeHg con-centrations are higher in autumn, in female, and in livers than those in spring, in male, in muscles respectively. The order of concentration degree in the frog organizations is as follows: liver>muscle>ovum>Fallopian tube. MeHg is themain form of mercury (Hg) existing in autumn, while inor-ganic Hg is in spring. There is no distinct difference ofCMeHg/CTHg between male and female, livers and muscles. The correlation between MeHg concentrations in the river sedi-ments and in the frog’s livers and muscles are significant, which is related to disorderly discharge of Hg.

太行林蛙地理分布厘定

太行林蛙Rana taihangensis目前已知分布在河南、河北、北京和辽宁等地。根据采自宁夏固原和山东青岛的标本,报道宁夏回族自治区和山东省太行林蛙新分布记录。根据采集自内蒙古通辽和辽宁阜新的太行林蛙标本对其进行形态学补充描述。基于不同地区蛙属Rana标本线粒体cyt b基因的分子系统发育分析结果显示,山西省和陕西省为此前尚未记录的太行林蛙分布区。由此可将太行林蛙在中国的分布范围增加4个省(自治区),目前太行林蛙的分布范围则包括河南省、河北省、北京市、天津市、辽宁省、山东省、山西省、陕西省、宁夏回族自治区和内蒙古自治区。

哈蟆油动物基原问题探讨

目的 :根据历史文献及现代在中国林蛙分类方面研究成果 ,确定哈蟆油的正品动物基原。方法 :查阅历代本草著作及相关历史文献资料与近代国内外对中国林蛙有关研究报道 ,进行整理、分析、总结。结果 :本草著作中记载的山蛤原动物为中国林蛙 ;中国林蛙是独立的种 ,而不是欧洲林蛙的一个亚种 ;中国林蛙有 4个亚种 ,分别为中国林蛙指名、兰州、康定、长白山亚种 ;哈什蟆的原动物为中国林蛙长白山亚种。结论 :中国林蛙长白山亚种为哈蟆油正品动物基原。

中国东北地区林蛙属物种的分类学研究(两栖纲:蛙科)

用比较分类学方法和数值分类学方法相配合，对我国东北产林蛙属物种多样性和原各命名物种的有效性及与中国林蛙Ｒａｎａｃｈｅｎｓｉｎｅｎｓｉｓ的关系进行了探讨，结果认为：地方名为哈士蟆的原中国林蛙的东北居群应属于我国蛙类的一新纪录东北林蛙Ｒ．ｄｙｂｏｗｓｋｉ，而Ｒ．ｓｅｍｉｐｌｉｃａｔａ（Ｎｉｋｏｌ’ｓｋｉ，１９１８）和Ｒ．ｃｈｅｎｓｉｎｅｎｓｉｓｃｈａｎｇｂａｉｓｈａｎｅｎｓｉｓ（魏刚等，１９９１ａ）为该种的同物异名；重新界定了东北林蛙和中国林蛙的分布区，提出中国林蛙仅限于中国分布。

Cu对中国林蛙蝌蚪的急性毒性

试验测定了不同温度条件下Ｃｕ２＋对中国林蛙蝌蚪的急性毒性．结果表明，在水温为９～１３℃、１８℃和２４℃条件下，Ｃｕ２＋对１０ｄ龄中国林蛙蝌蚪９６ｈ的ＬＣ５０分别为５．０１、３．８０和２．９９ｍｇ·Ｌ－１，致死阈浓度分别为４．０、２．５和１．６ｍｇ·Ｌ－１，而在９６ｈ内的无可观察效应浓度分别为２．５、１．６和１．０ｍｇ·Ｌ－１．１８℃条件下Ｃｕ２＋对２０ｄ龄中国林蛙蝌蚪的９６ｈＬＣ５０为２．１７ｍｇ·Ｌ－１．

不同温度条件下环境水分对中国林蛙幼体存活的影响

中国林蛙（ＲａｎａｃｈｅｎｓｉｎｅｎｓｉｓＤａｖｉｄ）主要生活于东北地区的森林中。由于过量捕捉和当地生境的破坏，野生资源已逐渐枯竭，不少地区已处于灭绝的边缘［１，２］。因此，开展中国林蛙的人工养殖不仅具有重要的经济价值，而且对保护生物多样性也有积极意...

温度变化对中国林蛙胚胎发育的影响

开展中国林蛙南移和人工养殖是一项很有意义的工作，而南方多变的气温是否影响其胚胎发育是一个尚未研究的课题。本文利用受精卵在不同温度的恒温培养箱间移换，研究了温度变化对中国林蛙早期胚胎发育的影响，结果表明：（１）快速升温对中国林蛙胚胎孵化率、正常率均有影响，且温差越大影响越大；缓慢升温对胚胎的影响与温差大小和升温起点有关，高起点的升温对胚胎的影响比低起点的升温更大，生产中快速升温以不超过２℃，慢速升温以不超过４℃为宜。（２）慢速降温幅度为４℃时，对胚胎发育无影响，为８℃时，有影响；快速降温幅度为４℃时，对胚胎孵化率和正常率均有影响，生产中慢速降温以不超过８℃，快速降温以不超过４℃为宜。（３）变温幅度相同时，变温速度越大，对胚胎的影响越大。（４）原肠期是中国林蛙胚胎发育的升温敏感期，神经管期为升温非敏感期，中国林蛙胚胎发育中对升温变化较降温敏感。（５）中国林蛙的胚胎发育不会因南方天气的异常而遭到严重的影响。

中国林蛙幼体适宜生存环境的探讨

本文通过人为设置六种不同的环境条件，研究了中国林蛙幼体对不同环境的适应性。结果表明：适宜的生存环境是水深低于蛙体长并设有栖息陆地，或者保持泥土湿润。水深高于蛙体长或环境干燥均对蛙不利。在适宜的环境条件下，幼蛙平均能存活达５３６６ｈ，最长可达２５ｄ。

中国林蛙蛙卵营养成分分析

对我国东北地区长白山麓的特有品种———中国林蛙的蛙卵中的水分、蛋白质、粗脂肪、灰分、总糖、游离氨基酸、氨基酸及微量元素进行了测试分析.结果表明:中国林蛙蛙卵中水分、蛋白质、粗脂肪、灰分、总糖、游离氨基酸的含量各为65.51、16.80、7.32、1.64、0.87、2.58g/100g,其中含有人体必需的微量元素5种,为开发利用这一资源提供了营养学依据.