中国林蛙消化管胚后发育的组织学观察

用解剖和组织切片技术对中国林蛙Rana chensinensis蝌蚪(23～46期)消化管的形态和显微结构变化进行观察比较.结果 表明,在中国林蛙幼体的胚后发育过程中,消化管长度从28期的37.0 mm逐渐增加至39期的95.3 mm,在变态过程中又缩短至22.0 mm.胃从42期开始膨大, 46期在位置和形态上与成蛙相似.消化管粘膜上皮逐渐完善,粘膜下层和肌层逐渐出现并加厚.胃腺、小肠腺以及杯形细胞在42～43期出现.说明林蛙消化管的组织结构特征与变态发育过程中食性的变化相适应.     

小弧斑姬蛙胚后发育的初步观察

初步观察记录了小弧斑姬蛙Microhyla heymonsi胚后发育的过程。在22.9～27.3℃的温度条件下,观察了其胚后发育全过程中的形态变化。该蛙胚后发育可划分为19期,历时35.7d。小弧斑姬蛙蝌蚪的头长、体长和后肢长度随发育时间的增加而增长。对其胚后发育各个阶段的后肢芽、趾、前肢的发育,以及尾的变化等方面做了系统的描述。讨论并比较了该蛙和其他蛙胚后发育的特点。     

昆嵛林蛙胚后发育的初步观察

昆嵛林蛙（Rana kunyuensis,Lu et Li2002）是新近在山东文登昆嵛山发现的蛙科（Ranidae）蛙属（Rana）林蛙群（Wlld-frogs）的一新种。我们在23.7-26.0℃的条件下,观察了其胚后发育全过程中的形态变化。该蛙胚后发育可划分为19期,历时约49d。唇齿式多为Ⅰ：1-1/Ⅲ、少数Ⅰ：1-1/Ⅱ：1-1,为昆嵛林蛙这一新种的确立提供了形态学依据。     

直链烷基苯磺酸钠对中国林蛙抗菌肽基因表达的影响

近年来由于生境丧失及环境污染等原因,中国林蛙野生种群急剧减少.为深入研究水体中普遍存在的阴离子表面活性剂直链烷基苯磺酸钠(linear alkylbenzene sulfonate,LAS)对中国林蛙生存的影响,通过急毒实验确定LAS对分别处于发育阶段26、32、42、46期的中国林蛙的半致死浓度.同时利用1、4、8 mg/L LAS处理不同发育阶段中国林蛙,研究LAS对中国林蛙存活率、发育状态的影响,并利用real-time PCR技术检测中国林蛙抗菌肽基因表达水平变化.结果表明,当处理时间为96 h,LAS浓度达到12.33 mg/L以上时会对不同发育阶段中国林蛙产生较为严重的致死效应.1~8 mg/L浓度范围的LAS虽然对中国林蛙存活和发育没有明显影响,但是改变了其抗菌肽基因表达模式.特别是中国林蛙胚后发育后期(32期)和变态期(42期),4~8 mg/L LAS会在不同程度上抑制prepropalustrin-2CE3、preprobrevinin-2CE1和preprotemporin-1CEd1抗菌肽基因表达.总体而言,高浓度LAS对中国林蛙免疫系统造成一定影响,可能使该物种更易受到致病菌的侵袭.研究结果为化学污染物对中国林蛙免疫系统影响积累了实验数据,同时为研究中国林蛙抗菌肽基因表达的调控通路提供一定的理论依据.     

高等植物胚胎的发育生物学研究——Ⅰ.水稻、小麦胚胎发育的生长模式

本文结合稻胚的发育模式描述了水稻开花后1至30天期间胚分化发育过程,并指出了胚分化早期胚柄部分与珠被等组织的密切关系。对水稻开花后6～30天及小麦开花后8～27天期间胚鲜重、干重、体积和细胞数的发育模式进行了测定。在此基础上计算了平均每个细胞鲜重和体积的变化,以及麦胚细胞鲜重、干重和体积的变化。此外还测定了水稻胚长度,胚乳鲜重和体积的变化。结果表明;稻胚体积、鲜重在开花后6～11天,干重在6～13天期间呈指数增长,小麦胚鲜重、干重和体积变化在开花后8～15天期间指数增长趋势亦明显,稻胚每个细胞的平均鲜重和体积在开花后7天以前较小,9天以后明显增大;小麦胚每个细胞的平均鲜重、干重、和体积在开花后10天以前较小,12天以后则明显增大。稻胚在开花后1～30天期间长度变化呈S形曲线。     

粤北山区饰纹姬蛙胚后发育的初步观察

初步观察记录了饰纹姬蛙Microhyla ornata胚后发育过程。饰纹姬蛙的卵采于广东韶关小坑国家森林公园,置于人工实验缸中自然孵化,在23.0～28.7℃水温条件下观察饰纹姬蛙蝌蚪胚后发育的过程。该蛙的胚后发育可以分为19期,历时31.1d,对其后肢芽、趾、前肢的发育,以及肛管及尾的变化等方面做了系统的描述,饰纹姬蛙蝌蚪的头长、体长、尾长和后肢长度随发育时间的增加而显著增长。     

介绍一种蛙类胚胎及蝌蚪发育的分期

以蛙类发育过程中的形态结构或生理特征为标志,对蛙类胚胎及胚后发育进行分期,便于对它们的发育进程进行描述,以及对发育与外界条件的关系进行比较研究。在恒温条件下培育蛙胚,可以作出能为今后重复验证的发育时刻表。比较不同蛙种的发育时,也应在相同的恒温条件下进行,才具可比性。分类学上描述蛙种时,蝌蚪也是被描述的对象。但过去描述蝌蚪,多数采用“当后肢长度若干毫米时,蝌蚪全长为若干毫米。”应用这一方法,很难与其他蛙种作比较,因为发育的水温不同和营养物丰歉不同,蝌蚪的生长速度差别很大,后肢与全长的比例关系也会产生变化。因此,制订一个比较适合多数蛙种通用、又易于掌握的蛙     

向日葵未受精胚珠培养时胚状体发生的显微观察

由开花前1—4天的向日葵子房中取出胚珠,在液体培养基上进行漂浮培养,诱导了未受精的卵细胞发育为单倍体的胚状体.亦诱导了珠被绒毡层产生胚状体。对两种胚状体的发生和发育过程及其形态发生特点作了显微观察与描述。     

补偿生长对中国林蛙肝脏生化组成的影响

在室内环境条件下研究了不同饥饿程度和再投喂后中国林蛙肝脏中蛋白质、糖原和脂肪含量的变化,并对中国林蛙在补偿生长过程中能量来源的变化进行了分析.结果 表明,饥饿状态下,中国林蛙的能量消耗主要来源于肝糖原,其次是蛋白质,即饥饿时中国林蛙首先动用糖原和蛋白质提供能量.     

发育中玉米胚结合蛋白(BiP)的动态变化及定位

Western blot检测表明,在玉米胚发育过程中结合蛋白(BiP)含量与胚可溶性蛋白含量变化一致,在授粉16d后BiP含量随发育而增加;对热激不敏感.组织化学免疫定位表明,在玉米胚发育的不同时期,BiP主要定位在胚芽端、初生维管组织和糊粉层中,提示胚在构建器官的同时,也为其功能执行准备了条件;热激不影响其定位.     

桓仁林蛙与东北林蛙蝌蚪的形态特征及比较

桓仁林蛙（Rana huanrenensis Liu,Zhang,＆ Liu,1993）为一种与东北林蛙（Rana dybowskii）同域分布的林蛙,但其分布区域较狭窄.通过对其蝌蚪的形态特征描述及测量相关的形态学量度并和东北林蛙比较,发现在体色、体型、尾部形态以及身体各部位量度比值等方面存在显著性差异.桓仁林蛙的蝌蚪外形与流水型的蝌蚪相类似,而东北林蛙的蝌蚪则属于典型的静水型蝌蚪.     

东北产三种林蛙血细胞的比较研究

应用Wright染色法对桓仁林蛙、东北林蛙和黑龙江林蛙的血细胞进行比较,将所得数据用SPSS软件统计。结果表明：桓仁林蛙、东北林蛙、黑龙江林蛙的血细胞形态、数量上存在差别,这可能与其生活环境和分类地位密切相关。     

Genetic relationships among Korean brown frog species (Anura,Ranidae), with special reference to evolutionary divergences between two allied species Rana dybowskii and R. huanrenensis

Allozyme analysis for 41 populations of brown frog species, Rana dybowskii, R. huanrenensis, and R. amurensis from Korea and three reference species (Chinese R. chensinensis and Japanese R. dybowskii and R. tsushimensis), were performed to clarify taxonomic status of Korean brown frogs. The level of average genetic differentiation (Nei's D) among local populations of each species in Korea was very low (D<0.01 2) and Korean and Japanese R. dybowskii also showed conspecific level of differentiation (D=0.070). Whereas, much larger, discrete genetic differences were detected in the interspecific comparisons (D>0.370). In the genetic relationships among five species examined, the 24 chromosome brown frogs (R. dybowskii, R. huanrenensis, and R. chensinensis) did not form a monophyletic group. Rana dybowskii with the chromosome number of 2n=24 was grouped together with R. amurensis with the chromosome number of 2n=26. The hypothesis of reversal change from 24 to 26 in Korean R. amurensis seems to better explain the phylogenetic relationships of east Asian brown frogs than the assumption of parallel reduction in chromosome number from 2n=26 to 24 in R. dybowskii and in the common ancestor of R. huanrenensis and R. chensinensis. The genetic, morphological, and reproductive divergences between Korean R. dybowskii and R. huanrenensis were compared.     

Phylogenetic relationships of the Chinese brown frogs (genus Rana) inferred from partial mitochondrial 12S and 16S rRNA gene sequences

Based on partial sequences of the 12S and 16S ribosomal RNA genes, we estimated phylogenetic relationships among brown frogs of the Rana temporaria group from China. From the phylogenetic trees obtained, we propose to include Rana zhengi in the brown frogs. Monophyly of the brown frogs was not unambiguously supported, but four well-supported clades (A, B, C, and D) always emerged, although relationships among them remained unresolved. Clade A contained brown frogs with 24 chromosomes and was split into two distinct subclades (Subclade A-1: R. chensinensis and R. huanrenensis; Subclade A-2: R. dybowskii). Polytomous relationships among populations of R. chensinensis and R. huanrenensis suggested the necessity of further taxonomic assessment. Rana kunyuensis proved to be the sister group to R. amurensis, and these two species formed Clade B. Clade C was composed of R. omeimontis and R. chaochiaoensis, and Clade D included R. sauteri, which has been placed in other ranid genera. These relationships did not change after adding published data, and monophyly of Subclade A-1, A-2, and other East Asian brown frogs with 24 chromosomes (R. pirica and R. ornativentris) was ascertained, though their relationships were unresolved. Clade C, together with R. japonica and R. longicrus, also formed a monophyletic group. Brown frogs related to Clades A and C were estimated to have dispersed from continental Asia to adjacent regions through multiple events.     

中国林蛙的分子系统关系

测定了 6种林蛙和用作外群的 2种侧褶蛙和 1种陆蛙的线粒体 12SrRNA基因序列 393bp。序列两两对位比较表明内外群间的位点替换率是 7 3%到 2 3 1% ,内群中物种间则为 0 0 %到 9 2 %。依据上述DNA序列 ,用距离法和最大简约法的系统发育分析表明 :①研究的 6种林蛙聚为一支 ,构成单系群 ,并有高的BPs值(90 %以上 )支持 ;② 6种林蛙可以分成 2个姐妹群 ,即中国林蛙、黑龙江林蛙和桓仁林蛙为一组 (BPs >94% ) ,峨眉林蛙、昭觉林蛙和镇海林蛙为一组 (BPs >5 0 % ) ;③昭觉林蛙与镇海林蛙有较近的亲缘关系 ;④中国林蛙的榆中种群与牡丹江种群间的遗传分化似乎达到了种级分化水平。     

Crucial stages of embryogenesis of Rana arvalis: Part 3. Modularity or developmental integrity

Static analysis of the morphometry of larvae of the postembryonic stages of development of Rana arvalis made it possible to estimate the extent and nature of correlations between developing structures. Considering a correlation as a manifestation of multiple physiological interactions between developing structures, we showed that the most likely scenario is the development of larval structures as a holistic process that is accompanied by an increase in the proportion of minor correlative interactions with an increase in the total number of structures. During the critical periods of development, an increase in the proportion of stronger correlative interactions between the structures that form at this stage of development is observed.     

Novel Rana keratin genes and their expression during larval to adult epidermal conversion in bullfrog tadpoles

The conversion of the larval to adult epidermis during metamorphosis of tadpoles of bullfrog, Rana catesbeiana, was investigated utilizing newly cloned Rana keratin cDNAs as probes. Rana larval keratin (RLK) cDNA (rlk) was cloned using highly specific antisera against Xenopus larval keratin (XLK). Tail skin proteins of bullfrog tadpoles were separated by 2-dimensional gel electrophoresis and subjected to Western blot analysis with anti-XLK antisera. The Rana antigen detected by this method was sequenced and identified as a type II keratin. We cloned rlk from tadpole skin by PCR utilizing primers designed from these peptide sequences of RLK. RLK predicted by nucleotide sequences of rlk was a 549 amino acid -long type II keratin. Subtractive cloning between the body and the tail skin of bullfrog tadpole yielded a cDNA (rak) of Rana adult keratin (RAK). RAK was a 433 amino acid-long type I keratin. We also cloned a Rana keratin 8 (RK8) cDNA (rk8) from bullfrog tadpole epidermis. RK8 was 502 amino acid-long and homologous to cytokeratin 8. Northern blot analyses and in situ hybridization experiments showed that rlk was actively expressed through prometamorphosis in larva-specific epidermal cells called skein cells and became completely inactive at the climax stage of metamorphosis and in the adult skin. RAK mRNA was expressed in basal cells of the tadpole epidermis and germinative cells in the adult epidermis. The expression of rlk and rak was down- and up-regulated by thyroid hormone (TH), respectively. In contrast, there was no change in the expression of RK8 during spontaneous and TH-induced metamorphosis. RK8 mRNA was exclusively expressed in apical cells of the larval epidermis. These patterns of keratin gene expression indicated that the expression of keratin genes is differently regulated by TH depending on the type of larval epidermal cells. The present study demonstrated the usefulness of these genes for the study of molecular mechanism of postembryonic epidermal development and differentiation.     

黑龙江林蛙胃肠道5-羟色胺细胞的胚后发生

采用亲和素-生物素复合物(ABC)免疫组织化学法,对黑龙江林蛙(Rana amurensis)胚后发育各时期胃肠道中5-羟色胺(5-HT)细胞的发生过程进行了研究。结果显示,黑龙江林蛙胃中的5-羟色胺阳性细胞最早在第38期被检测到,小肠和大肠最早出现在第41期。从41期开始,5-羟色胺细胞分布于胃、小肠和大肠中,在同一发育时期中,胃部分布密度最大。同一部位在不同发育时期5-羟色胺细胞的分布密度不同,其中胃内5-羟色胺细胞在38期和43期分布最多,分布密度分别为1.8±0.62和1.8±0.89,45期分布数量最少,分布密度为1.1±0.31;小肠中5-羟色胺细胞在41期和43期分布最多,分布密度均为1.2±0.41,在44期和45期分布最少,分布密度分别为0.7±0.49和0.7±0.47;大肠中5-羟色胺细胞在第42期分布最多,分布密度为1.2±0.41,在44期分布最少,分布密度为0.7±0.47。5-羟色胺细胞最早分布在上皮细胞之间,随着胃肠道发育的日渐完善,在上皮基部或腺泡上皮之间也有分布。细胞形态变化不明显,有圆形、椭圆形、锥体形、长梭形和三角形。推测5-羟色胺阳性细胞的出现时间、形态以及分布型是与黑龙江林蛙的个体生长发育需要、胃肠道各段的功能以及食性转变相适应的。     

辽宁老秃顶子自然保护区寄蝇科新种和新纪录(昆虫纲,双翅目)(英文)

系统地整理、分类和鉴定了辽宁老秃顶子自然保护区采集到的1500余号寄蝇科标本,其中发现并记述了3新种:祝氏喙寄蝇Mycteromyiella zhui,狭额毛瓣寄蝇Nemoraea angustifrons,桓仁突颜寄蝇Phasia huanrenensis;并报告3中国新纪录属(Mycteromyiella,寻寄蝇属Eliozeta、蛛寄蝇属Dicarca),8个中国新纪录种(三鬃菲寄蝇Phebellia triseta、毛菲寄蝇P.villica、加藤芙蕊寄蝇Phryno katoi、阿格筒腹寄蝇Cylindromyia agnieszkae、明寻寄蝇Eliozeta pellucens、高野短须寄蝇Linnaemya(Ophina)takanoi、敏阳寄蝇Panzeria laevigata、河蛛寄蝇Dicarca fluviatilis)。模式标本和其它标本均保存在沈阳师范大学昆虫标本馆。祝氏喙寄蝇,新种Mycteromyiella zhuisp.nov.(图1～7)本种外形近似于分布日本九州的缘喙寄蝇M.marginalis Shima,但区别于后者的主要特征为:额较窄,后面观肛尾叶较长,端部窄而尖,侧尾叶端部钝圆,明显宽于前种。正模♂,辽宁省桓仁县老秃顶子国家级自然保护区,2009-06-24～25,张春田采。副模15♂♂,采集地点和时间同正模,张春田和赵哲采。词源:新种名以辽宁省老秃顶子国家级自然保护区管理局祝业平科长姓氏命名,纪念他30多年来为辽宁省生物多样性保护事业所做的突出贡献。狭额毛瓣寄蝇,新种Nemoraea angustifronssp.nov.(图8～14)本种外形近似于高野毛瓣寄蝇N.takanoi(Baranov),但其别于后者的主要特征是体较小,单眼鬃细长且明显,雌、雄腹部第3背板均具4根中缘鬃和2～4对不规则的心鬃,雌性足均红黄色。正模♂,辽宁省桓仁县老秃顶子国家级自然保护区,2006-07-11,杨正卿采。副模:1♂,采集地点同正模,2006-06-01,冯立勇采;2♀♀,2009-06-25,赵哲和王强采。词源:新种名字来源于其外形特点具较窄的"angust"额"frons"。桓仁突颜寄蝇,新种Phasia huanrenensis sp.nov.(图15～21)本种外形近似于须突颜寄蝇群P.barbifrons-group中的罗氏突颜寄蝇P.rohdendorfi(Draber-Mońko),但区别于后者的主要特征是颊高较短,第1触角节较长,颜脊仅在下半部具鬃,下颚须端部和基部暗棕色,中部红黄色,前足胫节无前鬃,后胫具4～5根前背鬃,侧尾叶侧面观端部膨大。正模♂,辽宁省桓仁县老秃顶子国家级自然保护区,2006-05-30,冯立勇采。副模:4♂♂,采集地点同正模,1994-06-09,魏德采;37♂♂,5♀♀,2006-05-31和2006-06-01,张春田、刘家宇等采,6♂♂,1♀,2009-06-01,张春田、付超采。词源:新种名字源于标本采集地点辽宁省桓仁县。