裳卷蛾变形孢虫SSU rRNA核心序列的克隆与系统发育分析

[目的]利用分子生物学手段探索裳卷蛾变形孢虫的遗传发育地位.[方法]微孢子虫的SSUrRNA序列是构建系统发育进化树的重要工具.试验通过T-A克隆法对裳卷蛾变形孢虫(Vairimorphaceraces)SSU rRNA核心序列进行了克隆,并采用近邻法构建了系统发育进化树.[结果]克隆得到了长为1228bp的核苷酸序列(GenBank EU267796).系统发育分析结果表明:裳卷蛾变形孢虫与分离于小菜蛾(Plutella xylostella)的Vairimorpha sp.Germany(GenBank AF124331)和Vairimorphaimperyecta(GenBank AJ131645)相似性最高,它们在系统发育进化树中与寄主为鳞翅目昆虫的Nosema属聚为一类,与纳卡变形孢虫(Vairimorpha necatrix)为代表的Vairimorpha属为相邻集.[结论]结合其生物学特征,裳卷蛾变形孢虫确实为Vairimorph a属的成员,但根据系统发育分析归入Nosematidae科可能更为合适.     

自桑兰叶甲分离出的一种微孢子虫(Mic-Ⅰ)的研究

从广州郊区罗岗的桑园及菜地捕捉到的桑兰叶甲Mimastra cyanura Hope成虫体中分离到一种卵圆形的微孢子虫（简称Mic-Ⅰ）。孢子大小为(3.35±0.46)×(1.96±0.17) μm;孢子具单核、双核两种类型;极丝10～11圈;孢子表面抗原的血清学类型与家蚕微粒子虫Nosema bombycis孢子不同;在家蚕Bombyx mori体内以两种不同的生活史发育,发育过程符合变态孢虫属Vairimorpha微孢子虫发育特征。Mic-Ⅰ微孢子虫对家蚕具强病原性,对斜纹夜蛾Prodenia litura和小菜蛾Plutella xylostella也有感染能力。     

自桑兰叶甲分离出的一种微孢子虫(Mic-Ⅰ)的研究

从广州郊区罗岗的桑园及菜地捕捉到的桑兰叶甲Mimastra cyanura Hope成虫体中分离到一种卵圆形的微孢子虫（简称Mic-Ⅰ）。孢子大小为(3.35±0.46)×(1.96±0.17) μm;孢子具单核、双核两种类型;极丝10～11圈;孢子表面抗原的血清学类型与家蚕微粒子虫Nosema bombycis孢子不同;在家蚕Bombyx mori体内以两种不同的生活史发育,发育过程符合变态孢虫属Vairimorpha微孢子虫发育特征。Mic-Ⅰ微孢子虫对家蚕具强病原性,对斜纹夜蛾Prodenia litura和小菜蛾Plutella xylostella也有感染能力。     

南海石斑鱼苗种肠道微孢子虫病病原的鉴定

研究通过组织病理分析、超微结构观察以及分子特征分析对石斑鱼(Epinephelus spp.)苗种肠道微孢子虫病病原进行了鉴定。其为一肠孢虫属新种, 命名为石斑鱼肠孢虫(Enterospora epinepheli sp. n.), 专性寄生于细胞核内, 发育过程与肠孢虫属模式种黄道蟹肠孢虫(Enterospora canceri)一致。早期单核裂殖体通过一层简单的电子薄膜与宿主细胞核质隔离。随后, 单核裂殖体发育形成多核裂殖原质团。此时, 细胞核出现明显肥大, 有的甚至被裂殖子胀破。裂殖原质团进一步发育形成多核产孢体, 并开始出现许多高电子密度的囊泡状结构。这些与极丝及锚状盘有关的囊泡状结构聚集在藕核周围, 并组装形成微孢子虫特征性结构(挤出装置)前体。随后, 产孢体原生质团通过连续分裂形成一个个孢子母细胞。孢子母细胞与细胞核直接接触, 并直接发育形成成熟孢子。成熟孢子椭圆形, 孢子长(1.56±0.31) μm (1.07—1.96 μm), 宽(1.08±0.98) μm (0.93—1.28 μm)。 孢壁分为3层, 外壁电子密度高, 厚(15.51±0.95) nm (9.87—26.18 nm), 内壁为电子透明层, 较外层更厚(81.13±2.71) nm (57.16—110.81 nm), 最里面为孢质膜。极丝为同型极丝, 共5—6圈, 分2排排列。组织病理学分析发现该微孢子虫寄生于肠道上皮杯状细胞核内, 肠壁脱落的内容物中也发现大量的微孢子虫。序列比对发现该种与之前报道的石斑鱼肠道微孢子虫待定种(Microsporidium sp.)序列基本一致, 与其他相似性较高的种类的遗传距离在0.162—0.225。系统发育关系分析显示肠胞虫科的种类明显分为两支, 石斑鱼肠孢虫和肠孢虫属其他种类及毕氏肠胞虫聚为一个独立分支, 但不与该分枝中任何种类形成姊妹支。     

讷卡变形孢虫中国新纪录：微孢子门：布雷孢虫科

1983年从河南新乡棉铃虫幼虫分离出一株变形孢虫Vairimorpha sp., 经与美国产讷卡变形孢虫V. necatrix(Kramer, 1965)及从英国引进的印度谷螟变形孢虫V. plodiae (Kellen et Lindegren, 1968) 直接比较研究,确定它是讷卡变形孢虫。28℃染病玉米螟幼虫体内游离孢子形态特征如下表。孢子电镜图片表明它们的特征相同(图1—4)。孢子长度稍有不同,可以认为是地理生态不同所致。在20℃下三者发育周期各形态均与Pilly(J. Invertebr. )Patho1. 28: 177—183, 1976)的描述相同。     

中华球孢虫显微与亚显微观察

运用光镜和透射电镜对中华球孢虫(Sphaerospora chinensis)的结构与发育特点进行观察。结果表明,中华球孢虫呈卵圆形,孢质均匀。极囊和极丝都有透明中间层包围,极丝有粘性且易释放;孢子母细胞边缘有壳瓣形成体,是壳瓣细胞的发育起点,成熟孢子的壳瓣外周有突起的条纹,但两个壳瓣的条纹不尽相同,两个壳瓣之间以紧密连接的方式相连;孢质母细胞的发生属单孢子发育模式;随着细胞的成熟,球孢虫所含的内质网、微管等结构会退化乃至消失。文章还讨论了中华球孢虫极丝易释放性与其对寄生方式适应性的关系。重庆为该虫新分布区。     

楔形两极虫(黏体门,双壳目)的形态学重描述及其近缘种的分子系统学研究

两极虫属Myxidium和楚克拉虫属Zschokklella在形态上非常相似,形态鉴定界限模糊。为进一步厘清两者的分类学关系,本研究对采自涪江重庆市潼南县江段的楔形两极虫M.cuneiforme Fujita,1924进行了形态学重描述,并对其分子系统学进行了研究。楔形两极虫孢子壳面观呈长条形,中部稍凹陷或孢子一边突出,壳瓣上有6~8条与缝嵴平行的条纹;缝嵴直,缝面观呈梭形。孢子长12.5μm±0.3μm,宽5.4μm±0.3μm(n=20)。极囊2个,呈梨形,分布于孢子两极端;极囊长4.3μm±0.2μm,宽3.1μm±0.2μm(n=20),极丝细长且明显,盘曲5~6圈。以18S rDNA为分子标记,对楔形两极虫及其近缘种进行了保守区变异、遗传距离、序列相似度和分子系统发育分析,结果表明:两极虫属和楚克拉虫属均非单系发生,两属间物种相互交叉聚支;两极虫属与楚克拉虫属物种间具有很近的亲缘关系。本文基于形态和分子数据的研究结果支持将两极虫属和楚克拉虫属合并为一个属的观点。     

鲤肠道寄生岳阳碘泡虫新种的形态特征及系统发育分析

在洞庭湖岳阳地区开展鱼类寄生虫调查中,发现一种寄生于鲤Cyprinus carpio L.肠道的黏孢子虫。该黏孢子虫的孢囊呈白色,椭圆形,大小为(1.0±0.2) mm (0.8—1.2 mm)。成熟孢子具有壳瓣,壳面观近似圆形,后端有4—6个“V”形褶皱;缝面观呈纺锤形,缝脊直而粗;孢质均匀,含有一个嗜碘泡;孢子长(9.8±0.6) μm (9.6—10.0 μm),孢子宽(8.2±0.3) μm (8.0—8.5μm),孢子厚(7.3±0.1) μm (7.0—7.5 μm);2个极囊梨形,位于孢子顶端,大小相等,呈“八”字形;极囊长(4.4±0.4) μm (3.8—5.1 μm),宽(2.7±0.2) μm (2.2—3.2 μm),极丝4—5圈。该黏孢子虫与肠膜碘泡虫、丑陋圆形碘泡形态特征非常相似,但其极囊/孢子小于1/2;与文献已报道的鲤肠道寄生北京碘泡虫和鲤肠碘泡虫相比较,其在孢子形态、孢子和极囊大小方面分别存在明显差异。基于该黏孢子虫18S rDNA基因序列(GenBank登录号KY203795)比对分析,该黏孢子虫与山东碘泡虫相似率最高,仅为96%。系统发育分析发现,该黏孢子虫与山东碘泡虫、倪李碘泡虫、住心碘泡虫、Myxobolus encephalicus、Sphaerospora molnari、多涅茨尾孢虫和Henneguya zikaweiensis聚为独立分支,和其他已报道的黏孢子虫亲缘关系较远。综合形态学和18S rDNA基因序列数据,文章报道的鲤肠道寄生黏孢子虫为碘泡虫属一新物种,将其命名为岳阳碘泡虫。     

萨梅诺娃新佩雷斯虫中国新记录

研究报道了中国首例摇蚊微孢子虫, 结合各发育阶段形态特征、生态学特征及分子特征, 鉴定其为萨梅诺娃新佩雷斯虫Neoperezia semenovaiae Issi, et al. 2012, 系我国新记录。萨梅诺娃新佩雷斯虫寄生于羽摇蚊幼虫脂肪体组织, 导致其体表呈白浊状。成熟孢子呈卵圆形, 孢子长(5.7±0.2) μm (5.3—6.3 μm), 宽(3.7±0.1) μm (3.4—4.0 μm)。透射电镜观察显示各发育阶段均为离核, 发育不同步, 与宿主细胞质直接接触。早期发育阶段为高电子密度的多核裂殖体阶段, 经原生质团分裂形成单核或多核产孢体, 进一步发育为单核孢子母细胞。孢子母细胞形状不规则, 周围被内质网环绕, 并逐渐形成微孢子虫的典型结构如极丝、极质体和三层孢壁等。成熟孢子卵圆形, 离核, 细胞核较大, 位于孢子正中央, 被大量核糖体包围。极质体分为两部分, 前半部分为海绵状, 后半部分薄膜状。锚状盘位于孢子前端, 呈蘑菇状。孢壁三层, 外层为高电子密度层, 厚26.5—62.7 nm, 中间层为电子透明层, 厚151.8—236.1 nm, 里层为质膜层。同型极丝, 30—31圈, 分2—3列排列。扩增获得其小核糖体序列为1356 bp, 序列比较发现其与俄罗斯列宁格勒区羽摇蚊的N. semenovaiae相似性为99.1%。系统发育关系分析表明N. semonovaiae与Neoperezia、Bryonosema、Schroedera属种类聚为一独立进化枝, N. semonovaiae种群出现明显的地理分化。     

粉纹夜蛾微孢子虫的分离及其初步分类鉴定

从粉纹夜蛾(Trichoplussia ni,Tn)幼虫中分离纯化得到一种微孢子虫,孢子形状为卵圆形,大小为(2.832±0.486)μm×(1.947±0.317)μm.它能寄生粉纹夜蛾的大多数组织器官,在虫体内的发育过程具有微粒子属Nosema的分类特征,并初步命名为Nosema sp..     

On the Knowledge of Coprophilus (Zonyptilus) pennifer (Motschulsky, 1845) and Coprophilus (Zonyptilus) marginalis (Reitter, 1894) (Coleoptera,Staphylinidae)

Entomological Review - The type and other extensive material of Coprophilus (Zonyptilus) pennifer (Motschulsky, 1845) and some of its synonyms was examined. Coprophilus (Zonyptilus) marginalis...     

Electronic structures of collagen model polymers: (Gly-Pro)n, (Gly-Hyp)n, (Ala-Pro)n, (Ala-Hyp)n, (Gly-Pro-Gly)n, (Gly-Hyp-Gly)n, (Gly-Pro-Pro)n, and (Gly-Pro-Hyp)n

In order to investigate the relationship existing between the electronic structures of collagen and its biochemical functions in vivo, the semiempirical CNDO/2 SCF MO calculations were carried out on several model polymers of collagen, (Gly-Pro)_n, (Gly-Hyp)_n, (Ala-Pro)_n, (Ala-Hyp)_n, (Gly-Pro-Gly)_n, (Gly-Hyp-Gly)_n, (Gly-Pro-Pro)_n and (Gly-Pro-Hyp)_n. Geometries of the skeleton of these polymers were assumed to be the same as those of poly(l-proline) I (cis) and II (trans) and the calculations were performed only on infinite polymers in a single chain. The results show that the cis form is always more stable than the trans form for all the polymers treated. This energy difference between the cis and trans forms depends, for example, on the kind of amino acid residue, Gly or Ala, but this could not be seen in the Pro or Hyp residue. The flexibility or mobility of the collagen structure was explained using the energy difference between the cis and trans forms of the polymers, i.e. the cis-trans conversion of the collagen was discussed in connection with the energy difference. The reason why the collagen has the constitution of (Gly-Pro-Hyp)_n is briefly discussed.     

On the Synonymy of Coprophilus (Zonyptilus) pennifer (Motschulsky, 1845) (Coleoptera,Staphylinidae, Oxytelinae)

Entomological Review - The type material of several synonyms of Coprophilus (Zonyptilus) pennifer (Motschulsky, 1845) is revised. Both Coprophilus (Zonyptilus) striatipennis (Eppelsheim, 1894) and...     

Ecomorphology ofCrassostrea cucullata (Born, 1778) (Ostreidae) in a mangrove creek (Gazi,Kenya)

Variations of shell form and shell length were studied for oysters growing in the mangroves of Gazi Creek, Kenya, and related to different environmental factors.For the study of the form, Fourier analysis was performed on the circumference of 85 oysters. The resulting coefficients were compared among specimens using cluster analysis. The correspondence between this classification and substrate diameter is virtually perfect (only one misclassification out of 85 oysters). For the clusters based on height above chart datum, 9 specimens, all on intermediate height levels, were misclassified. Orientation with respect to tidal current had 13 misclassifications. Mangrove species seemed to influence form only marginally, if at all.In the study of the size ofCrassostrea cucullata, the length of 956 oysters, growing along two transects were measured, and correlated with several environmental factors. Oyster length was not related to substrate diameter or its orientation with respect to the main current. Length was not influenced by density up to a cover of 70%. For densities higher than 70%, there was a fairly strong negative correlation (r ² = 0.634,n = 217). Length was not correlated with height above bottom (base of the tree) for heights lower than 20 cm, while oysters growing closer to the bottom were smaller. The correlation with height above chart datum was negative but very low (r ² = 0.060,n = 957). However, if all measurements of oysters closer than 20 cm to the bottom, and all from a density of more than 70% cover are deleted from the data set, the correlation with height increased dramatically, the slope still being negative (r ² = 0.859,n = 543).     

Genetic polymorphism (ABO, Rh, Kell) in Kabul (Afghanistan)

ABO, rhesus and Kell blood group data on 1327 donors in Kabul are analysed by ethnic affinity and compared with existing data on Afghanistan peoples. Blood group frequencies are very similar in Pushtu and Tadjik, despite their different historical, linguistic, and cultural backgrounds. Inclusion of the small sample of Hazara in the analysis shows overall heterogeneity in rhesus D, E, and e frequencies, suggesting the existence of a broader pattern of genetic variation among the peoples of Afghanistan.     

Tachinid (Diptera, Tachinidae) parasitoids of sawflies (Hymenoptera, Symphyta)

A list of 31 genera of tachinid parasitoids of sawflies is given. Despite the ecological and habitual similarity of sawflies and Lepidoptera, utilization of sawflies by tachinids was not as successful from the evolutionary point of view as utilization of Lepidoptera. Parasitism of tachinids in sawflies is evidently secondary; it appeared repeatedly and originates from parasitism in Lepidoptera. Most genera of tachinids are polyphagous. Endoparasitic Hymenoptera are koinobionts similarly to tachinids. The direct contact of the parasitoid with active mobile host larva not only results in complication of behavior and its specialization, but also promotes formation of different morphological innovations faster than that in idiobionts.