利用线粒体COII基因序列对中国尾蛱蝶属系统分化的研究(鳞翅目:蛱蝶科)

利用线粒体C0II基因序列分析法,研究了我国尾蛱蝶属5种蝴蝶的系统分化.结果表明,在尾蛱蝶属5个种的12个样品中,405 bp长的COII片段有11.4%的位点为多态性位点,大部分的碱基改变是转换.各物种内不同个体间的差异明显小于不同物种间的差异,种内个体间的差异一般为0.5%～1.5%,各物种间的差异绝大多数在4%以上.利用最大似然性法构建的尾蛱蝶属聚类关系图显示,尾蛱蝶属蝴蝶分为两大分支,一支包括大二尾蛱蝶、二尾蛱蝶和忘忧尾蛱蝶,另外一个分支包括窄斑凤尾蛱蝶和黑凤尾蛱蝶聚在一起.在大二尾蛱蝶、二尾蛱蝶和忘忧尾蛱蝶这一分支中,大二尾蛱蝶和忘忧尾蛱蝶的亲缘关系较近,而二尾蛱蝶较远.这些分子系统学的结果均与形态学的结果相一致,是对形态分类的有力支持.     

中国尾蛱蝶属系统发育分析——基于雄性外生殖器形态特征

在对中国尾蛱蝶属雄性外生殖器解剖研究的基础上,选取了23个雄性外生殖器解剖特征,利用Paup4.0b10系统发育分析软件,分析探讨了中国尾蛱蝶属系统发育关系。结果表明:中国尾蛱蝶属9个种分为2个支系,一支系由凤尾蛱蝶Polyura arja、窄斑凤尾蛱蝶P.athamas及黑凤尾蛱蝶P.schreiber3种组成,另一支系由二尾蛱蝶P.narcaea、沾襟尾蛱蝶P.posidonius、忘忧尾蛱蝶P.nepenthes、异纹尾蛱蝶P.mutata、大二尾蛱蝶P.eudamippus及针尾蛱蝶P.dolon6种组成。两大支系均以渐次方式演化;凤尾蛱蝶与窄斑凤尾蛱蝶、大二尾蛱蝶与针尾蛱蝶各自互为姊妹群关系。     

基于线粒体CO Ⅰ基因的闪蛱蝶亚科系统发育关系研究

测定了重要林业害虫闪蛱蝶亚科Apaturinae 11属17种蝶类的线粒体细胞色素氧化酶Ⅰ的部分序列,并结合由GenBank下载的该亚科4种蝶类的相应序列进行分析,探讨了闪蛱蝶亚科各属间的系统发育关系。以玉杵带蛱蝶Athymajina,白斑眼蝶Penthema adelma,忘忧尾蛱蝶Polyura nepenthes和白带螯蛱蝶Charaxes bernardus作为外群,采用PAUP4.0b4a软件构建了闪蛱蝶亚科的MP和NJ分子系统树。虽然COI基因数据中第3位点的转换替换已达饱和,但由于这些位点含有大量系统发育信息,因而在数据统计分析时并没有将这些位点删除。同时通过对各分枝稳定性的比较,研究在简约分析中不同转换/颠换加权方式对假定所有特征具有相同权重的影响。分子系统树显示:NJ和不同转换/颠换加权方式下构建的MP系统树中闪蛱蝶亚科均有4个主要的聚类簇,该亚科系统树中存在许多置信度高且稳定的分枝,同时也存在一些因分枝置信度低且不稳定而使其分类地位不能确定的类群。在所构建的系统树中,由分子数据得到的蛱蝶亚科系统发育关系与传统分类学的基本一致,其中迷蛱蝶属Mimathyma为单系群;累积蛱蝶Lelecella limenitoides为明窗蛱蝶Dilipa fenestra的姐妹群且支持二者关系的置信度很高;支持将白斑迷蛱蝶Mimathyma schrenckii,迷蛱蝶M.chevana,夜迷蛱蝶M.nycteis,栗铠蛱蝶Chitoria subcaerulea,黄带铠蛱蝶C.fasciola,铂铠蛱蝶C.pallas,and银白蛱蝶Helcyra subalba等物种由闪蛱蝶属中移出的修订。     

线蛱蝶亚科蝶类部分类群线粒体COI基因的分子系统发生分析

以线粒体细胞色素氧化酶I（COI）基因作分子标记,对线蛱蝶亚科蝴蝶进行序列测定．序列分析的结果表明．经比对和处理后的序列总长度是645bp,其中有199个变异位点,147个简约信息位点;所编码的氨基酸序列中有18个变异位点,7个信息位点．A＋T平均含量为69．6％,G＋C平均含量为30．4％,碱基组成出现AT偏斜．以蛱蝶亚科及秀蛱蝶亚科物种为外类群,用NJ、MP及贝叶斯法重建了该亚科的系统发生树,探讨了它们主要类群间的系统发生关系．分子系统树显示,线蛱蝶亚科由以下3大支系：环蛱蝶族＋翠蛱蝶族、线蛱蝶族、丽蛱蝶族构成;其中,环蛱蝶族为单系群（NJ树也支持线蛱蝶族的单系性）;翠蛱蝶族与环蛱蝶族亲缘关系较近：丽蛱蝶族可能是该亚科较早分化出的一支．     

中国尾蛱蝶属雄性外生殖器比较形态学研究(鳞翅目,蛱蝶科,螯蛱蝶亚科)

对中国尾蛱蝶属Polyura9个种的雄性外生殖器进行了比较形态学研究,结果表明,雄性外生殖器种间存在明显差异,可作为种类鉴别依据;并根据雄性外生殖器特征编制了中国尾蛱蝶属的种检索表。     

基于线粒体Cyt b基因的线蛱蝶亚科的系统发育

基于线粒体Cytb基因部分序列,以波蛱蝶为外群,采用邻接法、最大简约法和贝叶斯法分别构建了中国线蛱蝶亚科10属25种蝶类的系统发育树,探讨了各主要类群间的系统发育关系。其结果表明,所有线蛱蝶亚科聚为两大枝:第一枝包括环蛱蝶属、菲蛱蝶属、蟠蛱蝶属和缕蛱蝶属,其中缕蛱蝶属与环蛱蝶族亲缘关系较近;第二枝包括丽蛱蝶属、穆蛱蝶属、线蛱蝶属、带蛱蝶属、律蛱蝶属和翠蛱蝶属,其中线蛱蝶属为非单系群,翠蛱蝶属和律蛱蝶属则为单系发生,并构成姐妹群。     

武铠蛱蝶线粒体基因组全序列测定和分析

【目的】了解闪蛱蝶亚科属间及种间的分子系统进化关系。【方法】采用PCR步移法对武铠蛱蝶 Chitoria ulupi 线粒体基因组全序列进行了测定和分析。基于线粒体基因组13个蛋白质编码基因的核苷酸序列构建了38种鳞翅目昆虫的系统发育树。【结果】分析结果表明,武铠蛱蝶线粒体基因组全长15 279 bp,包括13个蛋白质编码基因、22个tRNA基因、2个rRNA基因和一段长度为391 bp的A+T富含区,基因排列顺序与其他已知近缘种昆虫相同。武铠蛱蝶线粒体基因组中存在很高的A+T含量（79.9％）。13个蛋白质编码基因中,COII以TTG作为起始密码子,COI以CGA作为起始密码子外,其余均为昆虫典型的起始密码子ATN。COII和ND4基因使用了不完全终止密码子T,其余基因均以典型的TAA为终止密码子。在所测得的22个tRNA基因中,除tRNA^Ser(AGN)缺少DHU臂外,其余tRNA均能形成典型的三叶草结构。与其他多数鳞翅目昆虫一样,武铠蛱蝶的A+T富含区中有一段由ATAGAA引导的保守的多聚T结构,长度为21 bp,并散布着一些长短不一的串联重复单元。系统发育树结果显示,总科级别的系统发育关系为：卷蛾总科＋（凤蝶总科＋（螟蛾总科＋(夜蛾总科＋蚕蛾总科＋尺蛾总科)））;在蛱蝶科物种中,武铠蛱蝶与猫蛱蝶Timelaea maculate 亲缘关系最近。【结论】基于分子标记构建的鳞翅目昆虫系统发育关系与传统的形态学分类结果基本一致。     

基于线粒体Cyt b基因的线蛱蝶亚科的系统发育

基于线粒体Cyt b基因部分序列，以波蛱蝶为外群，采用邻接法、最大简约法和贝叶斯法分别构建了中国线蛱蝶亚科10属25种蝶类的系统发育树，探讨了各主要类群间的系统发育关系。其结果表明，所有线蛱蝶亚科聚为两大枝：第一枝包括环蛱蝶属、菲蛱蝶属、蟠蛱蝶属和缕蛱蝶属，其中缕蛱蝶属与环蛱蝶族亲缘关系较近；第二枝包括丽蛱蝶属、穆蛱蝶属、线蛱蝶属、带蛱蝶属、律蛱蝶属和翠蛱蝶属，其中线蛱蝶属为非单系群，翠蛱蝶属和律蛱蝶属则为单系发生，并构成姐妹群。     

基于线粒体16S rDNA序列探讨蛱蝶科(鳞翅目, 蝶亚目)主要分类群的系统发生关系

本文测定了蛱蝶科7亚科27种蛱蝶和斑蝶科2种蝴蝶的线粒体16S rRNA基因部分序列,并从GenBank中下载了6种蛱蝶的同源序列。以斑蝶科的幻紫斑蝶和绢斑蝶作外群,通过遗传分析软件对这些序列进行了比较分析,用邻接法和贝叶斯法重建了蛱蝶科的系统发育树,探讨了蛱蝶科主要类群间的系统发育关系。序列分析的结果显示:经比对处理后获得494bp长度序列,其中有可变位点206个,简约信息位点145个;A T平均含量78.4%,C G平均含量为21.6%,具A、T偏倚性。分子系统树显示:蛱蝶亚科并非单系群;蛱蝶族中眼蛱蝶属应移入斑蛱蝶族;闪蛱蝶和蛱蝶亚科与蛱蝶亚科具有较近的系统关系;结果支持豹蛱蝶和釉蛱蝶合为一亚科即釉蛱蝶亚科;支持将秀蛱蝶和蛱蝶亚科从线蛱蝶亚科中分离出来。     

华北山区干草地生境中耕作和放牧对网蛱蝶保育的作用

研究对象是中国河北省赤城县闫家坪村的两种共存网蛱蝶—大网蛱蝶和金堇蛱蝶, 这两种网蛱蝶以不同的集合种群类型续存. 在此区域内, 适宜网蛱蝶生存的栖息地斑块是干草地. 在这里主要的人为干扰是放牧和耕作. 研究结果表明: 斑块内是否嵌有耕地和在斑块内放牧对金堇蛱蝶的斑块占据性以及种群密度有交互作用, 金堇蛱蝶在内部嵌有耕地并且适度放牧的斑块上的占据性和种群密度最高. 斑块内是否嵌有耕地对两种网蛱蝶的斑块占据性都有显著影响, 两种网蛱蝶都喜欢占据内部嵌有耕地的斑块. 斑块内是否嵌有耕地对金堇蛱蝶的种群密度有显著影响, 而对大网蛱蝶种群密度没有影响. 但是两种网蛱蝶在田垄间的幼虫组的比例有逐年增加的趋势. 两种网蛱蝶都是在适度放牧的斑块内有最高的占据性和种群密度. 结果说明在该类型山区的小规模的耕种和适度放牧对蝴蝶的保育是有利的. 斑块内是否嵌有耕地和在斑块内放牧对两种网蛱蝶有不同的影响. 因此, 在对有人为活动的山区进行保育规划时, 要综合考虑其利弊以及不同物种的异同.     

Glacial effects on sequence divergence of mitochondrial COII of Polyura eudamippus (Lepidoptera: Nymphalidae) in China

Sequence divergence of mitochondrial COII was analyzed in 50 specimens belonging to five subspecies of Polyura eudamippus (Lepidoptera: Nymphalidae) collected from southern China. There were nine haplotypes in the 405 bp of partial COII sequence. Distribution of the five subspecies was primarily consistent with the spatial distribution of haplotypes. The K (st) statistic showed genetic differentiation among these subspecies, except between the pair of P. e. kuangtungensis and P. e. formosana, which were separated by the Taiwan Strait. This is consistent with the 10,000-year history of the Taiwan Strait, not long enough for detectable differentiation. The present distribution pattern of COII haplotypes of P. eudamippus should be shaped by the alteration of Pleistocene glaciations, and Yunnan might be the refugium of P. eudamippus in the ice age, judging from the abundant haplotypes remaining. There were two routes for P. eudamippus in the postglacial expansion, one northward to Sichuan, Chongqing, and Hubei and another eastward to the southeastern coast of mainland China and Taiwan Island. Because the haplotype of butterflies on Hainan Island (P. e. whiteheadi) was completely different from that of mainland China, it was estimated that butterflies on Hainan Island might be from the Indo-China Peninsula rather than from mainland China.     

RAPD分析与ITS序列分析在拟茎点霉分类鉴定上的应用

采用随机扩增多态性DNA(RAPD)技术和核糖体RNA基因(rDNA)转录间隔区(ITS)序列分析对22种拟茎点霉共34个菌株进行了系统发育研究。RAPD分析构建的UPGMA聚类图所反映的种间、种内关系与形态学分类结果基本一致,可以清楚地将分自7科寄主植物上的不同的种分别区分开来,但分自同科或同属寄主植物上的不同的种并不具有相近的亲缘关系。ITS序列分析结果不支持将Phomopsis mangiferae Ahmad和P. cytosporella Penz. et Sacc. 合并为同一个种的观点,同时还显示出P. mangiferae与P. psidii de Camara的亲缘关系非常近, 可能是异名同物;而为害木棉叶的拟茎点霉与杨梅枝枯病菌P. myricae Y.J.Huang et P.K.Chi之间的碱基差异亦属于种下不同菌株间的正常差别范围,很可能就是同一个种。对相同的供试菌株两技术所反映的亲缘关系趋势相同,表明两技术用于拟茎点霉的亲缘关系分析和种类鉴定均是可行的。     

RAPD分析与ITS序列分析在拟茎点霉分类鉴定上的应用*

采用随机扩增多态性DNA(RAPD)技术和核糖体RNA基因(rDNA)转录间隔区(ITS)序列分析对22种拟茎点霉共34个菌株进行了系统发育研究。RAPD分析构建的UPGMA聚类图所反映的种间、种内关系与形态学分类结果基本一致,可以清楚地将分自7科寄主植物上的不同的种分别区分开来,但分自同科或同属寄主植物上的不同的种并不具有相近的亲缘关系。ITS序列分析结果不支持将Phomopsis mangiferae Ahmad和P. cytosporella Penz. et Sacc. 合并为同一个种的观点,同时还显示出P. mangiferae与P. psidii de Camara的亲缘关系非常近, 可能是异名同物;而为害木棉叶的拟茎点霉与杨梅枝枯病菌P. myricae Y.J.Huang et P.K.Chi之间的碱基差异亦属于种下不同菌株间的正常差别范围,很可能就是同一个种。对相同的供试菌株两技术所反映的亲缘关系趋势相同,表明两技术用于拟茎点霉的亲缘关系分析和种类鉴定均是可行的。     

Lumping lumpsuckers: molecular and morphological insights into the taxonomic status of Eumicrotremus spinosus (Fabricius, 1776) and Eumicrotremus eggvinii Koefoed, 1956 (Teleostei: Cyclopteridae)

The genus Eumicrotremus comprises 16 lumpsucker species distributed in the Arctic and northern Atlantic and Pacific Oceans. The most common species in the North Atlantic is Eumicrotremus spinosus , described in 1776, and characterized partly by numerous bony tubercles on the head and body. Another Atlantic species, Eumicrotremus eggvinii , described in 1956, remained known only from a single specimen until additional specimens were recently recovered. To reassess the status of E. eggvinii , 21 meristic and 32 morphometric characters were analysed for a total of 83 specimens of E. spinosus and E. eggvinii . Mitochondrial (COI, COII and cyt- b ) and nuclear ( Tmo-4C4 ) genes were also sequenced for both species, along with Eumicrotremus derjugini . The results indicate that although E. spinosus and E. eggvinii are clearly separated by a considerable number of morphological characters, they in fact constitute a single, sexually dimorphic species. Thirteen specimens of E. eggvinii (including the holotype) and 59 E. spinosus could be sexed; all individuals of E. eggvinii turned out to be males and all E. spinosus were females. Identical DNA sequences were found in all E. eggvinii and E. spinosus for COI, COII and Tmo-4C4 , and a single shared synonymous substitution found in cyt- b . In contrast, E. spinosus , E. eggvinii and E. derjugini differed by 5·9% for COI and COII, 1·2% for Tmo-4C4 and 8·3% for cyt- b .     

云南边境地区果实蝇属种类DNA条形码鉴定

【目的】果实蝇属Bactrocera中有国际上重要的检疫性害虫, 基于形态的物种鉴定有一定的局限性。另一方面, 云南边境地区为东南亚地区实蝇入侵我国的重要通道。因此, 对该地区实蝇分子鉴定方法的研究对于该属物种的快速准确鉴定具有重要意义。本研究旨在探讨DNA条形码技术在果实蝇属物种鉴定中的有效性。【方法】使用线粒体基因COI和COII序列的通用引物对果实蝇属20个物种60份样品进行PCR扩增、测序和序列分析; 采取距离方法和建树方法评价2种序列的鉴别能力。【结果】COI和COII序列平均长度分别为682 bp和339 bp, 种内和种间遗传差异较大, 有较明显的遗传距离间隔（barcoding gap）, 鉴定成功率分别为91.2%和90.7%。另外, 分子系统树表明华实蝇亚属Sinodacus不是单系群。【结论】COI和COII序列均能够将绝大多数果实蝇属物种进行准确鉴别, 应用COI或COII序列进行果实蝇属物种鉴定具有一定的可行性。     

Species Delimitation of Pterygiella (Orobanchaceae), a Genus Endemic to Southwestern China on the Basis of Morphometric and Molecular Analyses

Pterygiella, a genus endemic to southwestern China, belongs to hemiparasitic rhinanthoid Orobanchaceae. Since P.bartschioides was segregated as an independent genus of its own, i.e. Xizangia, the genus was treated to include four species, P.duclouxii, P.nigrescens, P.cylindrica, and P.suffruticosa. However, the discriminative characters are obscure among these species, and the delimitation of the two varieties in P.duclouxii remains problematic. To revise the taxonomy of the genus, thirteen populations from the four species were collected for both morphometric and molecular systematic analyses. Principal component analysis (PCA) and cluster analysis via unweighted pair group method with an arithmetic average (UPGMA) were used. In addition, four chloroplast sequences (i.e., atpB rbcL, psbA trnH, trnD trnT and trnS trnG) and nuclear ITS region were studied. The results showed that Pterygiella includes only three species, namely P.nigrescens, P.duclouxii, and P.cylindrica, while P.suffruticosa should be merged into P.cylindrica as a variety. The previous delimitation of the two varieties within P.duclouxii was probably caused by imperfect analysis of morphological characters. Thus, a lectotype was designated for P.duclouxii and the plants formerly treated as P.duclouxii that bear eglandular stem hairs should be put into P.nigrescens, while the individuals with glabrous stem should remain in P.duclouxii. A new key to the three species in the genus reflecting the results of this study was presented, and a taxonomic revision was given.     

Glacial Effects on Sequence Divergence of Mitochondrial COII of Polyura eudamippus (Lepidoptera: Nymphalidae) in China

Sequence divergence of mitochondrial COII was analyzed in 50 specimens belonging to five subspecies of Polyura eudamippus (Lepidoptera: Nymphalidae) collected from southern China. There were nine haplotypes in the 405 bp of partial COII sequence. Distribution of the five subspecies was primarily consistent with the spatial distribution of haplotypes. The K _st statistic showed genetic differentiation among these subspecies, except between the pair of P. e. kuangtungensis and P. e. formosana, which were separated by the Taiwan Strait. This is consistent with the 10,000-year history of the Taiwan Strait, not long enough for detectable differentiation. The present distribution pattern of COII haplotypes of P. eudamippus should be shaped by the alteration of Pleistocene glaciations, and Yunnan might be the refugium of P. eudamippus in the ice age, judging from the abundant haplotypes remaining. There were two routes for P. eudamippus in the postglacial expansion, one northward to Sichuan, Chongqing, and Hubei and another eastward to the southeastern coast of mainland China and Taiwan Island. Because the haplotype of butterflies on Hainan Island (P. e. whiteheadi) was completely different from that of mainland China, it was estimated that butterflies on Hainan Island might be from the Indo-China Peninsula rather than from mainland China.     

Using RAPD for estimating genetic polymorphism in and phylogenetic relationships among species of the genus Lycopersicon (Tourn.) Mill

RAPD genome analysis of 53 species and cultivars of the genus Lycopersicon (Tourn.) Mill. revealed their high genetic polymorphism (Tourn.) Mill., based on which their phylogenetic relationships were inferred. In total, 248 polymorphic DNA fragments were amplified. Intraspecific polymorphism was maximum (79%) in L. peruvianum and minimum (9%) in L. parviflorum. In general, genome divergence among cross-pollinating tomato species was substantially higher than in self-pollinating species. An UPGMA dendrogram constructed from the RAPD patterns was consisted with the Lycopersicon phylogeny inferred from the molecular data of RFLP, ISSR, and microsatellite analyses and with a classification based on morphological characters. The relationships of taxa within the genus Lycopersicon are discussed.