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The immunoglobulin (Ig) joining (J) chain plays an important role in the formation of polymeric Igs and their transport into secretions. In the present study, the cDNA sequence of J chain has been cloned from the Chinese soft-shelled turtle (Pelodiscus sinensis) by reverse transcription (RT)-PCR and rapid amplification of cDNA ends (RACE). The cDNA sequence is 2347 bp in length and contains an open reading frame of 480 bp encoding 160 aa including the signal sequence. The deduced amino acid sequence has a high degree of homology with that of an already reported turtle J chain (80.7%), and of chicken (71.3%). By using real-time quantitative RT-PCR analysis, a significant up-regulation of J-chain transcripts was observed in spleen, kidney and blood of turtles injected with inactivated Aeromonas hydrophila, indicating the immune role of J chain in response to bacterial infection. (C) 2009 Elsevier B.V. All rights reserved.

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采用特殊声学调制的高重频激光技术对钢基体强化处理,使基体表面形成按一定几何规律排列的微米量级和微坑,使得基体的表面形貌、微观组织、铬层与基体的结合等特征均发生了本质的变化。它有效地增加了基体的表面积,有利于电沉积初期铬原子沿基体原有晶格的外延生长;有效地改善了铬层与基体的结合效果,提高了铬层抵抗复杂应力破坏的能力。

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牡丹复合体(Paeonia suffruticosa Andr. Complex)属芍药属牡丹组,为中国特有的落叶亚灌木,野生类型均为濒危种,仅局限分布于以秦岭为中心的较小区域。由于分布区有限,个体数量少,栽培历史长,育种广泛,种间极易杂交,网状进化的广泛发生,使得该复合体分类混乱。本文选取牡丹复合体6个野生种以及2个近缘类群,采用分别基于核酸印迹杂交和聚合酶链式反应的限制性酶切片段长度多态性分析,对细胞核核糖体基因片段ITS/18s的变异进行了分析,并且结合采用微卫星DNA指纹分析技术。选取18种内切酶对特异片段进行酶切消化。共得149个酶切位点,其中67个为变异位点,占45.0%。其中编码区(2.Okb,含18s,5.8s,26s)有突变位点29个,占该区段长度的1.4%;非编码区(490bp,含ITS-1,ITS-2)有突变位点38个,占该区段长度的7.8%。由此,可明显比较二者进化的保守程度和进化速率。两段间隔区的变异程度也存在差异。ITS-1为6.0%.ITS-2为9.9%。这说明构建系统树时二者的选用应得到综合考虑或加权。在复合体内不存在长度变异,即无缺失或插入发生,暗示了该复合体各种之间亲缘关系的紧密。根据Neighbor-joining法并计算遗传距离构建系统关系图,结果如下:(1)卵叶牡丹(神农架红花类群)与紫斑牡丹分化较早,考虑其与复合体内其它各种之间的遗传距离,支持将其定为新种的观点;(2)神农架白花类群与与卵叶牡丹亲缘关系非常相近,这一结果支持了来自其它分子和表型分析的结果;(3)延安牡丹与紫斑牡丹亲缘关系极近,但与矮牡丹关系较远,是否为上述两个种的杂交种,目前为止尚无充分的证据,作为存疑种处理;(4)川牡丹和矮牡丹进化关系密切,这一结果与ITS序列分析结果完全一致,加之其地理分布式样的不连续性说明了它们的古老和残存性质,这可进而推广至本复合体乃至整个牡丹组。我们认为现存的分布格局可能是地理与气候演化的产物,估计牡丹组的野牡丹复合体从本复合体分化出去的时间约为310 - 750万年前。由于基因间协调进化的不均一作用和该类群杂种的早期起源限制了核糖体基因在追溯其网状进化历程上的作用,这符合基因转换的梯度理论。最后讨论了nrDNA得到的基因树与其它的基因树和种系树之间的关系。

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稻属(Oryza L)属于禾本科的稻族,主要分布于世界的热带和亚热带地区。一般认为稻属包括20个左右的野生种和2个栽培种,我国有4-5种。迄今为止,对于稻属中种的数目和划分;稻属中所包括的种类是否为一单系类群;各基因组之闻的关系和起源以及稻属的起源等等,学术界仍然存有争议。本文评述了稻属分子系统学研究的进展,研究中存在的问题以及对未来稻属系统学研究的展望。针对稻属的分子系统学研究中所存在的问题,运用核糖体DNA转录间隔区序列测定、随机扩增多态性DNA(RAD)和SSR-anchoredPCR等分子生物学的手段对稻属中全部23个种的来自中国和世界不同地区的材料开展了分子系统学的研究。所得主要结果如下: 1.对产于中国的三种野生稻和栽培稻的二个亚种的核糖体DNA第一转录间隔区进行了序列测定。 DNA序列分别用PAUP程序进行分支分析和用MEGA程序中的UPGMA和Neighbor-joining进行聚类分析,结果表明核糖体DNA转录间隔区序列适合于进行稻属的分子系统学研究。 2.用荧光自动测序法和人工同位素测序法测定了稻属23个种和4个外类群的转录间隔区的序列。所有的DNA序列用Clustal V程序排阵后,再用PAUP程序进行分支分析。结果表明: A.稻属中的23个种组成四个主要分支,分别相当于Vaughan(1989)的四个种复合体。 B.所有AA基因组的种都在一个分支中。除了O.meridionalis与其它种的关系相对较远外,AA基因组种间有一定程度的分化,但不大。 C.尽管BB、BBCC、CC、CCDD和EE基因组的种都在一个分支中,但它们之间的分化是较为明显的。O.australiensis与该分支中的其它种的关系相对较远。以上两个分支的关系较为密切,它们组成一个自然类群,是稻属的核心部分。 D.O.ridleyi与O.longiglumis非常近缘,它们组成一个分支。 E.O.meyeriana与O.granulata有非常密切的关系。O.brachyantha与O.meyeriana近缘,而与AA基因组的关系较远。O,schlechter/与Leersia hexandra近缘,而与稻属中其它种的关系较远。作为外类群的Porteresia coarctata与稻属的a brachyantha非常近缘,这有可能说明将它作稻属中的一员的观点是正确的。 3.用RAPD和ISSR技术对稻属23介种的36份材料以及Porteresia coar ctata和Leersia hexandra各1份材料进行PCR扩增。16个10-mer RAPD随机引物共扩增出368个多态条带,5个锚定SSR引物共扩增出1 16个多态条带。RAPD和ISSR扩增出的多态条带合在同一数据矩阵中,用NTSYS-pc程序进行聚类分析。得出的结果与ITS序列分支分析的结果相似。 A.从表征图上亦可区分出四个群。所有的AA基因组的种聚在一起,O.nivara与O.rufipogon的关系密切,两个种的界限不清楚,所以认为它们是一个种:O.rufipogon.。 B.分布于中国的O.officinalis与O.minuta非常近缘,而菲律宾的四倍体O.officinalis则与同是基因组的O.eichingeri和O.rhizomatis关系密切。EE基因组的O.australiensis与Officinalis群保持一种松散的联系。 C。具FF基因组的O.brachyantha与O.schlechteri近缘,它们与Porteresiacoarctata和Leersia hexandra关系的密切程度要大于它们与稻属其它种的关系。 D。最后讨论了RAPD和ISSR用予植物属内的系统发育研究存在的问题。本研究结果说明RAPD和ISSR适合于进行属内近缘种亲缘关系的研究。

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结合野外生态调查与分子标记检测,本文探讨了显性遗传标记应用于居群遗传学研究时实验与数据分析需要注意的问题,并在此基础上对中国分布的疣粒野生稻(Oryza granulata Nees et Am. ex Watt.)居群遗传多样性与居群遗传结构进行了研究。然后从metapopulation结构动态、无性系生长和物种形成等角度研究了遗传结构的空间格局与生态学、系统学因素之间的相互作用,并将研究的空间尺度从聚群内(colony)、居群内、地区内推广到地区间和整个物种水平,反映了在不同空间等级上疣粒野生稻相异的进化模式。最后,综合上述结果,提出了保护疣粒野生稻的原则和策略。结果如下: 1.根据对分布于中国云南和海南33个分布点疣粒野生稻居群所做的野外生态学调查,该物种目前在中国的30个县(市)有分布,比1978-1982年全国野生稻普查时增加了3个县市(海南通什、云南思茅和勐腊)。疣粒野生稻具有较强的耐荫和抗旱能力,在群落总盖度为90-210%范围内生长良好。它是一种典型的适应中度干扰的物种,生长于有一定干扰的斑块状生境中。疣粒野生稻在群落内的分布格局为聚集型,其居群密度较小(1.13-2.95株/m2),依靠重力下落和动物传播种子。由于对热区资源的掠夺性开发,总计有12.9%的居群已因人为干扰而灭绝,83.9%的居群处于严重的危胁之下,处于濒危状态。对疣粒野生稻的破坏在不同地区间程度不同,生境恶化和放牧是造成其居群灭绝的最主要原因,对其进行保护已经迫在眉睫。在调查的基础上,本研究建立了含该物种34个居群共1109份个体样品的总DNA库,作为易位保护的一种措施,主要用于居群遗传学和保护生物学研究。 2.利用上述总DNA库中的材料,首先采用随机扩增多态(RAPD)对几类显性标记的居群遗传结构参数进行了比较。在衡量遗传多样性水平时,多态位点比率(PPB)会低估变异的程度,其价值不如Shannon多样性指数和Nei多样性指数。在计算个体之间的遗传关系时,Mantel检测表明17种相似性系数之间存在极显著的相关性。同时,基于Φst。遗传距离的分子方差分析(AMOVA)和基于Hardy-Weinberg平衡假设的Nei氏遗传距离分析的结果间具有显著的相关性,它们都适用于对疣粒野生稻居群遗传结构进行研究,且在使用后者时应对数据进行Lynch-Milligan矫正,剔除隐性基因型(0)频率小于3/N(N为样本总数)的条带数据,以矫正显性遗传方式对变异估计偏低的影响。此外,各类遗传结构分析参数之间的高度相关性也与疣粒野生稻居群内遗传多样性低,杂合体比率较低有密切关系。 利用RAPD和inter-简单重复序列(ISSR)对来自中国20个居群疣粒野生稻混合样品,以及海南(M5)和云南(M27)两个居群各20个植株的遗传多样性进行了检测。ISSR的实验稳定性优于RAPD,且总的来说它能检测到更多的遗传变异。前者与它在PCR反应时退火温度较高,引物.模板复合物较稳定有关;而后者则与其引物靶序列容易在细胞分裂中产生突变有关。Mantel检测结果表明,衡量样品间的遗传关系时,这两种标记的分析结果在物种水平存在极显著的相关性(r = 0.917, t = 12789),而在居群水平不相关(r < 0.200)。这不但与它们所扩增的相应基因组片断的变异方式及在居群内分辩率下降有关,同时也反映了疣粒野生稻居群内和居群间存在着不同的进化模式。 3.利用RAPD和ISSR标记,对中国20个居群疣粒野生稻混合样品的遗传多样性进行了研究。RAPD和ISSR分别扩增出209和122条PCR条带,其中各有64.1l%(134条带)和72.95%(89条带)为多态条带。基于Jaccard系数的UPGMA分析表明,同一地区内居群的遗传变异比较小。20个居群按来源聚为云南与海南两类,其间产生了一定程度的遗传分化。这种遗传多样性分布的特点可能与其起源、分布格局、交配系统和种子散布方式有关。此外,尽管混合取样会低估一定的遗传变异能力,也不能得到关于居群内的遗传结构情况,但它仍然是一种获取遗传多样性信息的高效方法,适用于对研究材料进行日常管理和评价。 4.按照居群取样的方法,利用RAPD对来自中国云南和海南的20个疣粒野生稻居群共396个植株进行了遗传结构分析。联合ISSR,对其中5个居群初步的分析表明该物种在居群内的遗传多样性水平很低,RAPD的多态位点比率(PPB)在居群内从4.52%到13.06%;而ISSR的PPB值在居群内从7.08%-26.55%。AMOVA分析表明,对于RAPD来说,云南与海南两地区之间遗传变异的量占总变异量的73.85%,地区之内占19.45%,而居群内仅占6.70%。对于ISSR,疣粒野生稻地区间,地区内和居群内遗传变异的分布比率分别为49.26%、38.070A和12.66%。UPGMA聚类将同一居群内的个体聚为一支,并将居群按来源分为云南和海南两类。由于疣粒野生稻在群落内的分布为典型的metapopulation格局,伴随各聚群(colony)在群落次生演替过程中周转(灭绝与定植)时发生的遗传漂变、建立者效应和居群内强烈的近交是造成其居群内遗传多样性极低的主要原因。 5.利用10个ISSR标记对中国4个疣粒野生稻居群内的无性系生长与基因型遗传多样性进行了分析。在小尺度取样(个体间隔1.0-1.5m)的情况下,所有居群中均检测到明显的无性系生长现象。参与无性系生长的个体百分比在各居群中从25%-60%不等,Simpson多样性指数表明疣粒野生稻居群内的基因型多样性保持在较高水平(0.837-0.958)。尽管如此,AMOVA对居群内遗传变异进行方差剖分的结果表明参与无性生长的个体所含有的变异量平均只占总变异量的16.7%。因此,疣粒野生稻居群内遗传变异的来源主要依靠有性生殖来维持。同时,处于人为中度干扰之下的疣粒野生稻居群不但个体密度较高,其居群遗传多样性也未因此而降低。 6.在假设RAPD在同一种及其近缘种内PCR产物同源性较高的前提下,利用该技术对来自世界的23份O. granulata和O. meyeriana样品进行了遗传多样性分析和系统学研究。在物种水平,O. granulata具有非常高的遗传多样性(多态位点比率达83.54%),表明该物种进化历史中存在大规模居群瓶颈效应的可能性较小。O.gramulata与O. meyeriana各居群的遗传分化与岛屿形成导致的地理隔离之间有密切的关系。基于Nei & Li遗传相似性系数,利用Neighbor-Joining和UPGMA聚类法构建的两个系统树并不完全一致。主坐标分析(PCoA)支持NJ法的结果:来自O.meyeriana的两个样品倾向于聚为一类,并获得bootstrap分析的支持,但它们的遗传变异范围并未超出O. granulata。因此,我们的结果支持将这两个种进行归并。 7.由于疣粒野生稻在物种水平的遗传多样性非常高,变异主要存在于各地区之间,因此要最大程度地维持该物种遗传多样性,使之不发生遗传侵蚀意味着保护应该针对整个物种的分布区进行。对于分布于中国的居群来说,一方面由于变异主要存在于云南和海南两地区之间,另一方面由于地区内和居群内的遗传多样性相对较低,因此,云南和海南应做为中国保护该物种的两个中心。此外,由于一定程度的人为干扰有利于为该物种创造适宜生境,增加其居群密度,且不会致使遗传变异能力下降,因此在实施就地保护时应充分考虑将其与当地的经济开发项目相结合,达到自然保护与地方经济可持续发展的目的。

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The Afrotheria, a supraordinal grouping of mammals whose radiation is rooted in Africa, is strongly supported by DNA sequence data but not by their disparate anatomical features. We have used flow-sorted human, aardvark, and African elephant chromosome painting probes and applied reciprocal painting schemes to representatives of two of the Afrotherian orders, the Tubulidentata (aardvark) and Proboscidea (elephants), in an attempt to shed additional light on the evolutionary affinities of this enigmatic group of mammals. Although we have not yet found any unique cytogenetic signatures that support the monophyly of the Afrotheria, embedded within the aardvark genome we find the strongest evidence yet of a mammalian ancestral karyotype comprising 2n = 44. This karyotype includes nine chromosomes that show complete conserved synteny to those of man, six that show conservation as single chromosome arms or blocks in the human karyotype but that occur on two different chromosomes in the ancestor, and seven neighbor-joining combinations (i.e., the synteny is maintained in the majority of species of the orders studied so far, but which corresponds to two chromosomes in humans). The comparative chromosome maps presented between human and these Afrotherian species provide further insight into mammalian genome organization and comparative genomic data for the Afrotheria, one of the four major evolutionary clades postulated for the Eutheria.

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The phylogenetic relationships among worldwide species of genus Ochotona were investigated by sequencing mitochondrial cytochrome b and ND4 genes. Parsimony and neighbor-joining analyses of the sequence data yielded congruent results that strongly indicated three major clusters: the shrub-steppe group, the northern group, and the mountain group. The subgeneric classification of Ochotona species needs to be revised because each of the two subgenera in the present classification contains species from the mountain group. To solve this taxonomic problem so that each taxon is monophyletic, i.e., represents a natural clade, Ochotona could be divided into three subgenera, one for the shrub-steppe species, a second for the northern species, and a third for the mountain species. The inferred tree suggests that the differentiation of this genus in the Palearctic Region was closely related to the gradual uplifting of the Tibet (Qinghai-Xizang) Plateau, as hypothesized previously, and that vicariance might have played a major role in the differentiation of this genus on the Plateau, On the other hand, the North American species, O. princeps, is most likely a dispersal event, which might have happened during the Pliocene through the opening of the Bering Strait. The phylogenetic relationships within the shrub-steppe group are worth noting in that instead of a monophyletic shrub-dwelling group, shrub dwellers and steppe dwellers are intermingled with each other. Moreover, the sequence divergence within the sister tars of one steppe? dweller and one shrub dweller is very low. These findings support the hypothesis that pikes have entered the steppe environment several times and that morphological similarities within steppe dwellers were due to convergent evolution. (C) 2000 Academic Press.

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The sequences of the mitochondrial ND4 gene (1339 bp) and the ND4L gene (290 bp) were determined for all the 14 extant taxa of the Drosophila nasuta subgroup The average A + T content of ND4 genes is 76.5% and that of ND4L genes is 83.5%. A total of 114 variable sites were scored. The ND4 gene sequence divergence ranged from 0 to 5.4% within the subgroup. The substitution rate of the ND4 gene is about 1.25% per million years. The base substitution of the genesis strongly transition biased. Neighbor-joining and parsimony were used to construct a phylogeny based on the resultant sequence data set. According to these trees, five, distinct mtDNA clades can be identified. D. niveifrons represents the most diverged lineage. D, sulfurigaster bilimbata and D. kepulauana form two independent lineages. The other two clades are the kohkoa complex and the albomicans complex. The Kohkoa complex consists of D. sulfurigaster sulfurigaster, D. pulaua, D. kohkoa, and Taxon-F. The albomicans complex can be divided into two groups: D. nasuta, D. sulfurigaster neonasuta, D. sulfurigaster albostrigata, and D.. albomicans from Chiangmai form one group; and D. pallidifrons, Taxon-I, Taxon-J, and D. albomicans from China form the other group. High genetic differentiation was found among D. albomicans populations. Based on our phylogenetic results, we hypothesize that D. niveifrons diverged first from the D, nasuta subgroup in Papua New Guinea about 3.5 Mya. The ancestral population spread to the north and when it reached Borneo, it diversified sequentially into the kohkoa complex, D. s. bilimbata, and D. kepulauana. About 1 Mya, another radiation occurred when the ancestral populations reached the Indo-China Peninsula, forming the albomicans complex. Discrepancy between morphological groupings and phylogenetic results suggests that the male morphological traits may not be orthologous. (C) 1999 Academic Press.

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Six sample specimens of Trachypithecus francoisi and 3 of T. leucocephalus were analyzed by use of allozyme electrophoresis and random amplified polymorphism DNA (RAPD) in order to clarify the challenged taxonomic status of the white-head langur. Among the 44 loci surveyed, only 1 locus (PGM-2) was found to be polymorphic. Nei's genetic distance was 0.0025. In total, thirty 10-mer arbitrary primers were used for RAPD analysis, of which 22 generated clear bands. Phylogenetic trees were constructed based on genetic distances using neighbor-joining and UPGMA methods. The results show that T. francoisi and T: leucocephalus are not monophyletic. T. francoisi from Guangxi, China and Vietnam could not be clearly distinguished, and they are not divided into 2 clusters. A t-test was performed to evaluate between genetic distances within and between T. leucocephalus and T. francoisi taxa groups. The statistical test shows that the taxa group within T: leucocephalus and T: francoisi does not significantly differ from that between T: leucocephalus and T: francoisi at the 5% level. Our results suggest that the level of genetic differentiation between T, leucocephalus and T. francoisi is relatively low. Recent gene flow might exist between T. francoisi and T. leucocephalus. Combining morphological features, geographical distribution, allozyme data, RAPD data, and mtDNA sequences, we suggest that the white-head langur might be a subspecies of T. francoisi.

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We investigated the phylogenetic relationships among most Chinese species of lizards in the genus Phrynocephalus (118 individuals, collected from 56 populations of 14 well-defined species and several unidentified specimens) using four mitochondrial gene fragments (12S rRNA, 16S rRNA, cytochrome b, and ND4-tRNA(LEU)). The partition-homogeneity tests indicated that the combined dataset was homogeneous, and maximum-parsimony (MP), neighbor-joining (NJ), maximum-likelihood (ML) and Bayesian (BI) analyses were performed on this combined dataset (49 haplotypes including outgroups for 2058 bp in total). The maximum-parsimony analysis resulted in 24 equally parsimonious trees, and their strict consensus tree shows that there are two major clades representing the Chinese Phrynocephalus species: the viviparous group (Clade A) and the oviparous group (Clade B). The trees derived from Bayesian, ML. and NJ analyses were topologically identical to the MP analysis except for the position of P. mystaceus. All analyses left the nodes for the oviparous group, the most basal clade within the oviparous group, and P. mystaceus unresolved. The phylogenies further suggest that the monophyly of the viviparous species may have resulted from vicariance, while recent dispersal may have been important in generating the pattern of variation among the oviparous species. (C) 2003 Elsevier Science (USA). All rights reserved.

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A fragment of mitochondrial DNA (mtDNA) control region (similar to700 bp) was sequenced in 104 individuals from 20 breeds (three Chinese domestic breeds, five recently derived breeds and 12 introduced breeds) of domestic rabbits, Oryctolagus cuniculus . Nineteen sites were polymorphic, with 18 transitions and one insertion/deletion, and eight haplotypes (A1, A2, A3, A4, A5, A6, A7 and A8) were identified. Haplotype A1 was the most common and occurred in 89 individuals. In the 25 Chinese rabbits, only haplotype A1 was observed, while four haplotypes (A1, A3, A5 and A6) were found in 26 recently derived individuals. Haplotype A2 was shared by seven individuals among three introduced strains. The other six haplotypes accounted for 0. 96-1. 92% of the animals. Combined with the published sequences of European rabbits, a reduced median-joining network was constructed. The Chinese rabbit mtDNAs were scattered into two clusters of European rabbits. These results suggest that the (so-called) Chinese rabbits were introduced from Europe. Genetic diversity in Chinese rabbits was very low.

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Fringillidae is a large and diverse family of Passeriformes. So far, however, Fringillidae relationships deduced from morphological features and by a number of molecular approaches have remained unproven. Recently, much attention has been attracted to mitochondrial tRNA genes, whose sequence and secondary structural characteristics have shown to be useful for Acrodont Lizards and deep-branch phylogenetic studies. In order to identify useful phylogenetic markers and test Fringillidae relationships, we have sequenced three major clusters of mitochondrial tRNA genes from 15 Fringillidae, taxa. A coincident tree, with coturnix as outgroup, was obtained through Maximum-likelihood method using combined dataset of 11 mitochondrial tRNA gene sequences. The result was similar to that through Neighbor-joining but different from Maximum-parsimony methods. Phylogenetic trees constructed with stem-region sequences of 11 genes had many different topologies and lower confidence than with total sequences. On the other hand, some secondary structural characteristics may provide phylogenetic information on relatively short internal branches at under-genus level. In summary, our data indicate that mitochondrial tRNA genes can achieve high confidence on Fringillidae phylogeny at subfamily level, and stem-region sequences may be suitable only at above-family level. Secondary structural characteristics may also be useful to resolve phylogenetic relationship between different genera of Fringillidae with good performance.

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作者进行了较广泛的样品采集,通过实验分离纯化培养得到多个鞘藻目种类的株系,并采用PCR技术新获得鞘藻目2属8个种类的部分28S rDNA序列,连同GenBank中的另两条序列,分析的物种涵盖了鞘藻目中的每个属。通过比较分析绿藻纲中包括此10条序列的共36个种类的同一基因序列,并选取Trebouxiophyceae中的椭圆小球藻(Chlorella ellipsoidea)和Fusochloris perforata作外类群,运用多种方法构建分子系统树,包括邻接法(Neighbor-Joining)、最大简