THE ROLE OF INTROGRESSIVE HYBRIDIZATION IN THE EVOLUTION OF THE GILA ROBUSTA COMPLEX (TELEOSTEI: CYPRINIDAE)

Abstract The extent and impact of introgressive hybridization was examined in the Gila robusta complex of cyprinid fishes using mitochondrial DNA (mtDNA) sequence variation. Lower Colorado River basin populations of G. robusta, G. elegans, and G. cypha exhibited distinct mtDNAs, with only limited introgression of G. elegans into G. cypha. The impact of hybridization was significant in upper Colorado River basin populations; most upper basin fishes sampled exhibited only G. cypha mtDNA haplotypes, with some individuals exhibiting mtDNA from G. elegans. The complete absence of G. robusta mtDNA, even in populations of morphologically pure G. robusta, indicates extensive introgression that predates human influence. Analysis of the geographic distribution of variation identified two distinctive G. elegans lineages; however, the small number of individuals and localities sampled precluded a comprehensive analysis. Analysis of haplotype and population networks for G. cypha mtDNAs from 15 localities revealed low divergence among haplotypes; however, significant frequency differences among populations within and among drainages were found, largely attributable to samples in the Little Colorado River region. This structure was not associated with G. cypha and G. robusta, as morphotypes from the same location are more similar than conspecific forms in other locations. This indicates that morphological and mtDNA variation are affected by different evolutionary forces in Colorado River Gila and illustrates how both hybridization and local adaptation can play important roles in evolution.     

PRELIMINARY STUDY ON MITOCHONDRIAL CYTOCHROME b DNA SEQUENCES AND PHYLOGENY OF FORMALIN FIXED SISORID FISHES (Teleostei:Siluriformes)

从９种科鱼类的福尔马林标本中获得了３３３ｂｐ的细胞色素ｂ基因片段的序列。这９个种分别代表科鱼类的８个属。３３３ｂｐ的ＤＮＡ序列经ＭＵＳＴ软件排序后,有１０１个变异位点,其中有３９个信息位点。序列在成对物种间的距离为８～４８。平均遗传距离为２４％～１４４％。简约分析产生了最大简约系统树,其步长是１６２（ＣＩ＝０７３５,ＲＩ＝０４９４）。在该系统树上,Ｂａｇａｒｉｕｓ是最原始的属,并与所有其他的物种形成姊妹群。其余８个属形成一个单系类群并分为二个姊妹群。尽管在形态上具有１３个离征,但在分子系统树上,鱼类并未形成一个单系类群。可能的原因是３３３ｂｐ序列中的星系信息位点太少;另外单从福尔马林浸制标本获得的ＤＮＡ序列的可靠性尚有待进一步验证     

迷蛱蝶属及相关物种细胞色素b基因序列及其分子系统学研究

迷蛱蝶属Mimathyma隶属于蛱蝶科Nymphalidae闪蛱蝶亚科Apaturinae,该属所包含的种类复杂,其分类学地位存在争议.本文对迷蛱蝶属、闪蛱蝶属Apatura和带蛱蝶属Athyma7个种共19个个体的线粒体DNA细胞色素b基因部分序列进行测定分析,并以花斑螯蛱蝶Charaxes kahruba (Moore)作为外群用PAUP软件构建MP和NJ分子系统树.结果显示迷蛱蝶Mimathyma chevana(Moore)、夜迷蛱蝶Mimathyma nycteis(Ménétriès)、白斑迷蛱蝶Mimathyma schrenckii (Ménétriès)和环带迷蛱蝶Mimathyma ambica Kollar形成1个聚类簇,支持Moore将这4个种由闪蛱蝶属移出并建立迷蛱蝶属的观点.同时,尽管迷蛱蝶在形态上与该属其余3种相似,但研究发现聚类簇Ⅰ中夜迷蛱蝶、白斑迷蛱蝶和环带迷蛱蝶首先相聚,然后再与迷蛱蝶聚在一起,表明迷蛱蝶与这3种亲缘关系较远.此外,本文的研究结果还显示迷蛱蝶属与闪蛱蝶属关系密切,而与带蛱蝶属的关系较远.     

质体醌重组的D1／D2／Cytb559复合物的荧光衰减动力学和光破坏作用

光系统Ⅱ反应中心Ｄ１／Ｄ２／Ｃｙｔｂ５５９ 在分离纯化过程中失去了电子受体ＱＡ 和ＱＢ,人工合成的质体醌可以与Ｄ１／Ｄ２／Ｃｙｔｂ５５９ 复合物发生重组。癸基质体醌（ＤＰＱ）与Ｄ１／Ｄ２／Ｃｙｔｂ５９９ 复合物的重组导致该复合物的荧光强度下降及发射光谱蓝移,同时两个与光化学活性相关的长寿命（２４ ｎｓ和７３ ｎｓ）荧光衰减组分占整个荧光的百分数下降,这些结果表明ＤＰＱ作为Ｐｈｅｏ－ 的电子受体,限制了Ｐ６８０＋ ·Ｐｈｅｏ－ 的电荷重组。ＤＰＱ 的加入对Ｄ１／Ｄ２／Ｃｙｔｂ５５９复合物中Ｃｈｌａ 分子的光破坏敏感性影响不大,但β－胡萝卜素在加入ＤＰＱ 之后可以被光照破坏,这个过程可能与β－胡萝卜素的生理功能相关。     

露螽属四种昆虫Cyt b基因序列及系统进化研究(直翅目,露螽科)

研究了露螽属4种昆虫线粒体细胞色素b基因(Cyt b)的部分序列,分析了核苷酸序列组成与变异及氨基酸差异.在得到的432bp序列中,A T约占66.9%,其中102个核苷酸位点发生了变异(约23.8%),从每个氨基酸密码子来看,第3位点的A T含量较高,为79.7%.Cyt b基因编码的144个氨基酸由19种氨基酸组成,有12个发生了变异,占氨基酸总数的8.33%,其中亮氨酸(Leu)与苯丙氨酸(Phe)的含量较高,谷氨酸(Glu)、赖氨酸(Lys)与精氨酸(Arg)的含量较低,无半胱氨酸(Cys).以日本纺织娘和中华螽斯为外群构建的NJ分子系统树显示,镰尾露螽与齿尾露螽是分化较晚的类群,其次是瘦露螽,黑角露螽是分化较早的类群.     

POPULATION STRUCTURE OF THE BOTTLENOSE DOLPHIN (TURSIOPS TRUNCATUS) AS DETERMINED BY RESTRICTION ENDONUCLEASE ANALYSIS OF MITOCHONDRIAL DNA

Abstract: Restriction fragment length polymorphisms of mitochondrial DNA (mtDNA) were used to test for population subdivision in the bottlenose dolphin (Tursiops truncatus). Atlantic and Pacific dolphin mtDNA samples exhibited distinctly different haplotypes (approximately 2.4% sequence divergence), indicating a lack of gene exchange. Within the Atlantic Ocean, mtDNA samples from the Gulf of Mexico and the Atlantic Coast were also found to be distinct, with a sequence divergence of approximately 0.6%. The Atlantic Coast–Gulf of Mexico dichotomy is consistent with patterns of genetic variation from other marine and coastal organisms from this region, and supports the hypothesized role of bio-geographic events in promoting the divergence of these and other forms. Regional differentiation was identified along the Atlantic Coast, whereas low sequence divergences among haplotypes and consistent haplotype frequencies across populations suggested considerable gene exchange among Gulf of Mexico populations. A highly divergent haplotype found in two individuals from two localities in the Gulf of Mexico is best explained by dispersal from either a distinct offshore Gulf stock or an unsampled Atlantic Coast stock. Additional samples are required to test for the existence of a distinct offshore race and, if it exists, to identify its distribution and contribution to population structure.     

CYTOGENETIC ANALYSIS OF CHROMOSOMAL INTERMEDIATES FROM A HYBRID ZONE BETWEEN TWO CHROMOSOME RACES OF THE SCELOPORUS GRAMMICUS COMPLEX (SAURIA,PHRYNOSOMATIDAE)

Underdominance for chromosomal rearrangements is the central assumption of several models of chromosomally based speciation including the cascade model, proposed for the Sceloporus grammicus complex. Several cytotypes of the S. grammicus complex hybridize at localities in central México. A hybrid zone between two of the most chromosomally divergent races (= cytotypes) of S. grammicus (F5, 2n = 34 and FM2, 2n = 44–46) was examined to assess the meiotic effects of heterozygosity at multiple chromosomes. Meiosis was examined in males heterozygous for “simple” Robertsonian fissions at chromosomes 1, 3, 4, and 6 and/or a pericentric inversion at chromosome 4. Analysis of synaptonemal complexes and chromosomal configurations at diakinesis showed trivalent formation in fission heterozygotes and heterosynapsis (lack of reverse-loop formation) in an inversion heterozygote. Analysis of metaphase II configurations revealed primarily balanced segregation and low levels of nondisjunction regardless of chromosomal background. The lack of underdominance associated with “simple” fission heterozygosity in this narrow hybrid zone contradicts the key premise of most chromosomally based models of speciation.     

ANALYSIS OF GAMETIC PROTOPLAST RELEASE IN THE CLOSTERIUM PERACEROSUM-STRIGOSUM-LITTORALE COMPLEX (CHLOROPHYTA)1

A biologically active glycoprotein (protoplast-release-inducing protein; PR-IP), which induces the release of gametic protoplasts from mating type minus (mt^-) cells of the Closterium peracerosum-strigosum-littorale complex, was prepared from a medium in which mt^- and mt⁺ cells had been previously incubated together. The process of PR-IP-inducing protoplast release was analyzed. Induction of protoplast release was dependent upon the duration of both PR-IP treatment and preincubation in nitrogen-deficient mating medium before PR-IP treatment. Low cell density in the preculture stage had a significant stimulative effect upon the induction of protoplast release. Light was necessary for protoplast release, especially just before PR-IP treatment. Chloramphenicol and 3-(4-chlorophenyl)-1,1-dimethylurea (CMU) exerted inhibitory effects on protoplast release, especially when they were applied to the preculture stage but not when they were applied to the protoplast-releasing stage after the PR-IP treatment. We suggest that preculture at a low cell density under continuous light conditions that may cause metabolic changes in the chloroplast is a very important stage for gametic protoplast release in this Closterium.     

DISTRIBUTION OF TWO TYPES OF EMILIANIA HUXLEYI (PRYMNESIOPHYCEAE) IN THE NORTHEAST ATLANTIC REGION AS DETERMINED BY IMMUNOFLUORESCENCE AND COCCOLITH MORPHOLOGY1

Culture strains of Emiliania huxleyi (Lohmann 1902) Hay et al. 1967 were placed into two groups designated E. huxleyi type A and type B on the basis of coccolith morphology and immunological properties of the coccolith polysaccharide. We studied the distribution of these types in the North Atlantic region using an indirect immunofluorescence assay with antisera directed against the coccolith polysaccharide of E. huxleyi type A and type B and epifluorescence microscopy. In field samples taken in the Northeast Atlantic Ocean, E. huxleyi type A was found exclusively. In contrast, type B was dominant in the North Sea. Scanning electron microscopy of the samples revealed the same unequal distribution of the two types as found with the immunofluorescent-labelling assay.     

脓疮草复合体的形态性状分析与分类修订

脓疮草复合体(Panzerina lanata complex)隶属于唇形科脓疮草属,原有6个合法学名。通过从内 蒙古、陕西、甘肃、宁夏及新疆等5个省或自治区的22个居群取样,进行野外和移栽后的对比观测及性 状分析,并结合多变量统计分析,从不同水平和角度对复合体进行了研究。结果表明,此复合体的根、茎 等都是环境可塑性很高的性状,叶次之,而花部、果实和种子性状的可塑性较小。以往划分复合体类群 的叶裂程度、茎叶被毛及萼齿形态等几个鉴别性状在天然居群中连续变化,以此划分的几个“种”,不过 是连续变异的极端个体。再结合地理分布的特征,将P．alaschanica、P．kansuensis和P. albescens归并 在P．1anata中,同样可证明P．argyracea不存在。对22个居群11个形态性状的统计学和主成分分析 与性状分析结果一致。研究表明,该复合体小坚果表面具疣与否、花冠超出萼筒的多少及花干后变色与 否等是其分种性状,以此分种标准该复合体可分为2种:Panzerina lanata(L．)Sojak和Panzerina parviflora(C．Y．Wu et H．W．Li)Y．Z.Zhao．     

PHYLOGENETIC IMPLICATIONS OF THE CAD COMPLEX FROM THE PRIMITIVE RED ALGA CYANIDIOSCHYZON MEROLAE (CYANIDIALES,RHODOPHYTA)1

The de novo pyrimidine biosynthetic pathway consists of six enzymes: carbamoyl‐phosphate synthetase II (CPS II), aspartate carbamoyltransferase (ACT), dihydroorotase (DHO), dihydroorotate dehydrogenase, orotate phosphoribosyltransferase, and orotidine‐5′‐monophosphate decarboxylase. The origin and organization of the first three enzymes differ markedly between Opisthokonta (Metazoa and Fungi) and the Amoebozoa and green plants. However, no information has been available regarding the characteristics of such genes in other photosynthetic eukaryotes. In this study, we examined the pyrimidine biosynthetic cluster in the primitive red alga Cyanidioschyzon merolae P. DeLuca et al. isolate 10D. Unlike the situation in green plants, the CPS II, ACT, and DHO of C. merolae were fused to form a single open reading frame (the CAD complex), as in the Opisthokonta and Amoebozoa. Phylogenetic analysis of the CPS domain sequences suggested that this red algal CAD complex did not result from a recent lateral gene transfer from Metazoa or Fungi but that the fusion of the three genes occurred before the divergence of Opisthokonta, Amoebozoa, and the red algae. These results cast doubt on the recent hypothesis that the Opisthokonta and Amoebozoa form a monophyletic group, based on the presence in both of the CAD complex.     

BIOCHEMICAL, PHYSIOLOGICAL, AND MOLECULAR ANALYSIS OF SEXUAL ISOLATION IN THE SPECIES COMPLEX CLOSTERIUM PERACEROSUM-STRIGOSUM-LITTORALE (CHLOROPHYTA)1

Closterium strains obtained from Japan ( NIES-64 and -65 ) and Nepal ( NIES-67 and -68 ) have been classified as the same taxonomic species; however, they are sexually isolated from each other. When NIES-64 and -65 cells were separately incubated in a medium in which both strains had previously been cultured together, release of protoplasts from both strains was observed. We suggest that factors responsible for the release of protoplasts from cells of both NIES-64 and -65 are produced in a mixed culture of these cells and function during conjugation. These factors, however, had no effect on the release of protoplasts from cells of strains NIFS-67 or -68. Alternatively, a protein that is responsible for the release of protoplasts from cells of NIES-68, called the protoplast-release-inducing protein ( PR-IP ), had no effect on the release of protoplasts from cells of strains NIES-64 or -65. When the media obtained from the culture of NIES-64 and -65 cells at various mixing ratios were analyzed by western blotting with antiserum to a 42-kDa subunit of PR-IP, no cross reaction was detected. In Southern hybridization analysis, no hybridizing band was observed when genomic DNAs of NIES-64 and -65 cells were probed with cDNAs encoding the two subunits of PR-IP. We suggest from these results that the factors responsible for the release of protoplasts from NIES-64 and -65 cells are not structurally similar to PR-IP. It is known that the release of PR-IP from NIES-67 cells can be induced by the action of another sex pheromone ( PR-IP inducer ) which is released by NIES-68 cells. In contrast, no protoplast-release-inducing activity was observed from either NIES-64 or -65 in a culture medium conditioned by opposite strains. We suggest that the conjugation systems employed by strains NIES-64/ NIES-65 and strains NIES-67 /NIES-68 differ, and we propose a possible mechanism of sexual isolation between these biological species .     

EVOLUTION OF MACROCYSTIS SPP. (PHAEOPHYCEAE) AS DETERMINED BY ITS1 AND ITS2 SEQUENCES1,

Macrocystis (Lessoniaceae) displays an antitropical distribution, occurring in temperate subtidal regions along western North America in the northern hemisphere and throughout the southern hemisphere. We used the noncoding rDNA internal transcribed spacer regions (ITS1 and ITS2) to examine relatedness among (1) Macrocystis and several genera of Laminariales, (2) four species of Macrocystis ( M. integrifolia Bory from the northern hemisphere, M. angustifolia Bory and M. laevis Hay from the southern hemisphere, and M. pyrifera [L.] C. Ag. from both hemispheres), and (3) multiple clones of several individuals. Of the taxa included in our phylogenetic analysis, the elk kelp, Pelagophycus porra (Lem.) Setch., was the sister taxon to Macrocystis spp. Macrocystis individuals from the southern hemisphere (representing three species) formed a strongly to moderately supported clade, respectively, when the ITS1 and ITS2 sequences were analyzed separately. No distinction was detected between the two species in the northern hemisphere. Thus, Macrocystis may be a monospecific genus ( M. pyrifera ). A northern-hemisphere-to-southern-hemisphere pattern of dispersal was inferred, because northern-hemisphere individuals were more diverse and displayed paraphyletic clades, whereas southern-hemisphere individuals were less diverse and formed a monophyletic clade. High intraindividual variation in ITS1 sequences was observed in one individual from Santa Catalina Island (CA), suggesting very recent and rapid mixing of genotypes from areas to the north and Baja California (Mexico) or introgressive hybridization with Pelagophycus.     

基于18S rRNA基因部分序列探讨四种蛙类多盘科吸虫分类地位

分析了寄生于蛙类膀胱的4种多盘虫科Polystomatidae吸虫:石林双睾吸虫Diplorchis shilinensis、锯腿树蛙多盘吸虫Polystoma carvirostris和分别寄生于昭觉林蛙Rana chaochiaoensis、华西雨蛙Hyla a.annectans的2未定种的分类地位,并利用18S rRNA基因部分序列进行了系统发育重建.结果表明:4种多盘科吸虫为多盘虫属Polystoma和双睾虫属Diplorchis的4个分类单元,其中2未定种为多盘虫属未记录种;肠管、生殖系统、后吸器、大钩和生殖棘等为多盘虫属属内种定种的可靠性状.基于18S rRNA基因序列,4种多盘科吸虫呈现以下进化关系:1)寄生于昭觉林蛙和华西雨蛙的多盘虫属2未定种进入欧非混合进化支;2)锯腿树蛙多盘吸虫位于多盘虫属进化支的最基部;3)石林双睾吸虫进入澳洲进化支.     

FEMALE FECUNDITY IN A HYBRID ZONE BETWEEN TWO CHROMOSOME RACES OF THE SCELOPORUS GRAMMICUS COMPLEX (SAURIA,PHRYNOSOMATIDAE)

Individuals of the F5 and FM2 cytotypes of the Sceloporus grammicus complex form a narrow zone of parapatric hybridization near Tulancingo, Hidalgo, Mexico. Reproductive parameters were examined among chromosomally parental and hybrid females to assess the degree to which reduced clutch size is correlated with the level of chromosomal heterozygosity. Although clutch size in the two parental groups was highly correlated with female body size, this was not the case for females with intermediate karyotypes. These females displayed increased levels of infertility manifested as smaller clutches and as inviable embryos. F₁ females produced the smallest average clutches and suffered the most precipitous fecundity loss (up to 75%). The number of heterozygous marker chromosomes and heterozygosity at chromosome 2 had significant effects on the number of viable embryos. Analysis of embryo karyotypes revealed the production of triploid offspring and an excess number of embryos heterozygous at chromosome 1. Differences in viability, among females heterozygous for the same number of chromosomes, suggest that genetic background of the female and/or sire may be an important factor in determining reproductive success.     

THE GENETIC STRUCTURE OF A HYBRID ZONE BETWEEN TWO CHROMOSOME RACES OF THE SCELOPORUS GRAMMICUS COMPLEX (SAURIA,PHRYNOSOMATIDAE) IN CENTRAL MEXICO

The F5 (2n = 34) and FM2 (2n = 44–46) chromosome races of the Sceloporus grammicus complex form a parapatric hybrid zone in the Mexican state of Hidalgo, characterized by steep concordant clines among three diagnostic chromosome markers across a straight-line distance of about 2 km. Here, we show that this zone is actually structured into local patches in which hybridization extends over an extremely irregular front. The distribution of hybrid-index (HI) scores across the transect reveals some hybridization at almost all localities mapped in a central 7 km × 3 km area. Pooling the central samples produces both a strong heterozygote deficit for all diagnostic markers and strong linkage disequilibria between all pairwise combinations of these (unlinked) markers. Moreover, a highly significant association exists between the habitat on which each individual was caught and its karyotype (F5 chromosomes are more likely to be found on oak). Analysis of genotype frequencies over a range of spatial scales shows that there is no significant heterozygote deficit or habitat association within local areas of less than about 200 m; however, there is significant linkage disequilibrium over the smallest scales (R = D (pquv)^1/2 = 0.29, support limits, 0.18–0.36) over 100 m. These patterns suggest that lizards mate and choose habitats randomly within local patches. This conclusion is supported by mark-recapture estimates of dispersal (≈ 80 m in a generation) and by inference of matings from embryo and maternal karyotypes. Closer examination of the two-dimensional pattern reveals a convoluted cline for all three markers, with a width of 830 m (support limits 770 m–930 m). This cline width, combined with the strength of local linkage disequilibrium, implies a dispersal rate of σ = 160 m in a generation and an effective selection pressure of 30% on each chromosome marker. The proportion of inviable embryos is greater in females from the center of the hybrid zone; this is caused by effects associated with both karyotype and location. The hybrid zone is likely to be maintained by selection against chromosomal heterozygotes, by other kinds of selection against hybrids, and by selection adapting the chromosome races to different habitats. The structure of the contact may be caused by both random drift and by selection in relation to habitat.     

繁殖于凤仙花上的缘毛果蝇种复合一新种记述(双翅目,果蝇科)(英文)

记述了采自云南昆明的毛果蝇属缘毛果蝇种复合1新种:亚平毛果蝇Hirtodrosophila yapingi sp.nov.。研究标本收藏于中国科学院昆明动物研究所标本馆、北海道大学博物馆及英国国家自然历史博物馆。亚平毛果蝇,新种Hirtodrosophila yapingi sp.nov.(图1～27)新种与报道自尼泊尔和中国云南的缘毛果蝇H.limbicostata(Okada)1966相似。两个种的区别在于腹部的斑纹和产卵瓣的形状。新种雌成虫腹部每节背板基部具浅色宽带,在第3和第4背板上被暗色带遮断;缘毛果蝇雌成虫腹部主要为暗棕色,各节背板基部中央具浅色斑,形成向腹中部下行的、中断的浅色纵带,并在靠近腹背板基部角处具小的较不明显的浅色区域。此外,新种雌成虫具明显较窄的产卵器(尤其是在端半部),产卵瓣端部不向内弯曲,而缘毛果蝇雌成虫产卵器宽得多,产卵瓣端部显著向内弯曲。正模♂,云南昆明筇竹寺,2006-08-22,户田正宪、高建军采。副模:7♂♂,6♀♀,同正模;3♂♂,3♀♀,2006-08-20,其余同正模;1♀,2007-09-16,高建军采,其余同正模。词源:新种以中科院院士、昆明动物研究所张亚平研究员之名命名,以感谢张先生对作者在果蝇分类及系统进化研究方面的支持。     

东北虎,云豹,金猫红细胞上C3b受体的测定

本文通过对健康的东北虎、云豹和金猫红细胞上C_(3b)受体和免疫复合物的检测,证实这三种动物的红细胞,除了具有免疫粘附,形成免疫复合物的花环(其花环率分别为7.00、6.50和5.67)外,还可通过其膜上的C_(3b)受体结合免疫复合物,形成C_(3b)受体的花环,其花环率分别为11.50、11.88和11.83。表明东北虎、云豹和金猫的红细胞具有较强的免疫功能。     

PHYLOGENY OF THE GENUS COLEOCHAETE (COLEOCHAETALES,CHAROPHYTA) AND RELATED TAXA INFERRED BY ANALYSIS OF THE CHLOROPLAST GENE rbcL1

The genus Coleochaete Bréb. is considered to be a key taxon in the evolution of green algae and embryophytes (land plants), but only a few of the approximately 15 species have been studied with molecular phylogenetic methods. We report here the sequences of the gene rbcL from six new cultures of Coleochaete and two of Chaetosphaeridium Klebahn. These sequences were combined with 32 additional sequences, and phylogenetic analyses were performed with maximum likelihood, distance optimality, and parsimony methods. Important subgroups within Coleochaete include two primary lineages, one marked by fully corticated zygotes and the other by naked or weakly corticated zygotes. In the first lineage there is a subclade with tightly joined filaments and distinctive (“T‐shaped”) cell division, an assemblage of strains that resembles the endophytic species Coleochaete nitellarum Jost, and a clade with loosely joined filaments and “Y‐shaped” cell divisions. Consistent with recent multigene phylogenies, these analyses support the monophyly of the Coleochaetales, place the Charales as the sister taxon to land plants, and indicate that Chaetosphaeridium is far more closely related to Coleochaete than to Mesostigma Lauterborn.