Molecular evolution of 5S rDNA region in Vigna subgenus Ceratotropis and its phylogenetic implications

The evolution of 5S rRNA gene unit (5S gene unit) was studied among the ten species belonging to Vigna subgenus Ceratotropis by sequencing and analyzing the intra- and inter-specific sequence heterogeneity. The 5S unit from these species ranged from 214 to 342 bp in length as a result of several indels in the intergenic spacer (IGS) region. A large deletion (>100 bp) was found specifically in the IGS of V. radiata accessions. IGS showed high sequence variation with more than 50% polymorphic and 35.4% parsimony informative sites. However, the coding region (5S gene) was highly conserved, both in length and in sequence. Intra-genomic and intra-specific divergence was observed among some species, which indicated that the 5S unit is evolving at different rates among the Vigna species. Most Vigna species harbored one type of 5S unit indicating complete homogenization among them. Vigna glabrescens, a tetraploid species, also showed single type of 5S rDNA from only one of the diploid progenitor indicating loss or homogenization of the other type. However, V. nakashimae and V. riukiuensis harbored multiple, diverse, ‘intra-genomic 5S types’ indicating that 5S rDNA is not completely homogenized by concerted evolution and is still evolving. In general, the phylogeny based on IGS sequences was in agreement with many of the earlier reports except some surprising observations such as, V. glabrescens clustered with V. mungo in section Ceratotropis and unlike most of the species, wild and cultivated types of V. umbellata were present in different subclusters. Presence of divergent 5S sequences in V. nakashimae and V. riukiuensis caused errors in phylogeny reconstruction at species level and suggested a horizontal ‘gene transfer’ as a result of inter-species hybridization. The comparative analysis showed that 5S IGS sequences have better phylogenetic utility than chloroplast DNA sequences, such as atpB-rbcL and is comparable to ITS1 and ITS2 in this respect.     

Non-concerted ITS evolution, early origin and phylogenetic utility of ITS pseudogenes in Pyrus

Molecular studies of 19 species of the genus Pyrus L. revealed different degrees of intra-individual polymorphism of the internal transcribed spacer (ITS 1, 5.8S rDNA and ITS 2) region due to the existence of putative non-functional copies (pseudogenes), putative recombinants and non-concerted evolution among functional copies. Different types of ITS pseudogenes displaying lower GC content and unstable secondary structure were preferentially amplified under normal PCR conditions. Functional ITS copies were successfully obtained in all investigated accessions under the modified PCR conditions. All pseudogenes were highly divergent from their corresponding functional copies and formed a monophyletic group in the phylogenetic tree based on all paralogs, indicating they were of relatively early origin. Functional ITS copies led to confused and poorly resolved phylogeny as a result of low sequence divergence, existence of unidentified ancient recombinants and a high degree of intra-individual functional ITS polymorphism, while certain types of pseudogenes and some relict pseudogenes offered more credible clues for the evolutionary history of Pyrus species.     

Early evolution in the Pinaceae

Nineteen species of structurally preserved ovulate cones of the Pinaceae are known from the Cretaceous. One of these belongs to Pinus, while the others contain anatomical features necessitating their classification in the organ genera Pityostrobus and Pseudoaraucaria. The six species of the latter group are very similar to one another and probably represent a natural, thought extinct, genus. By contrast, there is so much variety in the twelve Cretaceous species of Pityostrobus that when they are considered with respect to the uniformity of cone structure within each of the modern genera, each of the species of this organ genus may well reperesent an natural genus by itself. All expect one of these fossil forms contain features that are today characteristic of Pinus. This, combined with the Early Cretaceous occurrence of a structurally preserved Pinus cone, suggests that Pinus or something very close to it represents the phylogenetic centrum of the ancestral complex. Lack of cones showing distinct affinity with other modern genera supports this idea and further suggests that, while Pinus was in existence in the Early Cretaceous, other Recent genera of the Pinaceae may not have diverged from the complex until the Late Cretaceous or Early Tertiary.     

栽培种甘薯及其近缘野生种nrDNA ITS序列分析

采用核糖体DNA内转录间隔区(nrDNA ITS)序列比较分析了甘薯及其近缘野生种的遗传多样性及系统进化关系,首次报道了栽培种甘薯‘徐薯18’(Ipomoea batatas‘Xushu18’)及其近缘野生种I.triloba(DOM),I.cordatotriloba(MEX),I.nil(PER),I.nil(JPN),I.hederacea Jacq.(USA),I.hederacea Jacq.(HK)和种间杂交种67-1(I.batatas‘Xushu18’×I.hederacea Jacq.)及回交种(67-1×I.batatas‘Xushu18’)的nrDNA ITS序列。序列分析表明,栽培种甘薯及其近缘野生种nrDNA ITS序列长度为570~600bp。其中,ITS1序列为185~209 bp,GC含量为53.11%~61.83%;ITS2序列为214~226 bp,GC含量为61.21%~72.89%;5.8S序列均为165 bp,GC含量为54.55%~55.76%。此外,栽培种甘薯及其近缘野生种ITS序列信息位点均集中在ITS1和ITS2区;与其他甘薯属植物相比,I.wrightii ITS2的末端缺失了6~8个碱基。系统进化分析表明,栽培种甘薯‘徐薯18’(I.batatas‘Xushu18’)和野生种I.triloba、I.cordatotriloba、I.lacunosa、I.trifida的亲缘关系较近,与I.wrightii、I.pes-tigridis、I.grandifolia、I.nil、I.hederacea Jacq.、I.purpurea的亲缘关系较远;杂交后代与栽培种甘薯‘徐薯18’(I.batatas‘Xushu18’)亲缘关系较近,与野生种父本I.hederacea Jacq.的亲缘关系较远。     

A phylogenetic study ofSaxifraga sect.Saxifraga (Saxifragaceae) based on nrDNA ITS sequences

Parsimony analyses of 54 nrDNA ITS (Internal Transcribed Spacer) sequences ofSaxifraga sect.Saxifraga were performed. In addition to some unresolved clades, there is strong disagreement between the ITS phylogeny and previous classifications based primarily on morphology. The extensive cytological instability of sect.Saxifraga prevents previous cytotaxonomical results from resolving the incongruence between molecular and morphological data. Dissimilar topologies between chloroplast (matK) and nuclear (ITS) trees for eight species of sect.Saxifraga suggest that gene trees and the true species tree are not coincident. Recent and mid-term reticulation is proposed as an explanation for the incongruence between morphological, cytological, organellar, and nuclear data. Homogenization in multigene families, such as the ITS region, via concerted evolution may be the key to the interpretation of results based on ITS sequences within sect.Saxifraga. The use of organellar genes in a larger sample should help to determine whether extensive reticulation occurs in sect.Saxifraga, as has been documented in various genera of Saxifragaceae.     

Cell wall biochemistry in Archaea and its phylogenetic implications

Cellularization of a precellular ancestral condition of life must have occured separately for each of the three domains of life resulting in a protoarchaeon, a protobacterium and a protocaryon. Accordingly, a first cell, ancestral to the three domains of life has never existed. This statement is based on a retrospective evaluation of the distribution patterns of membrane lipids and of cell wall polymers with their different chemical nature and modes of biosynthesis in a global phylogenetic tree based on 16S rRNA sequence comparison.Dedicated to Professor Peter Sitte on the occasion of his 65th birthday.     

The evolution of SMC proteins: phylogenetic analysis and structural implications

The SMC proteins are found in nearly all living organisms examined, where they play crucial roles in mitotic chromosome dynamics, regulation of gene expression, and DNA repair. We have explored the phylogenetic relationships of SMC proteins from prokaryotes and eukaryotes, as well as their relationship to similar ABC ATPases, using maximum-likelihood analyses. We have also investigated the coevolution of different domains of eukaryotic SMC proteins and attempted to account for the evolutionary patterns we have observed in terms of available structural data. Based on our analyses, we propose that each of the six eukaryotic SMC subfamilies originated through a series of ancient gene duplication events, with the condensins evolving more rapidly than the cohesins. In addition, we show that the SMC5 and SMC6 subfamily members have evolved comparatively rapidly and suggest that these proteins may perform redundant functions in higher eukaryotes. Finally, we propose a possible structure for the SMC5/SMC6 heterodimer based on patterns of coevolution.     

The grass megagametophyte and its possible phylogenetic implications

The structure of the grasses megagametophyte is considered to be characteristic enough as to deserve a particular place in the megagametophyte typology. Furthermore, it is compared with those of other Monocotyledonous families to point out embryological affinities.Both are members of the Carrera del Investigador (Conicet, Argentina).     

Structural divergence among genomes of closely related baculoviruses and its implications for baculovirus evolution

Baculoviruses are members of a large, well-characterized family of dsDNA viruses that have been identified from insects of the orders Lepidoptera, Hymenoptera, and Diptera. Baculovirus genomes from different virus species generally exhibit a considerable degree of structural diversity. However, some sequenced baculovirus genomes from closely related viruses are structurally very similar and share overall nucleotide sequence identities in excess of 95%. This review focuses on the comparative analysis of partial and complete nucleotide sequences from two groups of closely related baculoviruses with broad host ranges: (a) group I multiple nucleopolyhedroviruses (MNPVs) from a cluster including Autographa californica (Ac)MNPV, Rachiplusia ou (Ro)MNPV, and Plutella xylostella (Plxy)MNPV; and (b) granuloviruses (GVs) from a cluster including Xestia c-nigrum (Xecn)GV and Helicoverpa armigera (Hear)GV. Even though the individual viruses in these clusters share high nucleotide sequence identities, a significant degree of genomic rearrangement (in the form of insertions, deletions, and homologous recombination resulting in allelic replacement) is evident from alignments of their genomes. These observations suggest an important role for recombination in the early evolution and biological characteristics of baculoviruses of these two groups.     

ITS regions in diploids of Aegilops (Poaceae) and their phylogenetic implications

The nucleotide sequences of the internal transcribed spacer (ITS) of nuclear ribosomal DNA in nine diploid species representing six sections of Aegilops were determined by direct sequencing of PCR-amplified DNA fragments. These sequences were aligned with two ITS sequences of additional species from Genbank. Sequence divergences were estimated using Kimura two-parameter model, and the phylogenetic analyses were performed using the maximum parsimony (MP) and the neighbor-joining (NJ) methods with PAUP and PHYLIP, respectively. The sequence divergences between the diploid species varied from 0.5% to 4.68%. The resulting MP tree and NJ tree showed relatively congruent phylogenetic relationships among these species, except Ae. caudata. Particularly, Ae. speltoides was basal within the two trees. The paraphyletic relationships between Ae. speltoides and two species of Sect. Sitopsis, and between Ae. uniaristata and two species of Sect. Comopyrum were supported strongly. The ITS data suggest that currently recognized sections within Aegilops should be reconsidered.     

粉拟青霉种内nrDNA ITS分析

通过对20株粉拟青霉(Paecilomyces farinosus)ITS1-5．8S-ITS2(rDNA)区域序列测定,确定了粉拟青霉ITS序列,而韩国学者测定的粉拟青霉ITS序列应为细脚拟青霉(P．tenuipes)的序列。序列比较发现,韩国2株未定名的拟青霉(Paecilomyces spp．)菌株(KACC40219、KACC40221)应为粉拟青霉。基于本研究构建的邻接树推断,粉拟青霉的有性型可能是一种虫草。粉拟青霉的起源应为单源的。不同的粉拟青霉菌株的ITS序列具有多态性,源于同一地区的菌株的ITS变异也较大。ITS序列的证据表明,粉拟青霉菌株间的差异与地理来源及寄主均无相关性。     

Allozyme differentiation in the Mediterranean firs (Abies, Pinaceae). A first comparative study with phylogenetic implications

Nineteen natural Mediterranean fir populations, belonging to eight species and to one natural hybrid (A. ×borisii-regis), were investigated by starch and polyacrylamide gel electrophoresis. A total of 31 alleles was scored at eight loci (IDH-B, ACP-A, PER-B, 6PGD-A, 6PGD-B, MNR-B, PGI-B, PGM-A. Great variation was observed in the heterozygosity among the population studied and ranged from 0.010 (A. pinsapo) to 0.328 (A. cephalonica). The interpopulation genetic diversity was about 26% of the total genetic diversity. From the dendrogram, new phylogenetic relationships were revealed. High affinity was observed between the Calabrian fir population and the one from north-west Greece as well as betweenA. equi-trojani grown in Asia Minor and the southern Greek populations. Species specific alleles were found inA. cilicica. From the findings of the present work, a new hypothesis concerning the taxonomy, distribution and evolution ofAbies species in the Balkan Peninsula is supported.     

Pollen morphology and its phylogenetic implications in the genus Picea

Pollen grains from 20 spruce species were examined using scanning electron microscopy, to study the phylogeny of pollen morphology in spruce. Morphological characteristics of the pollen were examined among different species based on statistical data and photographic observation. The mean pollen size of 20 spruce species ranged from 84.327 to 118.793.18 μm. Based on morphological photography, two zygomorphic bladders were distributed on the underside of the corpus and were found at the edge of the cap near the corpus surface. The bladder’s surface texture was smooth and had no distinct variations among species. Germinal furrows formed clear striates between the two bladders. According to clustering analysis, 20 spruce species were classified into three sections, namely, Castict, Schrenkiana and Picea, and the Picea section was classified into two subsections, Jezonensis and Picea. Corpus veins experienced notable differentiation among sections, and their evolutionary trend progressed through sections Castict, Schrenkiana and Picea. Within subsection Picea, parallel evolution occurred between the two subsections.     

中药蓼大青叶及其伪品的nrDNA ITS区序列的测定

中药蓼大青叶的基源为蓼科蓼属植物蓼蓝（Polygonum tinctorium Ait),同属植物的水蓼（辣蓼）,（P.hydropiperL.)的叶常作其伪品,本文采用PCR直接测序法,首次测定了它们的核糖体DNA ITS区序列,结果显示,两者充列长度（ITS1,5,8S,ITS2）均为646bp,序列间共有28个变异位点,本研究为产蓝植物的DNA分子鉴定提供了必要的序列资料。     

Nucleomorph genome diversity and its phylogenetic implications in cryptomonad algae

The relationship between phylogeny and nucleomorph genome size was examined in 16 strains of cryptomonad algae using pulsed‐field gel electrophoresis, Southern hybridization and phylogenetic analyses. Our results suggest that all cryptomonads examined in this study contain three nucleomorph chromosomes and their total genome size ranges from 495 to 750 kb. In addition, we estimated the plastid genome size of the respective organisms. The plastid genomes of photosynthetic strains were approximately 120–160 kb in size, whereas the non‐photosynthetic Cryptomonas paramecium NIES715 possesses a genome of approximately 70 kb. Phylogenetic analysis of the nuclear small subunit ribosomal DNA (SSU rDNA) gene showed that nucleomorph genome size varies considerably within closely related strains. This result indicates that the reduction of nucleomorph genomes is a rapid phenomenon that occurred multiple times independently during cryptomonad evolution. The nucleomorph genome sizes of Cryptomonas rostratiformis NIES277 appeared to be approximately 495 kb. This is smaller than that of Guillardia theta CCMP327, which until now was thought to have the smallest known nucleomorph genome size among photosynthetic cryptomonads.     

Analysis of Acorus calamus chloroplast genome and its phylogenetic implications

Determining the phylogenetic relationships among the major lines of angiosperms is a long-standing problem, yet the uncertainty as to the phylogenetic affinity of these lines persists. While a number of studies have suggested that the ANITA (Amborella-Nymphaeales-Illiciales-Trimeniales-Aristolochiales) grade is basal within angiosperms, studies of complete chloroplast genome sequences also suggested an alternative tree, wherein the line leading to the grasses branches first among the angiosperms. To improve taxon sampling in the existing chloroplast genome data, we sequenced the chloroplast genome of the monocot Acorus calamus. We generated a concatenated alignment (89,436 positions for 15 taxa), encompassing almost all sequences usable for phylogeny reconstruction within spermatophytes. The data still contain support for both the ANITA-basal and grasses-basal hypotheses. Using simulations we can show that were the ANITA-basal hypothesis true, parsimony (and distance-based methods with many models) would be expected to fail to recover it. The self-evident explanation for this failure appears to be a long-branch attraction (LBA) between the clade of grasses and the out-group. However, this LBA cannot explain the discrepancies observed between tree topology recovered using the maximum likelihood (ML) method and the topologies recovered using the parsimony and distance-based methods when grasses are deleted. Furthermore, the fact that neither maximum parsimony nor distance methods consistently recover the ML tree, when according to the simulations they would be expected to, when the out-group (Pinus) is deleted, suggests that either the generating tree is not correct or the best symmetric model is misspecified (or both). We demonstrate that the tree recovered under ML is extremely sensitive to model specification and that the best symmetric model is misspecified. Hence, we remain agnostic regarding phylogenetic relationships among basal angiosperm lineages.     

Bias in phylogenetic tree reconciliation methods: implications for vertebrate genome evolution

Background

Comparative genomic studies are revealing frequent gains and losses of whole genes via duplication and pseudogenization. One commonly used method for inferring the number and timing of gene gains and losses reconciles the gene tree for each gene family with the species tree of the taxa considered. Recent studies using this approach have found a large number of ancient duplications and recent losses among vertebrate genomes.

Results

I show that tree reconciliation methods are biased when the inferred gene tree is not correct. This bias places duplicates towards the root of the tree and losses towards the tips of the tree. I demonstrate that this bias is present when tree reconciliation is conducted on both multiple mammal and Drosophila genomes, and that lower bootstrap cut-off values on gene trees lead to more extreme bias. I also suggest a method for dealing with reconciliation bias, although this method only corrects for the number of gene gains on some branches of the species tree.

Conclusion

Based on the results presented, it is likely that most tree reconciliation analyses show biases, unless the gene trees used are exceptionally well-resolved and well-supported. These results cast doubt upon previous conclusions that vertebrate genome history has been marked by many ancient duplications and many recent gene losses.     

核糖体DNA ITS区序列在植物分子系统学研究中的价值

本文就近年来国内外有关被子植物核糖体ＤＮＡ中的内转录间隔区（ＩＴＳ）序列在植物属内,近缘属间乃至科内系统发育研究中的应用,结合作者在中国姜科山姜属（Ａｌｐｉｎｉａ Ｒｏｘｂ．）上的研究,对ＩＴＳ区序列在植物分子系统学研究中的价值作一简要综述,对其应用前景也进行了讨论。