DNA CONTENT AND EVOLUTION IN THE MICROSERIDINAE

Relative 2C nuclear DNA contents were microphotometrically determined from nuclei isolated from eight species of Microseris, four species of Agoseris, and Phalacroseris Bolanderi. The thirteen species are diploid (2n = 18) western North American members of the subtribe Microseridinae, tribe Cichorieae, of the family Compositae. A 7.7-fold variation in DNA content was detected. Phalacroseris has the highest DNA content and Agoseris heterophylla has the lowest. Within the genera Microseris and Agoseris, a 2.8- and 3.1-fold range in DNA content was detected. The higher values were from perennial species, and the lower values were from annual inbreeding species. Both evolutionary increases and decreases in nuclear DNA content have apparently occurred during the differentiation of the subtribe.     

Nuclear DNA Amounts in Palms (Arecaceae)

Nuclear DNA amounts are reported for 83 species and 53 genera of palms, covering all of the six subfamilies. 4C DNA contents range between 3.89 and 55.62 pg in diploids, showing an approximate 14.3-fold variation in genome size. Polyploids have DNA contents of up to 156.40 pg/4c which demonstrates a 40.2-fold variation. Diploids with high DNA contents occur in three subfamilies of palms (Coryphoideae, Calamoideae, Arecoideae), and seem to be further restricted to particular tribes or subtribes (Thrinacinae, Borasseae, Lepidocaryeae, Caryoteae, some subtribes of Areceae). Palms from the subfamilies Nypoideae and Phytelephantoideae have the lowest DNA amounts, followed by the Phoeniceae and the Corypheae: Livistoninae from the subfamily Coryphoideae. Although DNA amounts in some genera and subtribes are usually constant, e.g., in Phoenix, Phytelephas, the Livistoninae, Dypsidinae, diploid Butiinae), considerable variation occurs at the diploid level in some large and apparently actively evolving genera such as Chamaedorea, Pinanga, Cenoma and possibly Bactris. Formaldehyde fixation is recommended for palms, as conventional ethanol-acetic acid fixation has proved to be unsuitable for DNA estimation of Feulgen-stained nuclei by microdensitometry, since it can lead to errors up to 2.5-fold in extent. Chromosome counts are reported for 72 of the species studied, of which 42 are new.     

Conifer genome sizes of 172 species,covering 64 of 67 genera,range from 8 to 72 picogram

Nuclear genome size of conifers as measured by flow cytometry with propidium iodide was investigated, striving to collect at least a single species from each genus. 64 out of 67 genera and 172 species were measured. Of the 67 genera, 21 are reported here for the first time and the same is true for 76 species. This nearly doubles the number of measured genera and adds 50% to the number of analyzed species. Conifers have chromosome numbers in the range of n = (7)10–12(19). However, the nuclear DNA content (2C‐value) is shown here to range from 8.3 to 71.6 picogram. The largest genome contains roughly 6 × 10¹⁰ more base pairs than the smallest genome. Genome sizes are evaluated and compared with available taxonomic treatments. For the mainly (sub)tropical Podocarpaceae small genome sizes were found with a 2C‐value of only 8–28 pg, with 13.5 pg on average. For the Taxaceae 2C‐values from 23–60 pg were determined. Not surprisingly, the genus Pinus with 97 species (39 species measured here) has a broad range with 2C = 38–72 pg. A factor of 2 difference is also found in the Cupressaceae (136 species) with nuclear DNA contents in the range 18–35 pg. Apart from the allohexaploid Sequoia, ploidy plays a role only in Juniperus and some new polyploids are found. The data on genome size support conclusions on phylogenetic relationships obtained by DNA sequencing. Flow cytometry is applicable even to young plants or seeds for the monitoring of trade in endangered species.     

Genome Size Variation in Some Tropical Hardwoods

4-C DNA values of 36 tropical hardwood species belonging to 13 families show a range from 2.08 to 20.07 pg, a 9.64-fold difference. DNA per chromosome values show a 10-fold difference. The minimum range of variation (1.2-fold) is shown by Annonaceae and Sapindaceae while the maximum range (6.23-fold) is shown by Euphorbiaceae, and Fabaceae and Rutaceae show 1.7 and 1.9-fold differences, respectively. On an average, Euphorbiaceae and Rutaceae have been found to possess the largest and the smallest genome sizes respectively.     

Nuclear DNA amounts in pacific Crustacea

Nuclear DNA amounts have been determined for 42 species of crustaceans bringing the total number of species with known nuclear DNA content to over 70. Genome size in Crustacea varies over a 25-fold range with a modal value of 2 to 3 pg haploid being common in many groups. Both average genome size and the amount of variability among species are characteristic for certain groups. A trend towards small genomes is evident in advanced and specialized crustacean groups. Somatic polyploidy is a very pronounced feature of the Crustacea. The data suggest that evolution by polyploidy may be more common in crustaceans than earlier data had indicated. These features and the presence of very characteristic satellite fractions in the nuclear DNA recommend the Crustacea for further studies in evolutionary genetics.     

Molecular evidence for the phylogenetic position ofHanabusaya asiatica Nakai (Campanulaceae), an endemic species in Korea

The phylogenetic relationship of the Korean endemic genus,Hanabusaya, to other campanulaceous genera has been controversial since it was described by Nakai in 1911. Three genera of Campanuloideae,Symphyandra, Adenophora, andCampanula, have been considered closely related by various taxonomists on the basis of anther shape, gross morphology, and pollen characters, respectively. We have tested these competing taxonomic hypotheses using the internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA from 12 taxa representing 7 genera of Campanulaceae. The molecular phylogeny indicates strongly thatHanabusaya is more closely allied toAdenophora than toCampanula orSymphyandra. The phylogenetic affinity ofHanabusaya andAdenophora is supported by a 100% bootstrap value and a high decay index (13). The average sequence divergence value (Kimura’s 2-parameter method) betweenHanabusaya and theAdenophora species is 2.58. The value is significantly (about ten times) lower than the ones observed betweenHanabusaya and the species ofCampanula (average of 23.52) and betweenHanabusaya andSymphyandra (24.95). The ITS sequence phylogeny suggests that some morphological characters, such as fused anthers and corolla shape, are homoplastic in the Campanulaceae genera.     

核酸碱基组分与毛霉目的分类

本文对毛霉目中6科16属40个种共60株真菌DNA的G+C含量与分布作了系统的研究。在提取DNA时选用了Storck等人在Marmur提取细菌DNA方法基础上发展起来的真菌DNA提取法,经过一些修改后成功地提取了毛霉目真菌的DNA。经该法提取的DNA片段长、纯度高,在热变性时DNA的增色效应一般都大于35%。各个种的GC含量大致有一固定值。根据所测毛霉目16属各属真菌的平均GC含量,可将它们分为明显的三组:Gongronella, Haplosporangium, Mortierella及Syncephalastrum为一组,GC含量最高为46.0-49.8%; Cunninghamella单独成一组,GC含量最低只有28.8%;其他各属包括Absidia, Mucor, Rhizopus等GC含量介乎这两组之间,分布于34.9-41.9%。这一次序除Helicostylum与Circinella的数值低于他人的报道外,其余和文献报道值一致。一般属内GC含量变化小于10%,种内变化小于2%。所测得的G+C mol%对分Syncephalastrum monosporum Zheng et al.和Mortierella ramanniana (Moeller) Linneman的合理归属提供了佐证。对某些所测结果与文献报道值有出入的原因作了讨论和分析。对采用Mandel等1970年建议的公式GC=(Tm 0.1×SSC/50.2)-0.990来计算GC含量的依据也作了讨论。     

Quantitative karyology of some species ofLuzula

The dimensions of metaphase chromosomes and nuclear DNA contents were measured in eight species ofLuzula. The 2 C DNA contents ranged from 8.51 pg inL. purpurea to 0.55 pg inL. pilosa. Total chromosome volume shows a linear relationship with DNA content; however, the total chromosome length of the complement of the different species is approximately constant. Nucleolar volume and the number of chromocentres in the different species also show a relationship with DNA content. Taken together, these data suggest that while chromosome fragmentation could have generated the present-day range of chromosome numbers in the genus, there have also been changes in the total quantity of DNA with the result that species with similar chromosome numbers have different DNA contents. The relationships of DNA content with chromosome volume inLuzula and other genera are compared and the differences discussed.     

DNA content in species of Vernonia and Vernonanthura from South America: An approach to systematics and evolution of the Vernonieae (Asteraceae)

There are relatively few studies of DNA content in the Vernonieae (Asteraceae) tribe. The first studies were realized in the Lessingianthus genus and determined the DNA content of 25 species. After DNA content, ploidy level and the total karyotype were compared in 6 Chrysolaena species. The aim of this study was to present, for the first time, the DNA content values of Vernonanthura and Vernonia and to thereby expand knowledge of the Vernonieae tribe. A total of 19 natural populations belonging to the genera Vernonanthura and Vernonia were studied for the first time. The results were compared with other Vernonieae genera and with other Asteraceae tribes. Our results found that Vernonieae have the smallest range of 1C value variation in Asteraceae. Furthermore, there were differences in the DNA content of Vernonia and Vernonanthura. These results show that low DNA content and herbaceous habit in Vernonia are characters derived from the higher DNA content and woody habit present in Vernonanthura. These results could indicate a hybrid origin of one species and allow the determination of both the ploidy and chromosome number of other taxa. The results observed in Vernonanthura species showed a highly significant correlation between 1C-value and latitude.     

Genome size and polyploidy variation in the tropical hardwood genusTerminalia (Combretaceae)

Chromosome complements and 2C DNA amounts of six species ofTerminalia have been studied.Terminalia oliveri, T. myriocarpa andT. arjuna are diploid (2n = 24),T. chebula andT. bellirica are tetraploid (2n = 48),T. muelleri shows a triploid number (2n = 36). Two well demarcated groups of species are recognizable on the basis of chromosome length and 2C DNA values which range from 3.60 pg (T. oliveri) to 12.80 pg (T. bellirica) showing a 3.5-fold difference. Differences of DNA per basic genome or per chromosome are greatest (1.97-fold) betweenT. oliveri andT. arjuna. Two species groups (1)T. oliveri andT. chebula, and (2)T. myriocarpa, T. arjuna, T. muelleri, T. bellirica, therefore are well differentiated by DNA per basic genome, irrespective of polyploidy. The mean values of the two groups are 1.81 pg and 3.34 pg, respectively, showing a 1.84-fold difference. Within diploids and tetraploids there is 1.97-fold and 1.76-fold variation, respectively.     

Accelerated molecular evolution in Microtus (Rodentia) as assessed via complete mitochondrial genome sequences

Microtus is one of the most taxonomically diverse mammalian genera, including over 60 extant species. These rodents have evolved rapidly, as the genus originated less than 2 million years ago. If these numbers are taken at face value, then an average of 30 microtine speciation events have occurred every million years. One explanation for the rapid rate of cladogenesis in Microtus could be the karyotypic differentiation exhibited across the genus: diploid numbers range from 17 to 64. Despite the striking chromosomal variability within Microtus, phenotypic variation is unremarkable. To determine whether nucleotide substitution rates are also elevated in voles, we sequenced the entire mitochondrial DNA (mtDNA) genome of the Eurasian sibling vole (Microtus rossiaemeridionalis). We compared this genome to another previously sequenced vole mtDNA genome (Microtus kikuchii) and performed pairwise sequence comparisons with the mtDNA genomes of ten additional mammalian genera. We found that microtine mtDNA genomes are evolving more rapidly than any other mammalian lineage we sampled, as gauged by the rate of nucleotide substitution across the entire mtDNA genome as well as at each individual protein-coding gene. Additionally, we compared substitution rates within the cytochrome b gene to seven other rodent genera and found that Microtus mtDNA is evolving fastest. The root cause of accelerated evolution in Microtus remains uncertain, but merits further investigation. The mitochondrial genome sequence from this article has been deposited with the GenBank database under accession number DQ015676.     

TrnL-trnF sequence data imply paraphyly of Aquilaria and Gyrinops (Thymelaeaceae) and provide new perspectives for agarwood identification

The genera Aquilaria and Gyrinops (Thymelaeaceae, Malvales) are well known for the production of agarwood which is a highly wanted forest product of substantial economic value. The taxonomic status of Aquilaria and Gyrinops as separate genera is doubted as they are only distinguished by the number of stamens. We investigated their status by conducting phylogenetic analyses of DNA sequences from the plastid trnL-trnF spacer. Control of international trade of agarwood is currently hampered by the failure of traditional methods such as microscopy to identify samples to species level. We therefore evaluated the potential of molecular identification of agarwood by searching for species- and region-specific plastid DNA polymorphisms. DNA sequences were obtained from 31 Thymelaeaceae accessions encompassing 20 different species in six genera. Aquilaria and Gyrinops appear to be paraphyletic. Success in sequencing wood samples demonstrates that molecular markers provide new perspectives for agarwood identification.     

Nuclear DNA content of Vitis species,cultivars, and other genera of the Vitaceae

The nuclear DNA content was analyzed in Vitis species, hybrid cultivars, and genera of the Vitaceae using flow cytometry. Significant variation was found among Vitis species, hybrids, and other genera of the Vitaceae (Ampelopsis and Parthenocissus). DNA content was estimated to range from 0.98 to 1.05 pg/2C within V. labrusca (ns) and 0.86 to 1.00 pg/2C within V. vinifera (ns). Genotypes from Vitis and Parthenocissus were similar in nuclear DNA content (approximately 1.00 pg/2C) whereas they differed significantly from Ampelopsis (1.39 pg/2C). No correlation between DNA content and the center of origin of genotypes of the Vitaceae was noted. Based on the present study, the Vitis genome size is 475 Mbp, 96% of which is non-coding. Knowledge of DNA content is useful in order to understand the complexity of the Vitis genome and to establish a relationship between the genetic and physical map for map-based cloning.     

Cytophotometric studies on the DNA content in diploid and polyploid enchytraeidae (Oligochaeta)

The amount of DNA in nerve nuclei was determined by Feulgen cytophotometry in 45 species and cytotypes. The DNA value of spermatids, spermatogonial nuclei and nerve nuclei in newly hatched worms was determined in some species. The total variation in DNA content ranges from a relative value of 0.40 to 4.85. The DNA content in diploid species ranges from 0.40 to 1.12. In two genera (Lumbricillus and Enchytraeus) wide diversification in chromosome numbers at the diploid level is accompanied by wide variation in DNA values, whereas other genera and species groups (Mesenchytraeus, Henlea and Marionina), which are conservative in this respect, are closer in DNA values. In the genus Fridericia constancy in chromosome number is associated with wide variation in DNA values, this may be due to an increase in DNA taking place in old individuals. In 9 out of 10 cases nearly identical ratios are found between chromosome numbers and DNA values in polyploids and related diploids or lower polyploids. Differential polyteny is recorded in two cases.Dedicated to Professor Dr. J. Seiler on the occasion of his eightieth birthday.     

Morphological and molecular identification of Conilepia nigricosta,new to Korea,and Lithosia quadra with their phylogenetic placements within tribe lithosiina (Lepidoptera: Erebidae: Arctiinae)

Conilepia nigricosta (Leech 1888) is very similar to Lithosia quadra in the superficial appearance of females. We re‐examined their morphologies and analyzed their DNA barcode regions to assess their taxonomic status and phylogenetic relationships with other related genera. Our morphological study confirmed the diagnostic value of the wing venation and the genital structures in both sexes. Our analyses with DNA barcodes showed sufficient genetic divergences ranging from 6.21% to 7.16% between those two species. The phylogenetic relationship using the barcoding region for the members belonging to lithosiina showed that Conilepia and Lithosia are suggested as having strong monophyly compared to other genera.     

Congruence between molecular phylogeny and cuticular design in Echiniscoidea (Tardigrada,Heterotardigrada)

Although morphological characters distinguishing echiniscid genera and species are well understood, the phylogenetic relationships of these taxa are not well established. We thus investigated the phylogeny of Echiniscidae, assessed the monophyly of Echiniscus, and explored the value of cuticular ornamentation as a phylogenetic character within Echiniscus. To do this, DNA was extracted from single individuals for multiple Echiniscus species, and 18S and 28S rRNA gene fragments were sequenced. Each specimen was photographed, and published in an open database prior to DNA extraction, to make morphological evidence available for future inquiries. An updated phylogeny of the class Heterotardigrada is provided, and conflict between the obtained molecular trees and the distribution of dorsal plates among echiniscid genera is highlighted. The monophyly of Echiniscus was corroborated by the data, with the recent genus Diploechiniscus inferred as its sister group, and Testechiniscus as the sister group of this assemblage. Three groups that closely correspond to specific types of cuticular design in Echiniscus have been found with a parsimony network constructed with 18S rRNA data. © 2013 The Linnean Society of London     

FEULGEN MICROSPECTROPHOTOMETRIC STUDIES OF PANDORINA MORUM AND OTHER VOLVOCALES (CHLOROPHYCEAE)12

The nuclear DNA content during normal vegetative growth and division has been examined in three species of Volvocales, Chlamydomonas reinhardtii Dangeard, Pandorina morum Bory, and Volvox carteri f. nagariensis Iyengar. The results are consistent with the nuclear cycle reported in the literature for Eudorina. Nuclear DNA content does not increase during the prolonged cell growth phase. At the time of colony formation, nuclear DNA doubles, the nucleus divides, and this alternation continues until the final 2ⁿ complement of progeny nuclei is formed. The 4- and 8-nucleate stages of dividing gonidia of V. carteri have a nuclear DNA content in the same range as the somatic cells; they are not polyploid or polytene. Four normal clones of Pandorina, having 2, 5 or 12 chromosomes, all had similar amounts of DNA per nucleus, suggesting that the species has a nuclear genome of fairly constant size rather than consisting of many strains representing a polyploid series. One unique clone, a hybrid with double the chromosome number of either its parents, had twice as much DNA as the normal clones. The Feulgen spectrophotometric method is sufficiently sensitive to detect 2-fold differences in DNA content at the level of 2 × 10^?13 g of DNA /nucleus, and its use avoids the complications associated with the presence of organelle DNA.     

Genome size in Bulgarian Centaurea s.l. (Asteraceae)

Thirty-nine species and subspecies of the genera Centaurea, Colymbada, Psephellus and Cyanus (all included in Centaurea s.l.) including many rare and endemic taxa of preponderantly Bulgarian distribution have been investigated with Feulgen DNA image densitometry for holoploid and monoploid genome size (C- and Cx-values). Cyanus varies gradually 2.17-fold between 0.74 pg and 1.56 pg (1Cx). In the remaining taxa two major genome size groups are found, which differ about 1.8-fold in Cx-value. Low values occur in Centaurea subgenera Acrolophus, Solstitiaria, Phalolepis (0.77 pg to 0.90 pg, 1Cx) and Jacea (0.95 pg to 1.09 pg, 1Cx), high values in the genera Colymbada (1.65 pg to 1.93 pg, 1Cx) and Psephellus (1.79 pg, 1Cx, in P. marschallianus). Cx-values support a distinction of Colymbada from Centaurea. Genome size variation is discussed with regard to phylogeny, life form (annual versus perennial), polyploidy, chromosome basic numbers, altitude of occurrence and climate, endemism, and rarity.     

人为干扰对碧峰峡栲树次生林群落物种多样性及其优势种群生态位的影响

为进一步了解人为干扰对碧峰峡生态旅游区的生态环境及不同种群利用资源、占据生态空间的能力的影响,并为该区植物资源、大熊猫栖息地的保护及旅游可持续措施的制定提供依据,调查了人为干扰下碧峰峡栲树(Castanopsis fargesii)次生林物种多样性及其优势种群生态位的组成情况。结果表明:7200m2的样方中共记录到156个植物种,隶属于70科114属,其中乔木16科23属33种,灌木39科67属95种,草本38科42属47种;乔木层总体物种多样性水平在中度干扰下最低,在轻度及重度干扰下差异不大。灌木层总体物种多样性水平在中度干扰下最高,在轻度干扰下最低。而草本层物种多样性随着干扰强度的加深有明显降低趋势;随着人为干扰强度的增加,乔木层中优势种栲树的生态位宽度均为最大,与其他树种生态位重叠值在中度干扰下有所降低,枹栎(Quercus serrata)的生态位宽度会变小,山茶科(Theaceae)植物的生态位变宽,同时其生态位重叠值增大;在灌木层中,重度干扰下,菝葜(Smilax china)生态位宽度减小,栲树幼苗的生态适应范围也骤减,其他大多数物种在中度干扰下,生态位宽度最低;在草本层中,蕨类植物及皱叶狗尾草(Setaria plicata)在重度干扰下生态位宽度及重叠值均增高,其适应范围变广,种间竞争加剧。综合分析表明,群落各层次物种多样性水平对人为干扰的响应各不相同,其中人为干扰对灌木及草本层物种多样性影响最大。同时人为干扰会改变群落物种组成及种间原有的关系,特别是中度干扰会使群落间优势种地位增强,种间竞争关系加剧,导致其原有生境被破坏,不利于群落发展。因此强化景区保护及合理经营迫在眉睫。     

Variable Substitution Rates of the 18 Domain Sequences in Artemia Hemoglobin

The Artemia hemoglobin is a dimer comprising two nine-domain covalent polymers in quaternary association. Each polymer is encoded by a gene representing nine successive globin domains which have different sequences and are presumed to have been copied originally from a single-domain gene. Two different polymers exist as the result of a complete duplication of the nine-domain gene, allowing the formation of either homodimers or the heterodimer. The total population size of 18 domains comprising nine corresponding pairs, coupled with the probability that they reflect several hundred million years of evolution in the same lineage, provides a unique model in which the process of gene multiplication can be analyzed. The outcome has important implications for the reliability of local molecular clocks. The two polymers differ from each other at 11.7% of amino acid sites; however when corresponding individual domains are compared between polymers, amino acid substitution fluctuates by a factor of 2.7-fold from lowest to highest. This variation is not obvious at the DNA level: Domain pair identity values fluctuate by 1.3-fold. Identity values are, however, uncorrected for multiple substitutions, and both silent and nonsilent changes are pooled. Therefore, to determine the variability in relative substitution rates at the DNA level, we have used the method of Li (1993, J Mol Evol 36:96–99) to determine estimates of nonsynonymous (K _A ) and synonymous (K _S ) substitutions per site for the nine pairs of domains. As expected, the overall level of silent substitutions (K _S of 56.9%) far exceeded nonsilent substitutions (K _A of 6.7%); however, for corresponding domain pairs, K _A fluctuates by 2.3-fold and K _S by 1.7-fold. The large discrepancies reflected in the expressed protein have accrued within a single lineage and the implication is that divergence dates of different genera based on amino acid sequences, even with well-studied proteins of reasonable size, can be wrong by a factor well in excess of 2. Received: 4 June 1997 / Accepted: 17 December 1997