Nuclear DNA Amounts in Angiosperms--583 New Estimates

The amount of DNA in the unreplicated haploid nuclear genome(itsC-value) varies over 600-fold between angiosperm species.Information regarding this character is used in a strikinglywide variety of plant biological fields. Moreover, recent studieshave noted a significant need for more information about thisimportant aspect of genome biodiversity. Bennett and co-authorshave published four collected lists of nuclear DNA amounts inangiosperm species, compiled primarily for reference purposes,including the most recent in 1995 (Annals of Botany76: 113–176).Together they list estimates for over 2500 species which representabout 1% of the global angiosperm flora. Interest in angiospermgenome size has remained high, as shown by the recent publicationof many new estimates, creating a need for a fifth compilation.This paper presents a supplementary list of nuclear DNAC-valuesfrom 37 sources for 471 angiosperm species not listed in theaforementioned compilations, plus additional estimates for 113species already listed by them. It contains estimates for palm,orchid and tropical hardwood species which significantly improvesrepresentation of the global flora. Work is in hand to combinethe genome size data compiled in this and the aforementionedpapers into a unified database, and to present the informationin separate lists, with species in alphabetical and systematicorders, respectively. Meanwhile, the availability of DNAC-valuesfor angiosperm species can be checked on the World Wide Web(http://www.rbgkew.org.uk/cval/database1.html). Angiosperm DNA amounts; DNAC-values; nuclear genome sizes; reference lists; plant DNA database     

Nuclear DNA Amounts in Angiosperms and their Modern Uses--807 New Estimates

The DNA amount in the unreplicated haploid nucleus of an organismis known as its C-value. C-values differ about 1000-fold amongangiosperms and are characteristic of taxa. The data are usedin many biological fields, so they should be easily available.Values for 2802 angiosperm species (1%) were estimated during1950–1997, and five collected lists of C-values were publishedfor reference purposes during 1976–1997. Numbers of newangiosperm C-values published recently remained high, necessitatinga further supplementary list. This paper lists DNA C-valuesfor 807 angiosperm species from 70 original sources, including520 (75.2%) from sources published after 1996, and 691 for speciesnot included in any of the previous five lists. There is a continuingneed to estimate accurate DNA C-values for plant taxa, as shownin a workshop on this biodiversity topic sponsored by Annalsof Botany and held at Kew in 1997. Its key aim was to identifymajor gaps in our knowledge of plant DNA amounts and to recommendtargets and priorities for new work to fill them. A target ofestimating first C-values for the next 1% of angiosperm speciesin 5 years was set. The proportion of such C-values in the presentwork (85.6%) is very high; and the number being published (approx.220 per annum) has never been exceeded. In 1997, C-values werestill unknown for most (68%) families, so a target of completecoverage was set. This paper includes first C-values for 12families, but as less than 2% of such values listed here targetednew families, the need to improve familial representation remains.Copyright 2000 Annals of Botany Company Angiosperm DNA amounts, DNA C-values, nuclear genome sizes, plant DNA database     

First nuclear DNA amounts in more than 300 angiosperms

BACKGROUND AND AIMS: Genome size (DNA C-value) data are key biodiversity characters of fundamental significance used in a wide variety of biological fields. Since 1976, Bennett and colleagues have made scattered published and unpublished genome size data more widely accessible by assembling them into user-friendly compilations. Initially these were published as hard copy lists, but since 1997 they have also been made available electronically (see the Plant DNA C-values database http://www.kew.org/cval/homepage.html). Nevertheless, at the Second Plant Genome Size Meeting in 2003, Bennett noted that as many as 1000 DNA C-value estimates were still unpublished and hence unavailable. Scientists were strongly encouraged to communicate such unpublished data. The present work combines the databasing experience of the Kew-based authors with the unpublished C-values produced by Zonneveld to make a large body of valuable genome size data available to the scientific community. METHODS: C-values for angiosperm species, selected primarily for their horticultural interest, were estimated by flow cytometry using the fluorochrome propidium iodide. The data were compiled into a table whose form is similar to previously published lists of DNA amounts by Bennett and colleagues. KEY RESULTS AND CONCLUSIONS: The present work contains C-values for 411 taxa including first values for 308 species not listed previously by Bennett and colleagues. Based on a recent estimate of the global published output of angiosperm DNA C-value data (i.e. 200 first C-value estimates per annum) the present work equals 1.5 years of average global published output; and constitutes over 12 % of the latest 5-year global target set by the Second Plant Genome Size Workshop (see http://www.kew.org/cval/workshopreport.html). Hopefully, the present example will encourage others to unveil further valuable data which otherwise may lie forever unpublished and unavailable for comparative analyses.     

Nuclear DNA Amounts in Angiosperms

Bennett and Smith (Philosophical Transactions of the Royal Societyof London B274:227-274; B334: 309-345) and Bennett, Smith andHeslop-Harrison (Proceedings of the Royal Society of London,B216: 179-199) published lists of nuclear DNA amounts estimatedfor 1612 angiosperm species collected from 163 sources datedbetween 1951 and 1986. Subsequently, interest in genome sizein angiosperms and its significance has continued, and manynew DNA estimates were published during 1986-1994. Their inaccessibility,and the flow of enquiries for such information, shows that afurther compilation is needed. This paper presents a supplementarylist of nuclear DNA C-values for 105 sources for 899 angiospermspecies not listed in the above-mentioned compilations, plus284 additional estimates for 208 species already listed by them.The data are assembled primarily for reference purposes, withspecies listed in alphabetical order, rather than by any taxonomicscheme. Some advantages and limitations of flow cytometry, nowincreasingly used to quantify DNA C-values in plants, are reviewed.Recent reports regarding the occurrence and extent of intraspecificvariation in genome size are also discussed. While some examplesare real, others reflect technical shortcomings. Work has begunto combine the genome size data compiled in this and the above-mentionedpapers into a unified data base, and to present the informationin separate lists, with species in alphabetical and systematicorders, respectively. DNA C-values are now known for 1% of theworld's angiosperm flora, but improved representation of taxonomicgroups, geographical regions and plant life forms is urgentlyneeded.Copyright 1995, 1999 Academic Press Angiosperm DNA amounts, DNA C-values, nuclear genome sizes, intraspecific variation, flow cytometry, reference lists, genome size database     

Nuclear DNA amounts in angiosperms: progress, problems and prospects

BACKGROUND: The nuclear DNA amount in an unreplicated haploid chromosome complement (1C-value) is a key diversity character with many uses. Angiosperm C-values have been listed for reference purposes since 1976, and pooled in an electronic database since 1997 (http://www.kew.org/cval/homepage). Such lists are cited frequently and provide data for many comparative studies. The last compilation was published in 2000, so a further supplementary list is timely to monitor progress against targets set at the first plant genome size workshop in 1997 and to facilitate new goal setting. SCOPE: The present work lists DNA C-values for 804 species including first values for 628 species from 88 original sources, not included in any previous compilation, plus additional values for 176 species included in a previous compilation. CONCLUSIONS: 1998-2002 saw striking progress in our knowledge of angiosperm C-values. At least 1700 first values for species were measured (the most in any five-year period) and familial representation rose from 30 % to 50 %. The loss of many densitometers used to measure DNA C-values proved less serious than feared, owing to the development of relatively inexpensive flow cytometers and computer-based image analysis systems. New uses of the term genome (e.g. in 'complete' genome sequencing) can cause confusion. The Arabidopsis Genome Initiative C-value for Arabidopsis thaliana (125 Mb) was a gross underestimate, and an exact C-value based on genome sequencing alone is unlikely to be obtained soon for any angiosperm. Lack of this expected benchmark poses a quandary as to what to use as the basal calibration standard for angiosperms. The next decade offers exciting prospects for angiosperm genome size research. The database (http://www.kew.org/cval/homepage) should become sufficiently representative of the global flora to answer most questions without needing new estimations. DNA amount variation will remain a key interest as an integrated strand of holistic genomics.     

Climate and growth form: the consequences for genome size in plants

The adaptive significance of nuclear DNA variation in angiosperms is still widely debated. The discussion mainly revolves round the causative factors influencing genome size and the adaptive consequences to an organism according to its growth form and environmental conditions. Nuclear DNA values are now known for 3874 angiosperm species (including 773 woody species) from over 219 families (out of a total of 500) and 181 species of woody gymnosperms, representing all the families. Therefore, comparisons have been made on not only angiosperms, taken as a whole, but also on the subsets of data based on taxonomic groups, growth forms, and environment. Nuclear DNA amounts in woody angiosperms are restricted to less than 23.54 % of the total range of herbaceous angiosperms; this range is further reduced to 6.8 % when woody and herbaceous species of temperate angiosperms are compared. Similarly, the tropical woody dicots are restricted to less than 50.5 % of the total range of tropical herbaceous dicots, while temperate woody dicots are restricted to less than 10.96 % of the total range of temperate herbaceous dicots. In the family Fabaceae woody species account for less than 14.1 % of herbaceous species. Therefore, in the total angiosperm sample and in subsets of data, woody growth form is characterized by a smaller genome size compared with the herbaceous growth form. Comparisons between angiosperm species growing in tropical and temperate regions show highly significant differences in DNA amount and genome size in the total angiosperm sample. However, when only herbaceous angiosperms were considered, significant differences were obtained in DNA amount, while genome size showed a non-significant difference. An atypical result was obtained in the case of woody angiosperms where mean DNA amount of tropical species was almost 25.04 % higher than that of temperate species, which is because of the inclusion of 85 species of woody monocots in the tropical sample. The difference becomes insignificant when genome size is compared. Comparison of tropical and temperate species among dicots and monocots and herbaceous monocots taken separately showed significant differences both in DNA amount and genome size. In herbaceous dicots, while DNA amount showed significant differences the genome size varies insignificantly. There was a non-significant difference among tropical and temperate woody dicots. In three families, i.e., Poaceae, Asteraceae, and Fabaceae the temperate species have significantly higher DNA amount and genome size than the tropical ones. Woody gymnosperms had significantly more DNA amount and genome size than woody angiosperms, woody eudicots, and woody monocots. Woody monocots also had significantly more DNA amount and genome size than woody eudicots. Lastly, there was no significant difference between deciduous and evergreen hardwoods. The significance of these results in relation to present knowledge on the evolution of genome size is discussed.     

Nuclear DNA content in Sinningia (Gesneriaceae); intraspecific genome size variation and genome characterization in S. speciosa

The Gesneriaceae (Lamiales) is a family of flowering plants comprising >3000 species of mainly tropical origin, the most familiar of which is the cultivated African violet (Saintpaulia spp.). Species of Gesneriaceae are poorly represented in the lists of taxa sampled for genome size estimation; measurements are available for three species of Ramonda and one each of Haberlea, Saintpaulia, and Streptocarpus, all species of Old World origin. We report here nuclear genome size estimates for 10 species of Sinningia, a neotropical genus largely restricted to Brazil. Flow cytometry of leaf cell nuclei showed that holoploid genome size in Sinningia is very small (approximately two times the size of the Arabidopsis genome), and is small compared to the other six species of Gesneriaceae with genome size estimates. We also documented intraspecific genome size variation of 21%-26% within a group of wild Sinningia speciosa (Lodd.) Hiern collections. In addition, we analyzed 1210 genome survey sequences from S. speciosa to characterize basic features of the nuclear genome such as guanine-cytosine content, types of repetitive elements, numbers of protein-coding sequences, and sequences unique to S. speciosa. We included several other angiosperm species as genome size standards, one of which was the snapdragon (Antirrhinum majus L.; Veronicaceae, Lamiales). Multiple measurements on three accessions indicated that the genome size of A. majus is ~633 × 10? base pairs, which is approximately 40% of the previously published estimate.     

First nuclear DNA C-values for 18 eudicot families

BACKGROUND AND AIMS: A key target set at the second Plant Genome Size Workshop, held at the Royal Botanic Gardens, Kew in 2003, was to produce first DNA C-value data for an additional 1 % of angiosperm species, and, within this, to achieve 75 % familial coverage overall (up from approx. 50 %) by 2009. The present study targeted eudicot families for which representation in 2003 (42.5 %) was much lower than monocot (72.8 %) and basal angiosperm (69.0 %) families. METHODS: Flow cytometry or Feulgen microdensitometry were used to estimate nuclear DNA C-values, and chromosome counts were obtained where possible. KEY RESULTS: First nuclear DNA C-values are reported for 20 angiosperm families, including 18 eudicots. This substantially increases familial representation to 55.2 % for angiosperms and 48.5 % for eudicots. CONCLUSIONS: The importance of targeting specific plant families to improve familial nuclear DNA C-value representation is reconfirmed. International collaboration will be increasingly essential to locate and obtain material of unsampled plant families, if the target set by the second Plant Genome Size Workshop is to be met.     

Nuclear DNA amounts in angiosperms: targets, trends and tomorrow

BACKGROUND AND AIMS: The amount of DNA in an unreplicated gametic chromosome complement is known as the C-value and is a key biodiversity character of fundamental significance with many practical and predictive uses. Since 1976, Bennett and colleagues have assembled eight compilations of angiosperm C-values for reference purposes and subsequently these have been pooled into the Angiosperm DNA C-values Database (http://data.kew.org/cvalues/). Since the last compilation was published in 2005, a large amount of data on angiosperm genome size has been published. It is therefore timely to bring these data together into a ninth compilation of DNA amounts. Scope The present work lists DNA C-values for 2221 species from 151 original sources (including first values for 1860 species not listed in previous compilations). Combining these data with those published previously shows that C-values are now available for 6287 angiosperm species. KEY FINDINGS: Analysis of the dataset, which is by far the largest of the nine compilations published since 1976, shows that angiosperm C-values are now being generated at the highest rate since the first genome sizes were estimated in the 1950s. The compilation includes new record holders for the smallest (1C = 0·0648 pg in Genlisea margaretae) and largest (1C = 152·23 pg in Paris japonica) genome sizes so far reported, extending the range encountered in angiosperms to nearly 2400-fold. A review of progress in meeting targets set at the Plant Genome Size meetings shows that although representation for genera, geographical regions and some plant life forms (e.g. island floras and parasitic plants) has improved, progress to increase familial representation is still slow. In terms of technique it is now clear that flow cytometry is soon likely to become the only method available for plant genome size estimations. Fortunately, this has been accompanied by numerous careful studies to improve the quality of data generated using this technique (e.g. design of new buffers, increased awareness and understanding of problems caused by cytosolic inhibitors). It is also clear that although the speed of DNA sequencing continues to rise dramatically with the advent of next-generation and third-generation sequencing technologies, 'complete genome sequencing' projects are still unable to generate accurate plant genome size estimates.     

基于5 993个核基因的被子植物系统发育关系研究

系统发育关系的构建对被子植物分类及进化研究非常重要。长期以来,被子植物系统发育的研究,大多使用质体基因、线粒体基因或少数保守的单拷贝核基因。该研究从已注释基因组或转录组中搜集88种被子植物(包含58目)的核基因集;通过对其进行同源基因聚类及去旁系同源基因,获得了5 993个一对一的直系同源基因家族(即对于每个基因家族,每种植物最多一条序列,最少包含50个物种);使用截取各种不同数目基因集的DNA或氨基酸序列,采用串联法(concatenation)和溯祖法(coalescence),共构建了20棵进化树。比较这些进化树,虽然大部分结果支持APG IV中描述的被子植物主要支系之间的关系[(真双子叶植物,单子叶植物),木兰类植物],但真双子叶植物内部各目分支的演化关系与APG IV有一个很大的不同,即认为檀香目和石竹目是蔷薇类植物的姊妹群。基于这些进化树,估算了被子植物各目分支的分化时间,结果表明被子植物的起源时间为237.78百万年前(95%置信区间为202.6～278.08),与主流观点认为的225百万年至240百万年前一致。以上结果为构建进化树提供了一种可行性策略,这种方法允许使用基因数目更多而计算速度更快。     

Plant DNA barcodes and the influence of gene flow

Success of species assignment using DNA barcodes has been shown to vary among plant lineages because of a wide range of different factors. In this study, we confirm the theoretical prediction that gene flow influences species assignment with simulations and a literature survey. We show that the genome experiencing the highest gene flow is, in the majority of the cases, the best suited for species delimitation. Our results clearly suggest that, for most angiosperm groups, plastid markers will not be the most appropriate for use as DNA barcodes. We therefore advocate shifting the focus from plastid to nuclear markers to achieve an overall higher success using DNA barcodes.     

How much evolutionary history in a 10 x 10 m plot?

We use a fully dated phylogenetic tree of the angiosperm families to calculate phylogenetic diversity (PD) in four South African vegetation types with distinct evolutionary histories. Since the branch length values are in this case represented by the ages of plant lineages, PD becomes the cumulative evolutionary age (CEA) of assemblages. Unsurprisingly, total CEA increases with family and with species diversity and observed values are the same as expected from random sampling of family lists. However, when random sampling is done from species lists, observed CEAs are generally lower than expected. In vegetation types which have undergone recent diversification-grassland, fynbos and Nama-karoo-co-occurring species are more closely related than expected, but in subtropical thicket the observed CEAs are well described by random sampling. The use of CEA has great potential for assessing the age of biotic assemblages, particularly as the dating of genus and species-level phylogenies become more accurate.     

Many parallel losses of infA from chloroplast DNA during angiosperm evolution with multiple independent transfers to the nucleus

We used DNA sequencing and gel blot surveys to assess the integrity of the chloroplast gene infA, which codes for translation initiation factor 1, in >300 diverse angiosperms. Whereas most angiosperms appear to contain an intact chloroplast infA gene, the gene has repeatedly become defunct in approximately 24 separate lineages of angiosperms, including almost all rosid species. In four species in which chloroplast infA is defunct, transferred and expressed copies of the gene were found in the nucleus, complete with putative chloroplast transit peptide sequences. The transit peptide sequences of the nuclear infA genes from soybean and Arabidopsis were shown to be functional by their ability to target green fluorescent protein to chloroplasts in vivo. Phylogenetic analysis of infA sequences and assessment of transit peptide homology indicate that the four nuclear infA genes are probably derived from four independent gene transfers from chloroplast to nuclear DNA during angiosperm evolution. Considering this and the many separate losses of infA from chloroplast DNA, the gene has probably been transferred many more times, making infA by far the most mobile chloroplast gene known in plants.     

Quantifying effects of legal and non-legal designations of alien plant species on their control and profile

Invasive alien species (IAS) pose serious threats to native ecosystems worldwide. In some regions, laws and/or lists related to alien species have been made, but their effects on actual measures against alien species have been little studied. In Japan the IAS act, which came into force in 2005, lists many harmful species under the heading “Designated IAS”. The importation, domestication, sale or release of these species is tightly regulated. In addition, other species are named as alerted alien species (AASs) on the “alien species alert list” to raise public awareness. In this paper we examined the situation in Japan as case study that is exposed to threats from alien species under such laws in recent year to evaluate the effects of that for public interesting. We conducted a survey of the effects of the legal designation and other classification of alien plant species, examining a number of aspects, including research publications, eradications and public awareness. In our analysis, the degree of harmfulness of IASs was considered on the basis of weed risk assessment (WRA). The results show that legal classification of IASs has led to increases in research focused on the designated species, eradication efforts and public awareness. Similar results were also apparent for some AASs, but there was a great deal of variability among the many sub-categories of AASs. This may be partly because AAS sub-categories are based on different premises, and partly because the identification of AASs is not necessarily linked to legal controls that affect the behavior of individuals.     

Organization and variation of angiosperm mitochondrial genome

The mitochondrial genomes of angiosperms are the largest mitochondrial genomes so far reported and are highly variable in size among plant species. The comparative analysis of the angiosperm mitochondrial genomes at the nucleotide level has now become feasible for addressing long-standing questions, owing to the publication of five dicot and three monocot genomes. Whereas the identified genes and introns are rather well conserved, intergenic regions are highly variable in sequence, even between two close relatives. Promiscuous DNA and horizontally transferred sequence constitute part of the intergenic regions, but the origin of the majority of these regions is unknown. On the other hand, duplication and extensive rearrangement of preexisting sequences may be one of the explanations for the occurrence of unknown sequences. Functional aspects of the mitochondrial genome, such as RNA editing and expression of unique open reading frames (ORFs), can be changed under certain nuclear genotypes.     

Phylogenetic analysis of 25 plant species representing 19 angiosperm families and one gymnosperm family based on 390 orthologous genes

High-throughput sequencing is fundamentally altering traditional phylogenetic classifications. While the Angiosperm Phylogeny Group (APG) III system based on chloroplast sequences has opened up a new era of angiosperm molecular classification, the use of nuclear genome sequences is more helpful for a precise phylogenetic analysis. However, such attempts have so far been applied to only a very limited number of angiosperm plant families. We constructed a phylogenetic tree of 25 plant species with well assembled genome sequences representing 19 angiosperm families and one gymnosperm family (as out group) using 390 orthologous genes. Our results strongly support the moving of Rhamnaceae to order Rosales from Rhamnales, abolishing Rhamnales, and establishing the new order Malpighiales in the APG III system. Our data also showed some characteristics inconsistent with the APG III classification and provided a reinterpretation of phylogeny for some of the families involved. We propose that Malpighiales should be placed in Malvids, not Fabids. The four monocot species representing four families were clustered together, indicating that monocot is a natural taxon. In summary, our results support almost all the APGIII treatments of the orders involved in this study and provided some surprising reinterpretations at levels beyond order.     

Evolutionary Dynamics of Microsatellite Distribution in Plants: Insight from the Comparison of Sequenced Brassica,Arabidopsis and Other Angiosperm Species

Despite their ubiquity and functional importance, microsatellites have been largely ignored in comparative genomics, mostly due to the lack of genomic information. In the current study, microsatellite distribution was characterized and compared in the whole genomes and both the coding and non-coding DNA sequences of the sequenced Brassica, Arabidopsis and other angiosperm species to investigate their evolutionary dynamics in plants. The variation in the microsatellite frequencies of these angiosperm species was much smaller than those for their microsatellite numbers and genome sizes, suggesting that microsatellite frequency may be relatively stable in plants. The microsatellite frequencies of these angiosperm species were significantly negatively correlated with both their genome sizes and transposable elements contents. The pattern of microsatellite distribution may differ according to the different genomic regions (such as coding and non-coding sequences). The observed differences in many important microsatellite characteristics (especially the distribution with respect to motif length, type and repeat number) of these angiosperm species were generally accordant with their phylogenetic distance, which suggested that the evolutionary dynamics of microsatellite distribution may be generally consistent with plant divergence/evolution. Importantly, by comparing these microsatellite characteristics (especially the distribution with respect to motif type) the angiosperm species (aside from a few species) all clustered into two obviously different groups that were largely represented by monocots and dicots, suggesting a complex and generally dichotomous evolutionary pattern of microsatellite distribution in angiosperms. Polyploidy may lead to a slight increase in microsatellite frequency in the coding sequences and a significant decrease in microsatellite frequency in the whole genome/non-coding sequences, but have little effect on the microsatellite distribution with respect to motif length, type and repeat number. Interestingly, several microsatellite characteristics seemed to be constant in plant evolution, which can be well explained by the general biological rules.     

Nuclear DNA ploidy in mammary carcinomas; using nuclear size as co-parameter reveals more complex patterns.

Static DNA measurements using nuclear size as a co-parameter have been made on cell populations from 106 consecutive patients with proper imprints from fresh tumour tissues or fine-needle aspirates. When DNA content is plotted against the nuclear size of each individual cancer cell in a scatter diagram, different patterns are found. These patterns indicate the presence of subpopulations, which may be overlooked using conventional flow cytometric analysis or other techniques relying only on the nuclear DNA content. The presence of these subpopulations might, in addition, interfere with a correct definition and estimation of the percentage of S-phase cells.     

Highly conserved low-copy nuclear genes as effective markers for phylogenetic analyses in angiosperms

? Organismal phylogeny provides a crucial evolutionary framework for many studies and the angiosperm phylogeny has been greatly improved recently, largely using organellar and rDNA genes. However, low-copy protein-coding nuclear genes have not been widely used on a large scale in spite of the advantages of their biparental inheritance and vast number of choices. ? Here, we identified 1083 highly conserved low-copy nuclear genes by genome comparison. Furthermore, we demonstrated the use of five nuclear genes in 91 angiosperms representing 46 orders (73% of orders) and three gymnosperms as outgroups for a highly resolved phylogeny. ? These nuclear genes are easy to clone and align, and more phylogenetically informative than widely used organellar genes. The angiosperm phylogeny reconstructed using these genes was largely congruent with previous ones mainly inferred from organellar genes. Intriguingly, several new placements were uncovered for some groups, including those among the rosids, the asterids, and between the eudicots and several basal angiosperm groups. ? These conserved universal nuclear genes have several inherent qualities enabling them to be good markers for reconstructing angiosperm phylogeny, even eukaryotic relationships, further providing new insights into the evolutionary history of angiosperms.