Average Genome Size Research Articles

Genome size in Dahlia Cav. (Asteraceae–Coreopsideae)

The genus Dahlia (Asteraceae–Coreopsideae) is monophyletic according to a recent DNA phylogeny (ETS and ITS of rDNA). Traditionally, the genus has been divided into sections, but these have been shown not to be monophyletic. We have studied variation in genome size (DNA C-values) in a sample of species to investigate the possible effects of secondary metabolites on flow cytometry and Feulgen densitometry, and to see whether genome size variation has any systematic or phylogenetic significance. Using a range of cultivars, secondary compounds from corollas were shown to have only minor effects on the Feulgen method; the floral pigments were found to be relatively inert and seemed to have been extracted on fixation with acetic methanol. Freshly expanded corollas showed apparent apoptotic DNA decay in epidermal cells, so need to be used with caution. Flow cytometric measurements with propidium iodide in some cultivars resulted in a very similar average genome size (2C = 8.62 pg) as compared with Feulgen densitometry (2C = 8.84 pg). Leaf cytosol of D. variabilis has a demonstrable inhibitory effect on propidium iodide fluorescence, which may explain some of the intraspecific variation of C-values observed. DNA 2C-values ranged from 3.30 pg in D. dissecta (2n = 34) to 9.62 pg in a D. variabilis cultivar (2n = 64). The D. variabilis cultivars had broadly similar C-values showing a 1.16-fold range between cultivars. Some of this variation probably results from technical variables and the extent of genuine variation is uncertain. The highest 2Cx-value occurred in one D. coccinea accession (2.47 pg, 2n = 32; x = 8). D. coccinea with 2n = 64 showed slightly reduced Cx-values compared to D. coccinea with 2n = 32. Artificially produced interspecific hybrids had C-values that corresponded closely with expectations from the measured values obtained from their parents.

Evolution of genome size in fishes: a phylogenetic test of the Hinegardner and Rosen hypothesis

Despite remarkable advances in genomic studies over the past few decades, surprisingly little is known about the processes governing genome evolution at macroevolutionary timescales. In a seminal paper, Hinegardner and Rosen (Am Nat 106:621-644, 1972) suggested that taxa characterized by larger genomes should also display disproportionately stronger fluctuations in genome size. Therefore, according to the Hinegardner and Rosen (HR) hypothesis, there should be a negative correlation between average within-family genome size and its corresponding coefficient of variation (CV), a prediction that was supported by their analysis of the genomes of 275 species of fish. In this study we reevaluate the HR hypothesis using an expanded dataset (2050 genome size records). Moreover, in addition to the use of standard linear regression techniques, we also conducted modern comparative analyses that take into account phylogenetic non-independence. Our analyses failed to confirm the negative relationship detected in the original study, suggesting that the evolution of genome size in fishes might be more complex than envisioned by the HR hypothesis. Interestingly, the frequency distribution of fish genome sizes was strongly skewed, even on a logarithmic scale, suggesting that the dynamics underlying genome size evolution are driven by multiplicative phenomena, which might include chromosomal rearrangements and the expansion of transposable elements.

Intra- and inter-specific variations in the copy number of two types of retrotransposons from the ectomycorrhizal basidiomycete Tricholoma matsutake

To explore intra- and inter-specific variations of the ectomycorrhizal basidiomycete Tricholoma matsutake that produces the fruit body "matsutake", we carried out real-time PCR analysis based on two types of retrotransposons, one designated marY1, which resembles a retrovirus carrying the long terminal repeat (LTR) and the other marY2N, which resembles mRNA carrying the polyadenylated tail. Calculation based on the average genome size of homobasidiomycetes (34 Mbp) shows that ca. 5.5% of the total genome of T. matsutake isolated from Asia is made up of these retrotransposons, whereas they occupy ca. 1.4% in the isolates from Morocco, ca. 0.8% in isolates from Mexico, and ca. 0.5% in Tricholoma magnivelare, the species which produces "American matsutake". Other Tricholoma spp. that produce fruit bodies similar to those of T. matsutake, such as T. bakamatsutake, T. fulvocastaneum, and T. robustum, carry them in the region less than 0.05% of their total genome. Copy number of LTR of marY1 is consistently and markedly higher than that of the coding regions of marY1 and marY2N. Data suggest that retrotransposons are deeply involved in evolution of the ectomycorrhizal symbiont.

Chromosomal variation in plant evolution

Chromosomal variation in plant evolution

The arbuscular mycorrhizal fungus Glomus intraradices is haploid and has a small genome size in the lower limit of eukaryotes

The genome size, complexity, and ploidy of the arbuscular mycorrhizal fungus (AMF) Glomus intraradices was determined using flow cytometry, reassociation kinetics, and genomic reconstruction. Nuclei of G. intraradices from in vitro culture, were analyzed by flow cytometry. The estimated average length of DNA per nucleus was 14.07 ± 3.52 Mb. Reassociation kinetics on G. intraradices DNA indicated a haploid genome size of ∼16.54 Mb, comprising 88.36% single copy DNA, 1.59% repetitive DNA, and 10.05% fold-back DNA. To determine ploidy, the DNA content per nucleus measured by flow cytometry was compared with the genome estimate of reassociation kinetics. G. intraradices was found to have a DNA index (DNA per nucleus per haploid genome size) of approximately 0.9, indicating that it is haploid. Genomic DNA of G. intraradices was also analyzed by genomic reconstruction using four genes (Malate synthase, RecA, Rad32, and Hsp88). Because we used flow cytometry and reassociation kinetics to reveal the genome size of G. intraradices and show that it is haploid, then a similar value for genome size should be found when using genomic reconstruction as long as the genes studied are single copy. The average genome size estimate was 15.74 ± 1.69 Mb indicating that these four genes are single copy per haploid genome and per nucleus of G. intraradices. Our results show that the genome size of G. intraradices is much smaller than estimates of other AMF and that the unusually high within-spore genetic variation that is seen in this fungus cannot be due to high ploidy.

Analysis of genetic identity of North American Anaplasma phagocytophilum strains by pulsed-field gel electrophoresis.

Biological and geographic heterogeneity of anthropozoonosis caused by Anaplasma phagocytophilum is poorly understood. Seven North American A. phagocytophilum strains were compared by PFGE. The average genome size was 1.58 Mbp, and restriction patterns were identical. New World strains of A. phagocytophilum have a large genome and a high degree of genetic uniformity.

Flow cytometric analysis of nuclear DNA content in Musa

Nuclear genome size variation was studied in Musa acuminata (A genome), Musa balbisiana (B genome) and a range of triploid clones differing in genomic constitution (i.e. the relative number of A and B genomes). Nuclear DNA content was estimated by flow cytometry of nuclei stained by propidium iodide. The A and B genomes of Musa differ in size, the B genome being smaller by 12% on average. No variation in genome size was found among the accessions of M. balbisiana (average genome size 537 Mbp). Small, but statistically significant, variation was found among the subspecies and clones of M. acuminata (ranging from 591 to 615 Mbp). This difference may relate to the geographical origin of the individual accessions. Larger variation in genome size (8.8%) was found among the triploid Musa accessions (ranging from 559 to 613 Mbp). This variation may be due to different genomic constitutions as well as to differences in the size of their A genomes. It is proposed that a comparative analysis of genome size in diploids and triploids may be helpful in identifying putative diploid progenitors of cultivated triploid Musa clones. Statistical analysis of data on genome size resulted in a grouping which agreed fairly well with the generally accepted taxonomic classification of Musa.

Construction of a Fibrobacter succinogenes genomic map and demonstration of diversity at the genomic level.

The genomic cleavage map of the type strain Fibrobacter succinogenes S85 was constructed. The restriction enzymes AscI, AvrII, FseI, NotI, and SfiI generated DNA fragments of suitable size distribution that could be resolved by pulsed-field gel electrophoresis (PFGE). An average genome size of 3.6 Mb was obtained by summing the total fragment sizes. The linkages between the 15 AscI fragments of the genome were determined by combining two approaches: isolation of linking clones and cross-hybridization of restriction fragments. The genome of F. succinogenes was found to be represented by the single circular DNA molecule. Southern hybridization with specific probes allowed the eight genetic markers to be located on the restriction map. The genome of this bacterium contains at least three rRNA operons. PFGE of the other three strains of F. succinogenes gave estimated genome sizes close to that of the type strain. However, RFLP patterns of these strains generated by AscI digestion are completely different. Pairwise comparison of the genomic fragment distribution between the type strain and the three isolates showed a similarity level in the region of 14.3% to 31.3%. No fragment common to all of these F. succinogenes strains could be detected by PFGE. A marked degree of genomic heterogeneity among members of this species makes genomic RFLP a highly discriminatory and useful molecular typing tool for population studies.

Mini-chromosomes in Fusarium sporotrichioides are mosaics of dispersed repeats and unique sequences

Variations in trichothecene patterns of 26 Fusarium sporotrichioides isolates from different plant and geographic origins showed no correlation with electrophoretic karyotype polymorphisms. When intact chromosomes were examined, interisolate karyotype differences were observed only in the mini-chromosome range. Further polymorphisms were revealed in Notl-digested samples. By summing the Notl fragments the average genome size of F. sporotrichioides was estimated to be 20.4 Mb. Mini-chromosomes shared common sequences with the larger ones; however, clones (RMS-1 and RMS-2) specific to these structures have also been found. These clones contained no coding region and no promising similarities were observed when they were compared to sequences held at GenBank. Mini-chromosomes in F. sporotrichioides constitute a mosaic composed of dispersed repeats and unique sequences. This mosaic structure was maintained in all noninterbreeding, genetically isolated strains examined.

Physical and genetic map of Streptococcus thermophilus A054

The three restriction endonucleases SfiI, BssHII, and SmaI were found to generate fragments with suitable size distributions for mapping the genome of Streptococcus thermophilus A054. A total of 5, 8, and 24 fragments were produced with SfiI, BssHII, and SmaI, respectively. An average genome size of 1,824 kb was determined by summing the total fragment sizes obtained by digestions with these three enzymes. Partial and multiple digestions of genomic DNA in conjunction with Southern hybridization were used to map SfiI, BssHII, and SmaI fragments. All restriction fragments were arranged in a unique circular chromosome. Southern hybridization analysis with specific probes allowed 23 genetic markers to be located on the restriction map. Among them, six rrn loci were precisely located. The area of the chromosome containing the ribosomal operons was further detailed by mapping some of the ApaI and SgrAI sites. Comparison of macrorestriction patterns from three clones derived from strain A054 revealed two variable regions in the chromosome. One was associated with the tandem rrnD and rrnE loci, and the other was mapped in the region of the lactose operon.

Comparison of the Sma I-Digested Chromosomes of Staphylococcus epidermidis and the Closely Related Species Staphylococcus capitis and Staphylococcus caprae

Pulsed-field gel electrophoresis was used to examine the chromosomal polymorphisms existing within and between four closely related members of the Staphylococcus epidermidis species group, S. epidermidis, Staphylococcus caprae, Staphylococcus capitis subsp. capitis, and S. capitis subsp. ureolyticus. SmaI was chosen as the restriction endonuclease for this study because it generated only a few well-separated chromosomal fragments. Each of the species and subspecies showed distinct SmaI digest patterns. The strains examined in this study were collected over a 20-year period from various geographical locations. The results indicate that DNA fragment patterns are unique to each species and subspecies and represent a reasonably stable component in the chromosome structure. S. caprae and S. capitis demonstrated considerable conservation in chromosome structure as indicated by the large numbers of conserved SmaI digest fragments. The polymorphisms found within each species appear to be linked to the species' character variability. The genome size of each Staphylococcus strain was extrapolated from the SmaI digest fragment pattern obtained by pulsed-field gel electrophoresis. The average genome size for S. epidermidis is 2,364 +/- 119 kb; for S. caprae strains from humans it is 2,600 +/- 157 kb and for S. caprae strains from goats it is 2,493 +/- 15 kb; for S. capitis subsp. capitis it is 2,456 +/- 71 kb; and for S. capitis subsp. ureolyticus it is 2,276 +/- 90 kb.

Comparison of Leuconostoc oenos Strains by Pulsed-Field Gel Electrophoresis

Pulsed-field gel electrophoresis of chromosomal DNA digested with NotI or SfiI was used to differentiate individual strains of Leuconostoc oenos. L. oenos isolates with 13 different restriction digest patterns were detected in New Zealand wines undergoing malolactic fermentation. The average genome size was estimated to be 1,800 kb.

Physical genome map of the unicellular cyanobacterium Synechococcus sp. strain PCC 7002

A physical restriction map of the genome of the cyanobacterium Synechococcus sp. strain PCC 7002 was assembled from AscI, NotI, SalI, and SfiI digests of intact genomic DNA separated on a contour-clamped homogeneous electric field pulsed-field gel electrophoresis system. An average genome size of 2.7 x 10(6) bp was calculated from 21 NotI, 37 SalI, or 27 SfiI fragments obtained by the digestions. The genomic map was assembled by using three different strategies: linking clone analysis, pulsed-field fragment hybridization, and individual clone hybridization to singly and doubly restriction-digested large DNA fragments. The relative positions of 21 genes or operons were determined, and these data suggest that the gene order is not highly conserved between Synechococcus sp. strain PCC 7002 and Anabaena sp. strain PCC 7120.

Differentiation of the subspecies of Campylobacter fetus by genomic sizing.

Campylobacter fetus subsp. fetus and C. fetus subsp. venerealis are currently differentiated by tolerance to glycine and by their epidemiology. Analysis of C. fetus DNA by pulsed-field gel electrophoresis, after digestion with the restriction endonucleases SmaI and SalI, was used to differentiate between the subspecies. All strains presently identified as C. fetus subsp. fetus had a genomic size of 1.1 Mb, whereas the majority of the C. fetus subsp. venerealis strains had a genomic size of 1.3 Mb. An additional group of strains, which were previously described as C. fetus subsp. venerealis biovar "intermedius" and were able to tolerate higher concentrations of glycine than the rest of the C. fetus subsp. venerealis strains, had an average genome size of 1.5 Mb. We suggest that pulsed-field gel electrophoresis may be useful as an additional aid in the differentiation of C. fetus strains at the subspecies level.

Genome size in parental and hybrid species of Bacillus (Insecta, Phasmatodea) from southeastern Sicily: a flow cytometric analysis

In Sicily five Bacillus taxa are found: two bisexuals, B. grandii (2n = 34, female; 2n = 33, male) and B. rossius (2n = 36, female; 2n = 35, male), and three thelytokous, B. atticus (2n = 34), B. whitei (2n = 35), and B. lynceorum (3n = 52). Their cytofluorometric 2C DNA values, in picograms, in order are 5.22 (♀) –4.31 (♂); 4.33 (♀) –3.89 (♂); 4.40; 4.53; and 6.80. These values represent the first flow cytofluorometric data on phasmatodean genome size. The highest value of B. grandii and the lowest one of B. rossius appear to be related to abundance and scarcity of C-heterochromatin, respectively, which is also rather low in B. atticus. The genome size of B. whitei agrees perfectly with its shown hybrid derivation from B. grandii and B. rossius. On the other hand the value of B. lynceorum is significantly lower than the one expected (7.38 pg) on the basis of its formerly suggested hybrid origin from the backcross B. whitei × B. grandii, but it appears much closer to, and not significantly different from, the value of 6.97 pg expected according to the newly suggested double allotriploid origin, mainly based on electrophoretic evidence, from B. atticus, B. rossius, and B. grandii. Finally, as far as Bacillus taxa are concerned, average stick-insect genome size appears to be just lower than that of other orthopteroid insects, and much lower (10 times) than that of the leaf insect Extatosoma tiaratum, obtained through static cytometry on Feulgen-stained DNA squashes.Key words: cytotaxonomy, DNA amount, hybrid speciation, parthenogenesis, stick insects.

Use of pulsed-field agarose gel electrophoresis to size genomes of Campylobacter species and to construct a SalI map of Campylobacter jejuni UA580

To determine the physical length of the chromosome of Campylobacter jejuni, the genome was subjected to digestion by a series of restriction endonucleases to produce a small number of large restriction fragments. These fragments were then separated by pulsed-field gel electrophoresis with the contour-clamped homogeneous electric field system. The DNA of C. jejuni, with its low G+C content, was found to have no restriction sites for enzymes NotI and SfiI, which cut a high-G+C regions. Most of the restriction enzymes that were used resulted in DNA fragments that were either too numerous or too small for genome size determination, with the exception of the enzymes SalI (5' ... G decreases TCGAG ... 3'), SmaI (5' .... CCC decreases GGG .... 3'), and KpnI (5' ... GGTAC decreases C .... 3'). With SalI, six restriction fragments with average values of 48.5, 80, 110, 220, 280, and 980 kilobases (kb) were obtained when calibrated with both a lambda DNA ladder and yeast Saccharomyces cerevisiae chromosome markers. The sum of these fragments yielded an average genome size of 1.718 megabases (Mb). With SmaI, nine restriction fragments with average values ranging from 39 to 371 kb, which yielded an average genome size of 1.726 Mb were obtained. With KpnI, 11 restriction fragments with sizes ranging from 35 to 387.5 kb, which yielded an average genome size of 1.717 Mb were obtained. A SalI restriction map was derived by partial digestion of the C. jejuni DNA. The genome sizes of C. laridis, C. coli, and C. fetus were also determined with the contour-clamped homogeneous electric field system by SalI, SmaI, and KpnI digestion. Average genome sizes were found to be 1.714 Mb for C. coli, 1.267 Mb for C. fetus subsp. fetus, and 1.451 Mb for C. laridis.

Nuclear DNA content variation in the ancient indigenous races of Mexican maize

SUMMARY From eight maize accessions representing the four ancient indigenous races of mexican maize the nuclear DNA content was determined by flow cytometry. No significant difference between accessions of the same race was observed. Significant differences were found among the races, with two clusters observed. The lowland races had an average genome size of 12·6 pg while the highland races showed an average genome size of 11·1 pg. A significant negative correlation between genome size and altitude was observed.

Genome size variation in the North American sunfish genusLepomis(Pisces: Centrarchidae)

SummaryGenome sizes (nuclear DNA contents) were documented spectrophotometrically from individuals of each of nine species of the North American centrarchid (sunfish) genusLepomis. The distributions of DNA values within and among the nine species were essentially normal and continuous, suggesting that changes in DNA quantity inLepomisare small in amount, involve both gains and losses of DNA, and are cumulative and independent in effect. Significant differences in mean genome size were found between individuals within populations in all nine species and between species. Nested analysis of variance and comparisons of average genome size difference or distance between individuals drawn from different levels of taxonomic organization revealed that the majority of genome size divergence inLepomisoccurs above the hierarchical level of individuals within populations. TheLepomisdata when compared to similar data from North American cyprinid fishes appear to suggest that: (i) genome size evolution in these fishes at least follows a continuous rather than a discontinuous mode; (ii) the general predictions of hypothetical models relating genome size variation as a function of organismal position along adaptive continua may be oversimplified, or not applicable to complex, higher eukaryotes; and (iii) changes in genome size in these fishes may be concentrated in speciation episodes.

Endosperm responses to irradiated pollen in apples

The cytological effects of pollen γ-irradiation at 50 and 100 krad on both embryo and endosperm development were studied in Malus × domestica. Fruit and seed set were reduced by increasing doses of pollen irradiation, while embryo sacs resulting from the treatments differed in number and morphology of endosperm nuclei and in the presence or absence of an embryo. Nuclear abnormalities, distinguished from normal nuclear behaviour in embryo sacs derived from unirradiated pollen, included enhanced numbers of polyploid restitution nuclei, bridges between nuclei, excluded metaphase chromosome fragments and disrupted mitotic synchrony. Generally, a high dose of pollen irradiation (100 krad) generated an all-or-nothing response in the embryo sac, either creating highly abnormal embryos and/or endosperms which aborted, or showing relatively normal development. Callus produced from excised cellular endosperm differed in average genome size, that derived from 100 krad pollen being smaller than that from unirradiated pollen.

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.