Genome size variation in some representatives of the genus <Emphasis Type="Italic">Tripleurospermum</Emphasis>

Genome size has been estimated by flow cytometry in 14 populations belonging to eight taxa (seven species, one of them with two varieties) of the genus Tripleurospermum. 2C nuclear DNA amounts range from 4.87 to 9.22 pg, and nuclear DNA amounts per basic chromosome set from 1.99 to 2.75 pg. Statistically significant differences depending on ploidy level, life cycle or environmental factors such as altitude have been found. Also, genome size is positively correlated with total karyotype length. The presence of rhizome is related to nuclear DNA content in these species.This work was supported by project BOS2001-3041-C02-01 of the Spanish government, and one of the authors (S.G.) received a predoctoral grant from the Spanish government.     

Spatial and temporal expression of the two sucrose synthase genes in maize: immunohistological evidence

Summary The DNAs of two diploid species of Gossypium, G. herbaceum var. africanum (A₁ genome) and G. raimondii (D₅ genome), and the allotetraploid species, G. hirsutum (A_h and D_h genomes), were characterized by kinetic analyses of single copy and repetitive sequences. Estimated haploid genome sizes of A₁ and D₅ were 1.04 pg and 0.68 pg, respectively, in approximate agreement with cytological observations that A genome chromosomes are about twice the size of D genome chromosomes. This differences in genome size was accounted for entirely by differences in the major repetitive fraction (0.56 pg versus 0.20 pg), as single copy fractions of the two genomes were essentially identical (0.41 pg for A₁ and 0.43 pg for D₅). Kinetic analyses and thermal denaturation measurements of single copy duplexes from reciprocal intergenomic hybridizations showed considerable sequence similarity between A₁ and D₅ genomes (77% duplex formation with an average thermal depression of 6 °C). Moreover, little sequence divergence was detectable between diploid single copy sequences and their corresponding genomes in the allotetraploid, consistent with previous chromosome pairing observations in interspecific F₁ hybrids.Journal paper No. 4461 of the Arizona Agricultural Experiment Station     

Divergent genome sizes reflect the infrafamilial subdivision of the neotropical woody Marcgraviaceae

Neotropical Marcgraviaceae comprise about seven genera and 130 species of lianas and shrubs. They predominantly occur in lowland or montane rainforests and are characterized by a variety of pollination systems. Early classifications subdivided Marcgraviaceae into subfamilies Marcgravioideae and Noranteoideae, a concept supported by molecular data. Using flow cytometry and chromosome numbers, we investigated the role of genome size and polyploidization in the evolution of Marcgraviaceae and how genome sizes are distributed between the proposed infrafamilial groups. To do this we determined genome sizes and chromosome counts for six genera and 22 species for the first time. Our study supports the subfamilial classification of the family, revealing contrasting genome sizes in Noranteoideae (2C = 5.5–21.5 pg) and Marcgravioideae (2C = 2.3–6.2 pg). Polyploidy is considered to be the main source of genome size variation as in each subfamily the higher nuclear DNA amounts were associated with higher ploidy. In addition, genome size changes independent of polyploidy were also observed in some genera, suggesting an additional role for changes in repetitive DNA abundance in the evolution of Marcgraviaceae. A high chromosome base number (x = 18; 2n = 36 to ～70) points to an undetected lower diploid level or to palaeopolyploidy. Marcgraviaceae show a remarkable (nine‐fold) variation in genome size, and several Noranteoideae have genome sizes among the highest reported for tropical woody angiosperms worldwide. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 177 , 1–14.     

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.     

Quantitative DNA variation between and within chromosome complements of Vigna species (Fabaceae)

Cytogenetical investigations, so far, on the organisation and evolution of the genomes of Vigna species have proved difficult due to small chromosome size, large chromosome number and uniformity in chromosome shape and size within and between the complements. In this investigation the nature and extent of DNA variation between thirteen diploid and one polyploid species have been estimated. The DNA variation between diploid species was small and species clustered around a mean value of 2.7 pg. The polyploid species had a greater DNA value of 4.95 pg. No significant variation in 2C DNA content was found between accessions of V. radiata. A comparison of the distribution of DNA among the chromosomes within complements has shown that the excess DNA acquired in evolution was distributed evenly in all chromosomes despite significant differences in chromosome size. The relative changes in chromatin area and DNA density which accompany evolutionary DNA variation was also compared.     

Flow cytometric measurement of nuclear DNA content inCapsicum (Solanaceae)

Nuclei were isolated from leaf tissue of differentCapsicum species and the relative fluorescence intensity was measured by flow cytometry after propidium iodide staining.Pisum sativum nuclei with known nuclear genome size (9.07 pg) were used as internal standard to determine nuclear DNA content of the samples in absolute units. The 2C DNA contents ranged between 7.65 pg inC. annuum and 9.72 pg inC. pubescens, and the general mean of the genus was 8.42 pg. These values correspond, respectively, to 1C genome size of 3.691 (C. annuum), 4.690 (C. pubescens) and 4.063 (general mean) Mbp. In general, white-flowered species proved to have less DNA, with the exception ofC. praetermissum, which displayed a 2C DNA content of 9.23 pg. It was possible to divide the studied species into three main groups according to their DNA content, and demonstrate differences in DNA content within two of the three species complexes established on the basis of morphological traits.     

Quantitative nuclear DNA changes inEleusine (Gramineae)

2C nuclear DNA amounts were determined in 30 collections belonging to 10 species ofEleusine. About a 2.5-fold variation in genome size is evident in the genus. The 2C DNA amount in the diploid species ranged from 2.50 pg inE. verticillata to 3.35 pg inE. intermedia. In contrast, the tetraploid species showed a range from 4.95 pg inE. africana to 6.13 pg inE. floccifolia. At intraspecific level 10 collections ofE. coracana, 6 ofE. indica, 4 ofE. africana, 2 ofE. tristachya, and 2 ofE. kigeziensis did not show any significant variation. However, 2 collections ofE. floccifolia, connected with polyploidy, displayed about 90% variation. Polyploid species showed approximately double the genome size of that of their corresponding diploids. An evolutionary increase in DNA amount is evident inE. coracana during the course of its origin and domestication fromE. africana.     

Flow cytometric analysis of nuclear genome of the Ethiopian cereal tef [Eragrostis tef (Zucc.) Trotter]

Flow cytometric analysis of nuclear DNA content was performed by using nuclei isolated from young leaf tissue of tef (Eragrostis tef). The method was very useful for rapid screening of ploidy levels in cultivars and lines of tef representing the phenotypic variability of this species in Ethiopia. The results of the analysis showed that all cultivars were tetraploid. Flow cytometry was also used to determine nuclear DNA content in absolute units (genome size) in four tef cultivars. Nuclei isolated from tomato (Lycopersicon esculentum, 2C=1.96 pg) were used as an internal reference standard. The 2C DNA content of individual tef cultivars ranged from 1.48 to 1.52 pg (1C genome size: 714 Mbp-733 Mbp), the differences among them being statistically nonsignificant. The fact that the nuclear genome of tef is only about 50% larger than that of rice should make it amenable for analysis and mapping at the molecular level.     

Genome size variation inCajanus cajan (Fabaceae): A reconsideration

A recent investigation of genome size in certain samples of the pigeonpea,Cajanus cajan, indicates values from 1.55 pg to 1.99 pg (1C level), which is 1.29-fold variation between accessions. In the present analysis those of these accessions which had particularly high or low DNA contents in that study were subjected to a reanalysis using propidium iodide and DAPI flow cytometry and Feulgen densitometry. Only minor differences in genome size, not more than 1.047-fold, were found with flow cytometry, and no significant differences were obtained with Feulgen densitometry. The previously reported genome size cannot be confirmed. It is about half as large and was determined in the present study as 0.825 pg (1C, propidium iodide flow cytometry,Glycine max as standard) and 0.853 pg (1C, Feulgen densitometry,Allium cepa andPisum sativum as standards), respectively.     

Karyotype and genome size variation in Ajuga L. (Ajugoideae–Lamiaceae)

Chromosome number changes and karyotype evolution play an important role in plant genome diversification and eventually in speciation. The genus Ajuga L. (Lamiaceae) has approximately 50 species distributed in temperate to subtropical regions. Four of these species are currently recognized in Korea (A. decumbens Thunb., A. multiflora Bunge, A. nipponensis Makino and A. spectabilis Nakai). Understanding the karyotype evolution in Ajuga has been hampered by the small size of their chromosomes and symmetrical karyotypes. Here we used classic Feulgen staining to establish chromosome numbers and construct karyotypes of the four species of Ajuga recognized in Korea and flow cytometry was used to study their variation in genome. The chromosome number of all investigated plants was 2n = 32. Still, the 2C DNA content ranged from 2.18 pg (A. decumbens) to 4.53 pg (A. multiflora). While the chromosome numbers were the same for all investigated species, the genome size variation could potentially be used as a taxonomic marker.     

Taxonomic implications of genome size for all species of the genus Gasteria Duval (Aloaceae)

Nuclear DNA content (2C) is used as a new criterion to investigate all species of the genus Gasteria Duval including the three recently described species Gasteria polita van Jaarsv., G. pendulifolia van Jaarsv. and G. glauca van Jaarsv.. The 122 accessions investigated have the same chromosome number (2n=2x=14), with exception of three tetraploid plants found. The nuclear DNA content of the diploids, as measured by flow cytometry with Propidium Iodide, is demonstrated to range from 32.8–43.2 pg. This implies that the largest genome contains roughly 10¹⁰ more base pairs than the smallest. Based on DNA content the species could be divided in five groups: G. rawlinsonii Oberm. with 32.8 pg, 13 mostly inland species with 34.3–36.0 pg, five coastal species with 36.5–39.0 pg and Gasteria batesiana Rowley with 43.2 pg. The thirteen species with 34.3–36.0 pg could be divided further, in a group of eight species occupying mainly very restricted areas with 34.3–35.1 pg and a second group of five species with 35.2–36.0 pg mainly occupying large areas. These five groups did not coincide very well with the two sections and four series of Gasteria based on a cladistic analysis by van Jaarsveld et al. (1994). Based on its long leafy branches, location in the centre of Gasteria species distribution and its by far lowest DNA content, G. rawlinsonii might be the most primitive member of the genus. Nuclear DNA content as measured by flow cytometry is shown to be relevant to provide additional information on the relationships between Gasteria species.     

Genome size evolution in Sapindaceae at subfamily level: a case study of independence in relation to karyological and palynological traits

Sapindaceae s.l. is a moderately large family of trees, shrubs and lianas. Current knowledge on genome size and how it varies in this family is scarce. This research aims to characterize the DNA content in 39 species of Sapindaceae, mainly in tribe Paullinieae s.s., by the analysis of the variation in genome size relative to karyotypic and palynological features. Nuclear DNA amount was measured by flow cytometry, and linear regression analyses were conducted to analyse the relationship between genome size variation and various karyotypic and palynological features. Genome size varied nine‐fold among species, ranging from 1C = 0.305 pg (Lophostigma plumosum) to 2.710 pg (Cardiospermum heringeri). The low regression coefficients obtained suggest that genome size mainly varies independently of karyotypic and palynological features. With regard to karyotype evolution, the constant chromosome number but variable genome size in Houssayanthus, Paullinia and Serjania suggest that structural changes mainly caused by changes in the amounts of repetitive DNA are more important than numerical change. In contrast, in Cardiospermum and Urvillea, variation in chromosome number and genome size supports the suggestion that numerical and structural changes are important in the karyotype evolution of these genera. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 174 , 589–600.     

Karyotypes, constitutive heterochromatin, and genomic DNA values in the blowfly genera Chrysomya, Lucilia, and Protophormia (Diptera: Calliphoridae).

The karyotypes and C-banding patterns of Chrysomya species C. marginalis, C. phaonis, C. pinguis, C. saffranea, C. megacephala (New Guinean strain), Lucilia sericata, and Protophormia terraenovae are described. All species are amphogenic and have similar chromosome complements (2n = 12), including an XY-XX sex-chromosome pair varying in size and morphology between species. Additionally, the C-banding pattern of the monogenic species Chrysomya albiceps is presented. The DNA contents of these and of further species Chrysomya rufifacies, Chrysomya varipes, and Chrysomya putoria were assessed on mitotic metaphases by Feulgen cytophotometry. The average 2C DNA value of the male genomes ranged from 1.04 pg in C. varipes to 2.31 pg in C. pinguis. The DNA content of metaphase X chromosomes varied from 0.013 pg (= 1.23% of the total genome) in C. varipes to 0.277 pg (12.20%) in L. sericata; that of Y chromosomes ranged from 0.003 pg (0.27%) in C. varipes to 0.104 pg (5.59%) in L. sericata. In most species, the corresponding 5 large chromosome pairs showed similar relative DNA contents. The data suggest that the interspecific DNA differences in most species are mainly due to quantitative variation of (repetitive) sequences lying outside the centromeric heterochromatin blocks of the large chromosomes. The results are also discussed with regard to phylogenetic relationships of some species.     

Genome size and genome evolution in diploid Triticeae species.

One of the intriguing issues concerning the dynamics of plant genomes is the occurrence of intraspecific variation in nuclear DNA amount. The aim of this work was to assess the ranges of intraspecific, interspecific, and intergeneric variation in nuclear DNA content of diploid species of the tribe Triticeae (Poaceae) and to examine the relation between life form or habitat and genome size. Altogether, 438 plants representing 272 lines that belong to 22 species were analyzed. Nuclear DNA content was estimated by flow cytometry. Very small intraspecific variation in DNA amount was found between lines of Triticeae diploid species collected from different habitats or between different morphs. In contrast to the constancy in nuclear DNA amount at the intraspecific level, there are significant differences in genome size between the various diploid species. Within the genus Aegilops, the 1C DNA amount ranged from 4.84 pg in A. caudata to 7.52 pg in A. sharonensis; among genera, the 1C DNA amount ranged from 4.18 pg in Heteranthelium piliferum to 9.45 pg in Secale montanum. No evidence was found for a smaller genome size in annual, self-pollinating species relative to perennial, cross-pollinating ones. Diploids that grow in the southern part of the group's distribution have larger genomes than those growing in other parts of the distribution. The contrast between the low variation at the intraspecific level and the high variation at the interspecific one suggests that changes in genome size originated in close temporal proximity to the speciation event, i.e., before, during, or immediately after it. The possible effects of sudden changes in genome size on speciation processes are discussed.     

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.     

黄藤染色体核型及基因组大小分析

为探究黄藤(Daemonoropsjenkinsiana)染色体核型和基因组的大小,采用体细胞染色体常规制片法与显微摄影技术相结合的方法,对黄藤染色体进行了核型分析,同时以番茄(Lycopersicon esculentum)为内标,应用流式细胞术对黄藤叶片基因组大小、DNA含量和DNA倍性进行了测定。结果表明,黄藤茎尖是理想的染色体制片材料;黄藤的染色体数为2n=24,核型公式为K(2n)=1M+17m+5sm+1st,核型类型为2C;核型不对称系数61.20%;黄藤的DNA含量为1.57 pg,基因组大小为1 539.53 Mb,黄藤的DNA倍性为二倍体(2n)。这是首次报道黄藤的核型和基因组大小,为深入开展黄藤属及其近缘属植物的核型和基因组比较分析提供了参考依据。     

Variation of Nuclear DNA Content in the Biflorus Species of Lonchocarpus (Leguminosae)

This represents the first study of nuclear DNA content in alarge sample (135 spp.) from a tropical arboreal genus, in whicha large proportion of the species were examined (42 spp., 31.1%).Somatic chromosome numbers and 4C-DNA values for 51 taxa ofLonchocarpus are reported. All taxa were diploid with 2 n =22,but their DNA content ranged from 1.92 to 2.86 pg 4C nucleus,corresponding to a 48.95% variation in genome size. In the 74collections studied, no correlation was observed between DNAcontent and habitat altitude. Variation in nuclear DNA contentwas analysed at the level of genus, subgenus, section and subsection.Variation in genome size was also studied within some species,either among widely separated populations or among differentintraspecific taxa. Very little variation in genome size wasdetected between populations, subspecies, and varieties of thesame species. The taxonomic implications of variation in nuclearDNA content are discussed.Copyright 2000 Annals of Botany Company Lonchocarpus (Leguminosae), DNA content, chromosome number.     

Flow cytometric analysis reveals different nuclear DNA contents in cultivated Fonio (Digitaria spp.) and some wild relatives from West-Africa

Nuclear DNA amounts of 118 cultivated fonio accessions representing 94 landraces collected from the major growing areas of West-Africa (Benin, Burkina Faso, Guinea, Mali and Togo) and eight accessions of four wild relatives were investigated by Laser flow cytometry. In cultivated species, average 2C-values ranged from 1.848 ± 0.031 pg for Digitaria iburua to 1.956 ± 0.004 pg for D. exilis. In D. exilis landraces the chromosome number was determined at 2n = 36. The closely related wild species D. longiflora and D. ternata showed similar 2C DNA contents of 1.869 ± 0.035 pg and 1.775 ± 0.070 pg, respectively. Distinctly larger genomes were identified for more distant species D. lecardii and D. ciliaris with 2.660 ± 0.070 pg and 2.576 ± 0.030 pg per 2C nucleus, respectively. Intra-specific variations were found to be slight and insignificant, suggesting genome size stability mainly within the cultivated gene pool. These results support the distance of cultivated fonio species D. exilis and D. iburua from D. lecardii and D. ciliaris as well as their close relationships with D. longiflora and D. ternata. Relevance of the results for ploidy level considerations in fonio millets is discussed.     

Genome Size in Sphagnum (Peat Moss)

Abstract: Genome size was determined in thirty Austrian species of Sphagnum, using Feulgen absorbance photometry conducted on a video-based image analysis system (CIRES), and for comparison on a scanning cytophotometer (Leitz MPV II) with strongly correlated results. Pisum sativum (1C = 4.42pg DNA) was used for internal standardization. Between species, two levels of ploidy, haploid and diploid, could be unambiguously identified (although this identification remains, strictly speaking, hypothetical, as long as exact parallel chromosome counts are not available). Twenty-six haploid species yielded values from 0.392 pg to 0.506 pg DNA (1C), and four diploid species (including two varieties of S. palustre) from 0.814 pg to 0.952 pg. The average ratio between levels was 1:2.049. Variation between species within sections was lower than between sections. In some cases significant differences between accessions of one species were found. The genome size of Sphagnum palustre presented here strongly deviates from one estimate of this species in the literature.     

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.