Estimation of relative DNA content in nuclei of races of Phytophthora megasperma f. sp. glycinea by quantitative fluorescence microscopy

1985 ◽  
Vol 27 (5) ◽  
pp. 614-616 ◽  
Author(s):  
F. S. Rutherford ◽  
E. W. B. Ward

The fluorochrome 4,6-diamidino-2-phenylindole was used as a stain for nuclear DNA in the fungus Phytophthora megasperma f. sp. glycinea. Measurements of DNA contents were made cytofluorometrically using known haploid and diploid strains of Aspergillus nidulans as standards. No significant differences between DNA levels in nine races of Phytophthora megasperma f. sp. glycinea were found. The results are consistent with genetical data that all nine races are diploid in the vegetative state.Key words: Phytophthora, ploidy, fungi, DNA content, fluorescence.

1983 ◽  
Vol 29 (9) ◽  
pp. 1179-1183 ◽  
Author(s):  
A. L. Franklin ◽  
W. G. Filion ◽  
J. B. Anderson

Armillaria mellea, a phytopathogenic fungus, is the only member of the Agaricales (Basidiomycetes) whose fertile vegetative phase in nature is thought to be diploid, rather than dikaryotic. To examine the vegetative ploidy of A. mellea, we used the DNA-binding antibiotic, mithramycin, for fluorometry of in situ nuclear DNA. The measurements of nuclear DNA content indicated that strains derived from single basidiospores of A. mellea were haploid and that strains derived from matings of isolates of single spores were diploid. These data confirm the results of earlier genetic experiments, which show haploidy and diploidy in unmated and mated strains, respectively. Nuclear DNA measurements in known haploid and diploid strains of Aspergillus nidulans confirmed the validity of our protocol.


2009 ◽  
Vol 57 (6) ◽  
pp. 524 ◽  
Author(s):  
Milene Miranda Praça ◽  
Carlos Roberto Carvalho ◽  
Carolina Ribeiro Diniz Boaventura Novaes

Previous flow cytometry (FCM) analyses delivered nearly equal mean values of nuclear 2C DNA content for Eucalyptus grandis Hill ex Maiden and E. urophylla S. T. Blake (1.33 pg and 1.34 pg, respectively), whereas E. globulus Labill. presented distinct mean values (1.09, 1.13 and 1.40). These differences have been attributed to the different methodological approach, utilised plant cultivar and presence of intrinsic metabolic compounds that affect fluorochrome fluorescence. In the present study, a FCM and image cytometry (ICM) design, following international consensus criteria, were adopted to reassess the nuclear DNA contents of the above-mentioned Eucalyptus species. Statistical analyses revealed either similar or discrepant nuclear DNA contents, depending on the standard species used and whether FCM or ICM was employed. Our results demonstrated that 2C DNA values obtained by FCM and ICM were most uniform when Solanum lycopersicum was used as a standard. Moreover, the values obtained for E. grandis and E. urophylla were close, but differed as much as 24.63% in relation to previous data, and E. globulus proportionally varied up to 25%. New DNA content values are suggested for these eucalypt species.


2010 ◽  
Vol 2010 ◽  
pp. 1-7 ◽  
Author(s):  
B. J. M. Zonneveld

Genome size (C-value) was applied anew to investigate the relationships within the genus Hepatica (Ranunculaceae). More than 50 samples representing all species (except H. falconeri), from wild and cultivated material, were investigated. Species of Hepatica turn out to be diploid (), tetraploid ( ), and a possible pentaploid. The somatic nuclear DNA contents (2C-value), as measured by flow cytometry with propidium iodide, were shown to range from 33 to 80 pg. The Asiatic and American species, often considered subspecies of H. nobilis, could be clearly distinguished from European H. nobilis. DNA content confirmed the close relationships in the Asiatic species, and these are here considered as subspecies of H. asiatica. Parents for the allotetraploid species could be suggested based on their nuclear DNA content. Contrary to the increase in genome size suggested earlier for Hepatica, a significant (6%–14%) loss of nuclear DNA in the natural allopolyploids was found.


A survey of work on meiotic duration in diploid plants shows that the duration is positively correlated with the DNA content per nucleus and with the mitotic cycle time. However, meiotic duration is not solely determined by the DNA content per nucleus but is also affected by chromosomal organization, DNA structure and the developmental pattern of the organism. Thus, in three polyploid plant species meiosis is much shorter and in three animal species it is much longer than would be expected in diploid plant species having corresponding DNA contents. Differences in meiotic duration in plant species are usually the result of proportional differences in all the stages of meiosis. Factors affecting the initiation, control and duration of meiosis are discussed. The consequences of changes in nuclear DNA content on developmental processes and the life cycle in plants are considered. It is suggested that DNA influences development in two ways, first directly through its informational content, and second indirectly by the physical mechanical effects of its mass independent of its informational content.


AoB Plants ◽  
2011 ◽  
Vol 2011 ◽  
Author(s):  
Naomi Phillips ◽  
Donald F. Kapraun ◽  
Amelia Gómez Garreta ◽  
M. Antonia Ribera Siguan ◽  
Jorde L. Rull ◽  
...  

Abstract Background and aims Brown algae are critical components of marine ecosystems around the world. However, the genome of only one species of the class has so far been sequenced. This contrasts with numerous sequences available for model organisms such as higher plants, flies or worms. The present communication expands our coverage of DNA content information to 98 species of brown algae with a view to facilitating further genomic investigations of the class. Methodology The DNA-localizing fluorochrome DAPI (4′,6-diamidino-2-phenylindole) and the red blood cell (chicken erythrocyte) standard were used to estimate 2C values by static microspectrophotometry. Principal results 2C DNA contents are reported for 98 species of brown algae, almost doubling the number of estimates available for the class. The present results also expand the reported DNA content range to 0.2–3.6 pg, with several species of Fucales and Laminariales containing apparent polyploid genomes with 2C = 1.8–3.6 pg. Conclusions The data provide DNA content values for 12 of the 19 recognized orders of brown algae spanning the breadth of the class. Despite earlier contentions concerning DNA content and the presence of oogamy, the present results do not support a correlation between phylogenetic placement and genome size. The closest sister groups to the brown algae have genome sizes on the order of 0.3 pg (e.g. Schizocladiophyceae), suggesting that this may be the ancestral genome size. However, DNA content ranges widely across the class.


Author(s):  
Murat Deveci ◽  
Özcan Yavaş ◽  
Nihan Sahin ◽  
Metin Tuna

The aim of this research is to determine ploidy levels of 53 spinach (Spinacia oleracea L.) accessions obtained from abroad sources by using flow cytometry. According to the results of the statistical analysis, the differences among DNA content of accessions were statistically important. The average DNA content of spinach accessions used in the study ranged between 2.225 pg/2C (Esfenaj accession) and 2.059 pg/2C (Matador and Godir accessions). The mean nuclear DNA of accessions shown to be relatively stable due to low standard deviation (0.003-0.096). Based on the mitotic chromosome analyses of the some plants with different DNA contents, mitotic chromosome number of the all plants analyzed in the study were determined as 2n=12. These results indicate that all the accessions used in the study are diploid although their DNA content is significantly vary. According to the clustering analysis of the 2C DNA contents results, it was observed that eight sub clusters were formed under two main clusters. The clustering of axioms was performed in a similar manner to the multiple comparison test. As a result, the data will save time and labor convenience in a further breeding studies deal with the same 53 spinach accessions of our study.


HortScience ◽  
2009 ◽  
Vol 44 (1) ◽  
pp. 44-48 ◽  
Author(s):  
Irene E. Palmer ◽  
Thomas G. Ranney ◽  
Nathan P. Lynch ◽  
Richard E. Bir

Rudbeckia L. are valuable nursery crops that offer broad adaptability and exceptional ornamental merit. However, there is little information on interspecific and interploid crossability and ploidy levels of specific cultivars. The objectives of this study were to determine the ploidy levels and relative DNA contents (genome sizes) of selected species and cultivars, to evaluate self-compatibility and crossability among species and ploidy levels, and to explore reproductive pathways in triploid R. hirta L. with the goal of facilitating future breeding endeavors and development of new hybrids. Reciprocal interspecific crosses were performed between R. hirta cultivars and R. fulgida Ait., R. missouriensis Engelm. ex C.L. Boynton & Beadle, and R. subtomentosa Pursh. as well as reciprocal interploid crosses among four R. hirta cultivars. A combination of relative DNA content analysis and chromosome counts was used to test for hybridity and to determine ploidy levels for selected species, cultivars, and interploid R. hirta F1 hybrids. Of the specific clones tested, R. subtomentosa and R. missouriensis were diploid, R. fuligida varieties were tetraploid, and R. hirta include both diploid and tetraploid cultivars. Mean 1Cx DNA content varied over 320% among species. The interploid R. hirta crosses produced triploids as well as pentaploids and hexaploids. Seedlings from open-pollinated triploid R. hirta appeared, based on diverse phenotypes and DNA contents, to be aneuploids resulting from sexual fertilization, not apomixis. Of the 844 seedlings from interspecific F1 crosses, only one individual, R. subtomentosa ×R. hirta, had a DNA content intermediate between its parents and was confirmed as the only interspecific hybrid. Although most taxa had low self-fertility, seedlings (with genomic sizes similar to their maternal parent) resulted after interspecific crosspollination, indicating that pseudogamy is one reproductive pathway in Rudbeckia species.


2014 ◽  
Vol 71 (3) ◽  
pp. 195-200
Author(s):  
Hanna Kuran ◽  
Kazimierz Marciniak

Mitotic activity and nuclear DNA content in endosperm of <em>Zea mays</em> cv. Złota Karłowa were examined. DNA content was cytophotometrically measured on squashed preparations after Feulgen procedure. Mitotic activity in endosperm was determined till the stage when embryo sack reached 4.5 mm in length. Some of mitotic figures show multiplied DNA content. Endosperm nuclei have various DNA contents which increase throughout endosperm development. DNA content enhancement indicates endoreduplication in progress. Some nuclei with high DNA content display changes in chromatin structure, which are expressed by the presence of strands and aggregates of chromatin characterised by high staining intensity. A conclusion has been drawn that mitotic divisions and the endoreduplication phase of nuclear DNA may occur simultaneously and dominate one over another at different phases of endosperm development.


Agronomy ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1339
Author(s):  
Piotr Kamiński ◽  
Agnieszka Marasek-Ciolakowska ◽  
Małgorzata Podwyszyńska ◽  
Michał Starzycki ◽  
Elżbieta Starzycka-Korbas ◽  
...  

Interspecific hybridization between B. oleracea inbred lines of head cabbage, Brussels sprouts, kale and B. taurica and inbred lines of rapeseed (B. napus L.) were performed aiming at the development of the new sources of genetic variability of vegetable Brassicas. Using conventional crossings and the embryo-rescue techniques the following interspecific hybrids were developed: 11 genotypes of F1 generation, 18 genotypes of F2 and F1 × F2 generations (produced after self- and cross-pollination of interspecific F1 hybrids), 10 plants of the BC1 generation (resulted from crossing head cabbage cytoplasmic male-sterile lines with interspecific hybrids of the F2 and F1 generations) and 8 plants of BC1 × (F1 × F2). No viable seeds of the BC2 generation (B. oleracea) were obtained due to the strong incompatibility and high mortality of embryos. The morphological characteristics during the vegetative and generative stages, pollen characteristics, seed development and propagation, nuclear DNA contents and genome compositions of interspecific hybrids were analyzed. All the interspecific F1 hybrids were male-fertile with a majority of undeveloped and malformed pollen grains. They showed intermediate values for morphological traits and nuclear DNA contents and had nearly triploid chromosomal numbers (27 to 29) compared with parental lines. The F2 generation had a doubled nuclear DNA content, with 52 and 56 chromosomes, indicating their allohexaploid nature. F2 hybrids were characterized by a high heterosis of morphological characteristics, viable pollen and good seed development. F1 × F2 hybrids were male-fertile with a diversified DNA content and intermediate pollen viability. BC1 plants were male-sterile with an intermediate nuclear DNA content between the F2 and head cabbage, having 28 to 38 chromosomes. Plants of the BC1 × (F1 × F2) generation were in majority male-fertile with 38–46 chromosomes, high seed set, high heterosis and intermediate values for morphological traits. The obtained interspecific hybrids are valuable as new germplasm for improving Brassica-breeding programs.


1971 ◽  
Vol 13 (3) ◽  
pp. 607-611 ◽  
Author(s):  
Michael D. Bennett ◽  
J. B. Smith

The 4C nuclear DNA content was estimated for 17 wild Hordeum species and five cultivated Hordeum vulgare varieties which were chosen to include examples varying greatly in geographical origin and in morphological and physiological characters. Nuclear DNA was measured on an integrating microdensitometer using prophase nuclei in Feulgen stained root-tip squashes. There were no significant differences in DNA content between any or the 15 diploid genotypes measured, and the seven polyploid genotypes all had nuclear DNA contents which were simple multiples of the diploid genotypes. This result, namely that the DNA content of diploid Hordeum species is invariable, differs from results obtained from diploid species in several other plant genera which differed greatly in nuclear DNA content.


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