Performance of isolated microspore-derived doubled haploids of wheat (Triticum aestivum L.)

1997 ◽  
Vol 77 (4) ◽  
pp. 549-554 ◽  
Author(s):  
Tianci Hu ◽  
Ken J. Kasha
Keyword(s):  
In Vitro Selection ◽  
Nuclear Dna ◽  
Chromosome Doubling ◽  
Microspore Culture ◽  
Doubled Haploids ◽  
Spontaneous Mutation ◽  
Nuclear Dna Content ◽  
Root Tip ◽  
Ploidy Levels ◽  
Dh Lines

Ploidy level, genetic stability and field performance of isolated microspore-derived wheat plants were evaluated. The ploidy levels of isolated microspore-derived wheat plants from cv. Chris and reciprocal crosses of Chris × Sinton were determined by two methods, namely chromosome counts of root tip cells and flow cytometric measurement of nuclear DNA content from leaves. Both methods gave similar results with the frequencies of spontaneous chromosome doubling and completely fertile plants among microspore-derived H0 plants of wheat being about 80% and 75%, respectively, based on two H0 populations. Only 1.7% were aneuploid and 16% were haploid. Spontaneous mutation frequencies were low with 1 of 124 Chris DH plants having a recessive mutant for lemma awns while three were grass-like plants in the H0 generation. The field-measured traits of microspore-derived DH lines were similar to the check by the third generation. Ninety-two percent and 70% of Chris DH lines had 1000-kernel weights and yields similar to the check, respectively. These results indicate that microspore-derived DH lines should have good potential for applications in plant breeding, in vitro selection, plant transformation and genetic studies. Key words: Wheat, microspore, culture, haploidy, performance, stability

A correlation of nuclear DNA content and thin-layer chromatographic patterns in resolving genome relationships in Avena

1972 ◽  
Vol 50 (7) ◽  
pp. 1529-1545 ◽  
Author(s):  
Koji Iiyama ◽  
William F. Grant
Keyword(s):  
Thin Layer ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Chromosomal Polymorphism ◽  
Root Tip ◽  
Coumaric Acid ◽  
Leaf Extracts ◽  
B Genome ◽  
Nuclear Deoxyribonucleic Acid

Relative amounts of nuclear deoxyribonucleic acid (DNA) from telophase root tip nuclei and thin-layer chromatographic patterns of alcohol-soluble compounds from dry leaves were determined for seven diploids: Avena clauda, A. pilosa, A. ventricosa, A. strigosa, A. hirtula, A. wiestii, A. longiglumis; four tetraploids: A. barbata, A. magna, A. abyssinica, A. vaviloviana; and four hexaploids: A. sterilis, A. fatua, A. byzantina, and A. sativa, in order to elucidate species relationships. Variation in nuclear DNA content was correlated with differences in genomic constitution; a few exceptions are considered to reflect chromosomal polymorphism. The average DNA value of the hexaploid species approximated the sum of the DNA value for A. magna and the theoretical value of the B genome. Chromatographic patterns showed distinct variations between species but little correlation between number of compounds and DNA content. Chromatographic patterns of hexaploids showed close similarity with those of diploids and tetraploids, except species with modified C genomes (A. clauda, A. pilosa, A. ventricosa) and A. longiglumis. It is considered that A. clauda, A. pilosa, A. ventricosa, and A. longiglumis did not participate in the evolution of polyploid taxa. From their chromatographic profiles, A. wiestii, A. abyssinica, A. vaviloviana, and A. byzantina are very closely related. Both A. magna and the AABB tetraploid species appear to share two genomes in common with the hexaploids. Hence, the genomic constitutions AADD and AABBDD have been proposed for A. magna and the hexaploids, respectively. Six compounds from ethanol leaf extracts of A. sativa were identified as three apigenins, luteolin, ferulic acid, and p-coumaric acid.

Haploidy and plant breeding

1981 ◽  
Vol 292 (1062) ◽  
pp. 499-507 ◽  
Keyword(s):  
Plant Breeding ◽  
Doubled Haploids ◽  
Chromosome Complement ◽  
Basic Research ◽  
Breeding Value ◽  
Additive Variance ◽  
Ploidy Levels ◽  
Breeding Programmes ◽  
Dh Lines

A haploid is an organism that looks like a sporophyte, but has the chromosome complement of a reduced gamete. There are several ways in which haploids can occur or be induced in vivo : spontaneously, mostly associated with polyembryony, and through abnormal processes after crosses, like pseudogamy, semigamy, preferential elimination of the chromosomes of one parental species, and androgenesis. In the crops described, haploids are or are near to being used in basic research and plant breeding. The application of haploids in breeding self-pollinated crops is based on their potential for producing fully homozygous lines in one generation, which can be assessed directly in the field. Early generation testing of segregating populations is possible through haploids, because doubled haploids (DH) possess additive variance only. Haploids can also be applied in classical breeding programmes to make these more efficient through improved reliability of selection. The application of haploids in cross-pollinated crops is also based on a rapid production of DH-lines, which can be used as inbred lines for the production of hybrid varieties. By means of haploids all natural barriers to repeated selfing are bypassed. In autotetraploid crops there are two types of haploid. One cycle of haploidization leads to dihaploids; a second cycle produces monohaploids. The significance of dihaploids is in their greatly simplified genetics and breeding and in the possibility of estimation of the breeding value of tetraploid cultivars by assessing their dihaploids. The main drawback of dihaploids is their restriction to two alleles per locus. Also, after doubling, it is impossible to achieve tetra-allelism at many loci, the requirement for maximal performance of autotetraploid cultivars. Tetra-allelism can be obtained when improved dihaploids have a genetically controlled mechanism of forming highly heterozygous restitution gametes with the unreduced number of chromosomes. Monohaploids, after doubling or twice doubling, may lead to fully homozygous diploids and tetraploids. These are important for basic research, but not yet for practical application. Meiotic data of potato homozygotes at three ploidy levels are presented.

DNA content in Tradescantia

1985 ◽  
Vol 27 (6) ◽  
pp. 766-775 ◽  
Author(s):  
Arturo Martínez ◽  
Héctor D. Ginzo
Keyword(s):  
North America ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Life Forms ◽  
Chromosome Size ◽  
Polyploid Complex ◽  
Ploidy Levels ◽  
Vegetative Adaptation ◽  
The Relationship

There is a wide variation in the nuclear DNA content and chromosome size between the species belonging to the T. crassifolia and T. virginiana alliances (all the species but one are native to Central and North America). Also the DNA content per genome decreases when the ploidy level increases within the same specific polyploid complex with three ploidy levels (2x, 4x, and 6x). In contrast, no variation was found in the DNA content per genome between different ploidy levels in the T. fluminensis alliance (all the species are native to South America) where they range from 6x to 22x. Since all the species described here are perennials with various life forms, it was possible to analyze the relationship between the DNA content and their vegetative adaptation to the environment. The more specialized species (geophytes and hemicryptophytes) have a higher amount of DNA than the chamaephytes adapted to live in relatively more mesic regions. In the species living in Central and North America there is a positive correlation between the increase in DNA content and the latitude of their native regions.Key words: Tradescantia, DNA content, geographical distribution, life forms, polyploidy.

scholarly journals Karyotype analysis, DNA content and molecular screening in Lippia alba (Verbenaceae)

2011 ◽  
Vol 83 (3) ◽  
pp. 993-1006 ◽  
Author(s):  
Patrícia M.O. Pierre ◽  
Saulo M. Sousa ◽  
Lisete C. Davide ◽  
Marco A. Machado ◽  
Lyderson F. Viccini
Keyword(s):  
Genetic Variation ◽  
Dna Content ◽  
Chromosome Numbers ◽  
Rapd Markers ◽  
Nuclear Dna ◽  
Pollen Viability ◽  
Flow Cytometric Analysis ◽  
Nuclear Dna Content ◽  
Ploidy Levels ◽  
Lippia Alba

Cytogenetic analyses, of pollen viability, nuclear DNA content and RAPD markers were employed to study three chemotypes of Lippia alba (Mill.) (Verbenaceae) in order to understand the genetic variation among them. Different ploidy levels and mixoploid individuals were observed. This work comprises the first report of different chromosome numbers (cytotypes) in L. alba. The chromosome numbers of La2-carvone and La3-linalool chemotypes suggested that they are polyploids. Flow cytometric analysis showed an increase of nuclear DNA content that was not directly proportional to ploidy level variation. A cluster analysis based on RAPD markers revealed that La3-linalool shares genetic markers with La1-citral and La2-carvone. The analysis showed that the majority of genetic variation of La3-linalool could be a consequence of ixoploidy. ur data indicates that sexual reproduction aong those three chemotypes is unlikely and suggests the beginning of reproductive isolation. The results demonstrated that chromosome analysis, nuclear DNA content estimation and RAPD markers constitute excellent tools for detecting genetic variation among L. alba chemotypes.

Study of androgenesis and spontaneous chromosome doubling in barley ( Hordeum vulgare L.) genotypes using isolated microspore culture

2009 ◽  
Vol 57 (2) ◽  
pp. 155-164 ◽  
Author(s):  
D. Kahrizi ◽  
R. Mohammadi
Keyword(s):  
Chromosome Doubling ◽  
Microspore Culture ◽  
Doubled Haploids ◽  
Negative Relationship ◽  
Microspore Embryogenesis ◽  
Culture Technique ◽  
Hordeum Vulgare L ◽  
Spontaneous Chromosome ◽  
Barley Genotypes

This research aimed to study the androgenesis and spontaneous chromosome doubling of five barley genotypes using an isolated in vitro microspore culture technique, involving a completely randomized design (CRD) with three replications. Statistical analysis of embryogenesis and cytogenetic results showed that genotype had a significant effect on haploid embryogenesis (P<0.01) and on spontaneous chromosome doubling (P<0.05). The genotype Igri was found to have the highest potential to produce haploid embryos (1577 embryos from 100 anthers), followed by the genotypes Boyer/Rojo, Afzal/Turkman/Kavir, Ashar/Hebo and Agrigashar/Matico with 369, 304, 278 and 150 embryos from 100 anthers, respectively. The highest percentage of spontaneous chromosome doubling (76%) was observed for the genotype which had the lowest embryogenesis (Agrigashar/Matico) and the lowest (65%) for the genotype with the highest androgenic capacity (Igri). Microspore embryogenesis also showed comparatively higher genotypic (109.2) and phenotypic (109.5) coefficients of variation, heritability (99.62) and genetic advance (1206.77), indicating the pre-dominance of additive gene action in the control of this character in the material studied. Estimates of genetic parameters (PCV, GCV and heritability) for microspore embryogenesis were higher than for spontaneous doubled haploids. These results indicated that selection for androgenic capacity would be more effective than for spontaneous doubled haploids. The findings showed a negative relationship (r= −0.68) between embryogenesis and spontaneous chromosome doubling in the barley genotypes studied. All the large embryos used had high regenerability and good plantlet formation.

Nuclear DNA amount in pure species and hybrid willows (Salix): a flow cytometric investigation

1998 ◽  
Vol 76 (1) ◽  
pp. 157-165 ◽  
Author(s):  
Jérôme Thibault
Keyword(s):  
Flow Cytometry ◽  
Ploidy Level ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Natural Hybridization ◽  
Ploidy Levels ◽  
Key Words Flow Cytometry ◽  
Dna Values ◽  
Dna Amounts

Flow cytometry (FCM) has been used to estimate the nuclear DNA content of 11 Salix species and 5 hybrids. One hundred and sixty nine individuals were studied including 159 individuals from a sequence of 32 communities along a stretch of river in France and 10 individuals from French and English collections for comparison. Isolated nuclei were stained with propidium iodide. FCM was a significantly more practical and rapid technique than that of establishing the karyotype to survey many samples of Salix for variation in ploidy. The 2C DNA amounts for diploid species ranged from 0.76 to 0.98 pg, and tetraploid values ranged from 1.62 to 1.80 pg. The DNA values were consistent with the known ploidy levels. With the exception of a doubtful Salix xquercifolia, ploidy levels and DNA amounts of hybrids were intermediate compared with those of their parents. Intraspecific variation of nuclear DNA values including instrumental variation was low (i.e., 6-11% at the same ploidy level). FCM appeared to be an accurate tool for determination of Salix triploid hybrids. However, it remains limited concerning hybrids from crosses between species of the same ploidy level. Results suggest that natural hybridization might not be frequent in the communities studied, although they have been subject to disturbance. Previous overestimates of hybridization frequency in willows were probably due to misinterpretation of the effects of the environment on Salix spp. morphology; however, the extent and mechanisms of introgression in the genus remain to be further investigated. Key words: flow cytometry, Salix, hybridization, nuclear DNA content, riparian vegetation, disturbance.

scholarly journals Chromosome Number, Ploidy Level, and Nuclear DNA Content in 23 Species of Echeveria (Crassulaceae)

Genes ◽  
2021 ◽  
Vol 12 (12) ◽  
pp. 1950
Author(s):  
Guadalupe Palomino ◽  
Javier Martínez-Ramón ◽  
Verónica Cepeda-Cornejo ◽  
Miriam Ladd-Otero ◽  
Patricia Romero ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Chromosome Number ◽  
Ploidy Level ◽  
Dna Content ◽  
Chromosome Numbers ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Root Tips ◽  
Ploidy Levels ◽  
Dna Amounts

Echeveria is a polyploid genus with a wide diversity of species and morphologies. The number of species registered for Echeveria is approximately 170; many of them are native to Mexico. This genus is of special interest in cytogenetic research because it has a variety of chromosome numbers and ploidy levels. Additionally, there are no studies concerning nuclear DNA content and the extent of endopolyploidy. This work aims to investigate the cytogenetic characteristics of 23 species of Echeveria collected in 9 states of Mexico, analyzing 2n chromosome numbers, ploidy level, nuclear DNA content, and endopolyploidy levels. Chromosome numbers were obtained from root tips. DNA content was obtained from the leaf parenchyma, which was processed according to the two-step protocol with Otto solutions and propidium iodide as fluorochrome, and then analyzed by flow cytometry. From the 23 species of Echeveria analyzed, 16 species lacked previous reports of 2n chromosome numbers. The 2n chromosome numbers found and analyzed in this research for Echeveria species ranged from 24 to 270. The range of 2C nuclear DNA amounts ranged from 1.26 pg in E. catorce to 7.70 pg in E. roseiflora, while the 1C values were 616 Mbp and 753 Mbp, respectively, for the same species. However, differences in the level of endopolyploidy nuclei were found, corresponding to 4 endocycles (8C, 16C, 32C and 64C) in E. olivacea, E. catorce, E. juarezensis and E. perezcalixii. In contrast, E. longiflora presented 3 endocycles (8C, 16C and 32C) and E. roseiflora presented 2 endocycles (8C and 16C). It has been suggested that polyploidization and diploidization processes, together with the presence of endopolyploidy, allowed Echeveria species to adapt and colonize new adverse environments.

scholarly journals NUCLEAR DNA CONTENT IN BLUEBERRY DETERMINED BY FLOW CYTOMETRY

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 580e-580
Author(s):  
Rodomiro Ortiz ◽  
D.E. Costich ◽  
T.P. Meagher ◽  
N. Vorsa
Keyword(s):  
Flow Cytometry ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Haploid Genome ◽  
Dna Flow Cytometry ◽  
Dna Amount ◽  
Ploidy Levels ◽  
Polyploid Evolution ◽  
Haploid Genome Size

DNA flow cytometry was used to determine nuclear DNA content in diploid blueberry species, and 3x, 4x, 5x, and 6x ploidy levels. Relative fluorescence intensity of stained nuclei measured by flow cytometry was a function of the number of chromosome sets (X): Y = 3.7X – 2.3 (r2 = 95.1%). DNA flow cytometry should be useful for ploidy level determination in the seedling stage. A significant linear relationship was established between nuclear DNA content and number of chromosomes (x); DNA (pg) = 0.52 x1 (r2 = 99.8%). Based on this equation the haploid genome DNA amount (1C) was calculated as 0.62 ± 0.08 pg, with an approximate haploid genome size of 602 Mbp/1C. The results indicate that conventional polyploid evolution occured in the section Cyanococcus, genus Vaccinium: the increase in DNA was concurrent with increase in chromosome number. DNA content differences among 2x species were correlated with Nei's genetic distance estimates based on 20 isozyme markers. Most of the variation was among species (49%), with 26% between populations within species, and 25% within populations.

scholarly journals Flow cytometry-based determination of ploidy from dried leaf specimens in genomically complex collections of the tropical forage grass Urochloa s. l.

2021 ◽  
Author(s):  
Paulina Tomaszewska ◽  
Till K. Pellny ◽  
Luis Miguel Hernandez ◽  
Rowan A. C. Mitchell ◽  
Valheria Castiblanco ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Germplasm Collection ◽  
Forage Grass ◽  
Ploidy Levels ◽  
Sustainable Improvement ◽  
Tropical Forage ◽  
Relative Differences

We aimed to develop an optimized approach to determine ploidy for dried leaf material in a germplasm collection of a tropical forage grass group, including approaches to collect, dry and preserve plant samples for flow cytometry analysis. Urochloa (including Brachiaria, Megathyrus and some Panicum) tropical grasses are native to Africa and are now, after selection and breeding, planted worldwide, particularly in South America, as important forages with huge potential for further sustainable improvement and conservation of grasslands. The methods enable robust identification of ploidy levels (coefficient of variation, CV, typically <5%). Ploidy of some 353 forage grass accessions (ploidy range from 2 to 9), from international genetic resource collections, showing variation in basic chromosome numbers and reproduction modes (apomixis and sexual), were determined using our defined standard protocol. Two major Urochloa agamic complexes used in the current breeding programs at CIAT and EMBRAPA: the ' brizantha' and 'humidicola' agamic complexes are variable, with multiple ploidy levels and DNA content. U. brizantha has odd level of ploidy (x=5), and the relative differences in nuclear DNA content between adjacent cytotypes is reduced, thus more precise examination of this species is required. Ploidy measurement of U. humidicola revealed some aneuploidy.

scholarly journals MICROPROPAGATION AND PLOIDY STABILITY OF Lippia lacunosa Mart. & Schauer: AN ENDANGERED BRAZILIAN MEDICINAL PLANT

2019 ◽  
Vol 6 (1) ◽  
pp. 1-7
Author(s):  
Diego Pandeló José ◽  
José Marcello Salabert De Campos ◽  
Lyderson Facio Viccini ◽  
Emilly Ruas Alkimim ◽  
Marcelo De Oliveira Santos
Keyword(s):  
Flow Cytometry ◽  
Medicinal Plant ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Naphthalene Acetic Acid ◽  
Flow Cytometry Analysis ◽  
Ploidy Levels

Lippia lacunosa is a Brazilian savanna plant that belongs to the Verbenaceae family. It has been used in folk medicine as a treatment for different diseases. This species represents an endangered Brazilian medicinal plant, and this is the first report documenting a reliable protocol for the in vitro propagation and regeneration of L. lacunosa. Axenic explants were cultivated in MS medium containing different concentrations of naphthalene acetic acid (NAA) to induce root growth. The mean shoot length and the number of roots were highest with 0.06 mg·L-1 NAA. The highest number of buds in shoot regeneration was induced with 2 mg·L-1 6-benzylaminopurine (BA). To obtain a long-term culture, the dwarf shoots were elongated on MS media containing 0.5 mg·L-1 BA alternated with MS containing 2 mg·L-1 BA every 40 days. In the present protocol, the long-term shoots retained the ability to root even after long periods of BA treatment. In addition, we evaluated the nuclear DNA content and ploidy levels, including the occurrence of endopolyploidy, in long-term micropropagated plant leaves using flow cytometry analysis. The plants propagated in vitro over several years possessed nuclear DNA contents ranging from 2.940 to 3.095 pg, and no differences in DNA content were found among in vitro plants or between these plants and the control (L. lacunosa from a greenhouse with a DNA content of 3.08 pg). The flow cytometry analysis also demonstrated that there was no polyploidization. The present study will be useful for biotechnological approaches and provides the first estimate of the nuclear DNA content of this species using flow cytometry.

Sign in / Sign up

Export Citation Format

Share Document