Chromosome doubling via tuber disc culture in dihaploid potato as determined by confocal microscopy

Mozafari, J; Wolyn, David J.; Áli-Khan, S. T.

doi:10.1007/bf01088291

Cited by 8 publications

(8 citation statements)

References 17 publications

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“…Several authors have found that tetraploids can be induced by spontaneous chromosome doubling of plants regenerated through callus culture (Fleming et al 1992;Cardi et al 1993;Mozafari et al 1997). A possible disadvantage of this method could be the occurrence of somaclonal variation.…”

Section: Introductionmentioning

confidence: 98%

Oryzalin Treatment of Potato Diploids Yields Tetraploid and Chimeric Plants from which Euploids could be Derived by Callus Induction

Barandalla¹,

Ritter²,

Galarreta³

2007

Potato Res.

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Shoot apices from six diploid potato genotypes for polyploidisation were treated with oryzalin and colchicine solutions at different concentrations and incubation times. In addition to chimeric plants, diploids, one octoploid and completely ploidy doubled plants (4x) were regenerated in these experiments. Tetraploid plants were obtained for three of the six genotypes treated with oryzalin, and no tetraploid plants were obtained after colchicine treatments of any of the genotypes. The best result of 43% tetraploid plants was achieved using 28.8 μM oryzalin and 24 h of incubation with the L37 genotype. Moreover, we determined that doubled plants should not be selected on the basis of a single measurement and require additional ploidy checks in successive clonal generations. Some L37 plants that were initially identified as tetraploids by flow cytometry produced diploid or mixoploid plants in the next clonal generation. Morphological characters of doubled L37 plants were compared with the original genotype. The PVY resistance present in a parental clone was maintained in all regenerants and significant differences among ploidy variants were found for some quantitative characters. Molecular AFLP analyses did not show any differences among genotypes. Since polyploidizing substances may produce a large proportion of chimeric plants, we applied a method to increase the number of tetraploid plants. Callus culture of chimeric leaf explants was performed on regeneration medium and tetraploid shoots were obtained in four of the six genotypes tested. The best result of 40% tetraploid plants was achieved with explants from the L37 genotype. This method was successful in producing additional tetraploid genotypes from mixoploid plants obtained after oryzalin treatments.

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Section: Introductionmentioning

confidence: 98%

Oryzalin Treatment of Potato Diploids Yields Tetraploid and Chimeric Plants from which Euploids could be Derived by Callus Induction

Barandalla¹,

Ritter²,

Galarreta³

2007

Potato Res.

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“…Haploid plants must be chromosome doubled to restore fertility for use in plant breeding. The chromosome doubling of microsporederived plantlets and calli is a critical step in haploid breeding programmes (Mozafari et al, 1997).…”

Section: Introductionmentioning

confidence: 99%

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

Kahrizi

Mohammadi²

2009

Acta Agronomica Hungarica

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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.

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“…Determining the chloroplast number per cell in 3D reconstructions made from a series of confocal microscope images (Mozafari et al , 1997; Dinkins et al , 2001; Coate et al , 2012; Xu et al , 2012) can yield an unbiased estimate if SUR sampling is applied and a sufficient number of cells is analysed. However, this is a much more time-consuming approach than application of the disector method.…”

Section: Discussionmentioning

confidence: 99%

Unbiased estimation of chloroplast number in mesophyll cells: advantage of a genuine three-dimensional approach

Kubínová

Janáček

Lhotáková

et al. 2013

EXBOTJ

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Chloroplast number per cell is a frequently examined quantitative anatomical parameter, often estimated by counting chloroplast profiles in two-dimensional (2D) sections of mesophyll cells. However, a mesophyll cell is a three-dimensional (3D) structure and this has to be taken into account when quantifying its internal structure. We compared 2D and 3D approaches to chloroplast counting from different points of view: (i) in practical measurements of mesophyll cells of Norway spruce needles, (ii) in a 3D model of a mesophyll cell with chloroplasts, and (iii) using a theoretical analysis. We applied, for the first time, the stereological method of an optical disector based on counting chloroplasts in stacks of spruce needle optical cross-sections acquired by confocal laser-scanning microscopy. This estimate was compared with counting chloroplast profiles in 2D sections from the same stacks of sections. Comparing practical measurements of mesophyll cells, calculations performed in a 3D model of a cell with chloroplasts as well as a theoretical analysis showed that the 2D approach yielded biased results, while the underestimation could be up to 10-fold. We proved that the frequently used method for counting chloroplasts in a mesophyll cell by counting their profiles in 2D sections did not give correct results. We concluded that the present disector method can be efficiently used for unbiased estimation of chloroplast number per mesophyll cell. This should be the method of choice, especially in coniferous needles and leaves with mesophyll cells with lignified cell walls where maceration methods are difficult or impossible to use.

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Chromosome doubling via tuber disc culture in dihaploid potato as determined by confocal microscopy

Cited by 8 publications

References 17 publications

Oryzalin Treatment of Potato Diploids Yields Tetraploid and Chimeric Plants from which Euploids could be Derived by Callus Induction

Oryzalin Treatment of Potato Diploids Yields Tetraploid and Chimeric Plants from which Euploids could be Derived by Callus Induction

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

Unbiased estimation of chloroplast number in mesophyll cells: advantage of a genuine three-dimensional approach

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