Analysis of Nuclear DNA Content and Ploidy in Higher Plants

Galbraith, David W.; Lambert, Georgina M.; Macas, Jiří; Doležel, Jaroslav

doi:10.1002/0471142956.cy0706s02

Cited by 105 publications

(113 citation statements)

References 32 publications

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“…Kunnan (AB) showed a low DNA content, which is consistent with its interspecific genomic composition, M. balbisiana owning one of the smallest DNA content in the Eumusa section (unpublished results). These results are consistent with those of previous reports on Musa [22]. …”

Section: Flow Cytometrysupporting

confidence: 94%

In liquid medium colchicine treatment induces non chimerical doubled-diploids in a wide range of mono- and interspecific diploid banana clones

Bakry¹,

Reberdiere²,

Pichot³

et al. 2007

Fruits

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In liquid medium colchicine treatment induces non chimerical doubleddiploids in a wide range of mono-and interspecific diploid banana clones.Abstract --Introduction. The worldwide production of banana and plantain, which is estimated to 106 Mt·year -1 , relies on a narrow genetic basis. Thus, banana production is fragile regarding emergent diseases, and creation of new varieties of banana stands out as a real necessity for its sustainability. In banana, various improvement strategies aim to create triploid hybrids. The objective of this report is to present a simple in vitro methodology to induce stable tetraploid plants by colchicine treatment, usable as parent for obtaining triploid varieties. Materials and methods. Twenty-one diploid M. acuminata clones and three interspecific M. acuminata / M. balbisiana diploid clones were treated as proliferating culture in liquid medium with 1.25 mM of colchicine for 48 h. Plant screening has been performed by morphological identification in greenhouse, chromosome counts on root tips and flow cytometry on leave blades on vitroplants using propidium iodide staining. Results. Chromosome counts led to distinguish diploid and tetraploid plants but did not afford to detect chimeras. Flow cytometry allowed an early screening of a larger number of plants leading to detect rapidly chimerical plants. It was observed that some stable 2x / 4x cytochimeras are periclinal. Tetraploid clones were obtained with all diploid genotypes. General behaviour of tetraploids in the field was globally weaker than the corresponding diploids. Nevertheless, all the doubled-diploids flowered and crossed with diploid plants to obtain triploid progenies. Conclusion. This study has clearly shown that induction of stable doubled-diploid plants can be obtained from a wide range of genetically different bananas. These results open the way to the systematic use of doubled-diploids by banana breeding programs for the release of enhanced triploid varieties.Guadeloupe / Musa / tissue culture / colchicine / chromosome number / tetraploidy / chimeras La colchicine appliquée en milieu liquide induit des diploïdes doublés non chimériques dans une large gamme de clones de bananiers diploïdes mono ou interspécifiques.Résumé --Introduction. La production mondiale de bananes et plantains, qui est estimée à 106 Mt·an -1 , s'appuie sur une base génétique étroite. La production de bananes est donc fragile quant à la menace des maladies émergentes et la création de nouvelles variétés apparaît comme une réelle nécessité pour sa pérennité. Les diverses stratégies d'amélioration du bananier visent à créer des hybrides triploïdes. L'objectif de cet article est de présenter une méthode aisée de production de plantes tétraploïdes stables par traitement de colchicine appliqué in vitro, utilisables comme parents pour obtenir de nouvelles variétés hybrides triploïdes. Matériel et méthodes. Vingt-et-un clones diploïdes de M. acuminata et trois clones diploïdes interspécifiques M. acuminata / M. balbisiana ont été traités durant la ...

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Section: Flow Cytometrysupporting

confidence: 94%

In liquid medium colchicine treatment induces non chimerical doubled-diploids in a wide range of mono- and interspecific diploid banana clones

Bakry¹,

Reberdiere²,

Pichot³

et al. 2007

Fruits

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“…oleifera; Zea mays; Helianthus annuus; Pisum sativum; staining inhibitors Flow cytometry (FCM) is commonly used for plant genome size estimation, especially after Galbraith et al (1) developed a rapid procedure for nuclei isolation from intact plant tissues (2)(3)(4)(5)(6)(7). Despite a suggestion that all kinds of plant tissue are suitable for FCM analysis (2,8,9), usually young, rapidly growing leaves are used for 2C nuclear DNA content estimation (5). This material is easy to obtain and gives high resolution histograms.…”

Section: Discussionmentioning

confidence: 99%

Are seeds suitable for flow cytometric estimation of plant genome size?

2005

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Background: Nuclear DNA content in plants is commonly estimated using flow cytometry (FCM). Plant material suitable for FCM measurement should contain the majority of its cells arrested in the G 0 /G 1 phase of the cell cycle. Usually young, rapidly growing leaves are used for analysis. However, in some cases seeds would be more convenient because they can be easily transported and analyzed without the delays and additional costs required to raise seedlings. Using seeds would be particularly suitable for species that contain leaf cytosol compounds affecting fluorochrome accessibility to the DNA. Therefore, the usefulness of seeds or their specific tissues for FCM genome size estimation was investigated, and the results are presented here. Methods: The genome size of six plant species was determined by FCM using intercalating fluorochrome propidium iodide for staining isolated nuclei. Young leaves and different seed tissues were used as experimental material. Pisum sativum cv. Set (2C ϭ 9.11 pg) was used as an internal standard. For isolation of nuclei from species containing compounds that interfere with propidium iodide intercalation and/or fluorescence, buffers were used supplemented with reductants.

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“…The procedures for sample preparation and analysis according to DOLEŽEL (1997) andGALBRAITH et al (1998) were used. Unstressed leaves were collected from seedlings of each genotype in the greenhouse, wrapped in moistened paper tissue and put into plastic bags.…”

Section: Methodsmentioning

confidence: 99%

Polyploidy effects on frost tolerance and winter survival of garden pansy genotypes

Lagibo¹,

Kobza²,

Suchánková³

2005

Hort. Sci. (Prague)

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Pansies (Viola × wittrockiana Gams) are a commercially important ornamental plant, grown and marketed during autumn and spring (ADAMS et al. 1997).Two hexadecaploid (16x) genotypes were obtained by induced polyploidy in octoploid garden pansy cultivars (AJALIN et al. 2002). The hexadecaploids were selfed in advanced generations (LAGIBO, KOB-ZA 2004a,b) and used also for further breeding in order to exploit the induced desirable traits such as improved compactness and general vigour effect in plant height and in flower size. As a result, the genotypes with 2n = 10x, 2n = 12x, and 2n = 14x ploidy levels were produced from crosses and backcrosses which are included in this study.According to LEVIN (1983), polyploidy, induced or natural, may greatly alter the cytological, genetic and physiological characteristics, and it often alters resistance to cold. Moreover, the effect of polyploidy on frost tolerance may vary from species to species and in some plants it has been demonstrated to show an increase or decrease with higher ploidy level. For instance, in Brassica campestris, autotetraploids were more resistant to frost than their diploid counterparts (CHOUDHURY et al. 1968). On the other hand, GORAL et al. (1964) found lower resistance of tetraploids to frost in Trifolium repens and higher resistance of tetraploids in Trifolium pratense.In this study, in addition to induced polyploidy, since pansies are commonly planted in autumn for spring flowering, frost tolerance is one of the most important factors to be assessed in new genotypes. Therefore, this study was conducted to identify ABSTRACT: This study was conducted to interpret the differences in frost tolerance and winter survival between 8x, 10x, 12x, 14x, and 16x ploidy levels of garden pansy (Viola × wittrockiana Gams) genotypes grown in the field conditions. Plants of each genotype were analyzed for their ploidy levels using flow cytometry. The chlorophyll fluorescence parameters were measured with portable chlorophyll fluorometer in the greenhouse and in the field at different time intervals. Increased frost stress generally reduced the fluorescence values in all genotypes. However, the genotypes differed significantly in their responses to frost as they were exposed to minimum temperatures of 1°C to -7.7°C in the field. Based on the percentage reduction in F V /F M values against -7.7°C temperature the hexadecaploids were ranked as sensitive to intermediate followed by 12x (sensitive), and genotypes with 10x and 14x ploidy levels were tolerant as the controls. The winter survival rate of hexadecaploids was by 7 to 9% lower than in the controls followed by the genotype with 12x and both genotypes with 10x and 14x ploidy levels were about equal to the controls. On the other hand, the content of photosynthetic pigments (chlorophyll a, b and total carotenoids) was the highest in hexadecaploids and tended to increase with increasing ploidy level. Further, the results gave insight that chlorophyll fluorescence could be applied directly in the field conditions ...

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Analysis of Nuclear DNA Content and Ploidy in Higher Plants

Cited by 105 publications

References 32 publications

In liquid medium colchicine treatment induces non chimerical doubled-diploids in a wide range of mono- and interspecific diploid banana clones

In liquid medium colchicine treatment induces non chimerical doubled-diploids in a wide range of mono- and interspecific diploid banana clones

Are seeds suitable for flow cytometric estimation of plant genome size?

Polyploidy effects on frost tolerance and winter survival of garden pansy genotypes

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