Flow Cytometric Determination of Nuclear Replication Stage in Seed Tissues

Bino, R.J.

doi:10.1006/anbo.1993.1097

Cited by 115 publications

(76 citation statements)

References 17 publications

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“…However, estimation of absolute DNA content using dry seeds has not been reported. Our results confirmed previous observations (19,21) that tissues of different ploidy levels are present in dry seeds of some species (especially in endospermic ones), which makes FCM estimation of genome size difficult. However, the use of specific seed tissues, usually the radicles, which, like young leaves, contain most cells in the G 0 /G 1 phase of the cell cycle, facilitates interpretation of the histograms, and makes this plant material suitable for DNA content measurement by FCM.…”

Section: Discussionsupporting

confidence: 91%

“…However, seeds of some species contain an endosperm in addition to the embryo. This is a storage tissue, which in diploid species is triploid (formed by the fusion of two haploid nuclei from the female gametophyte and one haploid nucleus from the male gametophyte), composed often of regularly divided cells and ones that are endoreduplicated (e.g., up to 192C in maize) (19,21,24). Additional peaks on FCM histograms, representing endosperm cells, can overlap the G 0 /G 1 peak of the species used as a reference (internal) standard (species of known genome size processed simultaneously with a target species) and thus confuse genome size estimations.…”

Section: Discussionmentioning

confidence: 99%

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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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Section: Discussionsupporting

confidence: 91%

Section: Discussionmentioning

confidence: 99%

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

2005

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“…The embryos mainly contained cells with a 2C and 4C DNA content although a small pick for 8C DNA was observed. Earlier researches also showed that embryo tissues consist mostly of cells with 2C DNA (Fagus silvatica, Pinus nigra, Cichorium endive) or 2C and 4C DNA (Rahanus sativus) although there are species whose embryos contains some endopolyploid cells (Spinacia oleracea, Cucumis sativus) (Bino et al 1993;Gilissen et al 1993). The additional 8C DNA pick for C. album embryos indicated that endopolyploidization took place in a small number of cells at the early stages of tissue differentiation during embryo development.…”

Section: Genome Of C Albummentioning

confidence: 99%

Comparative cytogenetic analysis of diploid and hexaploid Chenopodium album Agg

2011

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Two cytotypes of Chenopodium album, diploid (2n=2x=18) and hexaploid (2n=6x=54), were analysed using flow cytometry and a FISH experiment. The genome size was indicated as 1.795 pg for the diploid and 3.845 pg for the hexaploid plants which suggested genome downsizing in the evolution of hexaploid cytotype. Double FISH with 25S rDNA and 5S rDNA allowed three to five homologue chromosome pairs to be distinguished depending on the cytotype. The variation in size and number of rDNA sites between the polyploid C. album and its putative diploid ancestor indicated that rDNA loci underwent rearrangements after polyploidization.Flow cytometry measurements of the relative nuclear DNA content in the somatic tissue of C. album revealed extensive endopolyploidization resulting in tissues comprising a mixture of cells with a different DNA content (from 2C to 32C) in varying proportions. The pattern of endopolyploidy was characteristic for the developmental stage of the plant and for the individual organ. Polysomaty was not observed in the embryo tissues however endopolyploidization had taken place in most tested organs of seedlings. The endopolyploidy in diploid and hexaploid C. album was compared to find any relationship between the pattern of polysomaty and polyploidy level in this species. This revealed that polyploid plants showed a decline in the number of endocycles as well as in the frequency of endopolyploidy cells compared to diploid plants.

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“…This suggests that the majority of cells were retained in G 1 phase, while a small number were in G 2 phase of the cellular cycle. This is a prevalent characteristic in orthodox seeds (Bino et al 1993, Castro et al 2000, Vázquez-Ramos and Sánchez 2003, Gendreau et al 2008, although there are reports of dispersed orthodox seeds with high 4C DNA contents (approximately 40%) (Faria et al 2005, Guimarães et al 2011. High 2C DNA contents have also been observed in the mature seeds of intermediate and recalcitrant species, such as Castanea sativa (Bino et al 1993), Inga vera (Faria et al 2004) and Coffea arabica , indicating that the relationship between tolerance to desiccation and events inherent to the cellular cycle, is weak.…”

Section: Discussionmentioning

confidence: 99%

Physiological and biochemical changes during the loss of desiccation tolerance in germinating Adenanthera pavonina L. seeds

Soares

Dias

Faria

et al. 2015

An. Acad. Bras. Ciênc.

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We investigated the loss of desiccation tolerance (DT) in Adenanthera pavonina seeds during germination. Seeds were subjected to imbibition for 0, 24, 36, 48, 60 and 81 h, then dried to their initial moisture content (13%), rehydrated and evaluated for survival (resumption of growth and development of normal seedlings) and membrane system integrity (electrolyte leakage). Embryonic axes of seeds subjected only to imbibition during the same early time periods were used to investigate the electrophoretic patterns of heat-stable proteins and the relative nuclear DNA content. In A. pavonina seeds, DT remained unchanged until 36 h of imbibition (resulting in germination and 82% normal seedlings), after which it was progressively lost, and seeds with a protruded radicle length of 1 mm did not withstand dehydration. The loss of desiccation tolerance could not be related to either membrane damage caused by drying or the resumption of the cell cycle during germination. However, the decrease in heat-stable protein contents observed throughout germination may be related to the loss of DT in A. pavonina seeds.

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Flow Cytometric Determination of Nuclear Replication Stage in Seed Tissues

Cited by 115 publications

References 17 publications

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

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

Comparative cytogenetic analysis of diploid and hexaploid Chenopodium album Agg

Physiological and biochemical changes during the loss of desiccation tolerance in germinating Adenanthera pavonina L. seeds

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