Revisiting the DNA C-values of the genome size-standards used in plant flow cytometry to choose the “best primary standards”

Fontes, Milene Miranda Praça; Carvalho, Carlos Roberto; Clarindo, Wellington Ronildo; Cruz, Cosme Damião

doi:10.1007/s00299-011-1026-x

Cited by 66 publications

(52 citation statements)

References 37 publications

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“…1c) was identical to that evidenced by Clarindo et al (2013). Praça-Fontes et al (2011) compared the DNA content measurement of eight plants widely used as primary standards in FCM approaches by applying each separately as primary standard, in a cascade-like manner. In conclusion, these authors reported that S. lycopersicum is a suitable primary reference standard for FCM assessments in plants.…”

Section: Fcm and Cytogenetic Analysesmentioning

confidence: 99%

“…These materials were used as source of leaf fragments for somatic embryo induction, based on method of van Boxtel and Berthouly (1996), as well as for FCM assessments. Seeds of Solanum lycopersicum L. Stupické (2C=2.00 pg, Praça-Fontes et al 2011) were kindly supplied by Dr. Jaroslav Doležel (Experimental Institute of Botany, Czech Republic), and used as primary standard for FCM analysis.…”

Section: Plant Materialsmentioning

confidence: 99%

“…The mean nuclear 2C value was determined for each sample (in pg) by multiplying the mean ratio by the 2C value of the primary standard (S. lycopersicum, 2C=2.00 pg; Praça-Fontes et al 2011).…”

Section: Fcm and Cytogenetic Analysesmentioning

confidence: 99%

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Regeneration of Allotriploid <i>Coffea</i> Plants from Tissue Culture: Resolving the Propagation Problems Promoted by Irregular Meiosis

2016

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Summary Híbrido de Timor (HT) is a natural interspecific hybrid of Coffea arabica L. with Coffea canephoraPierre ex Froehner that has played a substantial role in breeding programs as source of resistance genes. The original HT plant is represented by the anorthoploid accession CIFC 4106 , an allotriploid plant with 3x=33 chromosomes and 2C=2.10 pg. From this HT plant, other accessions have been obtained and used in crossings with C. arabica to provide resistant plants to the main pathogens. However, each HT accession and their derivates show a particular karyotype due to irregular anaphasic disjunction, which results in a few reproductive cells with an unbalanced number of chromosomes. Regarding this fact and the crop relevance of HT, this study aimed to develop a direct somatic embryogenesis system for the clonal propagation of this germplasm, using the first HT plant as explant donor. To accomplish this purpose, disinfested leaf explants of greenhouse-cultivated HT CIFC 4106 plants were inoculated in medium supplied with 0.001 g L 1 6-benzylaminopurine. Somatic embryogenesis process was asynchronous, with distinct developmental stages occurring simultaneously. Besides, emergence of secondary embryos from primary ones was observed. Mature embryos were germinated, and well-rooted embryos were selected for plantlet regeneration. The nuclear DNA content and the karyotype showed that allotriploidy was conserved in all regenerated plantlets. Considering these results, the direct somatic embryogenesis protocol adopted in the present work was imperative for the accurate clonal propagation of HT, being relevant for multiplication and conservation of elite accessions that show an irregular meiosis.

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Section: Fcm and Cytogenetic Analysesmentioning

confidence: 99%

Section: Plant Materialsmentioning

confidence: 99%

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Regeneration of Allotriploid <i>Coffea</i> Plants from Tissue Culture: Resolving the Propagation Problems Promoted by Irregular Meiosis

2016

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“…Myrsine parvifolia was collected in a forest remnant located in Guarapari municipality, ES, Brazil (20°36'15"S – 40°25'27"W), characterized as coastal sandy plains vegetation (Restinga) at sea level. Leaves and fruits of Solanum lycopersicum L. and Pisum sativum L. (internal standards for flow cytometry – FCM, 2C = 2.00 pg and 2C = 9.16 pg, respectively, Praça-Fontes et al 2011) were supplied by Dr. Jaroslav Doležel (Experimental Institute of Botany, Czech Republic).…”

Section: Methodsmentioning

confidence: 99%

“…The suspensions were processed and stained following Otto (1990) and Praça-Fontes et al (2011) and analyzed with the flow cytometer Partec PAS II/III (Partec GmbH). Myrsine genome size was measured by multiplying the 2C value of the internal standard using the fluorescence intensity corresponding to G 0 /G 1 nuclei peak.…”

Section: Methodsmentioning

confidence: 99%

First karyotype description and nuclear 2C value for Myrsine (Primulaceae): comparing three species

Carvalho¹,

Amaral-Silva²,

Spadeto³

et al. 2017

CCG

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Cytogenetic studies in Primulaceae are mostly available for herbaceous species, and are focused on the chromosome number determination. An accurate karyotype characterization represents a starting point to know the morphometry and class of the chromosomes. Comparison among species within Myrsine, associating these data with the nuclear 2C value, can show changes that led the karyotype evolution. Here, we studied three Myrsine species [Myrsine coriacea (Swartz, 1788) Brown ex Roemer et Schultes, 1819, Myrsine umbellata Martius, 1841 and Myrsine parvifolia Candolle, 1841] that show different abilities to occupy the varied types of vegetation within the Brazilian Atlantic Forest. Cytogenetic characterization showed some individuals with 2n = 45 chromosomes for Myrsine parvifolia and Myrsine coriacea, with most individuals of the three species having 2n = 46. The first karyograms for Myrsine were assembled and presented morphologically identical and distinct chromosome pairs. In addition, differences in the mean 2C nuclear value and chromosome morphometry were found. Therefore, the first description of the Myrsine karyotype has been presented, as well as the nuclear 2C value. The procedures can be applied to other Myrsine species for future investigations in order to better understand its effects on the differential spatial occupation abilities shown by the species in Brazilian Atlantic Forest.

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Reference standards for flow cytometric estimation of absolute nuclear DNA content in plants

et al. 2021

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The estimation of nuclear DNA content has been by far the most popular application of flow cytometry in plants. Because flow cytometry measures relative fluorescence intensities of nuclei stained by a DNA fluorochrome, ploidy determination, and estimation of the nuclear DNA content in absolute units both require comparison to a reference standard of known DNA content. This implies that the quality of the results obtained depends on the standard selection and use. Internal standardization, when the nuclei of an unknown sample and the reference standard are isolated, stained, and measured simultaneously, is mandatory for precise measurements. As DNA peaks representing G1/G0 nuclei of the sample and standard appear on the same histogram of fluorescence intensity, the quotient of their position on the fluorescence intensity axis provides the quotient of DNA amounts. For the estimation of DNA amounts in absolute units, a number of well‐established standards are now available to cover the range of known plant genome sizes. Since there are different standards in use, the standard and the genome size assigned to it has always to be reported. When none of the established standards fits, the introduction of a new standard species is needed. For this purpose, the regression line approach or simultaneous analysis of the candidate standard with several established standards should be prioritized. Moreover, the newly selected standard organism has to fulfill a number of requirements: it should be easy to identify and maintain, taxonomically unambiguous, globally available, with known genome size stability, lacking problematic metabolites, suitable for isolation of sufficient amounts of nuclei, and enabling measurements with low coefficients of variation of DNA peaks, hence suitable for the preparation of high quality samples.

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Revisiting the DNA C-values of the genome size-standards used in plant flow cytometry to choose the “best primary standards”

Cited by 66 publications

References 37 publications

Regeneration of Allotriploid <i>Coffea</i> Plants from Tissue Culture: Resolving the Propagation Problems Promoted by Irregular Meiosis

Regeneration of Allotriploid <i>Coffea</i> Plants from Tissue Culture: Resolving the Propagation Problems Promoted by Irregular Meiosis

First karyotype description and nuclear 2C value for Myrsine (Primulaceae): comparing three species

Reference standards for flow cytometric estimation of absolute nuclear DNA content in plants

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