A High Quality Chromosome Preparation from Cell Suspension Aggregates Culture of Coffea canephora

Clarindo, Wellington Ronildo

doi:10.1508/cytologia.71.243

Cited by 22 publications

(10 citation statements)

References 25 publications

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“…We considered these methodologies to be fundamental, as they supply more adequate chromosomic preparations in comparison with those yielded by the squashing technique (Carvalho and Saraiva 1993), usually applied in cytogenetic studies of ipecac (Assis and Giulietti 1999, Rout et al 2000, and Souza et al 2006). In accordance with Ohmido et al (1998), Guzzo et al (2000), Bauchan et al (2001), Clarindo and Carvalho (2006), Clarindo et al (2007) and Freitas et al (2007), the digital image analysis tools also contributed for providing high quality images of metaphasic cells and for discriminating subtle measurement differences in chromosomes.…”

Section: Cytogeneticssupporting

confidence: 58%

Karyotype and nuclear DNA content of Psychotria ipecacuanha: a medicinal species

Rossi

Clarindo

Oliveira³

2008

CYTOLOGIA

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Summary Psychotria ipecacuanha, ipecac, is a perennial, medicinal herb that grows in clusters in the understory of humid and shady areas of the Brazilian Atlantic and Amazon forests. Cytogenetic studies performed in this species have reported 2nϭ22 chromosomes, with limited karyotypic information. The present study was conducted in order to apply cytogenetic and flow cytometric tools in samples of ten ipecac populations, five from the Atlantic and five from the Amazon forest, with the purpose of characterizing their karyotype and measuring their 2C DNA content. The flow cytometry analyses evidenced two groups with distinct DNA amounts: MOZ population with 2Cϭ1.24 pg and GUA, TVI, TLM, VRB, RAP, MAN, RVE, BBU and PRA populations with 2Cϭ2.05 pg. Metaphasic chromosomes obtained from root apical meristems of GUA, TLM and TVI showed a karyotype consisting of 11 chromosome pairs, 4 metacentric (4, 5, 8 and 9) and 7 submetacentric (1, 2, 3, 6, 7, 10 and 11), with lengths varying from 3.97 to 2.53 mm. The secondary constriction was identified in the long arm of chromosome 6. Cytogenetic characterization also reveled that each chromosome pair 2-3; 4-5; 6-7; 8-9 and 10-11 is cytogenetically similar, with regard to total length, short and long arm size, and chromosomic class, suggesting that this species might be a tetraploid plant.

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Section: Cytogeneticssupporting

confidence: 58%

Karyotype and nuclear DNA content of Psychotria ipecacuanha: a medicinal species

Rossi

Clarindo

Oliveira³

2008

CYTOLOGIA

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“…This aspect was important for precise delineation of C. papaya chromosomes by digital segmentation and for karyogram assembly. The squashing technique, applied in C. papaya roots by Datta [14], has been considered unsuitable for obtaining high quality chromosome preparations, especially in species with small and morphologically similar chromosomes [7,[11][12][13]29]. CCD video camera was an appropriate tool for scattering the mitotic images showing well-individualized chromosomes.…”

Section: Resultsmentioning

confidence: 99%

“…Hsc-FA Banding Aged slides, 80-90 days at 30°C, were submitted to the Hsc-FA banding technique [1,11,39]. Slides were incubated in phosphate buffer (pH 6.5) at 85°C for 20 min and stained with 0.01% (w/v) acridine orange solution for 20 min.…”

Section: Methodsmentioning

confidence: 99%

Genome size, base composition and karyotype of Carica papaya L.

Araújo

Carvalho

Clarindo

2010

Nucleus

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Few data concerning Carica papaya DNA content and chromosomal characterization have been reported in specialized bibliography. Considering this fact, flow cytometry and cytogenetic methodologies were applied in order to improve the knowledge about the genome size, base composition and karyotype of C. papaya hermaphroditic plants. Flow cytometry results showed that hermaphroditic C. papaya genome size is relatively small (2 C=0.65 pg) and the base composition (63.49% AT and 36.51% GC) is typical of core dicots. All mitotic cells showed 2n=18 chromosomes, which were classified in metacentric or submetacentric. Despite small and homomorphic chromosomes, it has been made possible to resolve karyogram of hermaphroditic C. papaya. Through classical and banding techniques, the secondary constriction was observed in association to the chromosome 1 centromere. Here, we propose flow cytometry and cytogenetic protocols for studying the C. papaya genome. These methodologies may be useful as a basis for molecular cytogenetic techniques application.

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“…These small genome sizes are congruent with the similarly small sized chromosomes (1-3 μm length) formerly reported by Bouharmont (1963). High-quality chromosome plates were obtained from a cell suspension aggregate culture developed specifically for the two cultivated species C. canephora (Clarindo and Carvalho 2006) and C. arabica (Clarindo and Carvalho 2008). However, although karyograms have been obtained, their morphological identification, whatever the species considered, is currently difficult without additional diagnostic landmarks.…”

Section: Introductionmentioning

confidence: 99%

Physical mapping of rDNA and heterochromatin in chromosomes of 16 Coffea species: A revised view of species differentiation

et al. 2009

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The chromosome organization among 15 wild diploid Coffea species and cultivated tetraploid C. arabica was determined by fluorochrome banding (CMA, DAPI) and double fluorescence in-situ hybridization (FISH) of 5S and 18S rDNA achieved on the same chromosome plates. Two to five chromosome pairs (plus one putative chromosome B) are marked. Overall, there are two SAT-chromosome pairs for East African species and one for the Malagasy and the West and Central African species. 18S rDNA loci are telomeric and strongly marked the SAT-chromosome pairs. Generally, only one pericentromeric 5S rDNA locus characterized East African species, while an additional minor locus co-localized with the 18S rDNA-SAT locus for the Malagasy species and West and Central African species. A combination of rDNA FISH plus CMA and DAPI banding patterns enables identification of almost all the species, even those for which the genetic or botanical status is still being discussed. C. arabica clearly appears to be an allotetraploid species, including one genome from East Africa and one from West and Central Africa. However, since the minor 5S rDNA-SAT locus present in West/Central African genomes is not detected, two evolutionary hypotheses could be put forward for C. arabica. Considering only the diploid species, global trends are obvious in rDNA signal patterns, genome size variations, and geographic distribution of the species, but there are no clear evolutionary trends. However, complex interactions between these factors and environmental growing conditions exist, which have resulted in loss and gain of rDNA loci and probably also in copy repeat number variations in each rDNA family.

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A High Quality Chromosome Preparation from Cell Suspension Aggregates Culture of Coffea canephora

Cited by 22 publications

References 25 publications

Karyotype and nuclear DNA content of Psychotria ipecacuanha: a medicinal species

Karyotype and nuclear DNA content of Psychotria ipecacuanha: a medicinal species

Genome size, base composition and karyotype of Carica papaya L.

Physical mapping of rDNA and heterochromatin in chromosomes of 16 Coffea species: A revised view of species differentiation

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