Chromosome homologies between Citrus and Poncirus—the comparative cytogenetic map of mandarin (Citrus reticulata)

Abstract

Mandarins are among the main citrus species and constitute a morphologically diverse and taxonomically controversial group. Cytogenetic studies were limited to the determination of its karyotype formula and the localization of ribosomal DNA (rDNA) sites. The cytogenetic map of Poncirus trifoliata, established by fluorescent in situ hybridization (FISH) using single-copy bacterial artificial chromosomes (BACs) together with the localization of rDNA sequences and heterochromatin (CMA/DAPI banding pattern), represented an important milestone for the cytogenetic studies of the citrus species. In this work, the cytogenetic map of Citrus reticulata cv. Cravo, one of the pure species of the genus, was built using BACs from P. trifoliata, including the citrus tristeza virus resistance locus (Ctv), to detect homeologies and structural karyotypic changes between these species. Except for chromosomes 2 and 3, heterochromatin distribution and position of BACs were maintained in comparison to P. trifoliata, indicating a high synteny between both karyotypes and supporting a close phylogenetic relationship between these species. A break of colinearity was observed between BAC clones and the CMA+ bands (including rDNA loci) from chromosomes 2 and 3, which could be mainly explained by amplification and reduction of the satellite DNA sequences that constitute the heterochromatic bands of these chromosome pairs.

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Acknowledgments

We would like to thank Luiz Gustavo R. Souza and two anonymous referees for the valuable comments on the manuscript and Getúlio de Souza Vieira (Embrapa Cassava & Fruits) for his technical support in the field. This work was carried out with financial support from the Brazilian agency Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, 474330/2009-5) including a scholarship to S.C.S. (141121/2008-4) and a fellowship to A.P.-H (302953/2010-8).

Ethical standards

Experimental procedures comply with the current state and federal laws.

Conflict of interest

The authors declare that they have no conflict of interest.

Data archiving statement

The cytogenetic map has been submitted to and archived in the Citrus Genome Database (http://www.citrusgenomedb.org/tgg/data_submission).

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Correspondence to Andrea Pedrosa-Harand.

Additional information

Communicated by W.-W. Guo

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Supplemental Fig. S1

Flow cytometric DNA content (2C value) analysis of Citrus reticulata cv. Cravo using Raphanus sativus L. (2C = 1.11 pg; Doležel et al. 1992) as an internal reference standard. The coefficient of variation (CV) for mandarin ranged from 3.5 % to 4.3 %, while the standard CV ranged from 2.8 % to 3.8 %.

Supplemental Fig. S2

Mitotic metaphases from Citrus reticulata cv. Cravo stained with CMA/DAPI and hybridized in situ with 45S rDNA (ab) and different Poncirus trifoliata BACs (cf). In a and b, only CMA staining and Cy3 signals are shown, respectively, without superimposition with DAPI counterstained chromosomes. The inserts show the enlarged chromosomes pseudocoloured in gray and the BAC signals in red and green. The numbers identify the labelled chromosome pair in each case. Bar in f corresponds to 5 μm.

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da Costa Silva, S., Mendes, S., Soares Filho, W.d.S. et al. Chromosome homologies between Citrus and Poncirus—the comparative cytogenetic map of mandarin (Citrus reticulata). Tree Genetics & Genomes 11, 811 (2015). https://doi.org/10.1007/s11295-014-0811-4

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Keywords

  • BAC-FISH
  • Heteromorphism
  • Synteny
  • Heterochromatin
  • CMA/DAPI