Australian Systematic Botany Australian Systematic Botany Society
Taxonomy, biogeography and evolution of plants

Phylogenetic analysis of the subtribe Chloraeinae (Orchidaceae): a preliminary approach based on three chloroplast markers

M. Amelia Chemisquy A B and Osvaldo Morrone A
+ Author Affiliations
- Author Affiliations

A Instituto de Botánica Darwinion (CONICET, ANCEFN), Labardén 200, Casilla de Correo 22, B1642HYD San Isidro, Buenos Aires, Argentina.

B Corresponding author. Email:

Australian Systematic Botany 23(1) 38-46
Submitted: 2 June 2009  Accepted: 19 November 2009   Published: 17 February 2010


The systematic position and relationships between some South American terrestrial orchids, such as Bipinnula Comm. ex Juss., Chloraea Lindl., Gavilea Poepp. and Geoblasta Barb. Rodr., is unclear. These four genera have been grouped in the subtribe Chloraeinae by several authors. Previous phylogenetic studies of the group have included only a few species of Chloraea and Gavilea and not of Bipinnula or Geoblasta. Relationships among these four genera were explored and the monophyly of the subtribe Chloraeinae and the genera Chloraea and Gavilea were tested in this contribution. Molecular phylogenetic analyses were conducted, using the following three chloroplast markers: the matK–trnK intron, the atpB–rbcL spacer and the rpoC1 gene. Sequences were analysed under maximum parsimony and Bayesian inference. In all the analyses, Bipinnula, Chloraea, Gavilea and Geoblasta were grouped in a clade with high support, where Bipinnula, Geoblasta and Gavilea were nested inside Chloraea. Consequently, Chloraea was paraphyletic, whereas Gavilea turned out to be monophyletic with high values of support. The other species of tribe Cranichideae appeared as sister groups of the Chloraeinae. A more exhaustive taxonomic sampling is needed to resolve the systematic placement of the subtribe Chloraeinae and the internal relationships between the genera and species that form it.


We thank James Solomon and Alyse Rothrock (Missoury Botanical Garden) and Natalia Álvarez, Manuel Belgrano, Silvana Sede and Fernando Zuloaga (Instituto de Botánica Darwinion) for the tissue samples; Lone Aagesen and Liliana Giussani for critically reading the manuscript; Mariana Valente for helping with the figures; Victoria Gonzalez Eusevi for checking the English; Pancho Prevosti for helping with some analyses and for the stimulating discussion of the results and the methods. We also thank two anonymous reviewers and the editors for the comments that helped improve this manuscript. Field trips were possible thanks to the Myndel Botanica Foundation. CONICET provided financial support.


Ackerman JD, Williams NH (1981) Pollen morphology of the Chloraeinae (Orchidaceae: Diurideae) and related subtribes. American Journal of Botany 68, 1392–1402.
CrossRef |

Burns-Balogh P, Funk VA (1986a) A phylogenetic analysis of the Orchidaceae. Smithsonian Contributions to Botany 61, 1–71.

Burns-Balogh P, Funk VA (1986b) A phylogenetic analysis of the Orchidaceae: a summary. Lindleyana 1, 131–139.

Cameron KM (2004) Utility of plastid psaB gene sequences for investigating intrafamilial relationships within Orchidaceae. Molecular Phylogenetics and Evolution 31, 1157–1180.
CrossRef | CAS | PubMed |

Cameron KM (2006) A comparison and combination of plastid atpB and rbcL gene sequences for inferring phylogenetic relationships within Orchidaceae. Aliso 22, 447–464.

Cameron KM, Chase MW, Whitten WM, Kores PJ, Jarrell DC, Albert VA, Yukawa T, Hills HG, Goldman DH (1999) A phylogenetic analysis of the Orchidaceae: evidence from rbcL nucleotide sequences. American Journal of Botany 86, 208–224.
CrossRef |

Clements MA, Jones DL, Sharma IK, Nightingale ME, Garratt MJ, Fitzgerald KJ, Mackenzie AM, Molloy BPJ (2002) Phylogenetics of Diurideae (Orchidaceae) based on the internal transcribed spacer (ITS) regions of nuclear ribosomal DNA. Lindleyana 17, 135–171.

Correa MN (1968) Rehabilitación del género Geoblasta Barb. Rodr. Revista del Museo de La Plata. Sección Botánica 11, 69–74.

Correa MN (1969) Chloraea, género sudamericano de Orchidaceae. Darwiniana 15, 374–500.

Cribb PJ, Kores PJ (2000) The systematic position of Codonorchis (Orchidaceae: Orchidoideae). Lindleyana 15, 169–170.

Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin 19, 11–15.

Dressler RL (1981) ‘The orchids. Natural history and classification.’ (Harvard University Press: Cambridge, MA)

Dressler RL (1993) ‘Phylogeny and classification of the orchid family.’ (Dioscorides Press: Portland, OR)

Freudenstein JV, Rasmussen FN (1999) What does morphology tell us about orchid relationships? A cladistic analysis. American Journal of Botany 86, 225–248.
CrossRef |

Freudenstein JV, van den Berg C, Goldman DH, Kores PJ, Molvray M, Chase MW (2004) An expanded plastid DNA phylogeny of Orchidaceae and analysis of jackknife branch support strategy. American Journal of Botany 91, 149–157.
CrossRef | CAS |

Goldman DH, Freudenstein JV, Kores PJ, Molvray M, Jarrell DC, Whitten WM, Cameron KM, Jansen RK, Chase MW (2001) Phylogenetics of Arethuseae (Orchidaceae) based on plastid matK and rbcL sequences. Systematic Botany 26, 670–695.

Goloboff PA (1993) Estimating character weights during tree search. Cladistics 9, 83–91.
CrossRef |

Goloboff PA, Farris JS, Källersjö M, Oxelman B, Ramírez MJ, Szumik CA (2003) Improvements to resampling measures of group support. Cladistics 19, 324–332.
CrossRef |

Goloboff PA, Farris JS, Nixon K (2008a) TNT, a free program for phylogenetic analysis. Cladistics 24, 774–786.
CrossRef |

Goloboff PA, Carpenter JM, Arias JS, Miranda Esquivel DR (2008b) Weighting against homoplasy improves phylogenetic analysis of morphological data sets. Cladistics 24, 758–773.
CrossRef |

Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series 41, 95–98.

Hauman L (1920) Orchidées argentines. Anales de la Sociedad Científica Argentina 90, 95–154.

Hauman L (1922) La distribución geográfica del género Chloraea. Physis (Buenos Aires) 5, 293–295.

Hoehne FC (1940) Orchidaceas. Gêneros 1. Selenipedium – 1a. Triphora. Flora Brasílica 12, 1–254.

Johnson LA, Soltis DE (1994) matK DNA sequences and phylogenetic reconstruction in Saxifragaceae Systematic Botany 19, 143–156.
CrossRef |

Jones DL, Clements MA, Sharma IK, Mackenzie AM, Molloy BPJ (2002) Nomenclatural notes arising from studies into the tribe Diurideae (Orchidaceae). Orchadian 13, 436–468.

Källersjö M, Albert VA, Farris JS (1999) Homoplasy increases phylogenetic structure. Cladistics 15, 91–93.
CrossRef |

Kores PJ , Weston PH , Molvray M , Chase MW (2000) Phylogenetic relationships within the Diurideae (Orchidaceae): inferences from plastid matK DNA sequences. In ‘Monocots systematics and evolution’. (Eds KL Wilson, DA Morrison) pp. 449–456. (CSIRO: Melbourne)

Kores PJ, Molvray M, Weston PH, Hopper SD, Brown A, Cameron KM, Chase MW (2001) A phylogenetic analysis of Diurideae (Orchidaceae) based on plastid DNA sequence data. American Journal of Botany 88, 1903–1914.
CrossRef | CAS |

Kraenzlin F (1888) Orchidaceae herbariii Dom. J. Arechavaletae det. et descr. Botanische Jahrbücher für Systematik 9, 315–318.

Morgenstern B (2004) DIALIGN: multiple DNA and protein sequence alignment at BiBiServ. Nucleic Acids Research 32, W33–W36.
CrossRef | CAS | PubMed |

Müller K (2005) SeqState – primer design and sequence statistics for phylogenetic DNA data sets. Applied Bioinformatics 4, 65–69.
CrossRef | PubMed |

Pfeiffer L (1873) ‘Nomenclator Botanicus Vol. I.’ (Sumptibus Theodori Fischeri: Kassel)

Posada D, Crandall KA (1998) MODELTEST: testing the model of DNA substitution. Bioinformatics 14, 817–818.
CrossRef | CAS | PubMed |

Pridgeon A , Cribb PJ , Chase MW , Rasmussen FN (2001) ‘Genera Orchidacearum, Volume 3: Orchidoideae (Part one).’ (Oxford University Press: Oxford, UK)

Pridgeon A , Cribb PJ , Chase MW , Rasmussen FN (2003) ‘Genera Orchidacearum, Volume 3: Orchidoideae (Part two) – Vanilloideae.’ (Oxford University Press: Oxford, UK)

Ronquist F, Huelsenbeck JP (2003) MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 19, 1572–1574.
CrossRef | CAS | PubMed |

Salazar GA, Cabrera LI, Madriñán S, Chase MW (2009) Phylogenetic relationships of Cranichidinae and Prescottiinae (Orchidaceae, Cranichideae) inferred from plastid and nuclear DNA sequences. Annals of Botany 104, 403–416.
CrossRef | CAS | PubMed |

Simmons MP, Ochotorena H (2000) Gaps as character in sequenced-based phylogenetic analysis. Systematic Biology 49, 369–381.
CrossRef | CAS | PubMed |

Szlachetko DL (1995) Systema Orchidalium. Fragmenta Floristica et Geobotanica 3(Suppl.), 1–152.

Szlachetko DL, Margońska HB (2001) Genera et species Orchidalium. 4. Polish Botanical Journal 46, 123–125.

Szlachetko DL, Tukałło P (2008) Notes on subtribe Chloraeinae (Orchidaceae). Acta Societatis Botanicorum Poloniae 77, 111–116.

Whitten WM, Blanco MA, Williams NH, Koehler S, Carnevali G, Singer RB, Enadara L, Neubig KM (2007) Molecular phylogenetics of Maxillaria and related genera (Orchidaceae: Cymbideae) based on combined molecular data sets. American Journal of Botany 94, 1860–1889.
CrossRef | CAS |

Appendix 1.  Taxa and vouchers for species sampled
Species; geographic origin; collector and collection number (herbarium); atpB–rbcL GenBank accession number/matK–trnK GenBank accession number/rpoC1 GenBank accession number
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