Genetic Relationships Of Phragmipedium Species

LANKESTERIANA 7(3): 493-496. 2007.

MANUEL RODRÍGUEZ SALAS1,2, JORGE BENAVIDES RANILLA1 & JOSE R. ESPINOZA1
Molecular Biotechnology Unit, Laboratories for Research & Development (LID) Universidad Peruana Cayetano Heredia, Av. Honorio Delgado 430 Lima 31, Perú 2 Author for correspondence: manuel_rodsa@hotmail.com
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GENETIC RELATIONSHIPS OF PHRAGMIPEDIUM SPECIES (ORCHIDACEAE) USING AMPLIFIEDFRAGMENT LENGTH POLYMORPHISM (AFLP) ANALYSIS

ABSTRACT. The objective of this study is to evaluate the genetic relationships of eight Peruvian species of the genus Phragmipedium using amplified fragment length polymorphisms and a phenetic analysis (UPGMA). The analysis showed that the species are phenetically grouped in four large clusters. The first cluster includes species from theHimantopetalum (P. pearcei and P. richteri) and Lorifolia (P. boissierianum) sections. The second cluster comprised the species of the Micropetalum section (P. besseae and P. schlimii). The third one included P. kovachii in the Schluckebieria section. The fourth cluster contains species of the Phragmipedium (P. caudatum and P. wallisii) section. The results show that AFLP analysis is a powerful genetic toolfor the analysis of the taxonomy of orchids from the diverse and complex Phragmipedium genus. RESUMEN. El presente estudio tiene como objetivo evaluar la diversidad y distancia genética de 8 especies del género Phragmipedium usando la técnica de amplified fragment length polymorphisms (AFLP) y el método de análisis fenético (UPGMA). Las especies son agrupadas feneticamente en cuatro clustersmayores. El primer cluster comprende especies de las secciones Himantopetalum (P. pearcei y P. richteri) y Lorifolia (P. boissierianum). El segundo cluster incluye las especies de la sección Micropetalum (P. besseae y P. schlimii). El tercer cluster contiene a la especie de la sección Schluckebieria (P. kovachii). El cuarto cluster contiene a las especies de la sección Phragmipedium (P. caudatum y P.wallisii). Los resultados muestran que AFLP es una herramienta genética poderosa para el análisis taxonómico del diverso y complejo género Phragmipedium..

K EY WORDS : Orchidaceae, DNA markers, Phragmipedium, Phragmipedium kovachii, AFLP, Fingerprinting. The orchid genus Phragmipedium Rolfe comprises species distributed in six sections including; Phragmipedium, Himantopetalum, Platypetalum,Lorifolia, Micropetalum, and Schluckebieria.(Gruss 2003; Braem 2004). Concerns about the taxonomic placement of some species in this genus have frequently been raised in this genus, since some accessions show morphological characteristics that are intermediate between different species. Improvements in the understanding of taxonomic relationships are likely to occur with the extensive analysis ofmolecular markers that unravel the genetic relationships and allow us to produce a phylogeny of the new species, thus aiding the identification of varieties. Introduction

Analysis of the genetic diversity of endangered plant species is one of the key elements in the estimation of the viability of its populations, and in the establishment of efficient in situ as well as ex situ preservation. TheAFLP (amplified fragment length polymorphism) method (Vos et al. 1995) is a valuable DNAbased technique that informs about multiple polymorphic anonymous loci, generally in the nuclear genome of a species and can be used to determine genetic diversity in many plant species, particularly when there are few characterized molecular genetic markers (Mackil et al.1996; Segovia-Lerma et al. 2003; Nguyenet al. 2004) The objective of this study was to evaluate genetic distance among 8 species of genus Phragmipedium by using AFLP analysis

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LANKESTERIANA

P LANT MATERIAL . Plants were maintained under greenhouse conditions in the orchid collection at Peruanino Nursery (Vivero Peruanino) located in San Ramon, Region Junín. The plants were labeled by assigning an individual code in order...