DNA barcoding of perennial fruit tree species of agronomic interest in the genus Annona (Annonaceae).

Nerea Larranaga,José I Hormaza

doi:10.3389/fpls.2015.00589

Abstract

The DNA barcode initiative aims to establish a universal protocol using short genetic sequences to discriminate among animal and plant species. Although many markers have been proposed to become the barcode of plants, the Consortium for the Barcode of Life (CBOL) Plant Working Group recommended using as a core the combination of two portions of plastid coding region, rbcL and matK. In this paper, specific markers based on matK sequences were developed for 7 closely related Annona species of agronomic interest (Annona cherimola, A. reticulata, A. squamosa, A. muricata, A. macroprophyllata, A. glabra, and A. purpurea) and the discrimination power of both rbcL and matK was tested using also sequences of the genus Annona available in the Barcode of Life Database (BOLD) data systems. The specific sequences developed allowed the discrimination among all those species tested. Moreover, the primers generated were validated in six additional species of the genus (A. liebmanniana, A. longiflora, A. montana, A. senegalensis, A. emarginata and A. neosalicifolia) and in an interspecific hybrid (A. cherimola x A. squamosa). The development of a fast, reliable and economic approach for species identification in these underutilized subtropical fruit crops in a very initial state of domestication is of great importance in order to optimize genetic resource management.

Highlights

Annona L. is the type genus among the approximately 110 included in the Annonaceae (Chatrou et al, 2012), an angiosperm family within the Magnoliales in the Magnoliid clade (APG, 2009)
In order to have a fast and reliable method to discern between closely related Annona species with agronomic interest, in this work we developed species specific primers to unequivocally differentiate the seven most common agronomically interesting Annona species present in Central and South America (Annona cherimola, A. reticulata, A. squamosa, A. muricata, A. macroprophyllata, A. glabra and A. purpurea) based on newly sequenced data
The results obtained by distance analysis methods with 21 genotypes belonging to 7 agronomical important Annona species showed almost no intraspecific variation, which could be due to the limited number of samples analyzed

Summary

Introduction

Annona L. is the type genus among the approximately 110 included in the Annonaceae (Chatrou et al, 2012), an angiosperm family within the Magnoliales in the Magnoliid clade (APG, 2009). Different molecular markers based on both chloroplast and nuclear sequences have been used to infer phylogenetic relationships among species of the family. Additional molecular markers have been developed in the Annona genus mainly for fingerprinting and genetic diversity studies; isozymes (Ellstrand and Lee, 1987; Pascual et al, 1993; Perfectti and Pascual, 1998, 2004, 2005), randomly amplified polymorphic DNA (RAPDs) (Ronning et al, 1995), amplified fragment length polymorphism (AFLPs) (Rahman et al, 1998)

Methods

Results

Conclusion