Ruas 2015 (Conference Paper) Bromeliaceae

Cross-amplification of 10 microsatellites loci in bromeliad species from Pantanal and Chaco, Mato Grosso do Sul, Brazil

Author(s):—R. Ruas & G. Paggi in Benko-Iseppon, A.M.; Alves, M. & Louzada, R. (2015) An overview and abstracts of the First World Congress on Bromeliaceae Evolution. Rodriguésia 66(2): A1-A66.

Publication:— (2015).

Abstract:—The Bromeliaceae family have ca. 3,000 species, traditionally subdivided into the subfamilies Pitcairnioideae,Tillandsioideae and Bromelioideae, which are exclusively distributed in Americas. Bromeliads have been threatened by anthropic pressures due to predatory gathering and habitat destruction, being used as ornamental, forage, food and medical uses. Ironstone outcrops may be considered a peculiar feature of Corumbá and Ladário cities, which are located in the Pantanal wetlands of the state of Mato Grosso do Sul State (MS), Brazil. Chaco or ?Gran Chaco? is the term applied to seasonal vegetation, which is divided into an eastern sector, with a more humid climate, and a western, drier sector, whilst the submeridional lowlands include a large depression between these two areas. In Brazil, Chaco vegetation is restricted to the western border of MS, in Porto Murtinho. The bromeliads from the genera Aechmea, Bromelia, Deuterocohnia, Dyckia and Tillandsia, in these wetland regions, occur as terrestrial, rupicolous or epiphytic plants in these biomes. In this study, we used nine bromeliad species, Bromelioideae - Aechmea distichantha Lem., Bromelia balansae Mez, B. hieronyni Mez; Pitcairnioideae - Deuterocohnia meziana Kuntze ex Mez, Dyckia excelsa Leme, D. leptostachya Baker; Tillandsioideae -Tillandisa bulchlohii Rauh, T. loliaceae Mart. ex Schult. & Schult.f. and T. streptocarpa Baker in order to evaluate the cross-amplification rate of 10 microsatellites loci developed for other bromeliad species: Dyckia distachya, Dyckia marnier-lapostollei var. estevesii (Pitcairnioideae), Ananas comosus and Aechmea caudata (Bromelioideae). We conducted DNA extractions using the CTAB protocol in a sample of 2 individuals of each species (n = 18). We performed PCR reactions following the protocols previously developed by our research group. The PCR products were analyzed on 1% agarose gels stained withSyberGreen (Invitrogen) and visualized on a blue light transilluminator. For Pitcairnioideae species, 80% of primers amplified satisfactorily (4 from 5 primers), without any correlation with the primer original subfamily(Pitcairnioideae or Bromelioideae). For Bromelioideae species, on average, 60% of primers amplified satisfactorily (3 from 5 primers of Pitcairnioideae primers), and 87% of primers amplified satisfactorily (4.3 from 5 primers of Bromelioideae primers). For Tillandisioideae species, none of the primers amplified adequately. In B. balansae nine from 10 primers amplified with expected fragment sizes. Aechmea distichantha, D. meziana and D. excelsa presented 80% of amplification. A good set of polymorphic microsatellite loci are necessary to perform various studies concerning population genetics and conservation. Determining cross-amplificationprotocols for microsatellite loci in different native bromeliad species would have great relevance for research in different areas such as, population genetics (genetic diversity and structure), gene flow, that will produced data very important for diagnosis of natural populations viability and their conservation.

Keywords:—Bromeliaceae; Conservation; Wetlands.