PCR-RFLP analysis of non-coding regions of cpDNA in Araucaria angustifolia (Bert.) O. Kuntze

Abstract

The Araucaria angustifolia (Bert.) O. Kuntze, also named the "paraná pine" ('pinheiro-do-Paraná' in Portuguese), is a native conifer species naturally occurring in the Brazilian Tropical Atlantic Forest which in Brazil is mostly limited to the southern Brazilian states of Paraná, Santa Catarina and Rio Grande do Sul. Chloroplast DNA markers (cpDNA) are useful in populational genetic studies because of their low substitution rate and the uniparental transmission. The conservation of cpDNA genes between species has allowed the design of consensus chloroplast primers that have had a great impact on population genetics and phylogenetic studies. In this study we used the polymerase chain reaction technique combined with restriction enzyme fragment length polymorphism (PCR-RFLP) to characterize the genetic diversity of the chloroplast genome in nine natural A. angustifolia populations. Among the 141 trees surveyed we found 12 different cpDNA haplotypes and demonstrated that A. angustifolia has high levels of total diversity (h T = 0.612) and an average within-population diversity (h S ) of 0.441, suggesting the presence of high withinpopulation variation. The estimated genetic divergence could be helpful in designing breeding programs and species conservation strategies, although additional studies with a larger number of populations and trees is essential for a better understanding of gene flow and the inheritance of major Araucaria angustifolia traits.

Evidences of delayed size recovery in Araucaria angustifolia populations after post-glacial colonization of highlands in Southeastern Brazil

“…Shimizu and Higa 1980, Auler et al 2002, Sousa et al 2004, Mantovani et al 2006, Stefenon et al 2007, Schögl et al 2007). However, little is known about the relationship between historical demography and the current genetic structure of this species.…”

pH do Exsudato na Avaliação da Viabilidade de Sementes de Araucaria angustifolia

“…é a única do gênero nativa do Brasil, com grande relevância ecológica, econômica e social na sua região de ocorrência natural (Schlögl et al, 2007). Por isso, diversos esforços têm sido conduzidos a fim de propagar e conservar a espécie (Balbuena et al, 2011).…”

PCR‐RFLP and DAMD‐PCR genotyping for Salvia species

“…14 -16 Among the DNA-based methods, restriction fragment length polymorphism (RFLP) is one of the first molecular genetic techniques used in different organisms for genetic mapping and genotyping studies. 17,18 However, RFLP has several limitations in comparison with various polymerase chain reaction (PCR)-based DNA marker techniques. 19 RFLP requires high-quality DNA (degradation of the DNA can lead to inconclusive results), requires a considerable amount of DNA and usually produces a lower number of polymorphic markers in comparison with simple sequence repeat length polymorphism (SSRLP), amplified fragment length polymorphism (AFLP) and random amplified polymorphic DNA (RAPD) techniques.…”

“…20 -22 However, a number of laboratory techniques have been developed in recent years to improve RFLP techniques. 17,18 One of these methods involves the use of different DNA restriction endonuclease enzymes to digest DNA molecules of varying length depending on the presence or absence of restriction sites. The length variation in DNA fragments caused when the restriction enzymes digest their specific cutting sites is called RFLP, a method for amplifying and enriching the sample within a targeted region of DNA sequences.…”

“…However, a separation step is required to resolve the restriction fragments from one another and measure their lengths after the endonuclease enzymatic reactions. 17,18 SSRLP is the most promising molecular technique for identifying genotypes and molecular mapping studies. 13 It produces a high level of polymorphic and co-dominant markers.…”