Clones identification of Sequoia sempervirens (D. Don) Endl. in Chile by using PCR-RAPDs technique

Abstract

A protocol of polymerase chain reaction-random amplified polymorphic DNAs (PCR-RAPDs) was established to analyse the gene diversity and genotype identification for clones of Sequoia sempervirens (D. Don) Endl. in Chile. Ten (out of 34) clones from introduction trial located in Voipir-Villarrica, Chile, were studied. The PCR-RAPDs technique and a modified hexadecyltrimethylammonium bromide (CTAB) protocol were used for genomic DNA extraction. The PCR tests were carried out employing 10-mer random primers. The amplification products were detected by electrophoresis in agarose gels. Forty nine polymorphic bands were obtained with the selected primers (BG04, BF07, BF12, BF13, and BF14) and were ordered according to their molecular size. The genetic similarity between samples was calculated by the Jaccard index and a dendrogram was constructed using a cluster analysis of unweighted pair group method using arithmetic averages (UPGMA). Of the primers tested, 5 (out of 60) RAPD primers were selected for their reproducibility and high polymorphism. A total of 49 polymorphic RAPD bands were detected out of 252 bands. The genetic similarity analysis demonstrates an extensive genetic variability between the tested clones and the dendrogram depicts the genetic relationships among the clones, suggesting a geographic relationship. The results indicate that the RAPD markers permitted the identification of the assayed clones, although they are derived from the same geographic origin.

Genetic structure of coast redwood (Sequoia sempervirens [D. Don] Endl.) populations in and outside of the natural distribution range based on nuclear and chloroplast microsatellite markers

“…The reliable clone identification based on microsatellites in data set G ( S6 Fig ) confirmed results of previous studies based on allozyme and AFLP markers [ 27 , 91 ]. General difficulties to accurately genotype microsatellite markers in polyploid organisms excludes analyses based on their allele frequencies [ 35 , 92 – 95 ].…”

“…The presented study is the first study investigating coast redwood populations within and outside the natural distribution range with a high number of nuclear SSR markers and chloroplast SSRs. The new and significantly expanded marker set confirmed results of previous studies [ 27 , 28 , 31 ]. Surprisingly, neither the novel nuclear nor chloroplast microsatellite markers could improve resolution of population differentiation within the natural distribution range of coast redwood despite high polymorphism of the used microsatellite markers.…”

“…Additionally, these markers could be very efficient to assign introduced plants to their origin [4,20,21], but the detection power depends much on population differentiation [21,22]. Until now, previous coast redwood studies were not able to accomplish that based on the available allozyme and microsatellite markers [23][24][25][26][27][28][29][30][31].…”

Analysis of genetic diversity in banana cultivars (Musa cvs.) from the South of Oman using AFLP markers and classification by phylogenetic, hierarchical clustering and principal component analyses

“…Researchers have used several DNA fingerprinting techniques for the correct identification of plant species and cultivars. These DNA fingerprinting techniques have successfully used markers such as randomly amplified polymorphic DNA marker (RAPD) (Toral Ibañez et al, 2009;Ude et al, 2003;Gawel and Jarret, 1992), restriction fragment length polymorphism (RFLP) (Bhat et al, 1995), microsatellites (Grapin et al, 1998), inter simple sequence repeat (ISSR) (Godwin et al, 1997), and amplified fragment length polymorphism (AFLP) (Bhat et al, 2004;Ude et al, 2003). Among many researchers, AFLP is the marker technology of choice since it combines the reliability of classical restriction-based fingerprinting with the speed and convenience of polymerase chain reaction (PCR)-based marker techniques (Vos et al, 1995;Powell et al, 1996;Lu et al, 2002;Ude et al, 2002a;2002b).…”

Concentración de kinetina y tipo de explante en la multiplicación in vitro de Sequoia sempervirens (D. Don). Endl.

“…pertenece a la familia Taxodiaceae; especie poliploide (2n = 6x = 66) (Ahuja, 2005), originaria de la costa oeste de California y suroeste de Oregón, Estados Unidos (Olson et al, 1990). Se ha introducido como especie de ornato en: Nueva Zelanda (Davies et al, 2014), Brasil (Ribeiro et al, 2011), Rumania (Clapa et al, 2010), Chile (Toral Ibañez et al, 2009), Italia (Ozudogru et al, 2011) y España (Rojas- Vargas et al, 2021).…”