Genetic Analyses of Casuarinas Using ISSR and FISSR Markers

Abstract

Inter simple sequence repeat polymerase chain reaction (ISSR-PCR) was used for the genetic analysis of the six species of Allocasuarina, five species of Casuarina and 12 superior performing selections of C. equisetifolia L. We also fingerprinted C. equisetifolia L. selections using Fluorescent-ISSR-PCR (FISSR-PCR), an improvised ISSR-PCR assay. The ISSR analysis provided information on the frequency of various simple sequence repeats in the casuarina genome. The di-nucleotide repeats were more common, among which (CA) n and its complementary nucleotide (GT) n repeat motifs amplified relatively higher number of bands with an average of 6.0 ± 3.5 and 6.3 ± 1.8 respectively. Eleven species of casuarinas were amplified with 10 primers anchored either at 5¢ or 3¢ end. A total of 253 PCR products were obtained and all were polymorphic, out of which 48 were specific to Allocasuarina and 36 were specific to Casuarina genus. Genetic similarity among the species was 0.251. A UPGMA dendrogram grouped all the Casuarina species together. The 12 superior performing selections of C. equisetifolia L. produced 57 polymorphic ISSR markers while the FISSR assay revealed 105 polymorphic markers. The primer CRR(ATT) 4 distinguished all the selections. DNA profiles obtained with ISSR and FISSR assays would serve as a reference library for the establishment of clonal identity in casuarinas.

Discrimination of Calophyllum inophyllum L. Progenies Through ISSR Markers

“…Extensive studies have already been reported in many tropical trees viz., Pongamia pinnata (Shyam Sundar et al, 2014); Casuarinas (Yasodha and Kathirvel, 2004) ISSR techniques was used to study the genetic structure and relationship between species and populations. Similarly, DNA based ISSR techniques have been successfully utilized in many genera and species of forest trees (Mohapatra and Singhal, 2000) for diversity study.…”

Genetic variation of Casuarina equisetifolia subsp equisetifolia and C. equisetifolia subsp incana populations on the northern coast of Senegal

“…This conclusion supports the hypothesis that the plantation in this area was probably created in an effort to solve an immediate environmental problem such as windbreaks and sand-shifting control than for genetic variation. High genetic variation in C. equisetifolia provenances was reported by several studies by using morphological and molecular markers alone or in combination (Pinyopusarerk and Williams, 2000;Yasodha et al, 2004;Kamalakannan et al, 2006). Studies based on amplified fragment length polymorphism (AFLP) markers revealed a high genetic variation among provenances of C. equisetifolia (Huang et al, 2008) and other tropical trees such as Moringa oleifera (Muluvi et al, 1999) and Tectona grandis (Shrestha et al, 2005).…”

“…Because it is salt and drought tolerant, and fast growing, C. equisetifolia is also used as windbreaks and sand-shifting control in many parts of the world such as on the northern coast of Senegal (9700 ha) and Southern China (30,000 ha) (Mailly et al, 1994;Zhong et al, 2005). Due to its importance, there is a major effort underway focused on selecting and planting elite clones of C. equisetifolia in Australia and India, but the efficiency of this approach requires the genetic characterization of available genetic resources (Yasodha et al, 2004).…”

“…Other DNA markers such as ISSR (inter-simple sequence repeat), fluorescent-ISSR or combined with morphological traits were widely used for assessing the genetic diversity of C. equisetifolia. They showed polymorphism between clones of C. equisetifolia from Australia and suggested that morphological markers can be used in combination with molecular markers in genetic variability assessment and breeding programs (Yasodha et al, 2004;Kamalakannan et al, 2006). Genetic variation at the population level was also studied in a C. equisetifolia plantation in Taiwan by using random amplified polymorphic DNA (RAPD) markers.…”

Development of SCAR marker in Casuarina equisetifolia for species authentication

“…Seven ISSR primers were custom synthesized (SigmaAldrich, USA) for the study. The primers were selected based on the earlier report by Yasodha et al (2004) ( Table 4). PCR amplifications were performed in a 10 ll reaction volume containing about 30 ng of template DNA, 1.0 ll 109 PCR buffer (Bangalore Genei Ltd, India), 2.5 mM MgCl 2 , 0.4 mM dNTPs, 100 nM primer and 0.3 U Taq DNA polymerase (Bangalore Genei Ltd, India).…”