Abstract
The strength of sequence-related amplified polymorphism (SRAP) in comparison to random amplified polymorphic DNA (RAPD) in determining the genetic polymorphism among seven species of entomopathogenic nematodes (Steinernematidae) was tested. Nine RAPD and 12 SRAP primer pair’s combinations were used. The number of polymorphic bands and the polymorphism percentages was high in SRAP analysis (97 out of 107 bands were polymorphic (90.6%)) compared to RAPD (65 out of 89 bands were polymorphic (73%)). The highest number of RAPD bands was recorded for OPE-A-07 primer (14 band) while OPE-D-20 was scored the lowest band number (7 bands). The SRAP primers Me3-em2 registered the highest number of bands (13 bands) while Me3-em3and Me2-em2 showed the lowest band number (6 bands). The genotype-specific SRAP and RAPD markers for the different ENP species were recorded. The highest number of SRAP specific markers (7 markers) was scored for Steinernema glaseri, then 4 markers for S. abbasi 2 followed by S. riobrave (3 markers), 1 marker for S. abbasi 1, S. riobrave recorded the highest number of specific RAPD marker (4 specific markers) followed by S. carpocapsae DD 136 and S. glaseri (two specific markers), then only 1 specific marker for S. abbasi 1. Based on the data obtained from RAPD and SRAP analysis, the dendrogram was created to clarify the genetic distances among different species of the studied entomopathogenic nematodes. The present study indicated that SRAP was more powerful than RAPD analysis for detecting the genetic polymorphism among closely related species of nematodes. The genotype-specific markers detected from both RAPD and SRAP can be used in future biological control programs.
Highlights
Entomopathogenic nematodes (EPNs) are a promising candidate for biological control of insects due to their ability to find hosts, integrity to non-target organisms and the environment, high reproductive potential, capacity to be mass-produced and to be used with other agrochemical pesticides and fertilizers (Laznik and Trdan 2014)
Nine random random amplified polymorphic DNA (RAPD) primers were used to evaluate the genetic polymorphism among the seven EPN species which generated 65 bands, 89 (73%) of them were polymorphic
Padmanaban et al (2014) developed RAPD markers to discriminate the genetic diversity in 25 recovered EPNs (Steinernema spp. and Heterorhabditis spp.) and found that polymorphism percentage that recorded in all the strains was 12.31%
Summary
Entomopathogenic nematodes (EPNs) are a promising candidate for biological control of insects due to their ability to find hosts, integrity to non-target organisms and the environment, high reproductive potential, capacity to be mass-produced and to be used with other agrochemical pesticides and fertilizers (Laznik and Trdan 2014). RAPD has been used for the genetic variability and phylogenetic studies of different organisms including insects, EPNs, animals, and plants (Moghaieb et al 2004). RAPD analysis was used for the characterizations of a new species of Heterorhabditis from Hawaii (Gardner et al 1994). It has been shown to be more instructive than other PCR-based techniques in discovering genetic diversity (Budak et al 2004) and has been used to study the genetic diversity and relationships among several species (Abedian et al 2012)
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