Comparative Estimation of Genetic Diversity in Population Studies using Molecular Sampling and Traditional Sampling Methods.

Amr Tm Saeb,Satish Kumar David

doi:10.6026/97320630010347

Abstract

Entomopathogenic nematodes (EPN) are efficient biological pest control agents. Population genetics studies on EPN are seldom known. Therefore, it is of interest to evaluate the significance of molecular sampling method (MSM) for accuracy, time needed, and cost effectiveness over traditional sampling method (TSM). The study was conducted at the Mohican Hills golf course at the state of Ohio where the EPN H. bacteriophora has been monitored for 18 years. The nematode population occupies an area of approximately 3700 m2 with density range from 0.25-2 per gram soil. Genetic diversity of EPN was studied by molecular sampling method (MSM) and traditional sampling method (TSM) using the mitochondrial gene pcox1. The MSM picked 88% in compared to TSM with only 30% of sequenced cox 1 gene. All studied genetic polymorphism measures (sequence and haplotype) showed high levels of genetic diversity of MSM over TSM. MSM minimizes the chance of mitochondrial genes amplification from non target organisms (insect or other contaminating microorganisms). Moreover, it allows the sampling of more individuals with a reliable and credible representative sample size. Thus, we show that MSM supersedes TSM in labour intensity, time consumption and requirement of no special experience and efficiency.

Highlights

Entomopathogenic nematodes (EPN) and their symbiotic bacteria possess the ability to kill insect pests and are efficient biological pest control agents [1]
Suggested molecular sampling method (MSM) is more efficient in acquiring the nematode target gene compared. This implies that MSM is more cost effective to be used in molecular population genetic studies compared with traditional sampling method (TSM)
Traditional Sampling Method (Dissection for Individual Nematodes) (TSM) A total number of 50 individual first generation hermaphroditic nematodes were collected from the infected larvae and Genetic Diversity Evaluation Table 1 shows a comparative analysis between polymorphic sites achieved via MSM and TSM

Summary

Introduction

Entomopathogenic nematodes (EPN) and their symbiotic bacteria possess the ability to kill insect pests and are efficient biological pest control agents [1]. In order to investigate the genetic diversity between the molecular sampling method (MSM) and traditional sampling method (TSM), the pcox1 genes was isolated, sequenced and analyzed. For comparison using the TSM, a total number of 50 individuals of first generation hermaphroditic nematodes were collected by dissection from the infected larvae and subjected to the molecular genetics analysis following Blouin et al [12].

Results

Conclusion