Entomopathogenic nematodes (EPNs) are a group of biological control agents that are characterized by their ability to search for hosts, safety to non-target insects and environment, and their ability to be used combined with agricultural chemicals. The objectives of this study were to isolate EPNs from agricultural soil in Egypt and study their virulence against the great wax moth, Galleria mellonella L. (Lepidoptera: Pyralidae), for further use in biological control program. Two out of 20 soil samples collected from orchards cultivated with olives and mango were positive for the presence of EPNs, using the Galleria baiting technique. The positive soil samples were sandy clay loam. Sequencing of the internal transcribed spacer (ITS) region indicated that the isolates obtained belong to Heterorhabditis indica. The ITS sequences were submitted to the National Center for Biotechnology Information (NCBI) and registered under the accession nos. MH553167 and MK300683. The efficacy of the isolates was tested on G. mellonella, using different nematodes’ concentrations. Using 50 IJs/larvae from H. indica Aborawash and ERSAG2 showed 100 and 86% mortality rate after 48 h, respectively. The penetration rate reported in dead G. mellonella was 40% at H. indica Aborawash, while it was 35% in case of ERSAG2.
Background
Entomopathogenic nematodes (EPNs) are a group of nematode families, have the ability to search for their hosts, and are considered as promising biological control candidates for insect pests, providing protection to non-target organisms and the environment.
Results
This study was conducted to isolate indigenous EPN isolates from Egyptian agricultural soils for further use in biological control programs and study their genetic polymorphism among the previously isolated isolates under accession no. MH553167 and MK300683 and the new isolate (MH496627), using the start codon targeted (SCoT) marker. One out of 15 soil samples obtained from a banana cultivated field was positive for the presence of EPNs, using the Galleria baiting method. Morphological analysis and sequencing of the internal transcribed spacer (ITS) region suggested that the isolate obtained belongs to Heterorhabditis indica. The sequence of the ITS was submitted to the National Center for Biotechnology Information (NCBI) and registered under accession no. MH496627. Ten SCoT primers were used in the study; the polymorphic bands were 68 out of 76 with 89% as polymorphism percentage. The highest numbers of bands were 10 bands generated by SCoT 1 and SCoT 18 while SCoT 48 and SCoT 60 recorded the lowest band number (5 bands).
Conclusions
The present study is considered as a preliminary study to demonstrate the effectiveness of the SCoT marker for the first time in assessing genetic relationships in EPNs.
Background
Entomopathogenic nematodes (EPNs) are promising alternatives since they have many characteristics as a biological control agent against insect pests.
Results
Among indigenous EPN isolated, adapted to local environmental conditions by employing the Galleria baiting strategy, only one sample was positive for the presence of EPNs. The new isolate was identified at the species level using DNA sequencing of the internal transcribed spacer region and the Basic Local Alignment Search Tool search of GenBank showed that the isolate had a high similarity (99%) with that sequence available for Heterorhabditis taysearae. The pathogenicity of the EPN isolate was tested against the cotton leaf worm, Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae), using different concentrations (60, 90, 120 and 150 IJs/larvae). Data showed that 150 IJs/larvae caused 100% mortality rate, followed by 120 IJs/larvae (90%), while 60 IJs/larvae showed the lowest rate (60%) after 72 h.
Conclusions
The present study indicated that the native isolate of EPN could be recommended against S. littoralis as an efficient tool in its control programs.
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