Browsing by Author "Wachira, P.M."
Now showing 1 - 3 of 3
- Results Per Page
- Sort Options
Item Biocontrol Potential of Four Indigenous Entomopathogenic Nematodes From Kenya(Canadian Center of Science and Education, 2021-09-15) Ngugi, C.N.; Wachira, P.M.; Mbaka, J.N.; Okoth, S.; Haukeland, S.; Thuranira, E.G.; Kenya Agricultural and Livestock Research Organisation ; University of Nairobi ; International Center of Insect Physiology and EcologyGlobally, vegetable production including tomato is constrained by pests and diseases. The tomato leafminer, Tuta absoluta (Meyrick.), is a major pest of tomato, mainly managed using chemical pesticides. There is a need for integrated pest management (IPM), approaches that are human and environmentally friendly. The study aimed at molecular identification and virulence evaluation of four entomopathogenic nematodes (EPNs) against T. absoluta larvae. The DNA was extracted, sequenced, and phylogeny analyzed. The EPNs virulence was evaluated using the insect baiting technique at different nematode concentrations (0-Control, 100, 150, 200, and 250 infective juveniles/ml of distilled water). Larval mortality data were collected for five days. Molecular identification resulted in nucleotide sequence lengths of 877-895 base pairs (bp). All the isolates were found to be Steinernema species of EPNs. The % similarity of the isolates and their close relatives ranged between 82-100%. The isolates closely matched Steinernema spp. AY230184 (Sri Lanka); AY230186 (Kenya), JN651414 (Ethiopia), and MW151701 (Kenya). The sequences were deposited to Genbank as Steinernema sp. Isolate Kalro 75 (MW150871), Steinernema sp. Isolate Kalro S86 (MW150872), Steinernema sp. isolate Kalro97 (MW150873) and Steinernema sp. Isolate KalroR52 (MW150874). All the EPN isolates were virulent against T. absoluta. Isolate Kalro R52 and Kalro S86 recorded the highest mortality of 76±7.5%; 72±15.0% respectively in the 48 h. There was a significant difference (P < 0.001), between all the isolates and the Control. The four EPNs were virulent and have bio-control potential against, T. absoluta. Research on their virulence against T. absoluta in the field is recommended.Item Characterization and Pathogenicity of Symbiotic Bacteria Associated with Entomopathogenic Nematode: Steinernema Species KALRO(East African Journal of Science, Technology and Innovation, 2021-09-25) Ngugi, C.N.; Wachira, P.M.; Mbaka, J.N.; Okoth, S.; Mangua, P.W.; Kenya Agricultural and Livestock Research Organization (KALRO) ; University of NairobiThe soil inhibiting entomopathogenic nematodes (EPNs), in the family Steinernematidae and Heterorhabditidae, are useful insect biological control agents. They have been used in the management of economically important crop pests. The EPNs are mutually associated with symbiotic bacteria genus Xenorhabdus and Photoharbdus respectively. The study aimed to isolate, characterize and evaluate the pathogenicity of symbiotic bacteria associated with EPN Steinernema sp. Kalro (Accession MW151701). The EPN Steinernema sp. Karlo was multiplied using the insect baiting technique. Its bacteria symbiont was isolated and characterized based on microscopic, biochemical, and physiological features like Gram staining, urease, motility test, and glucose fermentation test). Molecular identification and phylogenetic analysis were performed on 16S rDNA nucleotide sequence. Pathogenicity of the bacteria isolate was evaluated against Tuta absoluta larvae with mortality data recorded after 24 and 48hours of exposure to the bacterial cell suspension. The bacteria were found to be motile and glucose fermentation positive. Sequence analysis of 16S rDNA region resulted in 1500bp sequence with maximum similarity of between 97 and 98.93%, with Xenorhabdus spp Accessions from Genbank. It closely matched to Xenorhabdus sp. My8NJ with 98.93% similarity (Accession AB507811.1). Mean percent larval mortality of 68±4.9 and 88±8.0 in the lowest cell suspension was observed in 24 and 48h of exposure. It’s concluded that, the symbiotic bacteria associated with Steinernema sp. Kalro is Xenorhabdus sp. strain Kalro Genbank Accession MW245845. The bacteria is a potential biological control agent against Tuta absoluta larvae. Further classification of the bacteria to species level and pathogenicity trials in the screen house and field are recommended.Item Characterization and Pathogenicity Test of Entomopathogenic Nematode Steinernema Species-Kalro(Canadian Center of Science and Education, 2021-02-15) Ngugi, C.N.; Wachira, P.M.; Mbaka, J.N.; Okoth, S.; Haukeland, S.; Mburu, H.M.; Adhiambo, M.; Sirengo, D.K.; Kenya Agricultural and Livestock Research Organisation ; University of Nairobi ; International Center of Insect Physiology and EcologyEntomopathogenic nematodes (EPNs) are worldwide soil-dwelling insect parasitic nematodes. They are potential pest bio-control agents a key component of Integrated Pest Management (IPM) programs. This study aimed to characterize and evaluate the pathogenicity of an EPN isolate from Kenya. The nematode was isolated from soils using insect bait technique and both morphological and molecular identification was performed. Efficacy of the isolate was evaluated against Tomato leafminer larvae (Tuta absoluta Meyrick.) using dose-based treatments of 0-control, 100, 150, 200, and 250 infective juveniles (IJs/ml). Morphological analysis revealed body length (L) of 835(659-987) µm and 1781 (1297-2097) µm from fresh IJs and males respectively. Males lacked a mucron. The isolate was characterized by the partial sequence length of 877 bp of the ITS region. Blastn results indicated the EPN isolate had a similarity match of 81-92% with Afro-tropical Steinernema species. It matched with Steinernema sp. (AY230186.1) from Kenya at 92% and Sri Lanka (AY230184.1). Phylogenetic analysis placed the isolate together with Steinernema sp. (AY230186.1) and (AY230184.1) with a bootstrap value of 100%. Maximum mean larval mortality (80%; 96%) was achieved 24 and 48 h post-treatment at concentration 150 IJs/ml. All nematode concentrations achieved over 50% mean mortality after 24 h period. There was a significant difference (P = 0.001) between doses 150 and 200 IJs/ml. From the study, it was concluded that the nematode isolate was Steinernema sp now referred to as Steinernema sp. Kalro (Genebank Accession MW151701). The EPN has the potential for development as a biological control agent against T. absoluta.
