Article Per Year (5 Year)
p-Index From 2020 - 2025
0.444
P-Index
This Author published in this journals
All Journal AGRIVITA, Journal of Agricultural Science Jurnal Perlindungan Tanaman Indonesia
Takemoto, Shuhei
Unknown Affiliation
Agriculture, Biological Sciences & Forestry

Published : 2 Documents Claim Missing Document
Claim Missing Document
Check
Articles

Found 2 Documents
Search

 1 
Specific Primer Designing for Quantitative PCR (qPCR) of Entomopathogenic Fungi Isaria fumosorosea from Soil Samples Saragih, Syaiful Amri; Takemoto, Shuhei; Sato, Hiroaki; Kamata, Naoto
Jurnal Perlindungan Tanaman Indonesia Vol 27, No 1 (2023)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jpti.77867

Abstract

Entomopathogenic fungi are important component for regulation of pests in ecosystem. Isaria fumosorosea, as one of the entomopathogenic fungi, was reported to successfully controlled the outbreaks of forest defoliators attacked larch (Larix kaempferi) plantation in Furano, Japan and beech (Fagus crenata) forest in Hachimantai, Japan. Instead of semi-cultured method, in this paper, a culture-independent method based on DNA using qPCR was developed for specific detection and quantification of I. fumosorosea directly from soil DNA extract using specific primer. The primer IFU5821F/IFU6061R was designed and found to be the best primer pair for I. fumosorosea. Standard soil DNA was obtained with strong relationship and good fitting with five levels (R2= 0.989, E = 0.58). I. fumosorosea could not be detected from all soil samples which was possibility caused by low density of the fungi. The qPCR was likely a rapid and specific method to detect the fungus from soil. 
Specific Primer Design for Detection and Quantification of Entomopathogenic Fungi Metarhizium anisopliae using Quantitative PCR (qPCR) in Soil and Cocoon Samples Saragih, Syaiful Amri; Takemoto, Shuhei; Kamata, Naoto
AGRIVITA Journal of Agricultural Science Vol 47, No 1 (2025)
Publisher : Faculty of Agriculture University of Brawijaya in collaboration with PERAGI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.17503/agrivita.v47i1.4644

Abstract

The information on the abundance and dynamics of entomopathogenic fungus Metarhizium anisopliae on the soil is limited by the existence of insect hosts. In this study, to detect and quantify specifically for M. anisopliae from extract of soil DNA, a culture-independent approach based on DNA and employing quantitative Polymerase Chain Reaction (qPCR) was developed. Primer pairs were designed and tested for their specificity to get a specific primer pair. The best primer pair was determined to be MA6071F/MA6218R. Two standard curves were created using 10 concentration levels (101 to 1010) by qPCR. Standard curves for genomic DNA showed a strong relationship and good fitting (R2 > 0.980). Six levels were obtained to generate standard genomic DNA (R2= 0.98, E = 1.05). Eight levels (R2= 0.9854, E = 0.91) were created for standard soil DNA. By qPCR, M. anisopliae was not found in all soil samples, possibly due to the samples' low fungal density. However, 13 dead cocoon samples out of 80 showed positive for M. anisopliae. To successsfully detect and quantify M. anisopliae in soil, the method of DNA extraction and soil sampling should be enhanced.
Co-Authors Kamata, Naoto Sato, Hiroaki Syaiful Amri Saragih