A Real-Time qPCR Assay to Quantify Ophiocordyceps sinensis Biomass in Thitarodes Larvae

Wei Lei; Shaosong Li; Qingyun Peng; Guren Zhang; Xin Liu

doi:10.1007/s12275-013-2241-7

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A Real-Time qPCR Assay to Quantify Ophiocordyceps sinensis Biomass in Thitarodes Larvae: Wei Lei , Shaosong Li , Qingyun Peng , Guren Zhang , Xin Liu; Journal of Microbiology 2013;51(2):229-233.
DOI: https://doi.org/10.1007/s12275-013-2241-7
Published online: April 27, 2013

Food and Health Engineering Research Center of State Education Ministry, School of Life Sciences, Sun Yat-sen University, Guangzhou, P. R. China

Corresponding author: Xin Liu , Tel: +86-20-84112299,

Received: 8 May 2012 • Accepted: 21 December 2012

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Abstract

Ophiocordyceps sinensis, an entomogenous fungus parasitic in the larvae of moths (Lepidoptera), is one of the most valuable medicinal fungi, and it only distributed naturally on the Tibetan Plateau. The parasitical amount of O. sinensis in various tissues of the host Thitarodes larvae has an important role in study the occurrence and developmental mechanisms of O. sinensis, but there no an effective method to detect the fungal anamorph. A real-time quantitative PCR (qPCR) system, including a pair of species-specific ITS primers and its related program, was developed for O. sinensis assay with high reliability and efficiency. A calibration curve was established and exhibited a very good linear correlation between the fungal biomass and the CT values (R2=0.999419) by the qPCR system. Based on this method, O. sinensis was detected rapidly in four tissues of its host caterpillars, and the results were shown as following: the maximum content of O. sinensis parasitized in the fat-body, and next came bodywall; both of them were much larger than that observed in the haemolymph and intestinal-wall. Taken together, these
results
show that qPCR assays may become useful tools for study on developmental mechanism of O. sinensis.

Keywords: O. sinensis;real-time quantitative PCR;caterpillars;internal transcribed spacer;DNA

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A Real-Time qPCR Assay to Quantify Ophiocordyceps sinensis Biomass in Thitarodes Larvae

J. Microbiol. 2013;51(2):229-233. Published online April 27, 2013

DOI: https://doi.org/10.1007/s12275-013-2241-7

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