Inhibitory Effects of PLM-WE1 Formulated from Extract of Phellinus linteus Mycelium against Plant Viruses Infection and Identification of Active Compound

Pepper mild mosaic virus(PMMoV) and Cucumber mosaic virus (CMV) are important pathogens in various vegetable crops worldwide. We have found that hot water extract of Phellinus linteus mycelium strongly inhibit PMMoV and CMV infection. Based on these results, the inhibitor named as 'PLM-WE1' formulated from extract of Phellinus linteus mycelium was tested for its inhibitory effects on PMMoV and CMV infection to each local lesion host plant (Nicotiana glutinosa: PMMoV, Chenopodium amaranticolor: CMV). Pretreatment effect of PLM-WE1 against infections of each virus (PMMoV and CMV) to local host plant was measured to be 99.2% to PMMoV and 80.3% to CMV, and its permeability effect was measured to be 45.0% to PMMoV and 41.9% to CMV. Duration of inhibitory activity of PLM-WE1 against PMMoV infection on N. glutinosa was maintained for 3 days at 75% inhibition level and CMV infection on C. amaranticolor maintained for 3 days at 62% inhibition level. Inhibitory effects on systemic host plants of PLM-WE1 were measured to be 75~85% to PMMoV and 75% to CMV. Under electron microscope, PMMoV particles were not denatured or aggregated by mixing PLM-WE1. It is suggested that the mode of action of PLM-WE1 differ from that of inactivation due to the aggregation of viruses. The methanol extract of P. linteus mycelium was sequentially partitioned with haxane, ethyl acetate, BuOH and $H_2O$. The $H_2O$ fraction was showed high activity than the other fractions. The active compound was isolated with a partial acid hydrolysis, fractional precipitation with ethanol. The inhibitory effect of the precipitate isolated from 70% ethanol fraction was 99.1% to PMMoV and 88.0% to CMV. The structure of isolated compound was determined by $^1H$-NMR and $^{13}C$-NMR. This compound was identified as a polysaccharide consisting alpha or beta-glucan.

목질진흙버섯(Phellinus linteus) 균사체 추출물 제제 PLM-WE1의 식물 바이러스에 대한 감염억제 효과 및 활성성분의 동정

목질진흙버섯(Phellinus linteus)을 액체 배양한 균사체의 열수 추출물을 이용하여 PMMoV 및 CMV에 대한 감염억제효과를 검정한 결과, PMMoV 및 CMV에 높은 감염억제효과가 발견되었다. 이 결과를 토대로, P. linteus 균사체의 열수추출물을 주성분으로 제제화한 PLM-WE1의 효능 평가를 위하여, PMMoV 및 CMV의 국부감염 기주식물에서 PLM-WE1의 바이러스 접종전 처리효과, 엽이면 처리시의 감염억제효과 및 PLM-WE1의 지속효과 실험을 수행하였다. 그 결과, PLM-WE1(처리농도 10 mg/ml)의 바이러스 접종전 처리시의 감염억제율은 PMMoV 99.2%와 CMV 80.3%이었으며, 본 제제(농도 5 mg/ml)의 엽이면 처리시의 감염억제율은 PMMoV 45.0%와 CMV 41.9%, 약효지속성은 처리 3일 후까지 PMMoV 75%와 CMV 62% 수준으로 감염억제 효과를 보였다. 바이러스 전신감염기 주식물을 이용한 감염억제효과는 PMMoV 75~85%와 CMV 75%이었다. PLM-WE1제제의 바이러스 감염억제 기작을 해석하기 위하여, 정제한 PMMoV에 제제를 혼합하여 전자현미경으로 관찰한 결과, 바이러스 입자의 형태적 변화는 관찰되지 않아, PLM-WE1의 바이러스 감염억제기작은 바이러스입자의 응집(aggregation) 작용과는 다른 기작에 의한 것임이 추정되었다. 목질진흙버섯 균사체로부터 식물바이러스 감염억제 성분의 분리 및 동정을 위하여 유기용매분획, 산 가수분해 및 에탄올 분별침전을 실시하여 고활성 분획을 수득하였다. 이 분획물을 $^1H$-NMR 및 $^{13}C$-NMR을 이용하여 분석한 결과, $\alpha$- 또는 $\beta$-glucan이 결합된 다당류(polysaccharide)로 확인되었다.