본 연구는 국내산 침엽수 톱밥추출물을 이용하여 버섯의 푸른곰팡이병 방제약제 원료를 개발하고자 시도 되었으며, 푸른곰팡이병의 원인균의 하나인 Trichoderma 속 곰팡이를 대상으로 항균활성을 시험하였다. 1,000 ppm 농도의 낙엽송 열수추출물은 Trichoderma 속 균주에 대하여 최고 20.6%의 항균활성을 나타냈다. 1,000 ppm 농도의 미송 열수추출물은 T. aggressivum에 대하여 60.3%, T. harzianum 52.2%, T. atroviride 46.8%, T. viride 46.1%의 높은 항균활성을 나타내었으며 T. koningii에서는 36.2%의 항균활성을 나타냈다. 미송 열수추출물이 낙엽송 열수추출물보다 Trichoderma 속 균주에 대하여 보다 효과적인 항균화합물을 함유하고 있다고 판단되었다. 낙엽송 열수추출물로부터 분리된 n-hexane 분획물은 Trichoderma 속에 대하여 68.5% ~ 79.9%의 높은 항균활성을 나타냈다. 열수추출물로부터 분리된 미송의 n-hexane 분획물은 T. aggressivum에 대하여 68.5%, T. atroviride에 대하여 71.4%, T. harzianum에 대하여 71.9%, T. koningii에 대하여 75.7%, T. viride에 대하여 82.3%의 높은 항균활성을 나타냈고, ethyl acetate 분획물이 n-hexane 분획물 다음으로 높은 항균활성이 나타내었다. Trichoderma 속 균주에 대한 낙엽송 및 미송 열수추출물의 항균활성은 미송 추출물이 낙엽송 추출물보다 높게 나타났지만, 낙엽송과 미송의 열수추출물로부터 분리된 n-hexane 분획물에 의한 항균활성 차이는 거의 없었다. 미송 열수추출물과 낙엽송 및 미송의 열수추출물로부터 획득된 n-hexane 분획물은 Trichoderma 속에 대한 항균원료로서 사용가능성을 확인할 수 있었다.

표고, 느타리, 양송이, 팽이등 버섯의 종류에 관계없이 발생되고 있는 푸른곰팡이병은 그린몰드(green mold)가 그원인으로 알려져 있으며 이들의 피해는 심각하다. 푸른 곰팡이병을 일으키는 곰팡이로는 Trichoderma, Gliocladium, Aspergillus, Penicillum 등이 현재 알려져 있다. 푸른곰팡 이 병원균은 균사생장 시 균상에 발생하여 버섯균의 균사 생장을 방해하거나 기생하여 버섯균사를 사멸시키며, 버 섯수확기에 병해를 발생시키고 있다. 따라서 본 연구는 버 섯의 수확을 저하시키는 Trichoderma를 억제하기 위하 여 수목추출물 2종을 혼합한 다음 그 효능을 평가하였다. Trichoderma에 대하여 항균력을 나타내는 유효 화합 물질을 확인하기 위하여 GC/MS를 이용하여 편백 휘발성 추출물을 분석한 결과, 50개의 테르펜계 화합물이 존재한 다는 것을 확인하였고 대부분의 물질이 동정되었다. 편백 휘발성 추출물의 주요 화학적 조성은 모노테르펜류인 sabinene 11.62%, limonene 4.31%, isobornyl acetate 6.23%, α-terpinyl acetate 11.75%였으며, 세스퀴테르 펜류로 elemol 11.08%, β-eudesmol 4.02%, epibicyclosesquiphellandrene 4.18%가 나타났다. 소나무와 편백 추출물의 혼합비율 10 : 1(g/g) 제제는 T. aggressivum에서 20.4%, T. atroviride에서 14.0%, T. harzianum에서 32.0%, T. koningii에서 26.8%, T. viride에서 19.0%의 항균활성을 나타냈다. Trichoderma 균주는 편백 추출물의 첨가량이 증가함에 따라 항균활성 이 감소하는 경향을 나타냈다. Trichoderma 균주는 편백 추출물보다는 소나무 열수추출물에 대하여 영향을 받았다.

The yields of the fractions of n-hexane-soluble, ethyl acetate-soluble and methanol-soluble from the P. densiflora sawdust were obtained 1.36%, 2.21% and 4.03% using organic solvents, respectively (Table 1). The antifungal activity of the n-hexane extracts against L. edodes at concentrations 125, 250, 500, 750 and 1000 ppm were 36.5%, 41.2%, 43.1%, 45.4% and 47.6%, respectively (Table 2). The percent of antifungal activity against L. edodes was the greatest for the n-hexane extracts, ranging from 36.5% to 47.6% at a concentrations of 125 ppm to 1000ppm, with the values for all concentrations significantly different from one another. In this experiment, the antifungal activity by the n-hexane extracts was the highest. This result supposed that the n-hexane extracts have not only many more inhibitory compounds against L. edodes than the ethyl acetate and methanol extracts but also the inhibitory compound may be more easily dissolved by n-hexane than by ethyl acetate or methanol.

The yields of the n-hexane-soluble, ethyl acetate-soluble, methanol-soluble and methanol-insoluble fractions from the powdered water extracts were 8.2%, 10.6%, 32.0% and 49.2%, respectively (Table 1). The antifungal activities of water extracts, n-hexane-soluble fraction, ethyl acetate-soluble fraction, methanol-soluble fraction and methanol-insoluble fraction against L. edodes were 23.5%, 26.2%, 41.5%, 25.4% and 2.5%, respectively, at 1000ppm (Figure3). The ethyl acetate-soluble fraction showed much higher antifungal activity than the other fractions. The antifungal activity of the ethyl acetate-soluble fraction against L. edodes at 1000 ppm showed a statistically significant difference in the fractions of n-hexane-soluble, ethyl acetate-soluble, methanol-soluble and methanol-insolub

1. Antifungal activity of water extracts The antifungal activity of the water extracts against L. edodes was proportional to the concentration of the water extracts. The antifungal activity of the water extracts at 1000 ppm was the highest among the tested concentration against L. edodes mycelia. The antifungal activity were significantly different from each concentration of the water extracts. Therefore, it can be concluded that the water extracts of the P. densiflora sawdust have inhibitory compounds to the growth of L. edodes mycelia. 2. Yield and antifungal activity of fractions from water extracts The powdered water extracts was further treated serially using organic solvents in order to separate the water extracts into the fractions of n-hexane-soluble, ethyl acetate-soluble, methanol-soluble and methanol-insoluble. The ethyl acetate-soluble fraction showed much higher antifungal activity than the other fractions. The antifungal activity of the ethyl acetate-soluble fraction against L. edodes at 1000 ppm showed a statistically significant difference in the fractions of n-hexane-soluble, ethyl acetate-soluble, methanol-soluble and methanol-insoluble. And, there was not significantly difference between the antifungal activities of the n-hexane-soluble and methanol-soluble fractions at 1000 ppm. 3. Antivungal activity of fractions of organic solvent extracts The fractions were isolated from each organic solvent by the silica-gel column chromatography and then the fractions were confirmed and divided on the values of Rf by the thin-layer chromatography. The fractions were produced six band of the n-hexane, nine band of the ethyl acetate and eight band of the methanol extracts. The HIII fraction from the n-hexane extracts showed the highest growth inhibition against L. edodes mycelium.

After having inoculated the mycelium of L. edodes on the block medium, the P. densiflora sawdust of the B and C groups were the slowest of their respective groups on day 169.4 and day 169.9 harvesting times. The first harvest of the fruiting body from the Q. variabilis sawdust of the control medium was obtained on day 162.9