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1 THE KOREAN JOURNAL OF MYCOLOGY Vl. 34, N 2, p79-83 December 2006 Cpyright 2006 by The Krean Sciety f Myclgy Printed in S. KOREA m ³, x ³ ³ w tšm z*á Á w yw Cmparisn f Shiitake Prductivity in Sawdust Media Accrding t the use f Sawdust, Plug-shaped and Liquid Spawn Bng-Hun Lee*, Wn-Chull Bak and Kab-Hee Yn Divisin f Wd Chemistry and Micrbilgy, Krea Frest Research Institute, Seul , Krea (Received September 29, 2006) ABSTRACT: This study was made t investigate the difference f the shiitake prductivity accrding t the use f sawdust, plug-shaped and liquid spawn. Cntaminatin f medium was cnfirmed by eye. There was n cntaminatin in the liquid spawn treatment. And, cntaminatin rates in the sawdust spawn and plug-shaped spawn were 6.7%, respectively. Prductivity f fruit-bdies was g frm sawdust spawn, g frm plug-shaped spawn and g frm liquid spawn until 3rd flushing f fruit-bdies. The number f fruit-bdies was 57.4 per medium f sawdust spawn, 64.3 plug-shaped spawn and 45.1 liquid spawn. At yield per medium, the percentage f media prducing ver 1,000 g was 13% in plug-shaped spawn and 7% in liquid spawn. Hwever, there was n medium prducing ver 1,000 g f fruit-bdies in sawdust spawn. Fruit-bdies f 0~900 g were yielded frm 79% f sawdust spawn treatment, 53% plug-shaped spawn and 33% liquid spawn treatment. When cmpared the prductivity f fruit-bdies, liquid spawn treatment appeared lwer than sawdust spawn and plug-shaped spawn treatment at 1st flushing, but appeared higher than the thers at 2nd and 3rd flushing. KEYWORDS: Liquid spawn, Plug-shaped spawn, Prductivity, Sawdust spawn, Shiitake ù 2004 x, tš 24,397m wš tš 1,921m w, mw 2,425 ƒ wù (, 2005). w tš m w 1% w tš 80% wš m wš m x % m wš y tš w ƒ (, 2006; Lin et al., 2000). ù tšm w» 1990,» ù» w k ù, ù m w š z» ³ w m ³ w šƒ (, 1991, 1994;, 1998). ù w ³ m ³ m» vlƒ w, *Crrespnding authr < bnghun90@naver.cm> ƒ w w», y w w x ƒ. š ³, j»,, ƒ» w, š ³ ³ y š (Kirchhff and Lelley, 1991). x m ³, x ³, ³, y w w ww ³ y w» w w. ³ w KFRI 491 ³ x w. (2000) šw ù m 77%, 79
2 80 THE KOREAN JOURNAL OF MYCOLOGY, 34(2), 2006 기울 19%, 설탕 3%, CaCO 0.6%, KNO 0.4% 비율로 혼합한 배지에 물을 첨가하여 함수율이 53%가 되도록 조 절했다. 배지는 무게 1,750 g, 길이 33 cm 정도 크기로 제 조되었으며, 살균의 용이 및 통기를 위해 봉지 앞쪽에 직 경 3 cm 정도의 filter를 부착했다. 제조된 배지는 121 C 에서 90분 동안 고압살균했다. 종균제조 접종 및 배양 톱밥종균: 신갈나무톱밥 80%, 미강 19%, 설탕 1% 비 율로 섞고 함수율을 65%로 조절한 후 1 l 용량의 종균병 에 800 ml씩 넣어서 고압살균(121 C, 90분) 했다. 살균이 끝난 배지는 무균상에서 균을 접종하고 23 C에서 30일 간 배양했다(Fig. 1A). 배지에의 접종은 칼로 봉지를 자르고 윗면 4군데에 스 프링봉 접종기를 이용해서 종균을 투입한 후 스카치테이 프로 밀봉하는 방법으로 실시하였다. 접종 후 배지는 23 C, 65~70%의 암배양 조건에서 일간 배양했으며, 암 배양 완료 후에는 배지를 명배양(약 200 lux, 형광등) 조 건에서 일간 배양했다. 실험은 15반복으로 진행되었다. 성형종균: 톱밥종균 과 같은 방법으로 제조, 살균, 접 종된 종균을 30일 배양한 후, 표고원목재배에서 사용하는 성형틀(남산엔지니어링)을 이용하여 성형종균을 만들었으 며, 6일간 배양한 후 접종에 사용하였다(Fig. 1B). 배지에의 접종은 봉지 윗면 4군데에 각 1개 씩의 성형 종균을 밀어 넣는 것으로 완료되었고, 배지의 배양은 톱 밥종균 과 동일한 조건에서 진행되었으며, 15반복으로 진 행되었다. 액체종균: PDB(Difc)와 antifamer(0.1%)를 섞은 배 지를 액체배양병에 넣은 후 121 C에서 20분 동안 고압살 균하였다. 살균 후 미리 배양한 균을 hmgeniger로 균질 화한 다음 배양병에 넣어 주었으며, air pump로 산소를 공급하였다. 그리고 NA(Difc) 배지를 사용하여 배양 중 세균에 의한 오염 여부를 확인하였다. 액체배양균은 23 C 에서 14일간 배양하였다(Fig. 1C). 배지에의 접종은 윗면 4군데에 각 5 ml씩의 액체종균을 투입하였고 배양은 톱밥종균 과 동일한 조건에서 진행되 었으며, 15반복으로 진행되었다. 오염조사 배지를 배양하는 과정 중에 각 종균 접종구별로 오염 여부를 확인하였다. 생산성 조사 버섯을 발생시키기 위해 경기도 양평에 소재한 일반 비 닐하우스 재배사를 이용하였다. 배지는 톱밥종균, 성형종 균, 액체종균 접종구 모두가 동시에 입상되었으며, 입상 당시 이들의 배양 일수는 일(암배양 일, 명배양 일) 이었다. 버섯 발생은 3주기까지 진행되었고 각 주기별 3 3, Fig. 1. Spawns inculated in varius media. A, sawdust spawn; B, plug-shaped spawn; C, liquid spawn. 로 수확이 완료된 배지는 15~20일 정도 휴양 후 다음 주 기의 발생작업에 들어갔으며, 수분 공급은 침봉주입 방법 (속이 빈 긴 관을 배지에 찌르고 고압의 물을 투입하는 방 법) 을 사용했다. 버섯 품질을 조사하기 위해서는 정상적 으로 갓을 형성하는 버섯들 중에서 크기별로 각 주기 당 10~15개씩 조사하였다. 그리고 버섯발생 기간 중 재배사 내부의 온도 및 습도 변화를 확인하기 위해 자기온습도계 (SATO, SIGMA-II) 를 사용하였다.
3 Cmparisn f Shiitake Prductivity in Sawdust Media Accrding t the use f Sawdust, Plug-shaped and Liquid Spawn Fig. 2. Sawdust medium cntaminated by Trichderma sp. 결과 및 고찰 오염조사 및 배지배양 중의 관찰 배양 중의 배지오염 여부를 조사한 결과, 톱밥종균과 성형종균 접종구의 배지오염율은 각각 6.7%였으며, 액체 종균 접종구에서는 오염이 없었다(Fig. 2, Table 1). 톱밥 종균과 성형종균 접종구에서 발생된 오염도 종균을 접종 한 부위가 아니라 봉지에 배지를 넣으면서 자주 부딪히는 봉지 결속부위에 뚫린 작은 구멍을 통한 오염이었으며, 오염균은 표고톱밥재배시 자주 발생하는 Trichderma sp. 인 것으로 확인되었다. 따라서 종균의 차이에 의한 오염 문제는 없을 것으로 판단되었다. 3가지 접종구 모두 배지 표면에 균사가 만연되는 데에 는 30여일 정도가 경과되었고 접종 후 암배양 여일이 경과한 다음 명배양(갈변작업) 에 들어갈 시점에는 대부 분의 배지 표면이 갈변되었으며, 암배양 및 명배양 후 입 상시점도 3가지 접종구 모두 동일하였다(Table 1). 따라서 접종구들 간에 있어서 배양 중 차이점은 거의 없는 것으 로 관찰되었다. 생산성 조사 3주기에 걸친 버섯발생 결과, 톱밥종균, 성형종균, 액체 종균 접종구에서 각각 g, g, g이 수확 되어 배지중량 대비 41%, 42%, 33%의 생산성을 보였으 며, 세 접종구 모두가 한 번에 집중 발생하는 경향을 띄었 다(Fig. 3, Table 2). 배지당 버섯은 톱밥종균과 성형종균 접종구에서 57.4개, 64.3개가 각각 발생했지만 액체종균 81 Fig. 3. Fruit-bdies appeared n sawdust medium inculated sawdust spawn at a first yield. 접종구에서는 45.1개가 발생해서, 톱밥을 사용한 종균들 과 액체종균 간의 차이를 확인할 수 있었다(Table 2). 우리나라의 등록균주인 산림 5호와 산림 10호는 배지중 량 대비 수확량이 각각 26~28%, 27%이고 일본에서는 표 고톱밥재배시 수확목표를 배지중량 대비 25%로 하고 있 으며, 일본의 대표적인 표고톱밥재배용 품종인 호켄 0 호와 3호의 경우에는 배지중량 대비 35~%까지 생산 할 수 있는 것으로 알려져 있다(산림청, 1995; 산림조합중 앙회, 2006; 農村文化社, 1999; 北硏食用菌類硏究所, 1993). 이런 점을 고려할 때, 액체종균 접종구에서의 수확 량이 적다고 할 수는 없지만 동일한 균주를 사용해서 종 균 종류만 달리했을 뿐인데도 불구하고 생산량이 다른 두 종균 접종구의 79% 정도에 머물렀다는 점은 액체종균 사 용에 신중을 기해야 한다는 의미로 받아들여야 한다. 반 면에 배지당 수확량을 비교해 볼 때, 톱밥종균 접종구에 서는 1,000 g 이상 수확된 배지가 없지만 성형종균과 액 체종균에서는 배지의 13%와 7%에서 1,000 g 이상이 수 확되었다(Fig. 4). 이 실험 결과에는 비록 액체종균이 버 섯수확량이 떨어지기는 해도 본 실험에서 사용한 버섯발 생 방법을 보완한 새로운 환경 조절 방법이 개발된다면, 배지중량의 57% 이상 수확도 가능하다는 의미를 포함하 고 있다. 또한 Fig. 4를 통해 한 가지 더 확인할 수 있는 데, 배지중량의 34% 이상에 해당하는 0~799 g의 버섯 을 수확한 배지 비율이 톱밥종균 접종구는 50%였고 성형 종균 접종구는 33%, 액체종균 접종구는 20%였다. 여기에 800~999 g 수확배지까지 포함하면 톱밥종균 접종구는 배 Table 1. Incubatin characteristics f sawdust media inculated sawdust, plug-shaped and liquid spawn Sawdust spawn Plug-shaped spawn Liquid spawn Mycelial grwth (darkness/days) Brwning (days) Ttal incubatin time (days) Cntaminatin rate (%)
4 82 THE KOREAN JOURNAL OF MYCOLOGY, 34(2), 2006 Table 2. Cultivatin characteristics in sawdust media inculating varius spawn Yield (until third) The number f fruit-bdies (until third) Weight (g) Pileus diameter (mm) Quality f fruit-bdies Pileus thickness (mm) Stem length (mm) Stem thickness (mm) Sawdust spawn ± 87.8a a 57.4 ± 9.6a ± 11.8a 6.1 ± 1.2a 1.8 ± 0.5a 4.0 ± 0.9a 1.3 ± 0.4a Plug-shaped spawn ± 137.4a 64.3 ± 16.5a 25.0 ± 17.9a 6.2 ± 1.6a 1.8 ± 0.4a 4.4 ± 1.1a 1.3 ± 0.4a Liquid spawn ± 154.9b 45.1 ± 11.4b 28.1 ± 20.1a 6.2 ± 1.4a 2.0 ± 0.8a 4.3 ± 1.0a 1.5 ± 0.4a a : Fllwed by LSD multiple rage test (p < 0.05). w 1» y 2» 3» y ùkû (Fig. 5). ³ p w, ³ w š y w ƒw.» s³ 7~22 C, s³ 70~ 93% (data nt shwn). y w» w y w» ¾ ³ m ³ ù x ³ w, yw d w m ³ w ƒw. Fig. 4. The rate f fruit-bdies classified accrding t the weight. SS, sawdust spawn; PSS, plug-shaped spawn; LS, liquid spawn. Fig. 5. The rate f fruit-bdies prduced by times. SS, sawdust spawn; PSS, plug-shaped spawn; LS, liquid spawn. 79% 0~999 g y x ³ 53%, ³ 33% ùkù. m ³ w, d w ƒ.» y w, ³ m ³, x ³ ³ w w» w w. y w, ³ š m ³ x ³ ƒƒ 6.7%. 3» e mw m ³, x ³, ³ ƒƒ g, g, g yw. m ³ x ³ 57.4, 64.3 ƒ w ³ 45.1 ƒ w. y m ³ 1,000 g y ƒ x ³ ³ 13% 7% ùkû. š m ³ 79%, x ³ 53%, ³ 33% 0~999 g y.» y w, ³ w 1» y 2» 3» y ùkû. š x ù e w tš s y. w wz 80: ù m w tš. w wz 83:
5 Cmparisn f Shiitake Prductivity in Sawdust Media Accrding t the use f Sawdust, Plug-shaped and Liquid Spawn 83,, ƒ x, x, z tš w». w 258y. w z (m«484y) » w (II) m. k,,, ½, tš». 158y. Pp k, û, ³ w tšm z. œwz 26: Œ hli ~(2000 ). ƒš ƒ sqtrlkig. Œ «. Kirchhff, B. and Lelley, J Investigatin f Shiitake (Lentinus eddes (Berk.) Sing.) bag-lg cultivatin t increase the yield in Germany. Science and Cultivatin f Edible Fungi, A. A. Balkema, Rtterdam, Netherlands. Pp Lin, F. C., Yang, X. M. and Wang, Z. W Cultivatin f the black ak mushrm Lentinula eddes in china. Science and Cultivatin f Edible Fungi, A.A.Balkema, Rtterdam, Netherlands. Pp
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