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1 Weed Turf. Sci. 4(3):219~224 Print ISSN , Online ISSN Research Article Weed & Turfgrass Science Weed & Turfgrass Science was renamed from both formerly Korean Journal of Weed Science from Volume 32 (3), 2012, and formerly Korean Journal of Turfgrass Science from Volume 25 (1), 2011 and Asian Journal of Turfgrass Science from Volume 26 (2), 2012 which were launched by The Korean Society of Weed Science and The Turfgrass Society of Korea founded in 1981 and 1987, respectively. 토양방선균유래 Herbicidin 의제초활성 원옥재 1 김영태 1 김재덕 2 최정섭 2 * 고영관 2 박기웅 1 * 1 충남대학교식물자원학과, 2 한국화학연구원친환경신물질연구센터 Herbicidal Activity of Herbicidin from a Strain of Soil Actinomycete Streptomyces scopuliridis Ok Jae Won 1, Young Tae Kim 1, Jae Deok Kim 2, Jung Sup Choi 2 *, Young Kwan Ko 2, and Kee Woong Park 1 * 1 Department of Crop Science, Chungnam National University, Daejeon 34134, Korea 2 Eco-friendly and New Materials Research Center, Korea Research Institute of Chemical Technology, P.O. Box 107, 141, Gajeong-ro, Yuseong-gu, Daejeon, 34114, Korea ABSTRACT. This study was conducted to evaluate the effect of herbicidin, new natural herbicidal substances, derived from soil actinomycetes Streptomyces scopuliridis. Several weed species were subjected to examine the germination inhibition and herbicidal activity at the concentration from 100 to 2,000 ppm. There was no selectivity in germination inhibition and herbicidal activity against crops. Germination of Echinochloa oryzoides, Digitaria ciliaris, Abutilon theophrasti and Amaranthus retroflexus was inhibited completely when 7.81 ppm of extract was treated in petri dish. Pre-emergence application of herbicidin in soil condition showed low inhibition against weeds. However, post application of herbicidin in green house resulted in the necrosis of weeds at the concentration of 2,000 ppm. A. retroflexus was sensitive to herbicidin at the low concentration of 62.5 ppm, whereas E. oryzoides was tolerant to lower concentration of herbicidin until it became withered at the concentration of 2,000 ppm. In conclusion, herbicidal substances derived from S. scopuliridis herbicidin, which is consisted with herbicidin A and B, have dominant effect on germination and growth inhibition. On the other hand, herbicidin was insufficient to control gramineous weeds. In future, it will be needed to develop the combination of herbicidin with other herbicide or compounds to control gramineous weeds as well. Key words: Germination, Herbicidin, Streptomyces scopuliridis, Weed control Received on August 20, 2015; Revised on September 3, 2015; Accepted on September 10, 2015 *Corresponding author: 1 Phone) , Fax) ; ) parkkw@cnu.ac.kr 2 Phone) , Fax) ; ) jschoi@krict.re.kr 2015 The Korean Society of Weed Science and The Turfgrass Society of Korea This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( which permits unrestricted noncommercial use, distribution, and reproduction in any medium, 서론 현재널리쓰이고있는유기합성제초제는인축에대한독성및토양잔류로인한환경오염에대한우려가있으나천연물제초제는이러한우려가적다 (Sekizawa and Takematsu, 1982). 미국및유럽등선진국에서는유기합성제초제의생산및사용량을규제하고있으며, 경제협력개발기구 (OECD) 회원국의경우 2004년생산량을기준으로 2013년까지 40% 를축소하는등세계적으로유기합성제초제의생산및사용을줄이는추세이다 (Kim, 2009). 따라서이러한유기합 성제초제의대안으로식물, 동물, 미생물에존재하는다양한 2차대사산물인천연물을기원으로하는제초제의개발이활발이이루어지고있다. 미생물기원제초활성물질은세균, 곰팡이, 방선균으로부터생산되는 2차대사산물을근간으로개발되고있다. 특히이중방선균은균사형태로생장하는그램양성토양미생물로 500가지이상의속으로분류되며다양한유효성분을지니고있다 (Joseph et al., 2012). 대표적으로 Streptomyces 속방선균으로부터분리된항생물질은전체항생물질의 45% 를차지하며 (Umezawa, 1982), 이들중 S. griseus에서분 219
2 220 원옥재 김영태 김재덕 최정섭 고영관 박기웅 리한 streptomycin은폐결핵치료제로개발되어이용되고있다 (Schatz et al., 1944). 또한 S. hygroscopicus로부터분리된 oligopeptide계제초활성물질인 bialaphos가상업화되었으며, 그밖에새로운미생물제초제의개발연구가현재까지활발히이루어지고있다 (Tachibana and Kaneko, 1986). 오늘날미생물분리및배양기술을통하여자연계에존재하는미생물중약 10% 미만이발견되어이용되고있으며앞으로도다양한미생물로부터신규물질의개발이이루어질것으로예측된다 (Iwai and Takahashi, 1992). 방선균이생산하는 2차대사산물을활용한제초활성물질로는 S. saganonensis가생산하는 herbicidin A와 B가 1976년에보고되었으며 (Arai et al., 1976), 이외에도 S. galilaeus가생산하는 homoalanosine (Fushimi et al., 1989), Kitasatospora phosalacinea가생산하는 phosalacine (Omura et al., 1984), S. hygroscopicus 가생산하는 hydantocidin 등이발견되었다 (Nakajima et al., 1991). 한편이러한방선균유래물질중 glufosinate-ammonium은일본에서 1984년 S. hygroscopicus 로부터발견된후비선택성제초제로개발하여현재전세계적으로사용되고있다 (Ebert et al., 1990). 국내의경우현재까지미생물, 자생식물, 식물의정유성분등으로부터천연제초활성물질탐색을위한연구가수행되어왔으나, 아직까지는상업화는이루어지지않고있다. 대표적으로 Streptomyces 속에서유래한 methoxyhygromycin (MHM) 은제초활성물질로서의가능성이있다고보고되었으나, 높은처리농도등의문제로실용화되지못하였다 (Lee et al., 2003). 토양방선균유래대사산물인 herbicidin은 herbicidin A 와 B가주성분인물질로, 2013년한국화학연구원의온실및포장조건에서살초력과작물선택성평가를통해비선택성천연제초제로서의가능성이제시된바있다 (Lee et al., 2013). Nucleoside계제초활성물질인 herbicidin A와 B 는모두벼 (Oryza sativa) 에대한약해가적었으나, 토마토 (Lycopersicon esculentu) 와무 (Raphanus sativus) 에서 75 ppm 의 herbicidin A와 300 ppm의 herbicidin B에서 % 의생장저해를보였다 (Arai et al., 1976). 잡초에대한약효시험결과화본과잡초보다광엽잡초에민감한반응을보여 herbicidin A와 B는 300 ppm에서피 (Echinochloa oryzoides) 의생육을각각 50-95% 와 80-95% 저해하고, 닭의장풀 (Commelina communis), 쇠비름 (Portulaca oleracea), 흰명아주 (Chenopodium album) 의생육은 100% 저해하였다. 한편전체적인잡초의생육저해효과는 herbicidin A가 herbicidin B 보다우수한것으로알려졌다. 따라서, 본연구는토양방선균유래대사산물 herbicidin 을이용하여다양한잡초와작물을대상으로처리농도에 따른발아, 생육저해효과및안전성등을조사하여새로운천연물제초제개발의가능성을확인하고자수행되었다. 재료및방법 시험약제 S. scopuliridis herbicidin ( 수탁번호 : KCTC 12156BT) 은한국화학연구원에서발굴한제초활성방선균으로 16S rrna 유전자비교조사에서 S. scopuliridis RB72 (T) 와 99.7% 의유사도를나타냈다. 실험에사용한 S. scopuliridis herbicidin 으로부터얻어진대사산물의제초활성물질은 herbicidin A (50%) 와 herbicidin B (5%) 로구성되어있다. Herbicidin의잡초에대한약효와작물에대한약해잡초에대한약효시험시험잡초는외떡잎식물인강아지풀 (Setaria viridis), 바랭이 (Digitaria cilialis), 피와쌍떡잎식물인소리쟁이 (Rumex crispus), 어저귀 (Abutilon theophrasti), 털비름 (Amaranthus retroflexus), 토끼풀 (Trifolium repens) 을대상으로하였다. 발아시험은패트리디쉬 (60 15 mm) 에여과지 (Advavtec No. 2, 55 mm) 2장을깔고그위에잡초종자를각각 20립씩올려놓았다. Herbicidin의처리농도는 25, 50, 100 ppm을 5ml 처리한후파라필름으로밀봉하여 3반복으로실시하였다. 이후 30 o C, 12시간광조건, 25 o C, 12시간암조건의생장상에서 1주일간발아를유도한뒤발아율을측정하였다. 경엽처리효과시험을위해유리온실 (25-35 o C) 에서 105구 (15 7) 트레이에수도용 ( 피 ) 과원예용 ( 그외잡초종 ) 상토를넣어준후 1구에 3-5개의종자를파종하였다. 물관리는트레이받침을이용하여저면관수하였으며, 2주간재배후평균 2-3엽기에 herbicidin (500, 1,000, 2,000 ppm) 에전착제인 DOS 70 (0.2%) 을혼합하여 100 L ha -1 의양으로분무기를이용하여처리하였다. Herbicidin 처리 2주후지상부를채취하여봉투에넣고건조오븐에서 72 o C, 72시간동안건조하였다. 이후건물중을측정하고, 무처리건물중대비처리구건물중의비율인방제가로각잡초에대한약효를평가하였다. 작물에대한약해시험시험작물은외떡잎식물인밀 (Triticum aestivum), 벼그리고쌍떡잎식물인고추 (Capsicum annuum), 배추 (Brassica campestris), 상추 (Lactuca sativa), 오이 (Cucumis sativus) 를대상으로하였다. 발아시험및경엽처리효과시험방법은잡초에대한약효시험의재배방법과처리시기, 처리농도, 조사방법과동일한방법으로수행하였다.
3 토양방선균유래 Herbicidin 의제초활성 221 잡초종별농도반응패트리디쉬에서의발아억제효과시험잡초는외떡잎식물인피, 바랭이와쌍떡잎식물인어저귀, 털비름을대상으로하여패트리디쉬 (60 15 mm) 에여과지 (Advavtec No. 2, 55 mm) 2장을깔고그위에잡초종자를각각 20립씩올려놓았다. Herbicidin 처리농도는 2,000 ppm을 1/2씩희석하여총 14개의농도와무처리로패트리디쉬에 herbicidin의농도별용액을각각 5ml씩처리한후파라필름으로밀봉하여 3반복으로실시하였다. 이후광조건 12시간에서 30 o C, 암조건 12시간에서 25 o C인생장상에서 7일간처리한후발아된종자수를조사하고농도에따른무처리대비발아율을구하였다. Herbicidin의농도그래프와 GR 50 ( 생장의 50% 를저해하는농도 ) 값은 Sigmaplot의 log-logistic 모델을이용한비선형회귀분석을사용하여구하였다 (Seefeldt et al., 1995). 토양처리에의한발아억제효과시험잡초는외떡잎식물인바랭이와피, 쌍떡잎식물인어저귀와털비름을대상으로하여유리온실 (25-35 o C) 에서 105 구 (15 7) 트레이에수도용 ( 피 ) 과원예용 ( 그외잡초종 ) 상토를넣어준후트레이 1구에 3-5개의종자를파종하였다. 실험은각각의육묘트레이를처리구로하여 3반복하였다. 파종직후충분한양의물을공급하였으며, 파종 1일후에 2,000 ppm 을기준으로 1/2로희석한 7개의농도를 1,000 L ha -1 의양으로토양에살포하여무처리와비교하였다. 처리 2일간물을공급하지않았으며, 이후물관리는트레이받침을통하여저면관수하였다. 처리 2주후지상부를채취하여봉투에넣고건조오븐에서 72 o C, 72시간동안건조시켰다. 이후건물중을측정한후무처리건물중대비처리구방제가로각잡초에대한발아억제및생육저해효과를평가하였다. 경엽처리에의한살초효과시험잡초는외떡잎식물인피, 바랭이와쌍떡잎식물인어저귀, 털비름을대상으로하여유리온실 (25-35 o C) 에서 105 구 (15 7) 트레이에수도용 ( 피 ) 과원예용 ( 바랭이, 어저귀, 털비름 ) 상토를넣어준후 1구에 3-5개의종자를파종하였으며, 물관리는트레이받침을통한저면관수로실시하였다. 실험은각각의육묘트레이를처리구로하여 7반복하였다. 2주간재배후평균 2-3엽기에 herbicidin을처리하였다. 처리농도는 2,000 ppm을기준으로 1/2씩희석하여총 14개의농도에전착제인 Tween 20 (0.2%) 을혼합하여 1,000 L ha -1 의양으로분무기를이용하여처리하고무처리와비교하였다. 처리 2주후지상부를채취하여봉투에넣어건조오븐에서 72 o C, 72시간동안건조시켰다. 이후건물중을측정하고무처리건물중대비처리구방제가로살초효과를평가하였다. 또한 herbicidin의농도그래프와 GR 50 ( 생장의 50% 를저해하는농도 ) 값은 Sigmaplot의 log-logistic 모델을이용한비선형회귀분석을사용하여구하였다 (Seefeldt et al., 1995). 결과및고찰 Herbicidin의잡초에대한약효와작물에대한약해잡초에대한약효시험토양방선균유래제초활성물질의잡초에대한발아억제및제초활성을조사한결과는다음과같다. 발아억제시험의경우 100 ppm 처리에서외떡잎식물은모두발아가억제되었으며, 쌍떡잎식물은토끼풀에서만무처리대비 75.0% 의발아억제를보였고, 나머지잡초초종은모두발아가 100% 억제되었다. 최저농도인 25 ppm에서피와강아지풀, 어저귀가추가로발아하였으나최종 80% 내외의발아억제 Table 1. Effect of pre-emergence and foliar applications on the herbicidal activity of herbicidin in several weed species. Herbicidin (ppm) Weed species ,000 2,000 Pre-emergence Post-emergence Herbicidal efficacy (%) Echinochloa oryzoides 90.0±5.1 z ± ± ±2.6 Setaria viridis 78.5± ± ± ± ±12.9 Digitaria ciliaris ± ± ±2.2 Abutilon theophrasti 79.8± ± ± ±2.4 Trifoloum repens 38.9± ± ± ± ± ±12.9 Amaranthus retroflexus ± ± ±0.4 Rumex crispus ± ± ±2.6 z Mean ± Standard error.
4 222 원옥재 김영태 김재덕 최정섭 고영관 박기웅 Table 2. Effect of pre-emergence and post-emergence applications of herbicidin on several crop species. Herbicidin (ppm) Crop species ,000 2,000 Pre-emergence Post-emergence Herbicidal efficacy (%) Oryza sativa 0±7.4 z 14.3± ± ± ± ±5.1 Triticum aestivum 84.4± ± ± ± ±2.0 Cucumis sativus ± ± ±4.9 Lactuca sativa 92 ± ± ± ± ±4.2 Brassicu rapa 74.7± ± ± ± ±1.2 Capsicum annuum 10.2± ± ± ± ± ±4.1 z Mean ± Standard error. Table 3. Effect of herbicid with different concentrations on emergence of weeds in greenhouse. Herbicid (ppm) Weed species ,000 2, Herbicidal efficacy (%) Echinochloa oryzoides 9.2±6.7 z 25.8± ± ± ± ± ±14.5 Digitaria ciliaris 6.1± ± ± ± ± ± ±3.3 Abutilon theophrasti 4.1± ± ± ± ± ± ±5.1 Amaranthus retroflexus 0.1± ± ± ± ± ± ±8.5 z Mean ± Standard error. 를보였으며, 토끼풀의경우 38.9% 의발아억제를나타냈다. 잡초의발아억제정도는쌍떡잎식물에서보다민감한반응을보였다. 제초활성시험은 2,000 ppm에서외떡잎식물인바랭이와쌍떡잎식물인털비름이각각 92.2% 와 98.6% 로민감한반응을보였으며, 그이외의잡초종은 70% 이상의제초활성을보였다. 특히바랭이와털비름은 500 ppm 에서도약 95% 이상의우수한방제효과를보였으며, 피 (60.3%) 와강아지풀 (53.2%) 의방제효과는다른잡초종에비해큰폭으로감소하였다 (Table 1). 이는 100 ppm의 herbicidin A에서피는 80-90% 생육저해를보인다는결과와크게다르게나타났다 (Kim et al., 1996). 이러한결과는처리당시잡초의생육차이및 herbicidin의유효성분의차이때문으로여겨진다. 이와같이다양한잡초초종을대상으로발아및살초효과를검증한결과 herbicidin은발아억제및경엽처리를위한제초제로의개발가능성을나타냈다. 작물에대한약해시험작물에대한발아억제및생육저해효과에대한 herbicidin 의약해시험결과는다음과같다. 100 ppm 농도처리시밀, 상추, 배추는 100% 의발아억제를보였으며, 벼와고추는각각 49.9% 와 69.5% 의발아억제를나타냈다. 반면오이는 발아억제를나타내지않았는데, 이는두꺼운종피로인하여 herbicidin의흡수가원활히이루어지지않았기때문이라여겨진다. 한편 25 ppm에서벼, 오이, 고추는 0-10% 수준의발아억제를보였으며, 나머지작물들은 70-90% 정도의발아억제를나타냈다. 생육에대한약해시험은 2,000 ppm 에서벼와오이가각각 49.3% 와 55.0% 의생육저해를보였으며, 이외의작물은모두 80% 이상의생육저해를나타냈다. 또한 500 ppm의농도에서도모든작물에대하여동일한정도의저해효과를보였다 (Table 2). 한편벼와오이에서는 50% 내외로잡초종들에비해낮은생육저해를나타냈지만실질적인선택성을나타내기에는불충분한결과를보였다. 이는경엽처리결과고추와토마토는심한약해를보이지만벼는비교적저해가덜하다는결과와도일치하였다 (Arai et al., 1976; Lee et al., 2013). 잡초종별농도반응패트리디쉬에서의발아억제효과 Herbicidin의잡초별발아억제농도를조사한결과피, 바랭이, 어저귀의경우 7.8 ppm, 털비름의경우 3.9 ppm까지 100% 의발아억제를보였다. GR 50 값은단자엽식물인피와바랭이에서 2ppm 정도로나타났으며, 쌍자엽식물인털
5 토양방선균유래 Herbicidin 의제초활성 223 Fig. 1. Effect of herbicidin at various concentration on germination of weeds in a growth chamber. 비름과어저귀의경우 1ppm 정도였다. 발아억제효과는모든잡초초종에서매우민감한반응을보였고, 쌍떡잎식물이보다더민감한발아억제반응을나타냈다 (Fig. 1). 토양처리에의한발아억제효과 Herbicidin의토양처리에따른잡초의발아및생육저해효과는외떡잎식물인바랭이와피의경우 2,000 ppm 처리에서각각 40% 와 28% 이하의생육저해를보였으나그이하의농도에서는외견상저해를보이지않았다. 반면쌍떡잎식물중어저귀는 2,000 ppm부터 125 ppm까지 50-60% 사이의생육저해를보였고, 털비름은 1,000 ppm까지 85% 내외의저해를보이고 500 ppm에서도 58.6% 의저해를나타냈다. 이는토양에 S. scopuliridis의에틸아세트분획물을 2,000 ppm의농도로처리시 % 의저해를보였다는결과와다른양상을나타냈다 (Lee et al., 2013). 한편본실험에서토양처리시쌍떡잎식물에서보다우수한생육저해효과를보였으나모든잡초초종에서완전한고사가나타나지않고추후생육이회복되어토양처리를통한적용은적합하지않은것으로여겨진다. 경엽처리에의한살초효과 Herbicidin의경엽처리후 1일이내에모든잡초에서잎의끝부분이꼬이거나말리는현상이관찰되었으며, 어저귀는엽에황변된반점이발생하였다. 약제처리 2주후에피는최고농도인 2,000 ppm에서도완전히고사하지않았으며, 1,000 ppm에서도역시경미한생육저해현상만을보였다. 바랭이와어저귀는 2,000 ppm에서고사하였으며, 500 ppm까지도생육저해를나타냈다. 털비름은가장민감한반응을보여 62.5 ppm에서도고사하였으며 1ppm에서 Fig. 2. Effect of various concentrations of herbicidin post application on four weed species. 도육안상저해되는모습이관찰되었다. 이는 S. scopuliridis 의에틸아세테이트분획물을 2,000 ppm의처리시 % 의생육저해를보인결과와유사하였으며, 1,000 ppm에서는완전방제가되지않았다는내용과도일치하였다 (Lee et al., 2013). 본실험의결과를종합하면 herbicidin을 2,000 ppm 의농도로경엽처리할경우많은종의잡초를방제할수있을것으로보이며피의경우도그보다높은농도로처리를하면방제가가능할것으로사료된다 (Fig. 2). 요 약 본연구를통해토양방선균유래제초활성물질을대상으로제초제로서의적용가능성여부를확인하기위하여수행되었다. 잡초에대한발아와경엽처리에대한약효시험결과, 종자발아는 100 ppm 수준으로처리하고경엽에는 2,000 ppm 수준으로처리해야저해효과를나타냈다. 작물에대한시험결과, 오이는발아억제가이루어지지않았으며벼는발아와생육모두 50% 내외의저해를보였다. 그외의모든시험작물초종은처리농도에따른민감한반응을보였다. 즉토양방선균으로부터발굴한천연제초활성물질은선택성제초제로의개발은어려워보이나비선택성제초제로의개발은가능할것으로보인다. 한편처리방법에따른농도별시험결과, 패트리디쉬에서실험의경우모든잡초종이농도에민감히반응하였으며, GR 50 값은 1-2 ppm 정도를보였다. 그리고토양처리효과에따른발아억제및생육저해효과는쌍떡잎식물에서보다민감한반응을보였고, 털비름의경우 1,000 ppm에서도 85% 의생육저해를보였다. 경엽처리에따른생육저해조사시 2,000 ppm 에서피는고사하지않았으나나머지초종은모두고사하
6 224 원옥재 김영태 김재덕 최정섭 고영관 박기웅 였고, 특히털비름은 6.25 ppm의농도에서도고사하였다. 경엽처리에따른대표적인반응으로바랭이는잎의꼬임현상을나타냈고, 어저귀는잎이변색되었다. 토양처리는그효과가경미하고일정기간경과후에재생되어방제효과가부족하였으며, 경엽처리의경우피를제외한잡초종등이 2,000 ppm에서방제가가능할것으로보인다. 또한모든결과를종합할때화본과잡초보다광엽잡초에서민감한발아억제및생육저해효과를볼수있었다. 따라서토양방선균유래제초활성후보물질은비선택성경엽처리제로서의개발이상대적으로유망할것으로판단되었다. 한편화본과잡초에대한저해효과가부족함으로추후다른제제와혼합함으로써화본과잡초도동시에방제할수있는기술의개발이필요할것이다. 주요어 : Herbicidin, Streptomyces scopuliridis, 발아, 잡초방제 Acknowledgement We would like to acknowledge the financial support from the R&D Convergence Program of NST (National Research Council of Science & Technology) of Republic of Korea. References Arai, M., Haneishi, T., Kitahara, N., Enikita, R. and Kawakubo, K Herbicidins A and B, two new antibiotics with herbicidal activity. I. Producing organism and biological activities. J. antibiot. 29(9): Ebert, E., Leist, K.H. and Mayer, D Summary of safety evaluation toxicity studies of glufosinate ammonium. Food Chem Toxicol. 28(5): Fushimi, S., Nishikawa, S., Mito, N., Ikemoto, M., Sasaki, M., et al Studies on a new herbicidal antibiotic, homoalanosine. J. Antibiot. 42(9): Iwai, Y. and Takahashi, T Seletion of microbial sources of bioactive compounds. Springer, pp In: Omura, S. (Ed.). The Search for Bioactive Compounds from Microorganisms. Spring-Verlag New York, USA. Joseph, B., Sankarganesh, P., Edwin, B.T. and Raj, S.J Endophytic streptomycetes from plants with novel green chemistry. Int. J. Biol. Chem. 6(2): Kim, J.C Research and development trends of the biological pesticide. Bioin-Special Zine. 10:1-20. (In Korean) Kim, W.G., Kim, J.P., Kim, C.J. and You, I.D A herbicidal nucleoside compound isolated from Streptomyces tubercidicus ME Kor. J. Appl. Microbiol. Biotechnol. 24(1): (In Korean) Lee, B.Y., Kim, J.D., Kim, Y.S., Ko, Y.K., Yon, G.H., et al Identification of Streptomyces scopuliridis KR-001 and its herbicidal characteristics. Weed Turf. Sci. 2(1): (In Korean) Lee, H.B., Kim, C.J., Kim, J.S., Hong, K.S. and Cho, K.Y A bleaching herbicidal activity of methoxyhygromycin (MHM) produced by an actinomycete strain Streptomyces sp. 8E-12. Lett. Appl. Microbiol. 36(6): Nakajimam, M., Kazuko, I., Takamatsu, Y., Kinoshita, T., Okazaki, T., et al Hydantocidin: a new compound with herbicidal activity from Streptomyces hygroscopicus. J. Antibiot. 44(3): Omura, S., Murata, M., Hanaki, H., Hinotozawa, K., Oiwa, R., et al Phosalacine, a new herbicidal abtibiotic containing phosphinothricin. fermentation, isolation. biological activity and mechanism of action. J. Antibiot. 37(8): Schatz, A., Bugie, E. and Waksman, S.A Streptomycin, a subastance exhibiting antibiotic activity against gram positive and gram negative bacteria. Exp. Biol. Med. 55: Seefeldt, S.S., Jensen, J.E. and Feurst, E.P Log-logistic analysis of herbicide dose-response relationships. Weed Tech. 9: Sekizawa, Y. and Takematsu, T How to discover new antibiotics for herbicidal use. pp In: Miyamoto, J. and Kearney, P.C. (Eds.). Natural products: Proceedings of the 5th International Congress of pesticide chemistry. Pergamon press, Kyoto, Japan. Tachibana, K. and Kaneko, K Development of a new herbicide, bialaphos. J. of pestic Sci. 11(2): (In Japanese) Umezawa, H Genetics, biosyntheses, actions & new substances: Proceedings, an International Conference on Trends in Antibiotic Research. Japan Antibiotics Research Association. Tokyo, Japan.
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