除草微生物禾长蠕孢的菌种改良与制剂的研究
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摘要
化学防治杂草给农业的发展做了较大的贡献，但伴随着环境污染、杂草抗性增加、农产品安全等问题。因此开发新型环保型除草剂、减少化学除草剂的使用是未来农业发展的趋势。发展微生物除草剂，实现杂草的生物防治，可以大幅度减少化学除草剂的使用，实现农业的可持续发展。
禾长蠕孢菌稗草专化型(Helminthosporium gramineum Rabenh f.sp．echinochloae，HGE)和弯孢菌(Curvularia lunata，CL)是中国水稻研究所杂草室从自然感病稗草上分离得到的两株病原真菌，它们是稗草的生防潜力菌株。HGE对稗草致病力强，但孢子产量低，而CL菌孢子产量高，但对稗草的致病力弱。本研究根据这两个菌株的特性，采用原生质体融合技术，将两者优良特性结合起来，培育出孢子产量提高、代谢产毒能力强、适合工业化发酵的优良工程菌株。并对探讨该菌株的发酵产毒素工艺、毒素微乳剂的制备技术，调查融合菌株的毒素田间防治杂草效果。获得以下主要结论：
1．确定了禾长蠕孢和弯孢菌的原生质体释放最佳条件。2％溶壁酶+2％蜗牛酶、2％纤维素酶+2％蜗牛酶分别为弯孢菌(CL)和禾长蠕孢菌(HGE)最佳溶菌酶组合；CL与HGE菌丝体释放原生质体最佳培养时间分别为16h、24h。原生质体经过液体培养基再生后，获得了较高的再生频率，CL和M1的原生质体再生率分别为14.70％±1.27％、11.11％±1.43％。
2．获得了较高的融合率。低浓度PEG甘氨酸溶液有利于弯孢菌与禾长蠕孢菌的原生质体融合，其融合率为2.6×10~(-3)和2.1×10~(-4)；通过对融合子生物学性状的评价，筛选出四株产毒素提高的融合子，其中融合子CM1—037毒素Ophiobolin A的合成能力较其亲本M1提高了34％；温室评价CL与M3之间的融合子，获得了两株孢子产量和生防潜力提高的融合菌株CM3—012、CM3—053，其孢子产量较其亲本M3分别提高了45.57％和17.72％、对稗草鲜重防效和干重防效分别提高了1.77％和4.89％、9.73％和12.55％。
3．RAPD指纹分析融合子与亲本的亲缘关系，验证了融合子遗传物质偏亲现象和融合过程中发生了遗传重组。
4．确定了融合子液体产毒素的摇瓶发酵条件为：菌株CM1—037；培养基为马铃薯200g，葡萄糖30g，麸皮20g,酵母膏1g，KH_2PO_4 2g,MgSO_4.7H_2O 1g；菌丝体接种量10ml/L。
5．确定了融合子CM1-037粗毒素的10％微乳剂剂型配方：10％粗毒素+15％助溶剂(8％9~#+7％5~#)+15％乳化剂(4.5％1~#+10.5％9~#)+2％氮酮+58％水。该制剂在大田条件下对丁香蓼、节节菜和水莎草的防治效果较好；结合低剂量化学除草剂(苄嘧磺隆+二氯喹啉酸)复合使用，该制剂对稗草和鸭舌草的防治效果达到80％以上。
The chemical weed-control had made large contribution to the development of agriculture.But the environmental pollution, increase of weed type with resistance against herbicide and safety of agricultural products came with the use of the chemical herbicide. Therefore, the new type herbicide had to be developed to reduce the usage of the tradiontal chemical herbicide, which was the trend of agriculture in future. Expending microherbicide, biologically control the weeds would reduce the usage of chemical herbicide, then to realize the sustainable development of the agriculture.
Helminthosporium gramineum Rabenh f. sp. echinochloae and Curvularia lunata were isolated by the China National Rice Research Institute, which were fungal pathogens of the barnyargrass from the natural infected weed plants and had potentials to develop into biological agents. HGE strains had high virulent to the barnyardgrass but low spore yield, while the CL strain had high spore yield but low virulent to the weed. The objectives of this study was to combine the advantages of the two strains into new ones by protoplast fusion, which provide higher spore yield and richer toxin, as well as to the favor of industrial production, then to study the procedures to make the microbe emulsion of the crude toxin, and to investigate its efficiency to control weeds in rice fields. The major results of the study showed as follows:
1. The optimal condition for protoplasts releasing was decided as that the most effective lytic enzyme mixture for CL was 2% lywallzyme plus 2% snaiiase and that for M1 was 2% cellulase plus 2% snaiiase, that the optimum incubation time was 16 h for CL and 24 h for M1. The protoplasts were regenerated in the liquid regeneration medium and gave a high regeneration frequency, which was 14.70%±1.27% for CL and 11.11%±1.43% for M1.
2. The protoplast fusion process gave a high fusion frequency, which was 2.6×10~(-3) between M1 and CL、2.1×10~(-4) between M3 and CL. There were four fusant strains that improved toxin production, especially for CM1-037, which gave a 1.34 times ophiobolin A production than its parent strain M1. And there were two fusant strains between M3 and CL, which gave a higher spore yield and improved biologolical control efficiency against barnyardgrass in greenhouse, and its fresh weight control efficiency and dry weight control efficiency of the fusatnt strain CM3-012 and CM3 - 053 were 1.77% and 4.89%, 9.73% and 12.55% respectively higher than their parent strain.
3. RAPD analysis on the relationship of fusant strains and the parent strains showed that genetic recombination occurred during protoplast fusion.
4. The condition for toxin production was that CM1-037 as strain, potato 200 g/L, glucose 30 g/L, wheat bran 20 g/L, yeast extract 1g/L, KH_2PO_4 2g/L, MgSO_4.7H_2O 1g/L as medium, and mycelia 10 ml/L as inoculums.
5. The 10% crude toxin microbial emulsion were that 10% crude toxin + 8% solvent 9~# + 7% solvent 5~#+4.5% emulsifying agent 1~# + 10.5% emulsifying agent 9~#+ 2% lanrocapran+58% water, which had a high efficiency control weeds such as false loosestrife, indian rotata, Sedge. When combination with the low usage of chemical herbicide ( bensulfuron methyl + quinclorac) , it improved the efficiency against barnyardgrass and Monochoria above 80%, against false pimpernel above 75%.
禾长蠕孢菌稗草专化型(Helminthosporium gramineum Rabenh f.sp．echinochloae，HGE)和弯孢菌(Curvularia lunata，CL)是中国水稻研究所杂草室从自然感病稗草上分离得到的两株病原真菌，它们是稗草的生防潜力菌株。HGE对稗草致病力强，但孢子产量低，而CL菌孢子产量高，但对稗草的致病力弱。本研究根据这两个菌株的特性，采用原生质体融合技术，将两者优良特性结合起来，培育出孢子产量提高、代谢产毒能力强、适合工业化发酵的优良工程菌株。并对探讨该菌株的发酵产毒素工艺、毒素微乳剂的制备技术，调查融合菌株的毒素田间防治杂草效果。获得以下主要结论：
1．确定了禾长蠕孢和弯孢菌的原生质体释放最佳条件。2％溶壁酶+2％蜗牛酶、2％纤维素酶+2％蜗牛酶分别为弯孢菌(CL)和禾长蠕孢菌(HGE)最佳溶菌酶组合；CL与HGE菌丝体释放原生质体最佳培养时间分别为16h、24h。原生质体经过液体培养基再生后，获得了较高的再生频率，CL和M1的原生质体再生率分别为14.70％±1.27％、11.11％±1.43％。
2．获得了较高的融合率。低浓度PEG甘氨酸溶液有利于弯孢菌与禾长蠕孢菌的原生质体融合，其融合率为2.6×10~(-3)和2.1×10~(-4)；通过对融合子生物学性状的评价，筛选出四株产毒素提高的融合子，其中融合子CM1—037毒素Ophiobolin A的合成能力较其亲本M1提高了34％；温室评价CL与M3之间的融合子，获得了两株孢子产量和生防潜力提高的融合菌株CM3—012、CM3—053，其孢子产量较其亲本M3分别提高了45.57％和17.72％、对稗草鲜重防效和干重防效分别提高了1.77％和4.89％、9.73％和12.55％。
3．RAPD指纹分析融合子与亲本的亲缘关系，验证了融合子遗传物质偏亲现象和融合过程中发生了遗传重组。
4．确定了融合子液体产毒素的摇瓶发酵条件为：菌株CM1—037；培养基为马铃薯200g，葡萄糖30g，麸皮20g,酵母膏1g，KH_2PO_4 2g,MgSO_4.7H_2O 1g；菌丝体接种量10ml/L。
5．确定了融合子CM1-037粗毒素的10％微乳剂剂型配方：10％粗毒素+15％助溶剂(8％9~#+7％5~#)+15％乳化剂(4.5％1~#+10.5％9~#)+2％氮酮+58％水。该制剂在大田条件下对丁香蓼、节节菜和水莎草的防治效果较好；结合低剂量化学除草剂(苄嘧磺隆+二氯喹啉酸)复合使用，该制剂对稗草和鸭舌草的防治效果达到80％以上。
The chemical weed-control had made large contribution to the development of agriculture.But the environmental pollution, increase of weed type with resistance against herbicide and safety of agricultural products came with the use of the chemical herbicide. Therefore, the new type herbicide had to be developed to reduce the usage of the tradiontal chemical herbicide, which was the trend of agriculture in future. Expending microherbicide, biologically control the weeds would reduce the usage of chemical herbicide, then to realize the sustainable development of the agriculture.
Helminthosporium gramineum Rabenh f. sp. echinochloae and Curvularia lunata were isolated by the China National Rice Research Institute, which were fungal pathogens of the barnyargrass from the natural infected weed plants and had potentials to develop into biological agents. HGE strains had high virulent to the barnyardgrass but low spore yield, while the CL strain had high spore yield but low virulent to the weed. The objectives of this study was to combine the advantages of the two strains into new ones by protoplast fusion, which provide higher spore yield and richer toxin, as well as to the favor of industrial production, then to study the procedures to make the microbe emulsion of the crude toxin, and to investigate its efficiency to control weeds in rice fields. The major results of the study showed as follows:
1. The optimal condition for protoplasts releasing was decided as that the most effective lytic enzyme mixture for CL was 2% lywallzyme plus 2% snaiiase and that for M1 was 2% cellulase plus 2% snaiiase, that the optimum incubation time was 16 h for CL and 24 h for M1. The protoplasts were regenerated in the liquid regeneration medium and gave a high regeneration frequency, which was 14.70%±1.27% for CL and 11.11%±1.43% for M1.
2. The protoplast fusion process gave a high fusion frequency, which was 2.6×10~(-3) between M1 and CL、2.1×10~(-4) between M3 and CL. There were four fusant strains that improved toxin production, especially for CM1-037, which gave a 1.34 times ophiobolin A production than its parent strain M1. And there were two fusant strains between M3 and CL, which gave a higher spore yield and improved biologolical control efficiency against barnyardgrass in greenhouse, and its fresh weight control efficiency and dry weight control efficiency of the fusatnt strain CM3-012 and CM3 - 053 were 1.77% and 4.89%, 9.73% and 12.55% respectively higher than their parent strain.
3. RAPD analysis on the relationship of fusant strains and the parent strains showed that genetic recombination occurred during protoplast fusion.
4. The condition for toxin production was that CM1-037 as strain, potato 200 g/L, glucose 30 g/L, wheat bran 20 g/L, yeast extract 1g/L, KH_2PO_4 2g/L, MgSO_4.7H_2O 1g/L as medium, and mycelia 10 ml/L as inoculums.
5. The 10% crude toxin microbial emulsion were that 10% crude toxin + 8% solvent 9~# + 7% solvent 5~#+4.5% emulsifying agent 1~# + 10.5% emulsifying agent 9~#+ 2% lanrocapran+58% water, which had a high efficiency control weeds such as false loosestrife, indian rotata, Sedge. When combination with the low usage of chemical herbicide ( bensulfuron methyl + quinclorac) , it improved the efficiency against barnyardgrass and Monochoria above 80%, against false pimpernel above 75%.
引文
1．包衍 罗江兰 张伟琼等．互米花草根际镰刀菌的分离鉴定及治病性测定．杂草科学，2006，2：6～9．
2．薄海波．蔗糖和还原糖测定方法的探讨．现代商检科技，1999，9(1)：52～54．
3．曹干，周学秋．近红外光谱法快速测定甘蔗蔗汁蔗糖分．甘蔗，2001，8(3)：20～25．
4．曹红，孟丽，储治国．鲜奶中蛋白质含量的快速测定．食品工业科技，2005，26(3)：174～176．
5．陈松茂，翁世伟主编．化工产品实用手册(三)[M]．上海：上海科学技术文献出版社，1989：678．
6．陈勇，倪汉文．稗草致病菌—尖角突脐孢菌菌株RAPD指纹图谱的分析．微生物学报，2003，43(4)：409～416．
7．陈勇强，但汉斌，郭富常．国外微生物除草剂的研究及应用现状。天津农业科学，1998，2(4)：5～9．
8．戴新宾，陈世国，强胜等．链格孢毒素对紫茎泽兰叶片光合作用的影响．植物病理学报．2004，34(1)：56～60．
9．段桂芳．稗草病原菌的生物学评价和利用研究[硕士学位论文]．北京：中国农业科学院研究生院，2002．
10．段桂芳，黄世文，颜秋生等．稗内脐蠕孢菌原生质体制备和再生，农业生物技术学报，2003，11(3)：245～248．
11．但汉斌，陈永强，魏雪生等．从杂草DBR中筛选微生物除草剂的研究．微生物雪通报，2002，29(4)：5～9．
12．杜海英，于宏伟，韩军等．原生质体诱变选育乳糖酶高产菌株．微生物学通报，2006，33(6)：48～51。
13．盖栋梁，邹晓，刘爱英等．紫红曲菌的诱变育种研究．莱阳农学院学报，2006，23(4)：293～296．
14．高昭远，干静娥．菟丝子的生物防除——“鲁保一号”的研究进展．生物防治通报，1992，8(4)：173～175．
15．甘志波．微生物原生质体融合：问题与对策．微生物学杂志，1996，16：46～50．
16．黄英，郭鹏飞，张翠竹等．由原生质体融合技术改良高温采油微生物．南开大学学报，2003，36(1)：109～113．
17．菅原二三男．利用植物病原菌微生物开发除草剂，农药译丛，1992，14(3)：40～43．
18．顾成波，赵长山．微生物及其天然产物防治杂草地发展及展望．农药科学与管理，2003，24(2)：19～21．
19．范树田，wood T．一种快速简便提取真菌染色体DNA的方法．遗传，1991，13(5)：33．
20．郝勃，阎淳泰，陈华癸．利用原生质体融合技术提高黑曲霉产酶活力的研究．微生物学通 报，1996，23(6)：332～335．
21．黄炳球，林韶湘．我国稻区稗草对禾草丹的抗性研究[J]．农药科学与管理，1993(1)：18～21．
22．黄炳球，林韶湘．我国稻田稗草对丁草胺的抗药性研究[J]．华南农业大学学报，1993，14(1)：103～108．
23．黄世文，段桂芳，余柳青等．三株病原真菌对稗草生防潜力的研究．植物保护学报，2001，28(4)：313～3117．
24．黄世文，余柳青．淡紫灰吸水链霉菌及紫外诱变菌株用于害物生防研究．农业生物技术学报，2000，8(1)：79～84．
25．王颖．氮源及碳源比影响两种真菌除草剂孢子形成．生物技术通报，1992，7：60．
26．李春光．除草微生物禾长蠕孢菌种诱变改良及分子鉴定[硕士学位论文]．北京：中国农业科学院研究生院，2006．
27．李迪，周勇军，刘小川等．中国部分稻种资源的化感控制杂草潜力评价．中国水稻科学，2004，18(4)：309～314．
28．李阜棣，喻子牛，何绍江主编．农业微生物学实验技术．北京：中国农业出版社1996：P312．
29．李海涛，王金信，杨合同．利用麦瓶草病原菌株WP—1开发微生物除草剂初探．生物农药与生物技术，2005，4(3)：23～27．
30．李越，虞磊，吴克等．荧光标记香菇原生质体融合菌株的鉴定，生物学杂志，2002，18(1)：21～23．
31．李利军，孔红星，伍时华等．蔗糖测定的紫外分光光度新方法研究急应用。分析科学学报，2003，19(4)：367～369．
32．李岷，吴绍熙．一种深部真菌早期诊断的新方法—测定真菌细胞壁成分(1—3)β—D—葡聚糖．国外医学皮肤性病分册，1998，24(6)：343～345．
33．李荣春，张丽梅．随机扩增多态DNA(RAPD)在野蘑菇杂交育种中的应用．微生物学通报，2001，28(5)：52～55．
34．李祥锴，安志东，朱非．林肯链霉菌双亲灭活原生质体融合的研究．氨基酸和生物资源，2001，23(4)：24～27．
35．李拥兵，黄华枝，黄炳球等．我国中部和南方稻区稗草对二氯喹啉酸的抗药性研究．华南农业大学学报：自然科学版，2002，23(2)：33～36．
36．廖庆生．稗草对丁草胺和禾草丹的抗药性研究。植物保护，2000，26(5)：17～19．
37．罗水忠，姜绍通，郑志等．同步辐射软X射线N_k对产L—乳酸米根霉的诱变效应．核农学报，2006，20(5)：375～378．
38．刘成勤．吸光光度法测定工业污水中NH_4~+—N含量．中国环境检测，1998，14(5)：15～16．
39．刘国振，李小兵．用RAPD方法对平菇、香菇属间原生质体融合子的研究．遗传，1995(5)：37～40．
40．刘素萍，梁爽，李福荣等．产金属硫蛋白酵母菌原生质体电融合研究．菌物系统，1998，17(4)：339～343．
41．卢燕回，黎起秦，林纬等．西瓜枯萎病生防菌枯草芽孢杆菌B11菌株高产拮抗物质的诱变选育．广西农业生物科学，2006，25(4)：300～304．
42．刘志海，朱全让．1980．鲁保一号菌．济南：山东科技出版社，1～179．
43．冷俊峰．无铅澄清旋光法测定赤砂糖和红糖中蔗糖的含量．甘蔗糖业，1995，4：46～49．
44．马立田，王式箴．高效液相色谱法测定低热量食品中的葡萄糖、果糖、蔗糖、麦芽糖醇和山梨糖醇的研究．食品与发酵工业，1998，24(4)：12～17．
45．孟盈，沙丽清．风干、烘干对不同热带森林土壤样品NH_4~+—N、NO_3—N测定结果的影响．农业环境保护，2001，20(5)：366～67．
46．宋安东，吴云汉，贾翠英等．离子注入对苹果酵母菌的诱变研究．郑州大学学报，2003，35(3)：48～63．
47．桑金隆，竺莉红，李孝辉等．离子注入诱变选育之江菌素产生菌．科技通报，2002，18(1)63～66．
48．单季先，陈悦．食品中蛋白质含量测定方法的探讨．中国卫生检验杂志， 2005，15(4)：453～454．
49．沈齐英，刘录，申林波．紫外诱变选育高效降酚微生物．环境科学与技术，2004，27(1)：82～85．
50．孙建义，翁晓燕．UV诱变微生物的突变参数的分析．生物数学学报，1996，11(1)：92～96．
51．谢志鹏，许志南，郑建明等．靛酚蓝反应测定发酵液中的氨态氮．浙江大学学报(工学版)，2005，39(3)：437～439．
52．陶黎明，顾学斌，王维炯等．微生物源黄酮类化合物的除草活性．农药，2006：45(3)：180～181．
53．唐启义，冯明光．实用统计分析及其计算机处理平台．北京：中国农业出版社出版，1997．
54．涂鹤龄．我国农田杂草研究和防治进展．农药，2001．40(3)：1～3．
55．万方浩，王韧．世界杂草生防的历史成就及我国杂草生防的现状与建议．生物防治通报，1991，7(2)：81～87。
56．王建荣，张曼夫．真菌高分子量核DNA的提纯．遗传， 1992，14(6)：29～30。
57．王婧，酿酒酵母原生质体菌株的筛选．大连轻工业学院学报，2002，21(4)：265～268．
58．王金霞，黄大年，杨炜等．稻瘟病菌原生质体制备和再生的研究．中国水稻科学，1992，6(1)：33～38．
59．王淑珍，白晨，范俊等．灵芝与糙皮侧耳原生质体融合子基因组RAPD分析．食用菌学报，2003，10(1)：1～5．
60．王强，何锦豪，李妙寿等．浙江省水稻田杂草发生种类及危害．浙江农业学报，2000，12(6)：317～324．
61．万佐玺．链格孢菌毒素对紫茎泽兰致病性及生物测定方法的研究．湖北民族学院学报(自然科学版)，2001，19(1)：20～23．
62．万佐玺，朱晶晶，强胜等．链格孢毒素对紫茎泽兰的致病机理．植物资源与环境学报，2001，10(3)：47～50．
63．翁艳军，赵作棋，徐建国等．青霉素产生菌的原生质体融合．中国抗生素杂志，2003，28(3)：129～130．
64．燕克勤，成亚利，朱宝成等．荧光标记紫孢侧耳和糙皮侧耳原生质体融合研究．食用菌学报，1997，4(4)：25～29．
65．叶丽娟，李强国．氨态氮测定方法的研究．衡阳师范学院学报：自然科学，2002，3(23)：18～19。
66．叶亚军，张敏，汤志良．抗稻瘟病菌株——诺卡氏菌A11的诱变育种．植物保护，2007：33(1)：33～36．
67．叶丽娟，朱辉，田敏．微生物来源的真菌细胞壁抑制剂的研究进展．国外医药抗生素分册，2005，1(26)：34～41．
68．袁德军，周启．农抗5102产生菌原生质体融合育种德研究Ⅳ．融合子FR—008德验证及其产生新活性物质的分离和鉴别．生物工程学报，1991，7(2)：142～147．
69．余荔华，刘祖同．米曲霉与黑曲霉原生质体中间电融杂交育种．清华大学学报，1994，34(6)：7～12．
70．余柳青，黄世文，徐正浩．稗草生物学研究进展．北京：21试剂植物保护的发展．中国科学技术出版社，1998，599～603．
71．赵航．微生物除草剂菌种的原生质体融合研究[硕士学位论文]．北京：中国农业科学院研究生院，2005．
72．赵航，周勇军，刘小川等．生物除草剂剂型研究进展．农药市场信息，2006，1：14～5．
73．张朝贤．我国农田杂草综合资力现状和任务．第六次全国杂草科学学术研讨会论文集．广西：广西民族出版社，1999，PP10～15．
74．张龙翔，张庭芳，李令媛等．生化实验方法和技术．北京：人民教育出版社，1981．
75．张天宇．胶胞炭疽菌菟丝子专化型．真菌学报，1985，4(4)：234～239．
76．张忠义，刘振林，陈辉．葡萄酒中总糖测定方法探讨．中国公共卫生，2005，21(11)：1398～1399．
77．张智维，杨辉，陈合．He—Ne激光诱变选育果胶酶高产菌株．食品工业科技，2006，27(10)：156～157．
78．张彭湃，2005．微生物菌种选育技术的发展与研究进展．生物学教学，30(9)：3～5．
79．周颖，叶非．生物源除草剂研究与使用进展．农药，2002，41(5)：7～17．
80．诸葛健，王正祥．1994．工业微生物实验技术手册．北京：中国轻工业出版社，464～467
81. Agbessi S, Beausejour J, Derg C. et al. Antagonistic properities of two recombinant strains of Streptomyces melanosporofaciens obtained by intraspecific protoplast fusion. Appl.Microbiol.Biot., 2003, 62:233-238.
82. Annamalai P and Lalithakumari D. Isolation and regeneration of protoplasts from mycelium of Drechslera oryzae. J.Plant Dis. Prot. 1991, 98:197-204.
83. Anne J, Peberdy J F. Conditions for induced fusion of fungal protoplasts in polgethylene glycol solutions. Arch Microbiol., 1975,105:201-205.
84. Auld B A, Hetherington S D, Smith H E. Advances in bioherbicide formulation .Weed Biology and Management, 2003, 3(2): 61—67
85. Auld B A. (Inventor) Bioherbicidal formulations. Australian Prrovisional Patent Application 2002952094.Patent Office, IP Australia, Canberra.Cheng S P and Cui Y B. Characteristics for the hybrid cell Foaz obtained from the protoplast fusion between yeast and photosynthetic bacteria in soybean processing wastewater. J. Environ Sci., 1998, 10 (3):365—371.
86. Balasubramanian N, Annie Juliet G, Srikalavivani P, et al. Release and regeneration of protoplasts from the fungus Trichothecium roseum. Can.J.Microbiol. 2003, 49 (4):263—267.
87. Barski G, Hebd C R. 1960.Seances Acid Sci. [M]. Oxford: Blackwell, 1825 — 1827.
88. Biologtami K S. Wide spread occurrence of toxigenic Alternaria in cereas and oilaseeds. Indian Phytopathol. 1995,48(2): 150-153.
89. Boyette C D, Tempton G E and Smith R J. Control of winged waterpromrose( Jussiaea decurrens) and northern jiontvetch (Aeschynomene virginica) with fungal pathogens. Weed Sc.i,1979, 27(5):497-501.
90. Boyete C D. Unrefined corn oil improves the mycoherbicidal activity of Colletotrichum truncatum for hemp sesbania (Sesbania exaltata) control. Weed Sci. 1994, 41:495—500.
91. Bowers R C. Commercialization of Collego -an industrialist's view. Weed Sci., 1986, 34: 24—25 (Suppl.1).
92. Cheng Shu-pei, Cui Yi-bin. Characteristics for the hybrid cell Foaz obtained from the protoplast fusion between yeast and photosynthetic bacteria in soybean processing wastewater. J. enviro sci., 1998, 10(3):365—371.
93. Chiaki I, Masanori O, Yoshiro O. Intergenus protoplast fusion between Streptomyces and Micromonospora with reference to the distribution of parental characteristics in the fusants. J.Biosci.Bioeng., 1999, 88 (2):143 —147.
94. Chiaki I, Yuko I, Takeshi K, et al. Isolation and characterization of the interspecific fusants from Streptomycetes obtained using a liquid regeneration method. Fisheries Sci., 2002, 68:395— 402.
95. Chittick A T, Ash G J, Kennedy R A, et al. Micro-encapsulation: an answer to formulation quandary? Proceedings of the international workshop, Bioherbicides: the Next Generation (27 April 2003), International Bioherbicide Group, 2003, Canberra.
96. Chung Y R. Identification of Exserohilum species, a fungal pathogen causing leaf blight of barnyardgrass (Echinochloa crusgalli). Korean J.Planol. 1990,6:429-433.
97. Ciotola M., Hallett S.G., Watson A.K. Impact of Tusarium oxysporum isolates M12-4A, upon seed germination of Striga hermothica in vitro. Sixth international Parasitic Weed symposium, 1996, pp871-875.
98. Connick W J Jr, Boyette C D, McAlpine J R. Formulation of mycoherbicides using a pasta-like process. Biol, control., 1991, 1:281—287.
99. Dai M H, Sara Z M, Thomas R, et al. Visualization of protoplast fusion and quantitation of recombination in fused protoplast of auxotrophic strains of Escherichia coli. Metabolic engineering. 2005, 7:45-52.
100. Daniel J T, Templeton G E, Smith R J JR. Biological control of northern jointvetch in rice with an endemic fungal disease. Weed sci., 1973,21:303—307.
101. Duan G F, Zhang Z B, Zhang J P, et al. 2007. Evaluation of crude toxin and metabolite produced by Helminthosporium gramineum Rabenh for the control of rice sheath blight in paddy fields. Crop Protection. 26:1036-1041.
102. Duan G F, Zhou Y J, Yuan Q S, et al. Phytotoxic ophiobolins produced by Helminthosporium Gramineum Rabenh, a potential bioherbicide for control of barnyardgrass (Echinochola Crus-Galli). NPAIJ, 2007.
103. Duke S O, Dayan F E and Rimandao A M. Natural products as tools for weed management. Proc.Japanese weed Sci Soc. (Suppl.). 1998,Pp.1—11.
104. Duke S Q, Lydon J. Herbicides from natural compounds. Weed Technology. 1987,1:122—128
105. Emge R G, Melching J S, Kingsolver C H. Epideniology of Puccinia chondrillina, a rust pathogen for the biocontrol o rush skeleton weed in the United States. Phytopathology, 1981,71:839-843.
106. Filippis L De, Hoffmann E, Hampp R. Identification of somatic hybrids of tobacco generated by electrofusion and culture of protoplasts using RAPD-PCR. Plant Science, 1996, 121:39-46.
107. Ferenczy L. 1980. Current microbiol [M]. London: Cambridge University Press , 555- 574.
108. Goto M. The relationship between Emmalocera sp. and barnyardgrass and its potential as a biological control. Integrated management of paddy and aquatic weeds in Asia, Proceedings of an international seminar, 1992, pp113—121.
109. Gohbara M, Yamaguchi K, Bay Petersen J. Biological agents for the control of paddy weeds in Japan. Integrated management of paddy and aquatic weeds in Asia. Proceedings of an international seminar, Tsukuba, Japan, 1994, pp184—194.
110. Hasan S. Specificity and host specialization of Puccinia chondrillina. Ann Appl Biol., 1972,72: 57—263.
111. Hasan S, Wapshere A J. The biology of Puccinia chondrillina a potential biological control agent of skeleton weed. Ann Appl Biol., 1973,74:325—332.
112. Imaizumi S, Nishino T, Miyabe K, Fujimori T, Yamada M. Biological control of annual bluegrass (Poa annual L.) with a Japanese isolate of Xanthomonas campestris pv.poae (JT-P482). Biological control, 1997,8:7-14.
113. Itoh k., Wang G X and Ohba S. Sulfonylures resisitance in Linderma micrantha, an Annual paddy weed in Japan. Weed Res., 1999, (39): 413-423.
114. Jackson M A, Shasha B S, Schialer D A. Formulation of Colletotrichum truncatum microsclerotia for improved biocontrol of the weed hempsesbania (Sesbania exaltata). Biological Control, 1996, 7: 107-113.
115. Kenney D, Devine S. The way it was developed-An industrialist's view. Weed Sci., 1986,34 (suppl.1):15-16.
116. Kishimoto K and Sawada H. Large-scale culture of the mildiomycin producer Streptoverticillium rimofaciens. Journal of Bioscience &Bioengineering. 1997, 75(4):245—253.
117. Kremer R J, Fatima M, Begonia T, et al. Characterization of rhizobacteria associated with weed seedlings. Applied Enviromental Microbiology, 1990,56:1649—1655.
118. Judit K, IIona P, Lajos F. A novel method for hybridization of Saccharomyces species without genetic markers. Can.J.Microbiol., 1998, 44 (10):959—964.
119. Lee GA. Integrated control of rush skeleton weed (Chondrilla juncea) in the Western US. Weed sci., 1986,34:2-6.
120. Lee H B, Kim C J, Hong K S, et al. A bleaching herbicidal activity of methoxyhygromycin(MHM) produced by an actinomycete strain Streptomyces sp. 8E-13, Letters in Applied Microbiology, 2003,36:387—391.
121. Lohuis H. Biological weeds control still in its fancy. PSP-Planzenchutz-Praxis, 1990,3:22—23.
122. Masao K, Tomoaki M, Kazunobu S, et al. Chromosomal deletion or rearrangement in chimeric hybrids of Saccharomycopsis fibuligera and Saccharomyces diastaticus obtained by cell fusion. Journal of Fermentation and Bioengineering. 1996, 81 (4):281—285.
123. McCullough H. A simple micro technique for the determination of blood ammonia and a note on the effect of exercise. Clinical Chemical Acta. 1968, 19:101 — 105.
124. Mohan B R, Sajeena A, Seetharaman K, et al .Advances in bioherbicides development—an overview. Crop Protection, 2003,22,253—260.
125. Mortenson K. The potential of an endemic fungus Colletotrichum gloeospioides f.sp. malvae for biological control of round-leaved mallow (Malvapusilla) and velvetleaf (Abutilon theophrastis). Weed Sci., 1992, 36: 473—478.
126. Mrinalini C and Lalithakumari D. Protoplast fusion: a biotechnological tool for strain improvement of Trichoderma sp. Curr. Rrend Life Sci., 1996, 21: 133—146.
127. Nakmura M, Ishibashi K. New antibiotic ophiobolin A produced by Ophiobolus miyabeanus. J.Agric. Chem. Soc, Japan. 1958, 32: 739—744.(in Japanese)
128. Ogina H. The registration of pesticides and establishment of safety evaluation methods for microbial pesticides. In: R. GBayot, et al. Process of international symposium on biological control and integrated management of paddy and aquatic weeds in Asia. October 20-23, 1992, Japan. pp425-437.
129. Okada Y. 1962. Exp cell [M]. New York: Academic Press, 98 - 107.
130. Quimby P C Jr, Zidack N K, Boyette C D, et al. A simple method for stabilizing and granulating fungi. Biocontrol Sci. Technol.,1999,(9):5-8.
131. Pesti M, Konszky E, Polga J, et al. 1979. Fifth international Protoplast Symposium. Baltimore: Williams & Wilkins, 54.
132. Rasmussen J O and Rasmussen O S. Characterization of somatic hybrids of potato by use of RAPD markers and isozyme analysis. Phys.Plant. 1995 , 93:357 - 364.Regulation and Guidelines: Critical issues in biological control. Proc, of a USDA/CSRS National Workshop. 10 -12June,1991.USA.
133. Resi E M. Multplication of Helminthosporium spp. on sensscent tissues of gtaminaceous weeds and soybean under natural conditions. Fitopatologia Brasileita.1985,10(3):643-648.
134. Rubinder K, Chadha B S, Singh S, et al. Amylase hyper-producing haploid recombinant strains of Thermomyces obtained by intraspecific protoplast fusion. Can.J.Microbiol., 000, 46 (7):669-673.
135. Saunders J A, Matthew B F, Miller P D. Plant gene transfer using electrofusion and electroporation. In electroporation and electrofusion in cell biology (Neumann.E, Sower A.E; Jordan C.A ). PP: 343-354,1989, Plenum press, New York and London.
136. Scheepens P C. Joint action of Cochliolus and atrazine on Echinochloa Crusgalli (L.) Beauv. Weed Res., 1987, 27: 43-47.
137. Seong H C, Chang S, Man J O, et al. Intergeneric protoplast fusion in Saccharomycopsis fibuligera and Saccharomyces cerevisiae. Journal of fermentation and bioengineering. 1997, 84 (2):158-161.Sugawara F, Strobel G, Strange R N, et al. Phytotoxins from the pathogenic fungi Drechslera maydis and Drechslera sorghicola. 1987, Proc.Natl.Acad.Sci., 84:3081 -3085.
138. Shabana Y M, Charudattan R, Devalerio J T, et al. An evaluationofhydrophilic polymers for formulating the bioherbicide agents Alternaria cassiae and A. eichhorniae,Weed Technol.,1997,(11):212-220.
139. Smith R J Jr. Biological control as components of integrated weed management for rice in United States. In: R. GBayot, et al. Process of international symposium on biological control and integrated management of paddy and aquatic weeds in Asia. October 20-23, 1992, Japan. p.335-351.
140. Spencer J F T, Spencer D M, Bizeau C, et al. The use of mitochondrial mutants in hybridation of industrial yeast strains.V: Relative parental contributions to the genomes of interspecific and intergeneric yeast hybrids obtained by protoplast fusion as determined by DNA reassociation. Curr. Genet., 1985, 9:623-625.
141. Suslow T V, Schroth M N. Role of deleterious rhizobacteria as minor pathogens in reducing crop growth. Phytopathology, 1982,72:111 —115.
142. Smith R J Jr. Biological control of Northern Jointvetch {Aeschynomene virginicd) in Rice {Oryza sativa) and soybeans {Glycine mar)—A Researcher's View. Weed Sci., 1986, 34(suppl.1):17—23.
143. Smitamana P, Watson A K. Screening of the selective biocontrol agents for weeds in Thailand. Abstracts of biological weed control in rice workshop, 10-12, may ,2000, IRRI.
144. Tanake H, et al. An attempt for development of bioherbicide for Eleocharis kuroquwai. In: R. GBayot. Process of international symposium on biological control and integrated management of paddy and aquatic weeds in Asia. October 20-23, 1992, Japan. pp381—392.
145. Teeradakorn S, Talawanich Y, Suzuki I, et al. Characterization of Streptomyces D3 derived from protoplast fusion between Streptomyces cyaneas 190-1 and Streptomyces griseoruber 42-9. World journal of microbiology & biotechnology. 1998, 14:565-570.86 Wang Z.Y., Nagel J., Potrykus I. and Spangenberg G Plants from cell suspension-derived protoplasts in Lolium species. Plant Sci., 1993, 94:179-193.
146. TeBeest D O, Templeton G E. Mycroherbicides progress in the biological control of weeds. Plant Dis.,1985, 69(1): 6-10.
147. TeBeest D O, Yang X B, Cisar C R. The status of biological control of weeds with plant pathogens. Annu. Rev. Phytopathol., 1992, 30: 637—657.
148. Trujillo E E, Aragaki M, Shoemaker R A. Infection, disease development, and axenic cultures of Entyloma compositarum, the cause of hamakua pamakani blight in Hawaii. Plant Dis., 1988,72: 355-357.
149. Tsukamoto H. Evaluation of fungal pathogens as biological agents for paddy weed, Echinochloa species by drop inoculation. Japan, Ann. Phytopathol. Soc, 1997, 63: 366—372.
150. Tsukamoto H. Effect of water management on mycoherbicidal activity of Exserohilum monoceras against Echinochlia orzicola. Japan, Ann. Phytopathol. Soc, 1998, 64: 526—631.
151. Vazquez F, Heluane H, Spencer J FT, et al. Fusion between protoplasts of Pichia Stipitis and isolated filamentous fungi myclei. Enzyme Microb.Tech., 1997, 21:32—38.
152. Watson A K. Current status of bioherbicide development and prospects for rice in Asia. In: R. GBayot. Process of international symposium on biological control and integrated management of paddy and aquatic weeds in Asia. October 20-23, 1992, Japan, p.367—379.
153. Yamaguchi K. Biological control of paddy weeds using fungal isolates. Proc International symposium on wild and weedy rices in agro-ecosystem. Ho Chi Minh, Vietnam, 1998, pp133— 136.
154. Watanabe H, Uchina A, Tachibana M. Application period of Drechslera monoceras, plant pathogen for the mycoherbicide, to control Echinochloa oryzicola vasing rice fields of Tohoku, Japan. The Proc I of the 18~(th) Asian-Pacific Weed Sci. Soc. Confi., Beijing, 2001, pp416—421.
155. Zhang W M, Watson A K. Host range of Exserohilum nomocoras, a potential bioherbicide for the control of Echinochloa species. Can. J. Bot.,1997, 75: 685—692.
2．薄海波．蔗糖和还原糖测定方法的探讨．现代商检科技，1999，9(1)：52～54．
3．曹干，周学秋．近红外光谱法快速测定甘蔗蔗汁蔗糖分．甘蔗，2001，8(3)：20～25．
4．曹红，孟丽，储治国．鲜奶中蛋白质含量的快速测定．食品工业科技，2005，26(3)：174～176．
5．陈松茂，翁世伟主编．化工产品实用手册(三)[M]．上海：上海科学技术文献出版社，1989：678．
6．陈勇，倪汉文．稗草致病菌—尖角突脐孢菌菌株RAPD指纹图谱的分析．微生物学报，2003，43(4)：409～416．
7．陈勇强，但汉斌，郭富常．国外微生物除草剂的研究及应用现状。天津农业科学，1998，2(4)：5～9．
8．戴新宾，陈世国，强胜等．链格孢毒素对紫茎泽兰叶片光合作用的影响．植物病理学报．2004，34(1)：56～60．
9．段桂芳．稗草病原菌的生物学评价和利用研究[硕士学位论文]．北京：中国农业科学院研究生院，2002．
10．段桂芳，黄世文，颜秋生等．稗内脐蠕孢菌原生质体制备和再生，农业生物技术学报，2003，11(3)：245～248．
11．但汉斌，陈永强，魏雪生等．从杂草DBR中筛选微生物除草剂的研究．微生物雪通报，2002，29(4)：5～9．
12．杜海英，于宏伟，韩军等．原生质体诱变选育乳糖酶高产菌株．微生物学通报，2006，33(6)：48～51。
13．盖栋梁，邹晓，刘爱英等．紫红曲菌的诱变育种研究．莱阳农学院学报，2006，23(4)：293～296．
14．高昭远，干静娥．菟丝子的生物防除——“鲁保一号”的研究进展．生物防治通报，1992，8(4)：173～175．
15．甘志波．微生物原生质体融合：问题与对策．微生物学杂志，1996，16：46～50．
16．黄英，郭鹏飞，张翠竹等．由原生质体融合技术改良高温采油微生物．南开大学学报，2003，36(1)：109～113．
17．菅原二三男．利用植物病原菌微生物开发除草剂，农药译丛，1992，14(3)：40～43．
18．顾成波，赵长山．微生物及其天然产物防治杂草地发展及展望．农药科学与管理，2003，24(2)：19～21．
19．范树田，wood T．一种快速简便提取真菌染色体DNA的方法．遗传，1991，13(5)：33．
20．郝勃，阎淳泰，陈华癸．利用原生质体融合技术提高黑曲霉产酶活力的研究．微生物学通 报，1996，23(6)：332～335．
21．黄炳球，林韶湘．我国稻区稗草对禾草丹的抗性研究[J]．农药科学与管理，1993(1)：18～21．
22．黄炳球，林韶湘．我国稻田稗草对丁草胺的抗药性研究[J]．华南农业大学学报，1993，14(1)：103～108．
23．黄世文，段桂芳，余柳青等．三株病原真菌对稗草生防潜力的研究．植物保护学报，2001，28(4)：313～3117．
24．黄世文，余柳青．淡紫灰吸水链霉菌及紫外诱变菌株用于害物生防研究．农业生物技术学报，2000，8(1)：79～84．
25．王颖．氮源及碳源比影响两种真菌除草剂孢子形成．生物技术通报，1992，7：60．
26．李春光．除草微生物禾长蠕孢菌种诱变改良及分子鉴定[硕士学位论文]．北京：中国农业科学院研究生院，2006．
27．李迪，周勇军，刘小川等．中国部分稻种资源的化感控制杂草潜力评价．中国水稻科学，2004，18(4)：309～314．
28．李阜棣，喻子牛，何绍江主编．农业微生物学实验技术．北京：中国农业出版社1996：P312．
29．李海涛，王金信，杨合同．利用麦瓶草病原菌株WP—1开发微生物除草剂初探．生物农药与生物技术，2005，4(3)：23～27．
30．李越，虞磊，吴克等．荧光标记香菇原生质体融合菌株的鉴定，生物学杂志，2002，18(1)：21～23．
31．李利军，孔红星，伍时华等．蔗糖测定的紫外分光光度新方法研究急应用。分析科学学报，2003，19(4)：367～369．
32．李岷，吴绍熙．一种深部真菌早期诊断的新方法—测定真菌细胞壁成分(1—3)β—D—葡聚糖．国外医学皮肤性病分册，1998，24(6)：343～345．
33．李荣春，张丽梅．随机扩增多态DNA(RAPD)在野蘑菇杂交育种中的应用．微生物学通报，2001，28(5)：52～55．
34．李祥锴，安志东，朱非．林肯链霉菌双亲灭活原生质体融合的研究．氨基酸和生物资源，2001，23(4)：24～27．
35．李拥兵，黄华枝，黄炳球等．我国中部和南方稻区稗草对二氯喹啉酸的抗药性研究．华南农业大学学报：自然科学版，2002，23(2)：33～36．
36．廖庆生．稗草对丁草胺和禾草丹的抗药性研究。植物保护，2000，26(5)：17～19．
37．罗水忠，姜绍通，郑志等．同步辐射软X射线N_k对产L—乳酸米根霉的诱变效应．核农学报，2006，20(5)：375～378．
38．刘成勤．吸光光度法测定工业污水中NH_4~+—N含量．中国环境检测，1998，14(5)：15～16．
39．刘国振，李小兵．用RAPD方法对平菇、香菇属间原生质体融合子的研究．遗传，1995(5)：37～40．
40．刘素萍，梁爽，李福荣等．产金属硫蛋白酵母菌原生质体电融合研究．菌物系统，1998，17(4)：339～343．
41．卢燕回，黎起秦，林纬等．西瓜枯萎病生防菌枯草芽孢杆菌B11菌株高产拮抗物质的诱变选育．广西农业生物科学，2006，25(4)：300～304．
42．刘志海，朱全让．1980．鲁保一号菌．济南：山东科技出版社，1～179．
43．冷俊峰．无铅澄清旋光法测定赤砂糖和红糖中蔗糖的含量．甘蔗糖业，1995，4：46～49．
44．马立田，王式箴．高效液相色谱法测定低热量食品中的葡萄糖、果糖、蔗糖、麦芽糖醇和山梨糖醇的研究．食品与发酵工业，1998，24(4)：12～17．
45．孟盈，沙丽清．风干、烘干对不同热带森林土壤样品NH_4~+—N、NO_3—N测定结果的影响．农业环境保护，2001，20(5)：366～67．
46．宋安东，吴云汉，贾翠英等．离子注入对苹果酵母菌的诱变研究．郑州大学学报，2003，35(3)：48～63．
47．桑金隆，竺莉红，李孝辉等．离子注入诱变选育之江菌素产生菌．科技通报，2002，18(1)63～66．
48．单季先，陈悦．食品中蛋白质含量测定方法的探讨．中国卫生检验杂志， 2005，15(4)：453～454．
49．沈齐英，刘录，申林波．紫外诱变选育高效降酚微生物．环境科学与技术，2004，27(1)：82～85．
50．孙建义，翁晓燕．UV诱变微生物的突变参数的分析．生物数学学报，1996，11(1)：92～96．
51．谢志鹏，许志南，郑建明等．靛酚蓝反应测定发酵液中的氨态氮．浙江大学学报(工学版)，2005，39(3)：437～439．
52．陶黎明，顾学斌，王维炯等．微生物源黄酮类化合物的除草活性．农药，2006：45(3)：180～181．
53．唐启义，冯明光．实用统计分析及其计算机处理平台．北京：中国农业出版社出版，1997．
54．涂鹤龄．我国农田杂草研究和防治进展．农药，2001．40(3)：1～3．
55．万方浩，王韧．世界杂草生防的历史成就及我国杂草生防的现状与建议．生物防治通报，1991，7(2)：81～87。
56．王建荣，张曼夫．真菌高分子量核DNA的提纯．遗传， 1992，14(6)：29～30。
57．王婧，酿酒酵母原生质体菌株的筛选．大连轻工业学院学报，2002，21(4)：265～268．
58．王金霞，黄大年，杨炜等．稻瘟病菌原生质体制备和再生的研究．中国水稻科学，1992，6(1)：33～38．
59．王淑珍，白晨，范俊等．灵芝与糙皮侧耳原生质体融合子基因组RAPD分析．食用菌学报，2003，10(1)：1～5．
60．王强，何锦豪，李妙寿等．浙江省水稻田杂草发生种类及危害．浙江农业学报，2000，12(6)：317～324．
61．万佐玺．链格孢菌毒素对紫茎泽兰致病性及生物测定方法的研究．湖北民族学院学报(自然科学版)，2001，19(1)：20～23．
62．万佐玺，朱晶晶，强胜等．链格孢毒素对紫茎泽兰的致病机理．植物资源与环境学报，2001，10(3)：47～50．
63．翁艳军，赵作棋，徐建国等．青霉素产生菌的原生质体融合．中国抗生素杂志，2003，28(3)：129～130．
64．燕克勤，成亚利，朱宝成等．荧光标记紫孢侧耳和糙皮侧耳原生质体融合研究．食用菌学报，1997，4(4)：25～29．
65．叶丽娟，李强国．氨态氮测定方法的研究．衡阳师范学院学报：自然科学，2002，3(23)：18～19。
66．叶亚军，张敏，汤志良．抗稻瘟病菌株——诺卡氏菌A11的诱变育种．植物保护，2007：33(1)：33～36．
67．叶丽娟，朱辉，田敏．微生物来源的真菌细胞壁抑制剂的研究进展．国外医药抗生素分册，2005，1(26)：34～41．
68．袁德军，周启．农抗5102产生菌原生质体融合育种德研究Ⅳ．融合子FR—008德验证及其产生新活性物质的分离和鉴别．生物工程学报，1991，7(2)：142～147．
69．余荔华，刘祖同．米曲霉与黑曲霉原生质体中间电融杂交育种．清华大学学报，1994，34(6)：7～12．
70．余柳青，黄世文，徐正浩．稗草生物学研究进展．北京：21试剂植物保护的发展．中国科学技术出版社，1998，599～603．
71．赵航．微生物除草剂菌种的原生质体融合研究[硕士学位论文]．北京：中国农业科学院研究生院，2005．
72．赵航，周勇军，刘小川等．生物除草剂剂型研究进展．农药市场信息，2006，1：14～5．
73．张朝贤．我国农田杂草综合资力现状和任务．第六次全国杂草科学学术研讨会论文集．广西：广西民族出版社，1999，PP10～15．
74．张龙翔，张庭芳，李令媛等．生化实验方法和技术．北京：人民教育出版社，1981．
75．张天宇．胶胞炭疽菌菟丝子专化型．真菌学报，1985，4(4)：234～239．
76．张忠义，刘振林，陈辉．葡萄酒中总糖测定方法探讨．中国公共卫生，2005，21(11)：1398～1399．
77．张智维，杨辉，陈合．He—Ne激光诱变选育果胶酶高产菌株．食品工业科技，2006，27(10)：156～157．
78．张彭湃，2005．微生物菌种选育技术的发展与研究进展．生物学教学，30(9)：3～5．
79．周颖，叶非．生物源除草剂研究与使用进展．农药，2002，41(5)：7～17．
80．诸葛健，王正祥．1994．工业微生物实验技术手册．北京：中国轻工业出版社，464～467
81. Agbessi S, Beausejour J, Derg C. et al. Antagonistic properities of two recombinant strains of Streptomyces melanosporofaciens obtained by intraspecific protoplast fusion. Appl.Microbiol.Biot., 2003, 62:233-238.
82. Annamalai P and Lalithakumari D. Isolation and regeneration of protoplasts from mycelium of Drechslera oryzae. J.Plant Dis. Prot. 1991, 98:197-204.
83. Anne J, Peberdy J F. Conditions for induced fusion of fungal protoplasts in polgethylene glycol solutions. Arch Microbiol., 1975,105:201-205.
84. Auld B A, Hetherington S D, Smith H E. Advances in bioherbicide formulation .Weed Biology and Management, 2003, 3(2): 61—67
85. Auld B A. (Inventor) Bioherbicidal formulations. Australian Prrovisional Patent Application 2002952094.Patent Office, IP Australia, Canberra.Cheng S P and Cui Y B. Characteristics for the hybrid cell Foaz obtained from the protoplast fusion between yeast and photosynthetic bacteria in soybean processing wastewater. J. Environ Sci., 1998, 10 (3):365—371.
86. Balasubramanian N, Annie Juliet G, Srikalavivani P, et al. Release and regeneration of protoplasts from the fungus Trichothecium roseum. Can.J.Microbiol. 2003, 49 (4):263—267.
87. Barski G, Hebd C R. 1960.Seances Acid Sci. [M]. Oxford: Blackwell, 1825 — 1827.
88. Biologtami K S. Wide spread occurrence of toxigenic Alternaria in cereas and oilaseeds. Indian Phytopathol. 1995,48(2): 150-153.
89. Boyette C D, Tempton G E and Smith R J. Control of winged waterpromrose( Jussiaea decurrens) and northern jiontvetch (Aeschynomene virginica) with fungal pathogens. Weed Sc.i,1979, 27(5):497-501.
90. Boyete C D. Unrefined corn oil improves the mycoherbicidal activity of Colletotrichum truncatum for hemp sesbania (Sesbania exaltata) control. Weed Sci. 1994, 41:495—500.
91. Bowers R C. Commercialization of Collego -an industrialist's view. Weed Sci., 1986, 34: 24—25 (Suppl.1).
92. Cheng Shu-pei, Cui Yi-bin. Characteristics for the hybrid cell Foaz obtained from the protoplast fusion between yeast and photosynthetic bacteria in soybean processing wastewater. J. enviro sci., 1998, 10(3):365—371.
93. Chiaki I, Masanori O, Yoshiro O. Intergenus protoplast fusion between Streptomyces and Micromonospora with reference to the distribution of parental characteristics in the fusants. J.Biosci.Bioeng., 1999, 88 (2):143 —147.
94. Chiaki I, Yuko I, Takeshi K, et al. Isolation and characterization of the interspecific fusants from Streptomycetes obtained using a liquid regeneration method. Fisheries Sci., 2002, 68:395— 402.
95. Chittick A T, Ash G J, Kennedy R A, et al. Micro-encapsulation: an answer to formulation quandary? Proceedings of the international workshop, Bioherbicides: the Next Generation (27 April 2003), International Bioherbicide Group, 2003, Canberra.
96. Chung Y R. Identification of Exserohilum species, a fungal pathogen causing leaf blight of barnyardgrass (Echinochloa crusgalli). Korean J.Planol. 1990,6:429-433.
97. Ciotola M., Hallett S.G., Watson A.K. Impact of Tusarium oxysporum isolates M12-4A, upon seed germination of Striga hermothica in vitro. Sixth international Parasitic Weed symposium, 1996, pp871-875.
98. Connick W J Jr, Boyette C D, McAlpine J R. Formulation of mycoherbicides using a pasta-like process. Biol, control., 1991, 1:281—287.
99. Dai M H, Sara Z M, Thomas R, et al. Visualization of protoplast fusion and quantitation of recombination in fused protoplast of auxotrophic strains of Escherichia coli. Metabolic engineering. 2005, 7:45-52.
100. Daniel J T, Templeton G E, Smith R J JR. Biological control of northern jointvetch in rice with an endemic fungal disease. Weed sci., 1973,21:303—307.
101. Duan G F, Zhang Z B, Zhang J P, et al. 2007. Evaluation of crude toxin and metabolite produced by Helminthosporium gramineum Rabenh for the control of rice sheath blight in paddy fields. Crop Protection. 26:1036-1041.
102. Duan G F, Zhou Y J, Yuan Q S, et al. Phytotoxic ophiobolins produced by Helminthosporium Gramineum Rabenh, a potential bioherbicide for control of barnyardgrass (Echinochola Crus-Galli). NPAIJ, 2007.
103. Duke S O, Dayan F E and Rimandao A M. Natural products as tools for weed management. Proc.Japanese weed Sci Soc. (Suppl.). 1998,Pp.1—11.
104. Duke S Q, Lydon J. Herbicides from natural compounds. Weed Technology. 1987,1:122—128
105. Emge R G, Melching J S, Kingsolver C H. Epideniology of Puccinia chondrillina, a rust pathogen for the biocontrol o rush skeleton weed in the United States. Phytopathology, 1981,71:839-843.
106. Filippis L De, Hoffmann E, Hampp R. Identification of somatic hybrids of tobacco generated by electrofusion and culture of protoplasts using RAPD-PCR. Plant Science, 1996, 121:39-46.
107. Ferenczy L. 1980. Current microbiol [M]. London: Cambridge University Press , 555- 574.
108. Goto M. The relationship between Emmalocera sp. and barnyardgrass and its potential as a biological control. Integrated management of paddy and aquatic weeds in Asia, Proceedings of an international seminar, 1992, pp113—121.
109. Gohbara M, Yamaguchi K, Bay Petersen J. Biological agents for the control of paddy weeds in Japan. Integrated management of paddy and aquatic weeds in Asia. Proceedings of an international seminar, Tsukuba, Japan, 1994, pp184—194.
110. Hasan S. Specificity and host specialization of Puccinia chondrillina. Ann Appl Biol., 1972,72: 57—263.
111. Hasan S, Wapshere A J. The biology of Puccinia chondrillina a potential biological control agent of skeleton weed. Ann Appl Biol., 1973,74:325—332.
112. Imaizumi S, Nishino T, Miyabe K, Fujimori T, Yamada M. Biological control of annual bluegrass (Poa annual L.) with a Japanese isolate of Xanthomonas campestris pv.poae (JT-P482). Biological control, 1997,8:7-14.
113. Itoh k., Wang G X and Ohba S. Sulfonylures resisitance in Linderma micrantha, an Annual paddy weed in Japan. Weed Res., 1999, (39): 413-423.
114. Jackson M A, Shasha B S, Schialer D A. Formulation of Colletotrichum truncatum microsclerotia for improved biocontrol of the weed hempsesbania (Sesbania exaltata). Biological Control, 1996, 7: 107-113.
115. Kenney D, Devine S. The way it was developed-An industrialist's view. Weed Sci., 1986,34 (suppl.1):15-16.
116. Kishimoto K and Sawada H. Large-scale culture of the mildiomycin producer Streptoverticillium rimofaciens. Journal of Bioscience &Bioengineering. 1997, 75(4):245—253.
117. Kremer R J, Fatima M, Begonia T, et al. Characterization of rhizobacteria associated with weed seedlings. Applied Enviromental Microbiology, 1990,56:1649—1655.
118. Judit K, IIona P, Lajos F. A novel method for hybridization of Saccharomyces species without genetic markers. Can.J.Microbiol., 1998, 44 (10):959—964.
119. Lee GA. Integrated control of rush skeleton weed (Chondrilla juncea) in the Western US. Weed sci., 1986,34:2-6.
120. Lee H B, Kim C J, Hong K S, et al. A bleaching herbicidal activity of methoxyhygromycin(MHM) produced by an actinomycete strain Streptomyces sp. 8E-13, Letters in Applied Microbiology, 2003,36:387—391.
121. Lohuis H. Biological weeds control still in its fancy. PSP-Planzenchutz-Praxis, 1990,3:22—23.
122. Masao K, Tomoaki M, Kazunobu S, et al. Chromosomal deletion or rearrangement in chimeric hybrids of Saccharomycopsis fibuligera and Saccharomyces diastaticus obtained by cell fusion. Journal of Fermentation and Bioengineering. 1996, 81 (4):281—285.
123. McCullough H. A simple micro technique for the determination of blood ammonia and a note on the effect of exercise. Clinical Chemical Acta. 1968, 19:101 — 105.
124. Mohan B R, Sajeena A, Seetharaman K, et al .Advances in bioherbicides development—an overview. Crop Protection, 2003,22,253—260.
125. Mortenson K. The potential of an endemic fungus Colletotrichum gloeospioides f.sp. malvae for biological control of round-leaved mallow (Malvapusilla) and velvetleaf (Abutilon theophrastis). Weed Sci., 1992, 36: 473—478.
126. Mrinalini C and Lalithakumari D. Protoplast fusion: a biotechnological tool for strain improvement of Trichoderma sp. Curr. Rrend Life Sci., 1996, 21: 133—146.
127. Nakmura M, Ishibashi K. New antibiotic ophiobolin A produced by Ophiobolus miyabeanus. J.Agric. Chem. Soc, Japan. 1958, 32: 739—744.(in Japanese)
128. Ogina H. The registration of pesticides and establishment of safety evaluation methods for microbial pesticides. In: R. GBayot, et al. Process of international symposium on biological control and integrated management of paddy and aquatic weeds in Asia. October 20-23, 1992, Japan. pp425-437.
129. Okada Y. 1962. Exp cell [M]. New York: Academic Press, 98 - 107.
130. Quimby P C Jr, Zidack N K, Boyette C D, et al. A simple method for stabilizing and granulating fungi. Biocontrol Sci. Technol.,1999,(9):5-8.
131. Pesti M, Konszky E, Polga J, et al. 1979. Fifth international Protoplast Symposium. Baltimore: Williams & Wilkins, 54.
132. Rasmussen J O and Rasmussen O S. Characterization of somatic hybrids of potato by use of RAPD markers and isozyme analysis. Phys.Plant. 1995 , 93:357 - 364.Regulation and Guidelines: Critical issues in biological control. Proc, of a USDA/CSRS National Workshop. 10 -12June,1991.USA.
133. Resi E M. Multplication of Helminthosporium spp. on sensscent tissues of gtaminaceous weeds and soybean under natural conditions. Fitopatologia Brasileita.1985,10(3):643-648.
134. Rubinder K, Chadha B S, Singh S, et al. Amylase hyper-producing haploid recombinant strains of Thermomyces obtained by intraspecific protoplast fusion. Can.J.Microbiol., 000, 46 (7):669-673.
135. Saunders J A, Matthew B F, Miller P D. Plant gene transfer using electrofusion and electroporation. In electroporation and electrofusion in cell biology (Neumann.E, Sower A.E; Jordan C.A ). PP: 343-354,1989, Plenum press, New York and London.
136. Scheepens P C. Joint action of Cochliolus and atrazine on Echinochloa Crusgalli (L.) Beauv. Weed Res., 1987, 27: 43-47.
137. Seong H C, Chang S, Man J O, et al. Intergeneric protoplast fusion in Saccharomycopsis fibuligera and Saccharomyces cerevisiae. Journal of fermentation and bioengineering. 1997, 84 (2):158-161.Sugawara F, Strobel G, Strange R N, et al. Phytotoxins from the pathogenic fungi Drechslera maydis and Drechslera sorghicola. 1987, Proc.Natl.Acad.Sci., 84:3081 -3085.
138. Shabana Y M, Charudattan R, Devalerio J T, et al. An evaluationofhydrophilic polymers for formulating the bioherbicide agents Alternaria cassiae and A. eichhorniae,Weed Technol.,1997,(11):212-220.
139. Smith R J Jr. Biological control as components of integrated weed management for rice in United States. In: R. GBayot, et al. Process of international symposium on biological control and integrated management of paddy and aquatic weeds in Asia. October 20-23, 1992, Japan. p.335-351.
140. Spencer J F T, Spencer D M, Bizeau C, et al. The use of mitochondrial mutants in hybridation of industrial yeast strains.V: Relative parental contributions to the genomes of interspecific and intergeneric yeast hybrids obtained by protoplast fusion as determined by DNA reassociation. Curr. Genet., 1985, 9:623-625.
141. Suslow T V, Schroth M N. Role of deleterious rhizobacteria as minor pathogens in reducing crop growth. Phytopathology, 1982,72:111 —115.
142. Smith R J Jr. Biological control of Northern Jointvetch {Aeschynomene virginicd) in Rice {Oryza sativa) and soybeans {Glycine mar)—A Researcher's View. Weed Sci., 1986, 34(suppl.1):17—23.
143. Smitamana P, Watson A K. Screening of the selective biocontrol agents for weeds in Thailand. Abstracts of biological weed control in rice workshop, 10-12, may ,2000, IRRI.
144. Tanake H, et al. An attempt for development of bioherbicide for Eleocharis kuroquwai. In: R. GBayot. Process of international symposium on biological control and integrated management of paddy and aquatic weeds in Asia. October 20-23, 1992, Japan. pp381—392.
145. Teeradakorn S, Talawanich Y, Suzuki I, et al. Characterization of Streptomyces D3 derived from protoplast fusion between Streptomyces cyaneas 190-1 and Streptomyces griseoruber 42-9. World journal of microbiology & biotechnology. 1998, 14:565-570.86 Wang Z.Y., Nagel J., Potrykus I. and Spangenberg G Plants from cell suspension-derived protoplasts in Lolium species. Plant Sci., 1993, 94:179-193.
146. TeBeest D O, Templeton G E. Mycroherbicides progress in the biological control of weeds. Plant Dis.,1985, 69(1): 6-10.
147. TeBeest D O, Yang X B, Cisar C R. The status of biological control of weeds with plant pathogens. Annu. Rev. Phytopathol., 1992, 30: 637—657.
148. Trujillo E E, Aragaki M, Shoemaker R A. Infection, disease development, and axenic cultures of Entyloma compositarum, the cause of hamakua pamakani blight in Hawaii. Plant Dis., 1988,72: 355-357.
149. Tsukamoto H. Evaluation of fungal pathogens as biological agents for paddy weed, Echinochloa species by drop inoculation. Japan, Ann. Phytopathol. Soc, 1997, 63: 366—372.
150. Tsukamoto H. Effect of water management on mycoherbicidal activity of Exserohilum monoceras against Echinochlia orzicola. Japan, Ann. Phytopathol. Soc, 1998, 64: 526—631.
151. Vazquez F, Heluane H, Spencer J FT, et al. Fusion between protoplasts of Pichia Stipitis and isolated filamentous fungi myclei. Enzyme Microb.Tech., 1997, 21:32—38.
152. Watson A K. Current status of bioherbicide development and prospects for rice in Asia. In: R. GBayot. Process of international symposium on biological control and integrated management of paddy and aquatic weeds in Asia. October 20-23, 1992, Japan, p.367—379.
153. Yamaguchi K. Biological control of paddy weeds using fungal isolates. Proc International symposium on wild and weedy rices in agro-ecosystem. Ho Chi Minh, Vietnam, 1998, pp133— 136.
154. Watanabe H, Uchina A, Tachibana M. Application period of Drechslera monoceras, plant pathogen for the mycoherbicide, to control Echinochloa oryzicola vasing rice fields of Tohoku, Japan. The Proc I of the 18~(th) Asian-Pacific Weed Sci. Soc. Confi., Beijing, 2001, pp416—421.
155. Zhang W M, Watson A K. Host range of Exserohilum nomocoras, a potential bioherbicide for the control of Echinochloa species. Can. J. Bot.,1997, 75: 685—692.