胶孢炭疽菌作为防除波斯婆婆纳的生物除草剂产业化相关技术的研究
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摘要
波斯婆婆纳(Veronica persica Poir)是危害小麦、油菜等夏熟作物田的世界性恶性杂草。胶孢炭疽菌婆婆纳专化型(Colletotrichum glorosporidies Penz．f．sp．veronicae S．Qiang et Q．Zeng)是在对该草生物防治研究过程中，从波斯婆婆纳植株上分离的一株自然致病菌。实验室研究结果表明，该菌株对波斯婆婆纳属杂草侵染力和寄主专化性强，对小麦、油菜等作物安全，具备发展为防除波斯婆婆纳的真菌除草剂的潜力。本文研究了胶孢炭疽菌作为生物除草剂防除波斯婆婆纳的产业化潜力，旨在加速该菌的产业化进程。
首先通过对波斯婆婆纳生防菌株胶孢炭疽菌婆婆纳专化型QZ—97a培养发酵方法、培养基的种类、pH和水分含量及培养时间的研究，试图发展该菌分生孢子大批量生产工艺。结果表明，液体和固体发酵均能用于该菌的大批量生产，而液-固体联合发酵可以获得最高的产孢量。多种固体基质可以作为培养基，麦麸豆粕最佳，当加水量在固体培养基千重的1.2倍，pH值范围为6.5-7.5，接种固体培养基6天后单位重量的干物质产孢量达到了8.30×10~9个／克。波斯婆婆纳粉对固体培养基的产孢有增效作用。
通过这种方法生产的干孢子粉每克含有大量的分生孢子(接近10~(10)个)，贮藏简便。将不同含水量的胶孢炭疽菌婆婆纳专化型菌株QZ-97a分生孢子粉置于4℃和室温下，密闭或开放贮存。每隔15天取样测定孢子的萌发率，每隔60天测定芽管长度。结果表明，4℃下密闭贮藏6个月，分生孢子萌发率和芽管长度无明显降低，室温贮藏分生孢子萌发率和芽管长度降低地也不明显。贮藏10个月后，各种贮藏孢子的活力都显著下降。总体上看，4℃、含水量为4％、密闭贮藏孢子活力最稳定，420天后，孢子萌发率为70.4％，芽管长度为贮藏60天时孢子芽管长度的43.2％。含水量为9.6％、开放室温下贮藏的孢子粉活力下降最大，相应的孢子萌发率和芽管长度值分别为35.5％和12.7％。因此，温度和孢子粉含水量是影响孢子货架期的两个主要因素。分生孢子粉对温度有一定的耐受性，随温度升高，敏感性增强，60℃以上分生孢子的活力下降显著，处理15分钟，孢子芽管长度下降50％。
胶孢炭疽菌孢子与化学除草剂间生物学相容性好。与6种常用除草剂相容性的测定结果显示，分生孢子在供试除草剂中均能够很好地萌发，即使在田间常规使用浓度
胶饱炭疽菌作为防除波斯婆婆纳的生物除草剂产业化相关技术的研究
下，袍子的萌发率也均大于78.3%.供试除草剂对袍子芽管生长的抑制随着除草剂浓
度上升而明显增大，田间常规使用浓度下，饱子芽管在使它隆、2甲4氛钠盐及两者
的复配剂中仍能够生长.除草剂药效生物测定表明，在1/10和2/10田间常规使用浓
度下，使它隆对波斯婆婆纳的防效最好.因此，使它隆是菌药混用较好的候选除草剂。
蔗糖、葡萄糖等营养物质能够促进该菌袍子的萌发.以煤油、卵磷脂和吐温为组
分的助剂2对袍子的萌发有抑制作用，但可以抑制波斯婆婆纳的生长势.助剂与袍子
协同作用能够增强袍子对4一6叶期杂草的防效，但助剂浓度升高，防效增高不显著。
低剂量的使它隆与该菌袍子及助剂复配使用，在不保湿的情况下，对波斯婆婆纳的防
效显著提高.并且随着使它隆使用浓度的提高，对波斯婆婆纳的致病进程加快，防效
增加.含助剂2%的105个/m1的袍子悬浮液与常规剂量15%使它隆复配可使4一6叶期
的波斯婆婆纳在自然条件下致死。
通过对几种接种方法进行比较研究，用离体叶片法检测炭疽菌袍子的活力是较有
效的方法.在12，卜时光照12小时黑暗交替条件下，损伤浸渍法接种的症状最为明显.
Birdseye speedwell (Veronica persica Poir) is a major weed worldwide that significantly reduces yield of many different crops. In the search for biological control methods for birdseye speedwell, Colletotrichum gloeosporidies Penz. f.sp. veronicae S.Qiang et Q.Zeng, isolated from naturally infected birdseye speedwell, was virulent to the Veronica species but was safety to such crops as wheat, rape, soybean, cotton and com under laboratory or greenhouse conditions, which had the potential to be developed as mycoherbicide for birdseye speedwell. The aim of this study was to evaluate and improve the some related techniques for the commercialization of Collectotrium gloeosporiodies f.sp. veronicae as a potential mycoherbicide for birdseye speedwell.
Firstly, the studies on fermentation methods, kinds, pH and water content of cultural media, and cultural time were conducted in order to develop a technique of mass-production of conidia of Collectotrichum gloeosporioides as a biological agent for Veronica persica. The research results showed that the conidia of this fungus could be mass-produced in both of submerged and solid substrate, but the highest harvest of conidia was made under the fermentation combined mycelia produced in submerged substrate and conidia sporulated in solid substrate. The wheat-bran and bean-cake substrate was the best one selected from the solid media tested, which all were suitable for the culture of this fungus. The highest yield at 8.30 x 109 conidia g-1 dry weight was obtained under pH 6.5-7.5 and water content equaled to 1.2 time dry-weight of solid media and after six-day culture. The addition of Veronica persica powder could promote to sporulate in solid media.
The raw conidia powder produced by combined liquid-solid substrate fermentation contains abundant conidia (nearly 1010 conidia g-1 dry weight) , and it can be stored with simple equipment. While the fresh conidial powder of Collectotrichum gloeosporioides Penz. f.sp. veronicae Sheng Qiang et Qing Zeng was stored at 4癈 and room temperature in open or close capsule, germination rates and length of germ tube of conidia were
examined at 15 and 60 days intervals, respectively. Results showed that there were no significant differences in germination rates and length of germ tube of conidia stored at 4 癈 and room temperature within 6 months. However, the germination rates and length of germ tube of conidia decreased significantly after storage for over 10 months at 4C or room temperature. In general, the conidia viability were most stable when the conidia powder with 4.0% water content was stored closely at 4C because the germination rates were 70.4% and the length of germ tube decreased only by 43.2% by the end of 420 days comparing to that by 60 days. Whereas, the conidia powder with 9.6% water content stored openly at room temperature were declined the most obviously, the corresponding values were 35.5% and 12.7% respectively. Therefore, storage temperature and water content of conidia were two key factors, influencing the shelf-life of Collectotrichum gloeosporioides f.sp. veronicae conidia. The conidia were heat-treated in oven at 40~60 C for 15~120 min to observe tolerance to high temperature of conidia. Results showed that the conidia powder was partly tolerant to temperature, while the susceptivity increased obviously with temperature rising. A regression analysis showed that the length of germ tube of conidia were decreased by 50% when heat-treated at 60C for 6 min comparing with that of control.
The compatibility of Collectotrichum gloeosporioides conidia with six widely used herbicides was investigated by comparing germination rates and germ tube length of conidia in liquid containing 0.1-1.0 fold of concentration recommended for field spray. The results showed that the conidia germinated well in herbicides tested, even at the field-spray concentration recommended, the germination rate was more than 78.3%. The results also showed that negative effect of herbicide on the growth of conidia germ tube increased signific
首先通过对波斯婆婆纳生防菌株胶孢炭疽菌婆婆纳专化型QZ—97a培养发酵方法、培养基的种类、pH和水分含量及培养时间的研究，试图发展该菌分生孢子大批量生产工艺。结果表明，液体和固体发酵均能用于该菌的大批量生产，而液-固体联合发酵可以获得最高的产孢量。多种固体基质可以作为培养基，麦麸豆粕最佳，当加水量在固体培养基千重的1.2倍，pH值范围为6.5-7.5，接种固体培养基6天后单位重量的干物质产孢量达到了8.30×10~9个／克。波斯婆婆纳粉对固体培养基的产孢有增效作用。
通过这种方法生产的干孢子粉每克含有大量的分生孢子(接近10~(10)个)，贮藏简便。将不同含水量的胶孢炭疽菌婆婆纳专化型菌株QZ-97a分生孢子粉置于4℃和室温下，密闭或开放贮存。每隔15天取样测定孢子的萌发率，每隔60天测定芽管长度。结果表明，4℃下密闭贮藏6个月，分生孢子萌发率和芽管长度无明显降低，室温贮藏分生孢子萌发率和芽管长度降低地也不明显。贮藏10个月后，各种贮藏孢子的活力都显著下降。总体上看，4℃、含水量为4％、密闭贮藏孢子活力最稳定，420天后，孢子萌发率为70.4％，芽管长度为贮藏60天时孢子芽管长度的43.2％。含水量为9.6％、开放室温下贮藏的孢子粉活力下降最大，相应的孢子萌发率和芽管长度值分别为35.5％和12.7％。因此，温度和孢子粉含水量是影响孢子货架期的两个主要因素。分生孢子粉对温度有一定的耐受性，随温度升高，敏感性增强，60℃以上分生孢子的活力下降显著，处理15分钟，孢子芽管长度下降50％。
胶孢炭疽菌孢子与化学除草剂间生物学相容性好。与6种常用除草剂相容性的测定结果显示，分生孢子在供试除草剂中均能够很好地萌发，即使在田间常规使用浓度
胶饱炭疽菌作为防除波斯婆婆纳的生物除草剂产业化相关技术的研究
下，袍子的萌发率也均大于78.3%.供试除草剂对袍子芽管生长的抑制随着除草剂浓
度上升而明显增大，田间常规使用浓度下，饱子芽管在使它隆、2甲4氛钠盐及两者
的复配剂中仍能够生长.除草剂药效生物测定表明，在1/10和2/10田间常规使用浓
度下，使它隆对波斯婆婆纳的防效最好.因此，使它隆是菌药混用较好的候选除草剂。
蔗糖、葡萄糖等营养物质能够促进该菌袍子的萌发.以煤油、卵磷脂和吐温为组
分的助剂2对袍子的萌发有抑制作用，但可以抑制波斯婆婆纳的生长势.助剂与袍子
协同作用能够增强袍子对4一6叶期杂草的防效，但助剂浓度升高，防效增高不显著。
低剂量的使它隆与该菌袍子及助剂复配使用，在不保湿的情况下，对波斯婆婆纳的防
效显著提高.并且随着使它隆使用浓度的提高，对波斯婆婆纳的致病进程加快，防效
增加.含助剂2%的105个/m1的袍子悬浮液与常规剂量15%使它隆复配可使4一6叶期
的波斯婆婆纳在自然条件下致死。
通过对几种接种方法进行比较研究，用离体叶片法检测炭疽菌袍子的活力是较有
效的方法.在12，卜时光照12小时黑暗交替条件下，损伤浸渍法接种的症状最为明显.
Birdseye speedwell (Veronica persica Poir) is a major weed worldwide that significantly reduces yield of many different crops. In the search for biological control methods for birdseye speedwell, Colletotrichum gloeosporidies Penz. f.sp. veronicae S.Qiang et Q.Zeng, isolated from naturally infected birdseye speedwell, was virulent to the Veronica species but was safety to such crops as wheat, rape, soybean, cotton and com under laboratory or greenhouse conditions, which had the potential to be developed as mycoherbicide for birdseye speedwell. The aim of this study was to evaluate and improve the some related techniques for the commercialization of Collectotrium gloeosporiodies f.sp. veronicae as a potential mycoherbicide for birdseye speedwell.
Firstly, the studies on fermentation methods, kinds, pH and water content of cultural media, and cultural time were conducted in order to develop a technique of mass-production of conidia of Collectotrichum gloeosporioides as a biological agent for Veronica persica. The research results showed that the conidia of this fungus could be mass-produced in both of submerged and solid substrate, but the highest harvest of conidia was made under the fermentation combined mycelia produced in submerged substrate and conidia sporulated in solid substrate. The wheat-bran and bean-cake substrate was the best one selected from the solid media tested, which all were suitable for the culture of this fungus. The highest yield at 8.30 x 109 conidia g-1 dry weight was obtained under pH 6.5-7.5 and water content equaled to 1.2 time dry-weight of solid media and after six-day culture. The addition of Veronica persica powder could promote to sporulate in solid media.
The raw conidia powder produced by combined liquid-solid substrate fermentation contains abundant conidia (nearly 1010 conidia g-1 dry weight) , and it can be stored with simple equipment. While the fresh conidial powder of Collectotrichum gloeosporioides Penz. f.sp. veronicae Sheng Qiang et Qing Zeng was stored at 4癈 and room temperature in open or close capsule, germination rates and length of germ tube of conidia were
examined at 15 and 60 days intervals, respectively. Results showed that there were no significant differences in germination rates and length of germ tube of conidia stored at 4 癈 and room temperature within 6 months. However, the germination rates and length of germ tube of conidia decreased significantly after storage for over 10 months at 4C or room temperature. In general, the conidia viability were most stable when the conidia powder with 4.0% water content was stored closely at 4C because the germination rates were 70.4% and the length of germ tube decreased only by 43.2% by the end of 420 days comparing to that by 60 days. Whereas, the conidia powder with 9.6% water content stored openly at room temperature were declined the most obviously, the corresponding values were 35.5% and 12.7% respectively. Therefore, storage temperature and water content of conidia were two key factors, influencing the shelf-life of Collectotrichum gloeosporioides f.sp. veronicae conidia. The conidia were heat-treated in oven at 40~60 C for 15~120 min to observe tolerance to high temperature of conidia. Results showed that the conidia powder was partly tolerant to temperature, while the susceptivity increased obviously with temperature rising. A regression analysis showed that the length of germ tube of conidia were decreased by 50% when heat-treated at 60C for 6 min comparing with that of control.
The compatibility of Collectotrichum gloeosporioides conidia with six widely used herbicides was investigated by comparing germination rates and germ tube length of conidia in liquid containing 0.1-1.0 fold of concentration recommended for field spray. The results showed that the conidia germinated well in herbicides tested, even at the field-spray concentration recommended, the germination rate was more than 78.3%. The results also showed that negative effect of herbicide on the growth of conidia germ tube increased signific
引文
1．陈涛，张友清，孙松柏等．生物农药检测及其原理．北京：农业出版社，1993．
2．董汉松，王智发．烟草赤星病致病菌测定方法的研究．山东农业大学学报．1989，20(4)：1～8．
3．方仲达．植病研究方法．北京：中国农业出版社，1998．
4．冯文煦．杂草与作物间的竞争关系概述．杂草学报．1990，4(2)：43-47
5．高昭远，干静娥，张立洁等．我国利用植物病原微生物防除杂草研究的回顾与现状．中国农学通报．1995，11(1)：31-37
6．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
7．郭凤根，李扬汉．园林菟丝子生防真菌喷施条件的研究．中国生物防治．2000，16(2)：81-83
8．郭水良，耿贺利．麦田波斯婆婆纳化除及其方案评价．农药．1998，37(6)：27-30
9．胡萃，王韧．国外应用微生物防治杂草的进展．生物防治通报．1987，3(1)：35-38
10．胡建平．使它隆防除麦田婆婆纳试验示范总结．杂草科学．1988，3(4)：20-21
11．黄世文，余柳青，Watson A K. 稗草病原菌 Alternative alternata 和 Curvularia lunata 的产孢 特性研究．面向21世纪中国农田杂草可持续治理．南宁：广西民族出版社，1999，170-176
12．黄世文，余柳青，Watson.A.k..影响链格孢菌生长及产孢的因子.中国生物防治．2001，17(1)：10-15．
13．黄颂禹，徐志康．婆婆纳的发生与防除初报．杂草科学．1988，2(4)：38
14．蒋玲秀，颜玉树．江苏省棉花苗期田间阔叶杂草发生调查研究．杂草学报．1995，9(2)：13-18
15．农向群，高松，邓春生等．白僵菌绿僵菌分生孢子对高温的耐受力．中国生物防．1999，15(3)：111-114
16．强胜(主编)．杂草学．北京：中国农业出版社，2001，144-155
17．强胜，郭爱民，李扬汉．大批量生产百日草链格孢菌孢子的技术．中国生物防治．1997，13(4)：169-172．
18．强胜．安徽沿江圩农区夏熟作物田杂草群落的研究．植物生态学与地植物学学报．1990，14(3)：213-219
19．强胜．链格孢菌作为真菌除草剂防除紫茎泽兰潜力的研究．1998，南京农业大学博士论文
20．强胜．生物除草剂的研究概况．杂草科学．1996，10(2)：10-14．
21．强胜．生物除草剂的最新研究进展．杂草科学．1997，11(4)：9．10
22．强胜．生物除草剂研究的历史、现状及展望．见：周大荣主编，植物保护21世纪展望．北京：中国科学技术出版社，1998，112-118
23．全国农田杂草考察组．中国大豆田主要杂草的分布与危害．杂草学报．1988，2(3)：18-27
24．全国农田杂草考察组．中国麦田主要杂草的分布与危害．杂草学报．1988，2(1)：14-23
25．沈萍，卫扬保，王静怡等．微生物学．北京：高等教育出版社，2001，82—86．
26．万方浩．杂草生物防治的传统方法及理论．生物防治通报．1992，8(3)：131-136
27．王金其，陆善庆．使它隆防除麦田阔叶杂草应用技术研究．杂草学报．1988，2(3)：37-40
28．徐志康，黄颂禹．麦田恶性杂草波斯婆婆纳的生物学特性与化学防除试验初报．江苏杂草科学．1985，2：19-21
29．许寿涛，应盛华，冯明光．十种常用农药与球孢白僵菌的生物学相容性．植物保护学报，2002，29(2)：158-162
30．严彪．绿麦隆与杀草丹混用防除婆婆纳试验．江苏杂草科学．1986，4：10-11
31．余柳青，黄世文，徐正浩．微生物除草剂的研究和开发进展．见喻子牛主编，微生物农药的及其产业化．北京：科学出版社，2000，256-259．
32．袁世斌．生物除草剂胶胞炭疽菌菟丝子专化型的研究．1999，南京农业大学硕士论文．
33．曾青，强胜．波斯婆婆纳生防真菌QZ—97a的生物学特性．面向21世纪中国农田杂草可持续治理．南宁：广西民族出版社．1999，170-176
34．曾青，强胜．低量化学除草剂对菌株QZ—97a侵染波斯婆婆纳的影晌．植物保护学报．2002，29(4)：361-365
35．曾青，强胜．波斯婆婆纳生防菌株QZ—97a的分离、鉴定与致病性研究．南京农业大学学报．2000，23(3)：21．24
36．曾青，强胜．胶胞炭疽菌婆婆纳专化型及其侵染过程．植物保护学报．2001，28(3)：279-284
37．曾青，强胜．胶胞炭疽菌婆婆纳专化型侵染波斯婆婆纳的影响因子的研究．应用生态学报．2002，13(7)：833-836
38．曾青．炭疽菌作为真菌除草剂防除波斯婆婆纳的潜力研究．1999，南京农业大学硕士论文．
39．张朝贤，胡详实．国外除草剂应用趋势及我国杂草科学研究现状和发展方向．植物保护学报．1997，24(3)：278-282
40．张玉聚，孙化田，王春生．除草剂及其混用与农田杂草化学防治 北京：中国农业科技出版社，2000，285．
41．张泽博．我国棉田杂草分类、分布和防除．杂草科学．1994，1：1-2
42．张宗俭，李扬汉．野燕麦生防真菌燕麦叶枯菌的分离培养及致病性研究．中国生物防治．1996，12(4)：171-173
43．周熙群．使它隆防除麦田阔叶杂草的效果．杂草科学．1991，1：17-18
44．Abbas H K, Egley G H. Influence of unrefined corn oil and surface-active agents on the germination and infectivity of Alternaria helianthi. Biocontrol Sci and Tech. 1996, 6(4): 531-538
45. Amsellem Z, Zidack N K, Quimby P C, et al. Long-term dry preservation of viable mycelia of two mycoherbicide organism. Crop protection. 18(10), 1999, 643-649.
46. Amsellen Z, Sharon A, Gressel J. Abolition of selectivity of two mycoherbicidal organisms and enhanced virulence of a virulent fungi by an invert emulsion. Phytopathology. 1991, 81: 925-929.
47. Auld B A. Mass production of fungi for biopesticides. Plant Protection Quarterly. 1993,8(1): 17-19
48. Auld B A. Economic aspects of biological weed control with plant pathogens. In Tebeest D. O. Microbial control of weed. New York Chapman and Hall, 1991, 262—273.
49. Auld B A, Smith H, Qiang S. Control of cocklebur with combination of Alternaria zinniae and low rates of Imazaquin. The Proceeding of 16th Asian-Pasific Weed Science Society Conference. Malasian Plant Protection Society. 1997, 345-347
50. Auld B A, Smith H E, and Qiang S. Control ofcocklebur with a combination of Alternaria zinniae and low rates of imazaquin. In "Proceedings of the 16th Asian-Pacific weed science society conference" (Anonymous. Ed.). Asian-Pacific Weed Sci. Soc., Kuala Lampur, Mylaysia. 1997, 345-357
51. Auld B A, Smith H & Qiang S. Control of cocklebur with a combination of Alternaria zinniae and low rates of imazaquin. The proc. of 16th Asian-Pacific weed science society conference, Malaysian plant protection society. 1997, pp. 345-347.
52. Bowers R C. Commercialization of Collego~(TM) -an industrialist's view. Weed Science. 1982, 34s:24-25
53. Bowers R C. Commercialization of microbial biological control agents. In: Charudattan, R. and H.L. Walker.(ed.) Biological Control of Weeds with Plant Pathogens. Wiley and Sons. New York. pp. 1982, 157-174.
54. Boyette C D, Quimby P C, Caesar A J, et al. Adjuvants, formulations and spraying systems for improvement of mycoherbicides. Weed Technology. 1996, 11(10): 637-644.
55. Boyette C D, Abbas H K, Smith R J. Invert emulsions alter host-specificity of biological fungi. Phytopathology. 1991, 71,126.
56. Boyette C D, Quimby P C, Bryson C T, et al. Biological control of hemp sesbania(sesbania exaltata) under field conditions with Colletotrichum truncatum formulated in inverted emulsion. Weed Science. 1993, 41:497-500.
57. Boyette C D, Quimby P C, Connick W J, et al. Progress in the production, formulation, and application of mycoherbicides, in Tebeest D O Ed. Microbial control of weed. New York Chapman and Hall, 1991,209-222.
58. Boyette C D, Smith R J, Abbas H K, et al. Bioherbicidal control of hemp sesbania (Sesbania exaltata) and northern jointvetch (Aeschynomene virginica) with anthracnose pathogen-invert formulations. Proc. South. Weed Science Soc. 1992, 45:293.
59. Boyette C D, Quimby P C, Caesar A J, et al. Adjuvants, formulations and spraying systems for improvement of mycoherbicides. Weed Technology. 1996, 11(10):637—644.
60. Browers R C. Commercialization of Collego~(TM):--an industrialist's view .Weed Science [J]. 1986, 34s: 24-25.
61. Chandramohan S, Charudattan R. Control of seven grasses with a mixture of three fungal pathogens with restricted host ranges. Biological Control. 2001, 22: 246——255.
62. Charudattan R, Dinoor A. Biological control of weeds using plant pathogens accomplishments and limitations. Crop Protection. 2000, 19: 691-695.
63. Charudattan R. Integrated Control ofWaterhyacinth (Eichhornia crassipes) with a pathogen, insects, and herbicides. Weed Science. 1986, 34s: 26-30.
64. Charudattan R. The mycoherbicide approach with plant pathogeus. In Tebeest D O. Microbial control of weed. New York Chapman and Hall, 1991, 24-57.
65. Churchill B W. Mass production of microorganisms for biological control. In: Charudattan, R. and H.L. Walker.(ed.) Biological Control of Weeds with Plant Pathogens. Wiley and Sons. New York. 1982, 139-156
66. Conneau M, Pasquette B. Metabolism of chlortoluron in tolerant species: possible role of cytochrome P-450 mono-oxygenases. Weed Research, UK. 1988, 28(1): 19-25
67. Dumas M T, Wood J E, Mitchell E G, et al. Control of stump sprouting of Populus tremuloides and P. grandidentata by inoculation with Chondrostereum purpureum.Biological control .1997, 10, 37-41
68. Ferrero A, Balsari P. Preliminary results of flame weeding in orchards. Weed Abstract. 1996, 45(1): 46
69. Gange A C, Brown V K. Effects of pesticides on the germination of weeds: implications for Manuplative experiments. Journal of Applied Ecology. 1992, 29(2): 30
70. Ghorbani R, Seel W, Litterick A, et al. Evaluation of Alternaria alternata for biological control of Amaranthus retrofiexus. Weed science, 2000, 48:474-480
71. Grunry A C, Mead A. Modelling the effect of weed seed distribution in the soil profile on seedling emergence. Weed Research. 1996, 36(5): 375-384
72. Gupta R, Mukerji K G Biological control of weeds with plant pathogens. In Upadhyay R K, Mukerji K G, Chamela B P. Biocontrol potential and its exploitation in sustainableagriculture. NewYork. Kluwer Academic/Plenum Publishers, 2000, 199-205
73. Hamill A S. Herbicide reduction in North America:In Canada, an Opportunity for motivation and growth in weed management. Weed Technology. 1994, 8:366-371
74. Heisel T, Andreasen C. Annual weed distributions can be mapped with kriging. Weed Research. 1996, 36(4): 325-337
75. Hoagland R E. Alternaria cassiae alters phenylpropanoid metabolism in sicklepod (Cassia obtusifolia). J. Phytopathol. 1990, 130, 177-187
76. Holm L, Pancho J V. A Geographical Atlas of World Weeds. New York: John Wiley & Sons, 1979.
77. Hosoda A, Hama H. Insecticide risistance of the cotton aphid, Aphis Goossypii Glover.Ⅲ.Host preference and organophosphorus susceptibility. Weed Abstract. 1994, 43(5) : 242
78. Jackson M A, Baruch S, David A S. Formulation of Colletotrichum truncatum microsclerotia for improved biocontrol of the weed hemp sesbania(sesbania exaltata). Biological control ,1996 (7),107-113
79. Kenney D S. Devine——the way it development——an industrialist's view. Weed Science. 1986, 34s: 15-16.
80. Khodayari K, Smith R J, Walke J T, et al. Applicators for a weed pathogen plus acifluorfen in soybeans. Weed Technology. 1987, 1:37-40.
81. Klein T A, Auld B A and Wang F. Evalution of oil suspension emulsion of Colletotrichum orbichlare as a mycoherbicide in field trials. Crop Protection, 1995, 14(3): 193-197
82. Wymore L A, Piorier C, and Watson A K. Colletotrichum coccodes a potential bioherbicide for control of velvet leaf (Abutition theophrasti), Plant Disease, 1988, 72:534-538
83. Mohanbabu R, Sajeena A, Seetharaman k, et al. Advances in bioherbicides development——an overiew. Crop Protection. 2002, Article In web.
84. Pulcher M, Hurle K. Weed flora and weed seed bank in wheat monoculture under different levels of crop protection. Weed Abstracts. 1984, 33(10): 341
85. Quimy P C, Boyette C D. Production and application of biocontrol agent[A]. in Mcwhort C G, Gebhardt M R. Methods of applying herbicides, Weed Science soc, Champaign IL, 1987. p.265-280
86. Quimy P C, Fulgham F E, Boyette C D,et al. New formulations nozzles boost efficacy of pathogens for weed control. Proc. Weed Science soc, 1988, 28-52.
87. Sansome G. Effect of crop population on the performance of herbicides in winter oilseed rape. Aspects of Applied Biology. 1990.23:257-262
88. Scacchi A, Bortolo R. Dectection, characterization, and phytotoxic activity of the nucleoside antibiotics, blasticidin S and 5-hydroxymethyl-blasticidin S. Journal of Plant Growth Regulation. 1992.11(1): 39-46
89. Smith R J. Jr. Integration of biological control agents with chemical pesticides.In Tebeest D O. Microbial control of weed. New York Chapman and Hall, 1991,
90. Stowell L J. Submerged fermentation of biological herbicides.In Tebeest D O. Microbial control of weed. New York Chapman and Hall, 1991, 225-261.
91. Tebeest D O, Templeton G E. Mycoherbicides: progress in the biological control weeds. Plant Disease.1985, 69 (2): 6—10.
92. Templeton G E, Tebeest D O, Smith R J. Biological weed control with myeoherbieides. Ann Rev Phytopathology.1979, 17:301-310.
93. Templeton G E. Status of weed control with plant pathogens.[a].in Charudattan R. and Walker H L. Biological control of weeds with plant pathogens. New York Wiley, 1982, 29-44
94. Templeton G E, Heiny D K. Improvement of fungi to enhance mycoherbicide potential, in Whipps J.M. and R.D. Lunsden eds. Biotechnology of Fungi for Improving Plant Growth. British Mycological Society, Cambridge University Press, Cambridge. pp. 1989. 127-151
95. Templeton G E. Weed control with pathogens: future needs and directions, in R.E. Hoagland, ed. Microbes and Microbial Products as Herbicides, ACS Symp. Set. 439. American Chemical Society, Washington, DC.pp. 1990.320-329
96. Vurro M, Ellis B E. Effect of fungal toxins on induction of phenylalanine ammonia-lyase activity in elicited cultures of hybrid poplar. Plant science 1997, 126,29-38
97. Vurro M, Zonno M C, Evidente A, et al. Enhancement of efficacy of Ascochyta caulina to control Chenopodium albumn by use ofphytotoxins and reduced rates of herbicides. BioControl. 2001.182-190.
98. Walker H L, Connick W J. Sodium alginate for production and formulation of mycoherbicides. Weed Science. 1983, 33: 333-338.
99. Womack J G, Eccleston G M, Burge M N. A vegetable oil-based invert emulsion for mycoherbicide delivery. Biological control, 1996(6), 23-28
100. Zhang W M, Watson. A K. Characterization of growth and conidia production of Exserohium monceras on different substrates. Biocontrol Sciences and Technology. 1997, 7: 75—86.
101. Zimdahl R L. Fundamentals of weed science. Academic Press. 2nd ed. San Diego. California: Academic Press, 1999, 247-269.
2．董汉松，王智发．烟草赤星病致病菌测定方法的研究．山东农业大学学报．1989，20(4)：1～8．
3．方仲达．植病研究方法．北京：中国农业出版社，1998．
4．冯文煦．杂草与作物间的竞争关系概述．杂草学报．1990，4(2)：43-47
5．高昭远，干静娥，张立洁等．我国利用植物病原微生物防除杂草研究的回顾与现状．中国农学通报．1995，11(1)：31-37
6．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
7．郭凤根，李扬汉．园林菟丝子生防真菌喷施条件的研究．中国生物防治．2000，16(2)：81-83
8．郭水良，耿贺利．麦田波斯婆婆纳化除及其方案评价．农药．1998，37(6)：27-30
9．胡萃，王韧．国外应用微生物防治杂草的进展．生物防治通报．1987，3(1)：35-38
10．胡建平．使它隆防除麦田婆婆纳试验示范总结．杂草科学．1988，3(4)：20-21
11．黄世文，余柳青，Watson A K. 稗草病原菌 Alternative alternata 和 Curvularia lunata 的产孢 特性研究．面向21世纪中国农田杂草可持续治理．南宁：广西民族出版社，1999，170-176
12．黄世文，余柳青，Watson.A.k..影响链格孢菌生长及产孢的因子.中国生物防治．2001，17(1)：10-15．
13．黄颂禹，徐志康．婆婆纳的发生与防除初报．杂草科学．1988，2(4)：38
14．蒋玲秀，颜玉树．江苏省棉花苗期田间阔叶杂草发生调查研究．杂草学报．1995，9(2)：13-18
15．农向群，高松，邓春生等．白僵菌绿僵菌分生孢子对高温的耐受力．中国生物防．1999，15(3)：111-114
16．强胜(主编)．杂草学．北京：中国农业出版社，2001，144-155
17．强胜，郭爱民，李扬汉．大批量生产百日草链格孢菌孢子的技术．中国生物防治．1997，13(4)：169-172．
18．强胜．安徽沿江圩农区夏熟作物田杂草群落的研究．植物生态学与地植物学学报．1990，14(3)：213-219
19．强胜．链格孢菌作为真菌除草剂防除紫茎泽兰潜力的研究．1998，南京农业大学博士论文
20．强胜．生物除草剂的研究概况．杂草科学．1996，10(2)：10-14．
21．强胜．生物除草剂的最新研究进展．杂草科学．1997，11(4)：9．10
22．强胜．生物除草剂研究的历史、现状及展望．见：周大荣主编，植物保护21世纪展望．北京：中国科学技术出版社，1998，112-118
23．全国农田杂草考察组．中国大豆田主要杂草的分布与危害．杂草学报．1988，2(3)：18-27
24．全国农田杂草考察组．中国麦田主要杂草的分布与危害．杂草学报．1988，2(1)：14-23
25．沈萍，卫扬保，王静怡等．微生物学．北京：高等教育出版社，2001，82—86．
26．万方浩．杂草生物防治的传统方法及理论．生物防治通报．1992，8(3)：131-136
27．王金其，陆善庆．使它隆防除麦田阔叶杂草应用技术研究．杂草学报．1988，2(3)：37-40
28．徐志康，黄颂禹．麦田恶性杂草波斯婆婆纳的生物学特性与化学防除试验初报．江苏杂草科学．1985，2：19-21
29．许寿涛，应盛华，冯明光．十种常用农药与球孢白僵菌的生物学相容性．植物保护学报，2002，29(2)：158-162
30．严彪．绿麦隆与杀草丹混用防除婆婆纳试验．江苏杂草科学．1986，4：10-11
31．余柳青，黄世文，徐正浩．微生物除草剂的研究和开发进展．见喻子牛主编，微生物农药的及其产业化．北京：科学出版社，2000，256-259．
32．袁世斌．生物除草剂胶胞炭疽菌菟丝子专化型的研究．1999，南京农业大学硕士论文．
33．曾青，强胜．波斯婆婆纳生防真菌QZ—97a的生物学特性．面向21世纪中国农田杂草可持续治理．南宁：广西民族出版社．1999，170-176
34．曾青，强胜．低量化学除草剂对菌株QZ—97a侵染波斯婆婆纳的影晌．植物保护学报．2002，29(4)：361-365
35．曾青，强胜．波斯婆婆纳生防菌株QZ—97a的分离、鉴定与致病性研究．南京农业大学学报．2000，23(3)：21．24
36．曾青，强胜．胶胞炭疽菌婆婆纳专化型及其侵染过程．植物保护学报．2001，28(3)：279-284
37．曾青，强胜．胶胞炭疽菌婆婆纳专化型侵染波斯婆婆纳的影响因子的研究．应用生态学报．2002，13(7)：833-836
38．曾青．炭疽菌作为真菌除草剂防除波斯婆婆纳的潜力研究．1999，南京农业大学硕士论文．
39．张朝贤，胡详实．国外除草剂应用趋势及我国杂草科学研究现状和发展方向．植物保护学报．1997，24(3)：278-282
40．张玉聚，孙化田，王春生．除草剂及其混用与农田杂草化学防治 北京：中国农业科技出版社，2000，285．
41．张泽博．我国棉田杂草分类、分布和防除．杂草科学．1994，1：1-2
42．张宗俭，李扬汉．野燕麦生防真菌燕麦叶枯菌的分离培养及致病性研究．中国生物防治．1996，12(4)：171-173
43．周熙群．使它隆防除麦田阔叶杂草的效果．杂草科学．1991，1：17-18
44．Abbas H K, Egley G H. Influence of unrefined corn oil and surface-active agents on the germination and infectivity of Alternaria helianthi. Biocontrol Sci and Tech. 1996, 6(4): 531-538
45. Amsellem Z, Zidack N K, Quimby P C, et al. Long-term dry preservation of viable mycelia of two mycoherbicide organism. Crop protection. 18(10), 1999, 643-649.
46. Amsellen Z, Sharon A, Gressel J. Abolition of selectivity of two mycoherbicidal organisms and enhanced virulence of a virulent fungi by an invert emulsion. Phytopathology. 1991, 81: 925-929.
47. Auld B A. Mass production of fungi for biopesticides. Plant Protection Quarterly. 1993,8(1): 17-19
48. Auld B A. Economic aspects of biological weed control with plant pathogens. In Tebeest D. O. Microbial control of weed. New York Chapman and Hall, 1991, 262—273.
49. Auld B A, Smith H, Qiang S. Control of cocklebur with combination of Alternaria zinniae and low rates of Imazaquin. The Proceeding of 16th Asian-Pasific Weed Science Society Conference. Malasian Plant Protection Society. 1997, 345-347
50. Auld B A, Smith H E, and Qiang S. Control ofcocklebur with a combination of Alternaria zinniae and low rates of imazaquin. In "Proceedings of the 16th Asian-Pacific weed science society conference" (Anonymous. Ed.). Asian-Pacific Weed Sci. Soc., Kuala Lampur, Mylaysia. 1997, 345-357
51. Auld B A, Smith H & Qiang S. Control of cocklebur with a combination of Alternaria zinniae and low rates of imazaquin. The proc. of 16th Asian-Pacific weed science society conference, Malaysian plant protection society. 1997, pp. 345-347.
52. Bowers R C. Commercialization of Collego~(TM) -an industrialist's view. Weed Science. 1982, 34s:24-25
53. Bowers R C. Commercialization of microbial biological control agents. In: Charudattan, R. and H.L. Walker.(ed.) Biological Control of Weeds with Plant Pathogens. Wiley and Sons. New York. pp. 1982, 157-174.
54. Boyette C D, Quimby P C, Caesar A J, et al. Adjuvants, formulations and spraying systems for improvement of mycoherbicides. Weed Technology. 1996, 11(10): 637-644.
55. Boyette C D, Abbas H K, Smith R J. Invert emulsions alter host-specificity of biological fungi. Phytopathology. 1991, 71,126.
56. Boyette C D, Quimby P C, Bryson C T, et al. Biological control of hemp sesbania(sesbania exaltata) under field conditions with Colletotrichum truncatum formulated in inverted emulsion. Weed Science. 1993, 41:497-500.
57. Boyette C D, Quimby P C, Connick W J, et al. Progress in the production, formulation, and application of mycoherbicides, in Tebeest D O Ed. Microbial control of weed. New York Chapman and Hall, 1991,209-222.
58. Boyette C D, Smith R J, Abbas H K, et al. Bioherbicidal control of hemp sesbania (Sesbania exaltata) and northern jointvetch (Aeschynomene virginica) with anthracnose pathogen-invert formulations. Proc. South. Weed Science Soc. 1992, 45:293.
59. Boyette C D, Quimby P C, Caesar A J, et al. Adjuvants, formulations and spraying systems for improvement of mycoherbicides. Weed Technology. 1996, 11(10):637—644.
60. Browers R C. Commercialization of Collego~(TM):--an industrialist's view .Weed Science [J]. 1986, 34s: 24-25.
61. Chandramohan S, Charudattan R. Control of seven grasses with a mixture of three fungal pathogens with restricted host ranges. Biological Control. 2001, 22: 246——255.
62. Charudattan R, Dinoor A. Biological control of weeds using plant pathogens accomplishments and limitations. Crop Protection. 2000, 19: 691-695.
63. Charudattan R. Integrated Control ofWaterhyacinth (Eichhornia crassipes) with a pathogen, insects, and herbicides. Weed Science. 1986, 34s: 26-30.
64. Charudattan R. The mycoherbicide approach with plant pathogeus. In Tebeest D O. Microbial control of weed. New York Chapman and Hall, 1991, 24-57.
65. Churchill B W. Mass production of microorganisms for biological control. In: Charudattan, R. and H.L. Walker.(ed.) Biological Control of Weeds with Plant Pathogens. Wiley and Sons. New York. 1982, 139-156
66. Conneau M, Pasquette B. Metabolism of chlortoluron in tolerant species: possible role of cytochrome P-450 mono-oxygenases. Weed Research, UK. 1988, 28(1): 19-25
67. Dumas M T, Wood J E, Mitchell E G, et al. Control of stump sprouting of Populus tremuloides and P. grandidentata by inoculation with Chondrostereum purpureum.Biological control .1997, 10, 37-41
68. Ferrero A, Balsari P. Preliminary results of flame weeding in orchards. Weed Abstract. 1996, 45(1): 46
69. Gange A C, Brown V K. Effects of pesticides on the germination of weeds: implications for Manuplative experiments. Journal of Applied Ecology. 1992, 29(2): 30
70. Ghorbani R, Seel W, Litterick A, et al. Evaluation of Alternaria alternata for biological control of Amaranthus retrofiexus. Weed science, 2000, 48:474-480
71. Grunry A C, Mead A. Modelling the effect of weed seed distribution in the soil profile on seedling emergence. Weed Research. 1996, 36(5): 375-384
72. Gupta R, Mukerji K G Biological control of weeds with plant pathogens. In Upadhyay R K, Mukerji K G, Chamela B P. Biocontrol potential and its exploitation in sustainableagriculture. NewYork. Kluwer Academic/Plenum Publishers, 2000, 199-205
73. Hamill A S. Herbicide reduction in North America:In Canada, an Opportunity for motivation and growth in weed management. Weed Technology. 1994, 8:366-371
74. Heisel T, Andreasen C. Annual weed distributions can be mapped with kriging. Weed Research. 1996, 36(4): 325-337
75. Hoagland R E. Alternaria cassiae alters phenylpropanoid metabolism in sicklepod (Cassia obtusifolia). J. Phytopathol. 1990, 130, 177-187
76. Holm L, Pancho J V. A Geographical Atlas of World Weeds. New York: John Wiley & Sons, 1979.
77. Hosoda A, Hama H. Insecticide risistance of the cotton aphid, Aphis Goossypii Glover.Ⅲ.Host preference and organophosphorus susceptibility. Weed Abstract. 1994, 43(5) : 242
78. Jackson M A, Baruch S, David A S. Formulation of Colletotrichum truncatum microsclerotia for improved biocontrol of the weed hemp sesbania(sesbania exaltata). Biological control ,1996 (7),107-113
79. Kenney D S. Devine——the way it development——an industrialist's view. Weed Science. 1986, 34s: 15-16.
80. Khodayari K, Smith R J, Walke J T, et al. Applicators for a weed pathogen plus acifluorfen in soybeans. Weed Technology. 1987, 1:37-40.
81. Klein T A, Auld B A and Wang F. Evalution of oil suspension emulsion of Colletotrichum orbichlare as a mycoherbicide in field trials. Crop Protection, 1995, 14(3): 193-197
82. Wymore L A, Piorier C, and Watson A K. Colletotrichum coccodes a potential bioherbicide for control of velvet leaf (Abutition theophrasti), Plant Disease, 1988, 72:534-538
83. Mohanbabu R, Sajeena A, Seetharaman k, et al. Advances in bioherbicides development——an overiew. Crop Protection. 2002, Article In web.
84. Pulcher M, Hurle K. Weed flora and weed seed bank in wheat monoculture under different levels of crop protection. Weed Abstracts. 1984, 33(10): 341
85. Quimy P C, Boyette C D. Production and application of biocontrol agent[A]. in Mcwhort C G, Gebhardt M R. Methods of applying herbicides, Weed Science soc, Champaign IL, 1987. p.265-280
86. Quimy P C, Fulgham F E, Boyette C D,et al. New formulations nozzles boost efficacy of pathogens for weed control. Proc. Weed Science soc, 1988, 28-52.
87. Sansome G. Effect of crop population on the performance of herbicides in winter oilseed rape. Aspects of Applied Biology. 1990.23:257-262
88. Scacchi A, Bortolo R. Dectection, characterization, and phytotoxic activity of the nucleoside antibiotics, blasticidin S and 5-hydroxymethyl-blasticidin S. Journal of Plant Growth Regulation. 1992.11(1): 39-46
89. Smith R J. Jr. Integration of biological control agents with chemical pesticides.In Tebeest D O. Microbial control of weed. New York Chapman and Hall, 1991,
90. Stowell L J. Submerged fermentation of biological herbicides.In Tebeest D O. Microbial control of weed. New York Chapman and Hall, 1991, 225-261.
91. Tebeest D O, Templeton G E. Mycoherbicides: progress in the biological control weeds. Plant Disease.1985, 69 (2): 6—10.
92. Templeton G E, Tebeest D O, Smith R J. Biological weed control with myeoherbieides. Ann Rev Phytopathology.1979, 17:301-310.
93. Templeton G E. Status of weed control with plant pathogens.[a].in Charudattan R. and Walker H L. Biological control of weeds with plant pathogens. New York Wiley, 1982, 29-44
94. Templeton G E, Heiny D K. Improvement of fungi to enhance mycoherbicide potential, in Whipps J.M. and R.D. Lunsden eds. Biotechnology of Fungi for Improving Plant Growth. British Mycological Society, Cambridge University Press, Cambridge. pp. 1989. 127-151
95. Templeton G E. Weed control with pathogens: future needs and directions, in R.E. Hoagland, ed. Microbes and Microbial Products as Herbicides, ACS Symp. Set. 439. American Chemical Society, Washington, DC.pp. 1990.320-329
96. Vurro M, Ellis B E. Effect of fungal toxins on induction of phenylalanine ammonia-lyase activity in elicited cultures of hybrid poplar. Plant science 1997, 126,29-38
97. Vurro M, Zonno M C, Evidente A, et al. Enhancement of efficacy of Ascochyta caulina to control Chenopodium albumn by use ofphytotoxins and reduced rates of herbicides. BioControl. 2001.182-190.
98. Walker H L, Connick W J. Sodium alginate for production and formulation of mycoherbicides. Weed Science. 1983, 33: 333-338.
99. Womack J G, Eccleston G M, Burge M N. A vegetable oil-based invert emulsion for mycoherbicide delivery. Biological control, 1996(6), 23-28
100. Zhang W M, Watson. A K. Characterization of growth and conidia production of Exserohium monceras on different substrates. Biocontrol Sciences and Technology. 1997, 7: 75—86.
101. Zimdahl R L. Fundamentals of weed science. Academic Press. 2nd ed. San Diego. California: Academic Press, 1999, 247-269.