玉米疯顶病病原菌检测和病害防治技术研究
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摘要
玉米疯顶病是一种毁灭性的玉米病害，由大孢指疫霉(Sclerophthora macrospora (Sacc.)Thirum.,Shaw et Naras)引起，曾在世界多个国家局部地区严重发生。该病于1902年在意大利首次报道，1974年在我国山东省首次发生，近年来其危害日趋严重，已扩展到17省(自治区、直辖市)的48个县市，成为影响很多地区玉米生产的突出问题之一。国内外的有关研究均表明，来自田间发病植株上所结的种子可以传播疯顶病，但从现实的角度考虑种子传播的可能性很小。本研究根据大孢指疫霉的形态特征和细胞壁组成特点，分别从常规染色方法和分子检测技术入手，试图阐明病害大面积突发和种子带菌之间的关系。在此基础上，研究病害发病条件及玉米的抗性，探讨玉米疯顶病的综合治理方法。主要研究结果如下：
1．应用碘—氯化锌染色法、碘液—硫酸染色法、几丁质染色法以及棉蓝乳酚油染色法对不同类群真菌进行了染色。根据病菌细胞壁的显色反应，4种染料都能有效地把细胞壁主要组成成分不同的两类菌区分开，但碘—氯化锌染色法为卵菌细胞壁特异性染料，显色迅速，易于观察，能有效鉴别玉米组织中的卵菌菌丝体。利用碘—氯化锌试剂对不同类型植株来源的种子及病穗苞叶进行特异性染色，结果在种皮、胚、胚乳中都检测到了大孢指疫霉菌丝体的存在，而病穗苞叶中则没有发现菌丝体，表明病菌可以侵染种子而造成种子带菌。
2．通过卵孢子富集、DNA提取方法的筛选，建立了从卵孢子中提取大孢指疫霉DNA的有效方法。利用通用引物(ITS1和ITS4)扩增、克隆和测序了大孢指疫霉ITS区段。以具有较高保守性的ITS区段序列为模板合成了3对特异性引物SM1、SM2、SM3。利用引物SM3对大孢指疫霉进行初步的分子检测，结果表明该引物能特异性地扩增出病原菌DNA而玉米组织则无扩增，说明引物SM3对大孢指疫霉具有专一性。
3．在温室和田间，采取不同湿度处理方式，研究病田土壤、植株病残体和种子在病害传播中的作用和病害诱发条件，但未能充分诱发疯顶病的发生，表明大孢指疫霉卵孢子具有复杂的休眠机制，萌发所需要的条件苛刻。玉米疯顶病的研究仍有许多难点需要攻克。
4．由于田间病害诱发不充分，因此在品种抗性的评价、杀菌剂防治效果试验和土壤湿度控制试验方面未获得预期效果。证明了一些玉米品种和自交系属于感病类型。目前对玉米疯顶病的控制措施主要有以下两点：把采取有效控制田间土壤水分作为预防玉米疯顶病的首选技术措施，同时避免种植已知的感病品种。
Crazy top caused by Sclerophthora macrospora, is a destructive maize disease, which is local economic important in many countries in the world. The first report of the disease on maize was published in Italy in 1902. Since it was first found in Shandong Province, China, in 1974, the disease expanded fastly and there were some reports from 17 provinces and 48 counties. In some maize production areas crazy top has becoming a major threat to production, such as in Ningxia and Xinjiang.
Maize crazy top is a seed-borne and seed-transmitted disease. Why did the disease expand to some of provinces during a short time although there were few seeds or no seeds harvested from infected plants. Based the morphologic characteristics and cell wall composition of Sclerophthora macrospora, this research tries to make clear of the relationship between mycelia in seeds and the spread of maize crazy top using techniques of hypha special stain method and molecular marker method. Also the conditions of disease development, maize resistance and disease control measureses were studied. The main results are as follows:
1. Four methods of mycelia stain were compared, including zinc-chlor-iodide, I2-H2SO4, chitin stain and cotton blue lactopheno, in Pythium inflatum, Phytophthora capsipi, Exserohilum turcicum, Bipolaris maydis and Fusarium moniliforme, which are in different taxonomic groups with celluloses or chitin in the cell wall. Results showed that using cell wall stain methods 5 fungi species can been distinguished in two group: oomycytes and true fungi. Zinc-chlor-iodide is specific to stain the mycelia of Oomycetes quickly and easily. Using the stain method of zinc-chlor-iodide the mycelia fractions in maize seeds and symptomized tissues, collected from infected plants, were detected. The pathogen mycelia fractions were found in the pericarp, endosperm and embryo of maize seeds and were not found in the plant tissues. The fact pointed that Sclerophthora macrospora can infect the seeds and is seed-borne pathogen.
2. By the collection of the oospores of Sclerophthora macrospora and comparison of methods of DNA extraction, an effective procedure of DNA extraction from the oospores of Sclerophthora macrospora was made. The internal transcribed spacer (ITS) region was amplified with the general primer (ITS1 and ITS4). The DNA segment amplified was cloned and sequenced. Based on the rRNA-ITS sequence three specific primers were designed and synthesized, including SMI, SM2 and SM3. SM3 primer was specific one for amplifing DNA of Sclerophthora macrospora and no amplification of DNA of maize tissues.
3. On glasshouse and in field the effects of the soil collected from crazy top field, infected plant tissues and seeds were evaluated for the disease development under different soil moisture conditions. Only few plants showed typical symptoms. The result showed that it is difficult to induce germination of oospores of Sclerophthora macrospora
4. There were no good results on identification of maize resistance, and using fungicides and culture method to control the disease. For reducing the disease severe controlling soil moisture is the effective measure as well as avoiding planting the susceptible varieties.
1．应用碘—氯化锌染色法、碘液—硫酸染色法、几丁质染色法以及棉蓝乳酚油染色法对不同类群真菌进行了染色。根据病菌细胞壁的显色反应，4种染料都能有效地把细胞壁主要组成成分不同的两类菌区分开，但碘—氯化锌染色法为卵菌细胞壁特异性染料，显色迅速，易于观察，能有效鉴别玉米组织中的卵菌菌丝体。利用碘—氯化锌试剂对不同类型植株来源的种子及病穗苞叶进行特异性染色，结果在种皮、胚、胚乳中都检测到了大孢指疫霉菌丝体的存在，而病穗苞叶中则没有发现菌丝体，表明病菌可以侵染种子而造成种子带菌。
2．通过卵孢子富集、DNA提取方法的筛选，建立了从卵孢子中提取大孢指疫霉DNA的有效方法。利用通用引物(ITS1和ITS4)扩增、克隆和测序了大孢指疫霉ITS区段。以具有较高保守性的ITS区段序列为模板合成了3对特异性引物SM1、SM2、SM3。利用引物SM3对大孢指疫霉进行初步的分子检测，结果表明该引物能特异性地扩增出病原菌DNA而玉米组织则无扩增，说明引物SM3对大孢指疫霉具有专一性。
3．在温室和田间，采取不同湿度处理方式，研究病田土壤、植株病残体和种子在病害传播中的作用和病害诱发条件，但未能充分诱发疯顶病的发生，表明大孢指疫霉卵孢子具有复杂的休眠机制，萌发所需要的条件苛刻。玉米疯顶病的研究仍有许多难点需要攻克。
4．由于田间病害诱发不充分，因此在品种抗性的评价、杀菌剂防治效果试验和土壤湿度控制试验方面未获得预期效果。证明了一些玉米品种和自交系属于感病类型。目前对玉米疯顶病的控制措施主要有以下两点：把采取有效控制田间土壤水分作为预防玉米疯顶病的首选技术措施，同时避免种植已知的感病品种。
Crazy top caused by Sclerophthora macrospora, is a destructive maize disease, which is local economic important in many countries in the world. The first report of the disease on maize was published in Italy in 1902. Since it was first found in Shandong Province, China, in 1974, the disease expanded fastly and there were some reports from 17 provinces and 48 counties. In some maize production areas crazy top has becoming a major threat to production, such as in Ningxia and Xinjiang.
Maize crazy top is a seed-borne and seed-transmitted disease. Why did the disease expand to some of provinces during a short time although there were few seeds or no seeds harvested from infected plants. Based the morphologic characteristics and cell wall composition of Sclerophthora macrospora, this research tries to make clear of the relationship between mycelia in seeds and the spread of maize crazy top using techniques of hypha special stain method and molecular marker method. Also the conditions of disease development, maize resistance and disease control measureses were studied. The main results are as follows:
1. Four methods of mycelia stain were compared, including zinc-chlor-iodide, I2-H2SO4, chitin stain and cotton blue lactopheno, in Pythium inflatum, Phytophthora capsipi, Exserohilum turcicum, Bipolaris maydis and Fusarium moniliforme, which are in different taxonomic groups with celluloses or chitin in the cell wall. Results showed that using cell wall stain methods 5 fungi species can been distinguished in two group: oomycytes and true fungi. Zinc-chlor-iodide is specific to stain the mycelia of Oomycetes quickly and easily. Using the stain method of zinc-chlor-iodide the mycelia fractions in maize seeds and symptomized tissues, collected from infected plants, were detected. The pathogen mycelia fractions were found in the pericarp, endosperm and embryo of maize seeds and were not found in the plant tissues. The fact pointed that Sclerophthora macrospora can infect the seeds and is seed-borne pathogen.
2. By the collection of the oospores of Sclerophthora macrospora and comparison of methods of DNA extraction, an effective procedure of DNA extraction from the oospores of Sclerophthora macrospora was made. The internal transcribed spacer (ITS) region was amplified with the general primer (ITS1 and ITS4). The DNA segment amplified was cloned and sequenced. Based on the rRNA-ITS sequence three specific primers were designed and synthesized, including SMI, SM2 and SM3. SM3 primer was specific one for amplifing DNA of Sclerophthora macrospora and no amplification of DNA of maize tissues.
3. On glasshouse and in field the effects of the soil collected from crazy top field, infected plant tissues and seeds were evaluated for the disease development under different soil moisture conditions. Only few plants showed typical symptoms. The result showed that it is difficult to induce germination of oospores of Sclerophthora macrospora
4. There were no good results on identification of maize resistance, and using fungicides and culture method to control the disease. For reducing the disease severe controlling soil moisture is the effective measure as well as avoiding planting the susceptible varieties.
引文
1 奥斯伯，等．精编分子生物学实验指南(颜子颖等译)．北京：科学出版社，1999，523
2 白金铠．中国东北霜霉菌．植物病理学报，1957，3：137-154
3 陈志，张继俊，胡润泽．玉米疯顶病在博乐发现．新疆农业科学，1997，(2)：65-66
4 戴芳澜．中国真菌总汇．北京：科学出版社，1979
5 何放亭，武红巾，陈子文．用荧光染色诊断植物类菌原体病害．植物保护，1994，20(5)：37
6 黄河，刘小勇，郑儒永．rDNA中ITS区序列在根霉属小孢根霉组分类上的应用．见：刘仪编．植物病害研究与防治．北京：中国农业科技出版社，1998．484-487
7 郝农，等．辽宁省首次发现玉米疯顶病．植物检疫，1996，10(3)：147
8 韩群营，等．武汉地区发现玉米疯顶病．植物检疫，2000，14(4)：217
9 焦志亮．玉米弯孢菌叶斑病病原菌遗传多样性分析和玉米种质抗病性鉴定．中国农业科学院硕士研究生学位论文．2000
10 李秉超等．有机化学．吉林科学技术出版社，1996
11 李常玉，蔡连恩，何萨丽．甘肃省天水市发现玉米霜霉病．植物检疫，1997，11(3)：164
12 鲁栋，程利群．铜陵市玉米疯顶病发生情况及防治措施．安徽农业科学，1999，27(4)：388-389
13 李大明，谢正元，沈积仁．玉米霜霉病病原及发病规律研究．甘肃农业科学，1993，(1)：34
14 陆家云．植物病原真菌学．北京：中国农业出版社，2001
15 罗畔池，孔令晓，霍志清．河北省玉米新病害—疯顶病．河北农业科技，1995，(增刊)：33
16 刘彦斌，等．1998年阳泉市首次发现玉米疯顶病．植保技术与推广，1999，19(6)：43
17 罗占忠，等．吴忠市玉米霜霉病发生情况调查．宁夏农林科技，1996，(4)：7-9
18 罗占忠，李彦录，刘江山．宁夏发生玉米霜霉病．植物保护，1994，(1)：51
19 罗占忠，等．玉米疯顶病症状类型及病原菌形态观察．植保技术与推广，2000，20(2)：9-10
20 罗占忠，等．玉米疯顶病传播途径的试验及调查．植物保护，1997，23(5)：33-34
21 彭先达，向喜，向启发．开县部分玉米爆发霜霉病．四川农业科技，1994，(6)：21
22 彭德良，S.Subbotin,M.Moens.小麦禾谷胞囊线虫(Heterodera avenae)的核糖体基因(rDNA)限制性片段长度多态性研究．植物病理学报，2003，33(4)：323-329
23 戚佩坤．玉米、高粱、谷子病原手册．西安：科学出版社，1978．
24 邱荣芳，杨山山，郝彦俊．新疆玉米新病害—疯顶病．新疆农业科学，1993，(2)：65-66
25 孙发仁．泰安地区玉米霜霉病(丛顶病)发生调查．植物保护，1981，(6)：31
26 孙运村，等．玉米疯顶病在喀什地区流行．植保技术与推广，1999，19(6)：42
27 魏景超．真菌鉴定手册．上海：上海科学技术出版社，1979．44．
28 王化波．中国大豆疫霉菌遗传多样性和大豆种质抗病性研究．中国农业科学院硕士研究生学位论文．2001
29 王士奎，王汉潜．几丁质及脱乙酰几丁质的微生物奖解作用．微生物学通报，1994，21(3)：180-183
30 王晓鸣，戴法超，朱振东．中国玉米疯顶病发生现状与病害控制策略．见：面向21世纪的植物保护发展战略．北京：中国科学技术出版社，2001．239-243
31 王晓鸣．玉米茎腐病病原菌致病竞争及病害综合防治技术研究．中国农业大学博士学位论文．2001
32 王圆，等．指疫霉属及所引致的玉米霜霉病．植物检疫，1995，9(5)：285-288
33 谢益书，等．宁夏灌区玉米霜霉病的诊断和发生调查．宁夏农林科技，1994，(2)：9-11
34 谢益书，等．宁夏玉米霜霉病发生规律的研究．宁夏农学院学报，1996，17(2)：1-6
35 谢益书，等．玉米霜霉病药剂防治的研究．宁夏农学院学报，1998，19(1)：5-9
36 许志刚．普通植物病理学(第二版)．北京：中国农业出版社，1997
37 竹铨煦．新疆莎车、和田等县发生玉米霜霉病．植物保护，1994，(1)：51
38 郑小波．疫霉菌及其研究技术．北京：中国农业出版社，1997
39 周永力，等．采用PCR-RFLP和RAPD对球壳孢目真菌系统学的研究．Myeosystem，1998，17(2)：160-166
40 朱振东，等．北京地区发现玉米疯顶病．植物保护，1998，24(1)：48
41 章正，等．烟草霜霉病卵孢子提取方法研究．植物病理学报，1996，(26)：223-230
42 张中义，等．中国指疫霉属Sclerophthora分类研究．西南农业大学学报，1987，9(2)：154-158
43 张中义，刘云龙，王英祥．大孢指疫霉三个新变种的研究．云南农业大学学报，1990，5(2)：79-85
44 张中义，等．植物病原真菌学．成都：四川科学技术出版社，1998
45 邹亚飞，简桂良，马存，等．棉花黄萎病菌致病型的AFLP分析．植物病理学报，2003，33(2)：135-141
46 Aronson J, Cooper B A, Fuller M S. Glucans of oomycete cell walls. Science, N.Y., 1967,155: 332-335
47 Aronson J M. The cell wall. In the Fungi: An Advanced Treatise, I, 1965, 49-76.Editors: G.C.Ainworth &A.S.Sussman. New York and London: Academic Press
48 Assigbetse, K.B., et al. Differentiation of Fusarium oxysporum f.sp. vasinfecturn Races on Cotton by Random Amplified Polymorphie DNA (RAPD) Analysis. Phytopathology, 1994, 84(6): 622-626
49 Bartinicki-Garicia S.(1970). Cell wall composition and other biochemical markers in fungal taxonomy. In Phytochemieal Phylogeny, 81-103. Editor:J.B.Harbome. New York and London: Academic Press
50 Bartnicki-Garcia S. Cell wall chemistry, morphogenesis, and taxonomy of fungi. Annual Review of Microbiology, 1968, 22:87-108
51 Burnett J H (1975). Fundamentals of Mycology (Second edition)
52 Catherine Rafin, et al. Restriction analysis of amplified ribosomal DNA of Pythium spp. isolated from soilness culture systems. Mycol. Res., 1995, 99(3): 277-281
53 Chioechetti A, Sciaudone L, Durando F, et al. PCR Detection of Fusarium oxysporum f.sp. basilici on Basil.Plant Disease,2001,85(6):607-611
54 Chiocchetti A, Ghignone S, Minuto A, et al. Identification of Fusarium oxysporum f.sp. basilici Isolated from soil,Basil seed, and plants by RAPD Analysis. Plant Disease, 1999, 83(6): 576-581
55 Cooke, D.E.L., et al. A molecular Phylogeny of Phytophthora and related oomycetes. Fungal Genetics and Biology, 2000, 30(1): 17-32
56 Deacon J W. Introduction to Modem Mycology. Volume 7. Basic Microbiology. Edetor:J.F. Wilkinson
57 Demoeden P H, JACKSON N. Infection and mycelial colonization of gramineous hosts by Sclerophthora macrospora. Phytopathology, 1980, 70(10): 1009-1013
58 Demoeden B P H, Jackson N. Enhanced germination of Sclerophthora macrospora oospores in response to various chemical and physecal treatmengs.Trans. Br. Mycol.Soc., 1981, 76(2): 337-341
59 Dey S K, Dhillon B S, Malhotra V V. Crazy top downy mildew of maize-a new record in Punjab. Current Science, 1986, 55(12): 577-578
60 Edel, C.,et al. Evaluation of restriction analysis of polymerase chain reaction (PCR)-amplified
ribosomal DNA for the identification of Fusarium species. Mycol. Res., 1996, 101(2): 179-187
61 Faggian, R., et al. Specific Polymerase Chain Reaction Primers for the Detection of Plasmodiophora brassicae in soil and water. Phytopathology, 1999, 89(5): 392-397
62 Fouly, H. M., et al. Detection of Gaeumannomyces graminis Varieties Using Polymerase Chain Reaction with Variety-Specific Palmers. Plant Disease, 2000, 84(9): 947-951
63 Grajai-Martin, M. J., et al. Use of Random Amplified Polymorphic DNA(RAPD) to Characterize Race 2 ofFusarium oxysporum f.sp.pisi. Phytopathology, 1993, 83 (6): 612-614
64 Guzman, P., et al. Detection and Differentiation of Phaeoisadopsis griseola Isolates with the Polymerase Chain Reaction and Group-Specific Primers. Plant Disease, 1999, 83(1): 37-42
65 Hannele L, et al. Peronospora sparsa on Cultivated Rubus arcticus and ITS Detection by PCR Based on ITS Sequences. Plant Disease, 1998, 82(12): 1304-1311
66 Jackson N., Dernoeden P H. Sclerphthora macrospora: The Infection of Yellow Tuft Diseases of Turf Grass. Plant Disease, 1980, 64(10): 915-916
67 John Webster. (1980), Introduction to Fungi (Second edition). Cambridge University Press
68 Jons V L. Crazy top of corn in North Dakota. Plant Disease, 1980, 64:103-104
69 http://www.fgsc.net/fgn44/weiland.html
70 Kageyama, K., et al. Detection of Pythium ultimum Using Polymerase Chain Reaction with Species-Specific Primers. Plant Disease, 1997, 81(10): 1155-1160
71 Kelly A, Alcala-Jimenez A R. Use of Genetic Fingerprinting and Random Amplified Polymorphic DNA to Characterize Pathotypes of Fusarium oxysporum f.sp. ciceris Infecting Chickpea. Phytopathology, 1994, 84(11): 1293-1298
72 Koehler B. Crazy top of corn. Phytopathology, 1939, 29:817-820
73 Lee H K, Tewari J P, Turkington T K. A PCR-Based Assay to Detect Rhynchosporum secalis in Barley Seed. Plant Disease, 2001, 85(2): 220-225
74 Levesque, C.A., et al. Development of a Species-Specific Probi for Pythium ultimum Using Amplified Ribosomal DNA. Phytopathology, 1994, 84(5): 474-478
75 McDonough E S. A cytological study of the developmeng of the oospores of Sclerospora macrospora Sacc. Trans. Wis. Acad. Sci, 1946, 38:211-218
76 McGee D C. Maize Disease. American Phytopathology Society Press. Minnesota, 1988
77 Muralidhara Rao B, Prakash H S, Shetty H S, et al. Downy mildew inoculum in maize seeds: Techniques to detect seed-borne inoculum of Peronosclerospora sorghi in maize. Seed Sci. & Technol. 1985, 13:593-600
78 Murillo, I., et al. The development of a rapid PCR assay for detection of Fusarium moniliforme. European Journal of Plant Pathology, 1998, 104:301-300
79 Navi S S, Singh S D. Detection of Sclerospora graminicola Mycelium in Infected Pearl Millet Leaves. International Sorghum and Millets Newsletter, 1999, 40:58
80 Nicholson,P., et al. Detection and quantification of Fusarium culmorum and Fusarium graminearum in cereals using PCR assays. Physiological and Molecular Plant Pathology, 1998, 53:17-37
81 Novaes-Ledieu M., Jimenez-martinez A. Chemical composition of hyphal wall of Phycomycetes. Journal of General Microbiology, 1967, 47: 237-245
82 Panabieres F, et al. Repetitive DNA polymorphism analysis as a tool for identifying Phytophora species [J]. Phytopathology, 1989, 79(10): 1105-1109
83 Payak M M, Renfro B L. Sangam Lal. Downy mildew diseases incited by Scletophthora. Indian
Phytopathology, 1970, 13:183-193
84 Peglion V La. farmazione dei conidi e la gcrminazione delle oospore della'Sclerospora macrospora'Sacc. Boll. della R. Staz. Patol Veg. Roma N.S., 1930, 10:153-164
85 Peyronel B. Gli zoosporangi nella Sclerospora macrospora. Boll. della R. Staz. Patol. Veg. Roma N.S., 1929, 9:353-357
86 Pryor B M, Gilbertson R S. A PCR-based Assay for Detection of Alternaria radicina on Carrot Seed. Plant Disease, 2001, 85(1): 18-23
87 Rathore R S, Siradhana B S, Mathur K. Transmission and location of Peronoselerospora hetoropogohi in maize seeds. Seed Sci. & Technol., 1987, 15:101-107
88 Saccardo PA. Fungi aliquot australienses. Hedwigia, 1890, 29:154-156
89 Salazar, O., et at. Primers based on specific rDNA-ITS sequences for PCR detection of Rhizoctonia solami, R. solani AG2 subgroups and ecological types, and binucleate Rhizoetonia. Mycological Research, 2000, 104(3): 281-285
90 Semeniuk G. Sclerophthora macrospora infection of three annual grasses by oospore and asexual inocula. Plant Disease Reporter, 1976, 60(9): 745-748
91 Semeniuk G. Occurrence and development of Sclerophthora maerospora on cereals and grasses in South Dakota. Phytopathology, 1964, 54:409-416
92 Shurtleff M C. Compendium of Corn diseases.American Phytopathology Society Press. Minnesota, 1980
93 Singh P J, Bedi P S. Factors affecting germination of sporangia of Sclerophthora macrospora, the incitant of downy mildew of wheat. Plant Disease Research, 1993, 9(2): 185-189
94 Suga, H., et at. Phylogenetic analysis of the Phytopathogenic fungus Fusarium solani based on the Rdna-ITS region. Mycol. Res., 2000, 104(10): 1175-1183
95 Sun M H, Ullstrup A J. Etiology of crazy top of cora. Phytopathology, 1971, 61(8): 883-919
96 Tanaka I. Phytophthora maerospora(Sacc.) S. Ito and I.Tanaka on wheat plant. Ann. Phytopath. Soc. Japan, 1940, 10:127-138
97 Thirumalaehar M J, Shaw C G. Narsimhan, M,.J. The sporangial phase of the downy mildew on Eleusine coracana with a discussion on the identity of Sclerospora maerospora Sacc. Bull. Torrey bot. C1, 1953, 80:299-307
98 Mosmann Tim. Rapid Colorimetric Assay for Cellar Growth and Survival: Application to Proliferation and Cytotoxicity Assays. Journal of Immunological Methods, 1983, 65:55-63
99 Trout C L, Ristaino J B, Madritch M, et al. Rapid Detection of Phytophthora infestans in Late Blight-Infected Potato and Tomato Using PCR. Plant Disease, 1997, 81(9): 1042-1048
100 Troutman Joseph L, Matejka Joseph C. Downy mildew of small grains and sorghum in Arizona. 1972, 56(9): 773-774
101 Ullstrup A J. Crazt top of maize. Indian Phytopathology, 1970, 13:250-261
102 Ullstrup Arnold J. Observation on crazy top of corn. Phytopathology, 1952, 42:675-680
103 Ullstrup A J, Sun M H. The prevalence of crazy top of corn in 1968. Plant Disease Reporter, 1969, 53(4): 246-250
104 Vandemark G J, Kraft J M, Larsen R C, et at. A PCR-Based Assay by Sequence-Characterized DNA Markers for the Identification and Detection of Aphanomyces euteiches. Phytopathology, 2000, 90(10): 1137-1144
105 Wang P H, et at. Detection of the low-germination-rate resting oospores of Pythium myriotylum from soil by PCR. Letters in Applied Microbiology, 2003, 36:157-161
106 Wang P H, et al. Molecular characterization of Pythium species based on RFLP analysis of the internal transcribed spacer region of ribosomal DNA. Physiological and Molecular Plant Pathology, 1997, 51:129-143
107 Whitehead M D. Pathology and pathological histology of Downy mildew, Sclerphthora macrospora, on six graminicolous hosts. Phytopathology, 1958, 48:485-493
108 Willits, D.A., et al. Polymerase Chain Reaction Detection of Ustilago hordei in Leaves of Susceptible and Resistant Barley Varieties. Phytopathology, 1999, 89(3): 212-217
2 白金铠．中国东北霜霉菌．植物病理学报，1957，3：137-154
3 陈志，张继俊，胡润泽．玉米疯顶病在博乐发现．新疆农业科学，1997，(2)：65-66
4 戴芳澜．中国真菌总汇．北京：科学出版社，1979
5 何放亭，武红巾，陈子文．用荧光染色诊断植物类菌原体病害．植物保护，1994，20(5)：37
6 黄河，刘小勇，郑儒永．rDNA中ITS区序列在根霉属小孢根霉组分类上的应用．见：刘仪编．植物病害研究与防治．北京：中国农业科技出版社，1998．484-487
7 郝农，等．辽宁省首次发现玉米疯顶病．植物检疫，1996，10(3)：147
8 韩群营，等．武汉地区发现玉米疯顶病．植物检疫，2000，14(4)：217
9 焦志亮．玉米弯孢菌叶斑病病原菌遗传多样性分析和玉米种质抗病性鉴定．中国农业科学院硕士研究生学位论文．2000
10 李秉超等．有机化学．吉林科学技术出版社，1996
11 李常玉，蔡连恩，何萨丽．甘肃省天水市发现玉米霜霉病．植物检疫，1997，11(3)：164
12 鲁栋，程利群．铜陵市玉米疯顶病发生情况及防治措施．安徽农业科学，1999，27(4)：388-389
13 李大明，谢正元，沈积仁．玉米霜霉病病原及发病规律研究．甘肃农业科学，1993，(1)：34
14 陆家云．植物病原真菌学．北京：中国农业出版社，2001
15 罗畔池，孔令晓，霍志清．河北省玉米新病害—疯顶病．河北农业科技，1995，(增刊)：33
16 刘彦斌，等．1998年阳泉市首次发现玉米疯顶病．植保技术与推广，1999，19(6)：43
17 罗占忠，等．吴忠市玉米霜霉病发生情况调查．宁夏农林科技，1996，(4)：7-9
18 罗占忠，李彦录，刘江山．宁夏发生玉米霜霉病．植物保护，1994，(1)：51
19 罗占忠，等．玉米疯顶病症状类型及病原菌形态观察．植保技术与推广，2000，20(2)：9-10
20 罗占忠，等．玉米疯顶病传播途径的试验及调查．植物保护，1997，23(5)：33-34
21 彭先达，向喜，向启发．开县部分玉米爆发霜霉病．四川农业科技，1994，(6)：21
22 彭德良，S.Subbotin,M.Moens.小麦禾谷胞囊线虫(Heterodera avenae)的核糖体基因(rDNA)限制性片段长度多态性研究．植物病理学报，2003，33(4)：323-329
23 戚佩坤．玉米、高粱、谷子病原手册．西安：科学出版社，1978．
24 邱荣芳，杨山山，郝彦俊．新疆玉米新病害—疯顶病．新疆农业科学，1993，(2)：65-66
25 孙发仁．泰安地区玉米霜霉病(丛顶病)发生调查．植物保护，1981，(6)：31
26 孙运村，等．玉米疯顶病在喀什地区流行．植保技术与推广，1999，19(6)：42
27 魏景超．真菌鉴定手册．上海：上海科学技术出版社，1979．44．
28 王化波．中国大豆疫霉菌遗传多样性和大豆种质抗病性研究．中国农业科学院硕士研究生学位论文．2001
29 王士奎，王汉潜．几丁质及脱乙酰几丁质的微生物奖解作用．微生物学通报，1994，21(3)：180-183
30 王晓鸣，戴法超，朱振东．中国玉米疯顶病发生现状与病害控制策略．见：面向21世纪的植物保护发展战略．北京：中国科学技术出版社，2001．239-243
31 王晓鸣．玉米茎腐病病原菌致病竞争及病害综合防治技术研究．中国农业大学博士学位论文．2001
32 王圆，等．指疫霉属及所引致的玉米霜霉病．植物检疫，1995，9(5)：285-288
33 谢益书，等．宁夏灌区玉米霜霉病的诊断和发生调查．宁夏农林科技，1994，(2)：9-11
34 谢益书，等．宁夏玉米霜霉病发生规律的研究．宁夏农学院学报，1996，17(2)：1-6
35 谢益书，等．玉米霜霉病药剂防治的研究．宁夏农学院学报，1998，19(1)：5-9
36 许志刚．普通植物病理学(第二版)．北京：中国农业出版社，1997
37 竹铨煦．新疆莎车、和田等县发生玉米霜霉病．植物保护，1994，(1)：51
38 郑小波．疫霉菌及其研究技术．北京：中国农业出版社，1997
39 周永力，等．采用PCR-RFLP和RAPD对球壳孢目真菌系统学的研究．Myeosystem，1998，17(2)：160-166
40 朱振东，等．北京地区发现玉米疯顶病．植物保护，1998，24(1)：48
41 章正，等．烟草霜霉病卵孢子提取方法研究．植物病理学报，1996，(26)：223-230
42 张中义，等．中国指疫霉属Sclerophthora分类研究．西南农业大学学报，1987，9(2)：154-158
43 张中义，刘云龙，王英祥．大孢指疫霉三个新变种的研究．云南农业大学学报，1990，5(2)：79-85
44 张中义，等．植物病原真菌学．成都：四川科学技术出版社，1998
45 邹亚飞，简桂良，马存，等．棉花黄萎病菌致病型的AFLP分析．植物病理学报，2003，33(2)：135-141
46 Aronson J, Cooper B A, Fuller M S. Glucans of oomycete cell walls. Science, N.Y., 1967,155: 332-335
47 Aronson J M. The cell wall. In the Fungi: An Advanced Treatise, I, 1965, 49-76.Editors: G.C.Ainworth &A.S.Sussman. New York and London: Academic Press
48 Assigbetse, K.B., et al. Differentiation of Fusarium oxysporum f.sp. vasinfecturn Races on Cotton by Random Amplified Polymorphie DNA (RAPD) Analysis. Phytopathology, 1994, 84(6): 622-626
49 Bartinicki-Garicia S.(1970). Cell wall composition and other biochemical markers in fungal taxonomy. In Phytochemieal Phylogeny, 81-103. Editor:J.B.Harbome. New York and London: Academic Press
50 Bartnicki-Garcia S. Cell wall chemistry, morphogenesis, and taxonomy of fungi. Annual Review of Microbiology, 1968, 22:87-108
51 Burnett J H (1975). Fundamentals of Mycology (Second edition)
52 Catherine Rafin, et al. Restriction analysis of amplified ribosomal DNA of Pythium spp. isolated from soilness culture systems. Mycol. Res., 1995, 99(3): 277-281
53 Chioechetti A, Sciaudone L, Durando F, et al. PCR Detection of Fusarium oxysporum f.sp. basilici on Basil.Plant Disease,2001,85(6):607-611
54 Chiocchetti A, Ghignone S, Minuto A, et al. Identification of Fusarium oxysporum f.sp. basilici Isolated from soil,Basil seed, and plants by RAPD Analysis. Plant Disease, 1999, 83(6): 576-581
55 Cooke, D.E.L., et al. A molecular Phylogeny of Phytophthora and related oomycetes. Fungal Genetics and Biology, 2000, 30(1): 17-32
56 Deacon J W. Introduction to Modem Mycology. Volume 7. Basic Microbiology. Edetor:J.F. Wilkinson
57 Demoeden P H, JACKSON N. Infection and mycelial colonization of gramineous hosts by Sclerophthora macrospora. Phytopathology, 1980, 70(10): 1009-1013
58 Demoeden B P H, Jackson N. Enhanced germination of Sclerophthora macrospora oospores in response to various chemical and physecal treatmengs.Trans. Br. Mycol.Soc., 1981, 76(2): 337-341
59 Dey S K, Dhillon B S, Malhotra V V. Crazy top downy mildew of maize-a new record in Punjab. Current Science, 1986, 55(12): 577-578
60 Edel, C.,et al. Evaluation of restriction analysis of polymerase chain reaction (PCR)-amplified
ribosomal DNA for the identification of Fusarium species. Mycol. Res., 1996, 101(2): 179-187
61 Faggian, R., et al. Specific Polymerase Chain Reaction Primers for the Detection of Plasmodiophora brassicae in soil and water. Phytopathology, 1999, 89(5): 392-397
62 Fouly, H. M., et al. Detection of Gaeumannomyces graminis Varieties Using Polymerase Chain Reaction with Variety-Specific Palmers. Plant Disease, 2000, 84(9): 947-951
63 Grajai-Martin, M. J., et al. Use of Random Amplified Polymorphic DNA(RAPD) to Characterize Race 2 ofFusarium oxysporum f.sp.pisi. Phytopathology, 1993, 83 (6): 612-614
64 Guzman, P., et al. Detection and Differentiation of Phaeoisadopsis griseola Isolates with the Polymerase Chain Reaction and Group-Specific Primers. Plant Disease, 1999, 83(1): 37-42
65 Hannele L, et al. Peronospora sparsa on Cultivated Rubus arcticus and ITS Detection by PCR Based on ITS Sequences. Plant Disease, 1998, 82(12): 1304-1311
66 Jackson N., Dernoeden P H. Sclerphthora macrospora: The Infection of Yellow Tuft Diseases of Turf Grass. Plant Disease, 1980, 64(10): 915-916
67 John Webster. (1980), Introduction to Fungi (Second edition). Cambridge University Press
68 Jons V L. Crazy top of corn in North Dakota. Plant Disease, 1980, 64:103-104
69 http://www.fgsc.net/fgn44/weiland.html
70 Kageyama, K., et al. Detection of Pythium ultimum Using Polymerase Chain Reaction with Species-Specific Primers. Plant Disease, 1997, 81(10): 1155-1160
71 Kelly A, Alcala-Jimenez A R. Use of Genetic Fingerprinting and Random Amplified Polymorphic DNA to Characterize Pathotypes of Fusarium oxysporum f.sp. ciceris Infecting Chickpea. Phytopathology, 1994, 84(11): 1293-1298
72 Koehler B. Crazy top of corn. Phytopathology, 1939, 29:817-820
73 Lee H K, Tewari J P, Turkington T K. A PCR-Based Assay to Detect Rhynchosporum secalis in Barley Seed. Plant Disease, 2001, 85(2): 220-225
74 Levesque, C.A., et al. Development of a Species-Specific Probi for Pythium ultimum Using Amplified Ribosomal DNA. Phytopathology, 1994, 84(5): 474-478
75 McDonough E S. A cytological study of the developmeng of the oospores of Sclerospora macrospora Sacc. Trans. Wis. Acad. Sci, 1946, 38:211-218
76 McGee D C. Maize Disease. American Phytopathology Society Press. Minnesota, 1988
77 Muralidhara Rao B, Prakash H S, Shetty H S, et al. Downy mildew inoculum in maize seeds: Techniques to detect seed-borne inoculum of Peronosclerospora sorghi in maize. Seed Sci. & Technol. 1985, 13:593-600
78 Murillo, I., et al. The development of a rapid PCR assay for detection of Fusarium moniliforme. European Journal of Plant Pathology, 1998, 104:301-300
79 Navi S S, Singh S D. Detection of Sclerospora graminicola Mycelium in Infected Pearl Millet Leaves. International Sorghum and Millets Newsletter, 1999, 40:58
80 Nicholson,P., et al. Detection and quantification of Fusarium culmorum and Fusarium graminearum in cereals using PCR assays. Physiological and Molecular Plant Pathology, 1998, 53:17-37
81 Novaes-Ledieu M., Jimenez-martinez A. Chemical composition of hyphal wall of Phycomycetes. Journal of General Microbiology, 1967, 47: 237-245
82 Panabieres F, et al. Repetitive DNA polymorphism analysis as a tool for identifying Phytophora species [J]. Phytopathology, 1989, 79(10): 1105-1109
83 Payak M M, Renfro B L. Sangam Lal. Downy mildew diseases incited by Scletophthora. Indian
Phytopathology, 1970, 13:183-193
84 Peglion V La. farmazione dei conidi e la gcrminazione delle oospore della'Sclerospora macrospora'Sacc. Boll. della R. Staz. Patol Veg. Roma N.S., 1930, 10:153-164
85 Peyronel B. Gli zoosporangi nella Sclerospora macrospora. Boll. della R. Staz. Patol. Veg. Roma N.S., 1929, 9:353-357
86 Pryor B M, Gilbertson R S. A PCR-based Assay for Detection of Alternaria radicina on Carrot Seed. Plant Disease, 2001, 85(1): 18-23
87 Rathore R S, Siradhana B S, Mathur K. Transmission and location of Peronoselerospora hetoropogohi in maize seeds. Seed Sci. & Technol., 1987, 15:101-107
88 Saccardo PA. Fungi aliquot australienses. Hedwigia, 1890, 29:154-156
89 Salazar, O., et at. Primers based on specific rDNA-ITS sequences for PCR detection of Rhizoctonia solami, R. solani AG2 subgroups and ecological types, and binucleate Rhizoetonia. Mycological Research, 2000, 104(3): 281-285
90 Semeniuk G. Sclerophthora macrospora infection of three annual grasses by oospore and asexual inocula. Plant Disease Reporter, 1976, 60(9): 745-748
91 Semeniuk G. Occurrence and development of Sclerophthora maerospora on cereals and grasses in South Dakota. Phytopathology, 1964, 54:409-416
92 Shurtleff M C. Compendium of Corn diseases.American Phytopathology Society Press. Minnesota, 1980
93 Singh P J, Bedi P S. Factors affecting germination of sporangia of Sclerophthora macrospora, the incitant of downy mildew of wheat. Plant Disease Research, 1993, 9(2): 185-189
94 Suga, H., et at. Phylogenetic analysis of the Phytopathogenic fungus Fusarium solani based on the Rdna-ITS region. Mycol. Res., 2000, 104(10): 1175-1183
95 Sun M H, Ullstrup A J. Etiology of crazy top of cora. Phytopathology, 1971, 61(8): 883-919
96 Tanaka I. Phytophthora maerospora(Sacc.) S. Ito and I.Tanaka on wheat plant. Ann. Phytopath. Soc. Japan, 1940, 10:127-138
97 Thirumalaehar M J, Shaw C G. Narsimhan, M,.J. The sporangial phase of the downy mildew on Eleusine coracana with a discussion on the identity of Sclerospora maerospora Sacc. Bull. Torrey bot. C1, 1953, 80:299-307
98 Mosmann Tim. Rapid Colorimetric Assay for Cellar Growth and Survival: Application to Proliferation and Cytotoxicity Assays. Journal of Immunological Methods, 1983, 65:55-63
99 Trout C L, Ristaino J B, Madritch M, et al. Rapid Detection of Phytophthora infestans in Late Blight-Infected Potato and Tomato Using PCR. Plant Disease, 1997, 81(9): 1042-1048
100 Troutman Joseph L, Matejka Joseph C. Downy mildew of small grains and sorghum in Arizona. 1972, 56(9): 773-774
101 Ullstrup A J. Crazt top of maize. Indian Phytopathology, 1970, 13:250-261
102 Ullstrup Arnold J. Observation on crazy top of corn. Phytopathology, 1952, 42:675-680
103 Ullstrup A J, Sun M H. The prevalence of crazy top of corn in 1968. Plant Disease Reporter, 1969, 53(4): 246-250
104 Vandemark G J, Kraft J M, Larsen R C, et at. A PCR-Based Assay by Sequence-Characterized DNA Markers for the Identification and Detection of Aphanomyces euteiches. Phytopathology, 2000, 90(10): 1137-1144
105 Wang P H, et at. Detection of the low-germination-rate resting oospores of Pythium myriotylum from soil by PCR. Letters in Applied Microbiology, 2003, 36:157-161
106 Wang P H, et al. Molecular characterization of Pythium species based on RFLP analysis of the internal transcribed spacer region of ribosomal DNA. Physiological and Molecular Plant Pathology, 1997, 51:129-143
107 Whitehead M D. Pathology and pathological histology of Downy mildew, Sclerphthora macrospora, on six graminicolous hosts. Phytopathology, 1958, 48:485-493
108 Willits, D.A., et al. Polymerase Chain Reaction Detection of Ustilago hordei in Leaves of Susceptible and Resistant Barley Varieties. Phytopathology, 1999, 89(3): 212-217
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