中国部分地区草坪丝核菌的融合群鉴定及生防木霉菌株的筛选
|
摘要
随着城市化进程的不断加快,草坪在城市绿化中的作用也越来越明显。然而由于不当的水肥管理和修剪,草坪丝核菌病害的发生也越来越严重。目前,在对病害的防治中,提倡综合防治,而生物防治,尤其是拮抗木霉在土传病害防治中的作用尤为显著。本研究对山东省青岛、东营、泰安、上海市、广东省广州等地的草坪丝核菌进行了系统研究并筛选出了对优势融合群具有明显拮抗作用的生防木霉菌株。具体结果有如下几个方面:
1.从山东省青岛、东营、泰安、上海市、广东省广州等地采集草坪丝核菌病害标本,采用常规组织分离法分离菌株获得48个丝核菌菌株。融合群测定结果表明,这些菌株分别属于多核丝核菌的WAG-Z、AG1-IA、AG4-HG-I融合群及双核丝核菌的AG-A、AG-P、AG-L融合群构成,其中WAG-Z群出现频率最高,占全部菌株的70.83%,为优势融合群;其次是AG-A群,占14.59%;再依次为AG1-IA群,占分离菌株的8.34%;AG4-HG-I、AG-P和AG-L群均有1个菌株,均占分离菌株的2.08%。另外AG-P群、AG-L群和AG-A群是国内外首次从草坪上分离得到的三个融合群。
2.分离获得的草坪丝核菌,其菌丝经DAPI染色后制成玻片,置于荧光显微镜下观察,发现草坪丝核菌的细胞核相大致分两种:多核和双核,其中AG1-IA融合群的菌株细胞核数目多为5-11个;AG4-HG-I融合群的菌株细胞核数目为3-10个;WAG-Z融合群的菌株核相较为复杂,有单核细胞、双核细胞及多核细胞,其中以多核为主;AG-P、AG-L和AG-A融合群的菌株均为双核。
3.利用5.8SrDNA-ITS区序列分析法,对草坪丝核菌不同融合群菌株进行了分子鉴定。结果表明,所有测试菌株其5.8S rDNA序列高度保守,ITS1、ITS2区存在不同程度的差异,隶属不同融合群或不同亚群的菌株其5.8S rDNA-ITS区序列存在较大的差异,而相同融合群或亚群菌株的一致性较高,利用此基因序列对丝核菌不同融合群或亚群的菌株可以进行有效区分和鉴定。
4.运用灰色关联度分析法,对10个初筛木霉菌株的生长速度、耐性、产孢量、抗生作用等8个主要性状指标进行综合评价和分析。结果表明,木霉菌株T05-037的灰色关联度ri=0.9064最大,其次是T05-066和T05-048菌株,其灰色关联度分别为0.8714和0.8712。表明T05-037、T05-066、T05-048这三个木霉菌株的综合性状与构建的理想的参考菌株接近程度大,是比较理想的拮抗丝核菌的木霉生防菌株。其中,T05-037菌株的综合评价结果最好。
5.本试验筛选获得的优良木霉菌株T05– 037,利用形态分类学的方法初步将其鉴定为绿色木霉(Trichoderma viride)。利用室内平板对峙方法评价木霉菌株T05-037对隶属不同融合群丝核菌的抑制作用。结果表明,T05-037木霉菌对供试的分属4个不同融合群的丝核菌菌株都有较强的抑制作用(抑制率为55.52%-64.49%),其中T05-037菌株对属于WAG-Z融合群的丝核菌菌株抑制率最高,说明木霉菌株T05–037对草坪丝核菌病害具有良好的生防潜力。
Turfgrasses are more and more impotant in the urban afforestation, and now the disease of turfgrasses caused by Rhizoctonia spp is a common and important disease. In this study, we indentified the anastomosis group obtained from turfgrass in Qingdao, Taian, Dongying of Shandong Province, Shanghai and Guangzhou of Guangdong Province. Also we selected a high efficient antagonistic Trichoderma isolate to the the major anastomosis group of turfgrass Rhizoctonia. The results gained are summarized as following :
1.Fourty-eight isolates were obtained from turfgrasses in several areas of China (including Qingdao, Taian, Dongying, Shanghai and Guangzhou). Anastomosis group indentification showed that the isolates belonged to multinucleat Rhizoctonia WAG-Z、AG1-IA、AG4-HG-I and binucleate Rhizoctonia AG-A、AG-P、AG-L.Of these, WAG-Z was the major anastomosis group(70.83% of total isolates),AG-A was the second anastomosis group ( 14.59% of total isolates) , AG1-IA was the third(8.34% of total isolates), there were all one strain of AG4-HG-I、AG-P and AG-L(2.08% of total isolates). AG-P、AG-L and AG-A were isolated for the first time from turfgrass in China.
2.Putting the mycelium on the slide after DAPI, obsorbed under the microscope. The results showed that there were multinucleate and binucleate of the nucleus of the mycelium cell of Rhizoctonia from turfgrass. There were 5-11 nucleus of AG1-IA and 3-10 nucleus of AG4-HG-I. The nucleus of WAG-Z were complex, and the main was multinucleat Rhizoctonia. AG-P, AG-L and AG-A were binucleate Rhizoctonia.
3.Phylogenetic analysis among isolates belonging to different AGs was studied based upon 5.8S rDNA-ITS sequences. The results obtained from nuclear and mitochondrial sequence analyses were similar. The sequence alignment of 5.8S rDNA-ITS revealed that the isolates of the same AGs had a high sequence similarity.The isolates of the different AGs had a low sequence similarity. So using 5.8sDNA-ITS sequence analysis was a effective method of the AGs identification.
4.There were 8 biocontrol characters of 10 Trichoderma isolates (including growth, inhibitionrate, amount of sporulation, tolerate to low temperature, tolerate to high temperature, tolerate to acid, tolerate to alkaline and Tolerate to Carbendazim).They were evaluated and analyzed by Grey related degree.The results showed that the biggest Grey related degree was T05-037, the second one was T05-066 and the third was T05-048. The biocontrol characters of these three isolates were approximative to the reference. T05-037 was the best among these isolates.
5.Using morphology analysis, we identified that isolate T05-037 that we selected in this study was Trichoderma viride. We studied the inhibition percentage of T05-037 to different anastomosis group by cultivated on the plate. The results showed that the isolate T05-037 is effectivity to the 4 isolates that belonged to different anastomosis group.The inhibitionrate to WAG-Z is the biggest. So we can conclude that T05-037 is potential to prevente the disease of turfgrass caused by Rhizoctonia spp.
1.从山东省青岛、东营、泰安、上海市、广东省广州等地采集草坪丝核菌病害标本,采用常规组织分离法分离菌株获得48个丝核菌菌株。融合群测定结果表明,这些菌株分别属于多核丝核菌的WAG-Z、AG1-IA、AG4-HG-I融合群及双核丝核菌的AG-A、AG-P、AG-L融合群构成,其中WAG-Z群出现频率最高,占全部菌株的70.83%,为优势融合群;其次是AG-A群,占14.59%;再依次为AG1-IA群,占分离菌株的8.34%;AG4-HG-I、AG-P和AG-L群均有1个菌株,均占分离菌株的2.08%。另外AG-P群、AG-L群和AG-A群是国内外首次从草坪上分离得到的三个融合群。
2.分离获得的草坪丝核菌,其菌丝经DAPI染色后制成玻片,置于荧光显微镜下观察,发现草坪丝核菌的细胞核相大致分两种:多核和双核,其中AG1-IA融合群的菌株细胞核数目多为5-11个;AG4-HG-I融合群的菌株细胞核数目为3-10个;WAG-Z融合群的菌株核相较为复杂,有单核细胞、双核细胞及多核细胞,其中以多核为主;AG-P、AG-L和AG-A融合群的菌株均为双核。
3.利用5.8SrDNA-ITS区序列分析法,对草坪丝核菌不同融合群菌株进行了分子鉴定。结果表明,所有测试菌株其5.8S rDNA序列高度保守,ITS1、ITS2区存在不同程度的差异,隶属不同融合群或不同亚群的菌株其5.8S rDNA-ITS区序列存在较大的差异,而相同融合群或亚群菌株的一致性较高,利用此基因序列对丝核菌不同融合群或亚群的菌株可以进行有效区分和鉴定。
4.运用灰色关联度分析法,对10个初筛木霉菌株的生长速度、耐性、产孢量、抗生作用等8个主要性状指标进行综合评价和分析。结果表明,木霉菌株T05-037的灰色关联度ri=0.9064最大,其次是T05-066和T05-048菌株,其灰色关联度分别为0.8714和0.8712。表明T05-037、T05-066、T05-048这三个木霉菌株的综合性状与构建的理想的参考菌株接近程度大,是比较理想的拮抗丝核菌的木霉生防菌株。其中,T05-037菌株的综合评价结果最好。
5.本试验筛选获得的优良木霉菌株T05– 037,利用形态分类学的方法初步将其鉴定为绿色木霉(Trichoderma viride)。利用室内平板对峙方法评价木霉菌株T05-037对隶属不同融合群丝核菌的抑制作用。结果表明,T05-037木霉菌对供试的分属4个不同融合群的丝核菌菌株都有较强的抑制作用(抑制率为55.52%-64.49%),其中T05-037菌株对属于WAG-Z融合群的丝核菌菌株抑制率最高,说明木霉菌株T05–037对草坪丝核菌病害具有良好的生防潜力。
Turfgrasses are more and more impotant in the urban afforestation, and now the disease of turfgrasses caused by Rhizoctonia spp is a common and important disease. In this study, we indentified the anastomosis group obtained from turfgrass in Qingdao, Taian, Dongying of Shandong Province, Shanghai and Guangzhou of Guangdong Province. Also we selected a high efficient antagonistic Trichoderma isolate to the the major anastomosis group of turfgrass Rhizoctonia. The results gained are summarized as following :
1.Fourty-eight isolates were obtained from turfgrasses in several areas of China (including Qingdao, Taian, Dongying, Shanghai and Guangzhou). Anastomosis group indentification showed that the isolates belonged to multinucleat Rhizoctonia WAG-Z、AG1-IA、AG4-HG-I and binucleate Rhizoctonia AG-A、AG-P、AG-L.Of these, WAG-Z was the major anastomosis group(70.83% of total isolates),AG-A was the second anastomosis group ( 14.59% of total isolates) , AG1-IA was the third(8.34% of total isolates), there were all one strain of AG4-HG-I、AG-P and AG-L(2.08% of total isolates). AG-P、AG-L and AG-A were isolated for the first time from turfgrass in China.
2.Putting the mycelium on the slide after DAPI, obsorbed under the microscope. The results showed that there were multinucleate and binucleate of the nucleus of the mycelium cell of Rhizoctonia from turfgrass. There were 5-11 nucleus of AG1-IA and 3-10 nucleus of AG4-HG-I. The nucleus of WAG-Z were complex, and the main was multinucleat Rhizoctonia. AG-P, AG-L and AG-A were binucleate Rhizoctonia.
3.Phylogenetic analysis among isolates belonging to different AGs was studied based upon 5.8S rDNA-ITS sequences. The results obtained from nuclear and mitochondrial sequence analyses were similar. The sequence alignment of 5.8S rDNA-ITS revealed that the isolates of the same AGs had a high sequence similarity.The isolates of the different AGs had a low sequence similarity. So using 5.8sDNA-ITS sequence analysis was a effective method of the AGs identification.
4.There were 8 biocontrol characters of 10 Trichoderma isolates (including growth, inhibitionrate, amount of sporulation, tolerate to low temperature, tolerate to high temperature, tolerate to acid, tolerate to alkaline and Tolerate to Carbendazim).They were evaluated and analyzed by Grey related degree.The results showed that the biggest Grey related degree was T05-037, the second one was T05-066 and the third was T05-048. The biocontrol characters of these three isolates were approximative to the reference. T05-037 was the best among these isolates.
5.Using morphology analysis, we identified that isolate T05-037 that we selected in this study was Trichoderma viride. We studied the inhibition percentage of T05-037 to different anastomosis group by cultivated on the plate. The results showed that the isolate T05-037 is effectivity to the 4 isolates that belonged to different anastomosis group.The inhibitionrate to WAG-Z is the biggest. So we can conclude that T05-037 is potential to prevente the disease of turfgrass caused by Rhizoctonia spp.
引文
1.晁龙军,单学敏,车少臣等, 2000.草坪褐斑病病原菌鉴定、流行规律及其综合控制技术的研究.中国草地,(4):42-47
2.陈海波,刘荣堂,杜广真,王代军,2002.草坪草褐斑病的研究进展和现状.草原与草坪,(3):10-14
3.陈延熙,张敦华,段霞渝等,1985.关于Rhizoctonia solani菌丝融合分类和有性世代研究.植物病理学报,15(3):139-144
4.郭润芳,史宝胜,高宝嘉,2001.木霉菌在植病生物防治中的应用.河北林果研究,16(3):294-298
5.郭润芳,刘晓光,高克祥等,2002.拮抗木霉菌在生物防治中的应用与研究进展.中国生物防治,18(4):180-184
6.黄江华,杨媚,周而勋等,2001.丝核菌细胞核染色技术的研究.仲恺农业技术学院学报,14 (4): 13-17
7.黄江华,杨媚,周而勋等,2002.丝核菌研究进展.仲恺农业技术学院学报,15 (1): 61-67
8.黄江华,杨媚,周而勋等,2002.广州地区10种作物立枯丝核菌菌丝融合群测定.仲恺农业技术学院学报,15(1):14-18
9.惠有为,潘亚妮,孙勇等,2004 .耐低温木霉TR165固态发酵条件的研究.西北大学学报, 34 (1) : 69- 73.
10.金月波,矫天育,李春艳,2008.草地早熟禾褐斑病病原菌的分离及发生规律的研究.辽宁农业职业技术学院学报,10(2):19-20
11.金月波,曲强,魏雅冬,2009.草地早熟禾褐斑病菌主要生物学特性与致病影响因素研究.沈阳农业大学学报,40(3)356-359
12.李华荣,1999.丝核菌的菌丝融合群及其遗传多样性研究的新进展.菌物系统,18(1):100-107
13.卢圣栋,1999.现代分子生物学试验技术.第2版.北京:中国协和医科大学出版社
14.蒙姣荣,张朝冲,李界秋等,2007.广西玉米纹枯病菌的菌丝融合群及其对杀菌剂的敏感性.中国农学通报,22 (7): 452-454
15.蒙姣荣,张超冲,李界秋等,2006.广西水稻纹枯病菌菌丝融合群鉴定初报.中国农学通报,22(6):327-329
16.倪合兵,2004.草坪褐斑病的识别与防治.安徽农学通报,10(5):64
17.钱振官,沈国辉,张繁琴等,2003.上海地区高羊茅草坪褐斑病的发生及防治研究.草原与草坪,(3):39-41
18.商鸿生,吕学农,1996.草坪早熟禾叶枯病的病原真菌鉴定.中国草地,4:36～39
19.石仁才,商鸿生,张敬泽,2007.草坪禾草丝核菌的核相研究.菌物学报,26(2):221-225
20.石仁才,商鸿生,张敬泽,2008.中国中部5省(市)草坪禾草立枯丝核菌的菌丝融合群研究.植物病理学报,38(2):147-152
21.宋晓妍,孙彩云,陈秀兰,2006.木霉生防作用机制的研究进展.中国农业科技导报,8(6):20-25
22.苏春丽,唐传红,张劲松等,2006.基于β-微管蛋白基因部分序列探讨灵芝属菌株的亲缘关系.菌物学报,25(3):439-445
23.孙芙蓉,吴悦明,徐玉芳等,2005. T95对灰葡萄孢的拮抗作用及田间防病研究.山东农业大学学报,36(2):213-216
24.王洪凯,刘开启,吴洵耻,1997.丝核菌分类研究进展.山东农业大学学报,28 (3): 375-382
25.王芊,2001.木霉菌在生物防治上的应用及拮抗机制.黑龙江农业科学,(1):41-43
26.辛雅芬,商金杰,高克祥,2005.拮抗木霉菌的生防机制研究进展.东北林业大学学报,33(4):88-91
27.邢楠,谢响明,何晓青,2005.核酸技术在立枯丝核菌分类与生态学研究中的应用.微生物学杂志,25 (2):93-97
28.杨合同,唐文华,李纪顺等,2004.绿色木霉LTR2菌株的紫外线诱变改良.中国生物防治, 20 (3) : 182 - 186.
29.杨合同,唐文华,M. Ryder,1999.木霉菌与植物病害的生物防治.山东科学,12(4):7-15
30.杨依军,王勇,杨秀荣等,2000.拮抗木霉菌在生物防治中的作用.天津农业科学,6(3):29-3
31.燕嗣皇,陆德清,1992.木霉拮抗齐整小核菌菌株筛选.贵州农业科学, (2) : 7 - 10.
32.姚彦坡,吕国忠,张淑金等,2006.草坪褐斑病菌和镰刀枯萎病菌拮抗木霉菌的筛选及效果研究.草原与草坪,(6):52-55
33.于金凤,伍恩宇,夏海波,2006.核糖体RNA基因在丝核菌分子系统学研究中的应用.菏泽学院学报,28(5): 103-107
34.于金凤,张修国,张天宇等, 2003.立枯丝核菌(Rhizoctonia solani Kühn)第一融合群遗传分化关系的研究.菌物系统,22(1):69-73
35.于新,田淑慧,徐文兴等,2005.木霉菌生防作用的生化机制研究进展.中山大学学报,44(2);86-90
36.张敦华,陈延熙,1986.双核丝核菌的菌丝融合分类.植物病理学报,16(3): 139-144
37.张敏,陶家凤,1993.丝核菌分离方法的研究.四川农业大学学报,11(2): 261-265
38.张穗,朱有釭,吴友三,1989.东北地区丝核菌菌群分类及对禾谷类作物的致病性.沈阳农业大学学报,20 (4) : 431-435
39.张敬泽,商鸿生,2003.草地早熟禾和高羊茅病原丝核菌的物种鉴定.草业学报,12(4):31-34
40.赵蕾,1999.木霉菌的生物防治作用及其应用.生态农业研究,7(1):66-68
41.朱廷恒,邢小平,孙顺娣,2004.木霉T97菌株对几种植物病原真菌的拮抗作用机制和温室防治试验.植物保护学报,31(2):139-144
42. Aghajani MA, Alizadeh A, and Rahimian H, 2008. First report of brown patch on bristle basket grass in Iran. Plant Pathology , 57(2): 384-384
43. Aoyagi T, Kageyama K, and Hyakumachi M, 1998. Characterization and survival of Rhizoctonia solani AG2-2 LP associated with large patch disease of zoysia grass. Plant Disease, 82:857-863
44. Boysen M, Borja M, Moral C, et al., 1996. Identification at strain level of Rhizoctonia solani AG4 isolates by direct sequence of asymmetric PCR products of the ITS regions. Current Genetics, 29:174-181
45. Burpee LL, 1980. Rhizoctonia cerealis causes yellow patch of turfgrass. PlantDisease, 64: 1114-1116
46. Burpee LL, 1992. Biology of Rhizoctonia Species associated with turfgrasses. Plant Disease, 16(2): 112-117
47. Carling DE, Baird EE, Gitaitis RD, et al., 2002. Characterization of AG-13, a newly reported anastomosis group of Rhizoctonia solani. Phytopathology, 92(8): 893-899
48. Chao LJ, Shan XM, Che SC, 2000. Study on causal agent, epidemic lawand integrated control techniques of Turfgrass Brown Patch Disease. Glassland of China. 4: 42-27 (in Chinese)
49. Chet I, Inbar J, Hadar I, 1997 .Fungal antagonists and mycoparasites[M]//Berlin, Wicklow DT, S?derstr?m B. The Mycota IV: Environmentaland microbial relationships: 165-184.
50. Cubeta MA., Echandi E, Abernethy T, et al., 1991. Characterization of anastomosis groups of binucleate Rhizoctonia species using restriction analysis of an amplified ribosomal RNA gene. Phytopathology, 81(11): 1395-1400
51. ELAD Y,2003. Biocontrol of foliar pathogens:mechanisms and application.Commun Agric Appl Biol Sci, 68: 17 - 24.
52. Hayakawa T, Toda T, Ping Q, et al., 2006. A new subgroup of Rhizoctonia solani AG-D, AG-D III, obtained from Japanese zoysia grass exhibiting symptoms of a new disease. Plant Disease. 90: 1389-1394
53. Haygood RA, and Martin SB, 1990. Characterization and pathogenicity of species of Rhizoctonia associated with centipede- grass and St. Augustinegrass in South Carolina. Plant Disease. 74:510-514.
54. Herr LJ, and MM Fulton, 1995. Rhizoctonia solani AG1-IA and AG2-2 IIB cause brown patch on tall fescue and creeping bentgrass in Ohio. Plant Disease. 79:1186.
55. Howell C R, 1998.The role of antibiosis in biocontrol[M] / /HARMAN G E,KUB ICEK C P. Trichoderma and Gliocladium, Vol. 2. Enzymes, biological control and commercial applications. London: Taylor & Francis: 173- 184.
56. Hsiang T, and Dean J D, 2001. DNA Sequencing for anastomosis grouping ofRhizoctonia solani isolates from Poa Annua . International Turfgrass Society.9:674-678
57. Hurd B, and MP Grisham, 1983. Rhizoctonia spp. Associated with brown patch of St. Augustinegrass. Phytopathology 73:1661-1665.
58. Julian MC, Acero J, Salazar O, et al., 1999. Mating typecorrelated molecular markers and demonstration of heterokaryosisin the phytopathogenic fungus Thanatephorus cucumeris (Rhizoctonia solani) AG1-IC by AFLP DNA fingerprinting analysis. Journal of Biotechnology, 67(1): 49-56
59. Kuninaga S, Yokosawa R, 1985. DNA base homology in Rhizoctonia solani Kühn VII Genetic Relatedness between AG-BI and other anastomosis groups. Annual phytopatholoy Society Japan, 51: 133-138
60. Kuninaga S, Yokosawa R, 1983. DNA base homology in Rhizoctonia solani Kühn III genetic relatedness within AG-3, AG-5, AG-7 and AG-BI. Annual phytopatholoy Socity Japan, 49: 648-652
61. Kuninaga S, Natsuaki T, Takeuchi T, 1997. Sequence variation of the rDNA ITS regions within and between anastomosis groups in Rhizoctonia solani. Current Genetics, 32: 237-243
62. Lees AK, Cullen DW, Sullivan L, et al., 2002. Development of conventional and quantitative real-time PCR assays for the detection and identification of Rhizoctonia solani AG-3 in potato and soil. Plant Pathology, 51 (3): 293-302
63. Lilja A, Hietala AM, Karjalainen R, 1996. Idetification of a uninucleate Rhizoctonia spp. By pathogenicity, hyphal anastomosis and RAPD analysis. Plant Pathology, 45: 997-1006
64. Liu ZL, Domier LL, Sinclair JB, 1995. Polymorphism of genes coding for nuclear 18S rRNA intinctiveness of anastomosis group 10 from other groups in the Rhizoctonia solani species complex. Applied and Environmental Microbiology, 61(7): 2659-2664
65. Martin SB, CL Campbell, and LT Lucas,1983. Horizontal distribution and characterization of Rhizoctonia spp. in tall fescue turf. Phytopathology 73:1064-1068.
66. Martin SB, and LT Lucas, 1984. Characterization and pathogenicity of Rhizoctonia spp. and binucleate Rhizoctonia-like fungi from turfgrasses in North Carolina. Phytopathology 74:170-175.
67. Matsumoto M, 2002. Trials of direct detection and identification of Rhizoctonia solani AG 1 and AG 2 subgroups using specifically primed PCR analysis. Mycoscience, 23 (2) :185-189
68. Matsumoto M, Furuya N, Takanami Y, et al., 1997. Rapid detection of Rhizoctonia species, causal agents of rice sheath diseases, by PCR-RFLP analysis using an alkaline DNA extraction method. Mycoscience, 38 (4) :451-454
69. Matsumoto M, Furuya N, Takanami Y, et al., 1996. RFLP analysis of the PCR-amplified 28S rDNA in Rhizoctonia solani. Mycoscience, 37 (3) : 351-356
70. Monte E, 2001.Understanding Trichoderma between biotechnology and micro2 bial ecology. IntMicrobiol, 4: 1 - 4
71. Oniki M, Kobayahsi K, Araki T, et al., 1986. A new disease of turfgrass caused by binucleate Rhizoctonia, AG-Q. Ann. of the Phyto. Socie. of Japan, 52: 850-853
72. Piper CV, and Coe HS, 1919. Rhizoctonia in lawns and pastures. Phytopathology 9:89-95.
73. Salazar O, Julian MC, et al., 2000. Primers based on specific rDNA-ITS sequences for PCR detection of Rhizoctonia solani, R.solani AG2 subgroups and ecological types, and binucleate Rhizoctonia. Mycological Research, 104(3): 281-285
74. Sneh B, Burpee L, Ogoshi A, 1991. Identification of Rhizoctonia species. American Phytopathological Society, St. Paul, Minnesota. 18-78
75. Takamatsu S, Nakano M, Yokota H, et al., 1998. Detection of Rhizoctonia solani AG-2-2Ⅳ, the causal agent of large patch of zovsiagrass, using plasmid DNA as a probe. Annals of the Phytopathological Society of Japan, 64(5): 451-457
76. Takamatsu S, Nakano M, Kunasakdakul K, et al., 1999. Cloning and sequenceanalysis of the plasmid DNA found in Rhizoctonia solani AG-2-2 LP isolate and its potential use for fungal detection. Mycoscience, 20 (1): 29-33
77. Toda T, Hyakumachi M, Suga H, 1999(a). Differentiation of Rhizoctonia AG-D isolates from turfgrass into subgroups I and II based on rDNA and RAPD analyses. European Journal of Plant Pathology 105: 835–846
78. Toda T, Nasu H, Kageyama K, 1998. Hyakumachi-M Genetic identification of web-blight fungus (Rhizoctonia solani AG1) obtained from European pear using RFLP of rDNA-ITS and RAPD analyses. Research Bulletin of the Faculty of Agriculture, Gifu University, 63:1-9
79. Toda T, Hyakumachi M, Arora DK, 1999(b). Genetic relatedness among and within different Rhizoctonia solani anastomosis groups as assessed by RAPD, ERIC and REP-PCR. Microbiological Research, 154(3):247-258
80. Yan W, Xu Y, Li YZ, Chi XZ, 2003. Study on tall fescue brown patch in Shanghai.Journal of Shanghai Jiao Tong University(Agriculture Science), 21: 71-74(in Chinese)
81. Zhang M, and Dernoeden PH, 1995. Facilitating anastomosis grouping of Rhizoctonia solani isolates from cool season turfgrasses. HortScience, 30: 1260-1262
82. Zhang JZ, Shang HS, 2003. Identification of Rhizoctonia species associated with Kentucky bluegrass and Tall fescue. Acta Prataculturae Sinica. 12(4):31-34 (in Chinese)
83. Zhang M, and PH Dernoeden. 1995. Facilitating anastomosis grouping of Rhizoctonia solani isolates from cool season turfgrasses. HortScience 30:1260-1262.
2.陈海波,刘荣堂,杜广真,王代军,2002.草坪草褐斑病的研究进展和现状.草原与草坪,(3):10-14
3.陈延熙,张敦华,段霞渝等,1985.关于Rhizoctonia solani菌丝融合分类和有性世代研究.植物病理学报,15(3):139-144
4.郭润芳,史宝胜,高宝嘉,2001.木霉菌在植病生物防治中的应用.河北林果研究,16(3):294-298
5.郭润芳,刘晓光,高克祥等,2002.拮抗木霉菌在生物防治中的应用与研究进展.中国生物防治,18(4):180-184
6.黄江华,杨媚,周而勋等,2001.丝核菌细胞核染色技术的研究.仲恺农业技术学院学报,14 (4): 13-17
7.黄江华,杨媚,周而勋等,2002.丝核菌研究进展.仲恺农业技术学院学报,15 (1): 61-67
8.黄江华,杨媚,周而勋等,2002.广州地区10种作物立枯丝核菌菌丝融合群测定.仲恺农业技术学院学报,15(1):14-18
9.惠有为,潘亚妮,孙勇等,2004 .耐低温木霉TR165固态发酵条件的研究.西北大学学报, 34 (1) : 69- 73.
10.金月波,矫天育,李春艳,2008.草地早熟禾褐斑病病原菌的分离及发生规律的研究.辽宁农业职业技术学院学报,10(2):19-20
11.金月波,曲强,魏雅冬,2009.草地早熟禾褐斑病菌主要生物学特性与致病影响因素研究.沈阳农业大学学报,40(3)356-359
12.李华荣,1999.丝核菌的菌丝融合群及其遗传多样性研究的新进展.菌物系统,18(1):100-107
13.卢圣栋,1999.现代分子生物学试验技术.第2版.北京:中国协和医科大学出版社
14.蒙姣荣,张朝冲,李界秋等,2007.广西玉米纹枯病菌的菌丝融合群及其对杀菌剂的敏感性.中国农学通报,22 (7): 452-454
15.蒙姣荣,张超冲,李界秋等,2006.广西水稻纹枯病菌菌丝融合群鉴定初报.中国农学通报,22(6):327-329
16.倪合兵,2004.草坪褐斑病的识别与防治.安徽农学通报,10(5):64
17.钱振官,沈国辉,张繁琴等,2003.上海地区高羊茅草坪褐斑病的发生及防治研究.草原与草坪,(3):39-41
18.商鸿生,吕学农,1996.草坪早熟禾叶枯病的病原真菌鉴定.中国草地,4:36～39
19.石仁才,商鸿生,张敬泽,2007.草坪禾草丝核菌的核相研究.菌物学报,26(2):221-225
20.石仁才,商鸿生,张敬泽,2008.中国中部5省(市)草坪禾草立枯丝核菌的菌丝融合群研究.植物病理学报,38(2):147-152
21.宋晓妍,孙彩云,陈秀兰,2006.木霉生防作用机制的研究进展.中国农业科技导报,8(6):20-25
22.苏春丽,唐传红,张劲松等,2006.基于β-微管蛋白基因部分序列探讨灵芝属菌株的亲缘关系.菌物学报,25(3):439-445
23.孙芙蓉,吴悦明,徐玉芳等,2005. T95对灰葡萄孢的拮抗作用及田间防病研究.山东农业大学学报,36(2):213-216
24.王洪凯,刘开启,吴洵耻,1997.丝核菌分类研究进展.山东农业大学学报,28 (3): 375-382
25.王芊,2001.木霉菌在生物防治上的应用及拮抗机制.黑龙江农业科学,(1):41-43
26.辛雅芬,商金杰,高克祥,2005.拮抗木霉菌的生防机制研究进展.东北林业大学学报,33(4):88-91
27.邢楠,谢响明,何晓青,2005.核酸技术在立枯丝核菌分类与生态学研究中的应用.微生物学杂志,25 (2):93-97
28.杨合同,唐文华,李纪顺等,2004.绿色木霉LTR2菌株的紫外线诱变改良.中国生物防治, 20 (3) : 182 - 186.
29.杨合同,唐文华,M. Ryder,1999.木霉菌与植物病害的生物防治.山东科学,12(4):7-15
30.杨依军,王勇,杨秀荣等,2000.拮抗木霉菌在生物防治中的作用.天津农业科学,6(3):29-3
31.燕嗣皇,陆德清,1992.木霉拮抗齐整小核菌菌株筛选.贵州农业科学, (2) : 7 - 10.
32.姚彦坡,吕国忠,张淑金等,2006.草坪褐斑病菌和镰刀枯萎病菌拮抗木霉菌的筛选及效果研究.草原与草坪,(6):52-55
33.于金凤,伍恩宇,夏海波,2006.核糖体RNA基因在丝核菌分子系统学研究中的应用.菏泽学院学报,28(5): 103-107
34.于金凤,张修国,张天宇等, 2003.立枯丝核菌(Rhizoctonia solani Kühn)第一融合群遗传分化关系的研究.菌物系统,22(1):69-73
35.于新,田淑慧,徐文兴等,2005.木霉菌生防作用的生化机制研究进展.中山大学学报,44(2);86-90
36.张敦华,陈延熙,1986.双核丝核菌的菌丝融合分类.植物病理学报,16(3): 139-144
37.张敏,陶家凤,1993.丝核菌分离方法的研究.四川农业大学学报,11(2): 261-265
38.张穗,朱有釭,吴友三,1989.东北地区丝核菌菌群分类及对禾谷类作物的致病性.沈阳农业大学学报,20 (4) : 431-435
39.张敬泽,商鸿生,2003.草地早熟禾和高羊茅病原丝核菌的物种鉴定.草业学报,12(4):31-34
40.赵蕾,1999.木霉菌的生物防治作用及其应用.生态农业研究,7(1):66-68
41.朱廷恒,邢小平,孙顺娣,2004.木霉T97菌株对几种植物病原真菌的拮抗作用机制和温室防治试验.植物保护学报,31(2):139-144
42. Aghajani MA, Alizadeh A, and Rahimian H, 2008. First report of brown patch on bristle basket grass in Iran. Plant Pathology , 57(2): 384-384
43. Aoyagi T, Kageyama K, and Hyakumachi M, 1998. Characterization and survival of Rhizoctonia solani AG2-2 LP associated with large patch disease of zoysia grass. Plant Disease, 82:857-863
44. Boysen M, Borja M, Moral C, et al., 1996. Identification at strain level of Rhizoctonia solani AG4 isolates by direct sequence of asymmetric PCR products of the ITS regions. Current Genetics, 29:174-181
45. Burpee LL, 1980. Rhizoctonia cerealis causes yellow patch of turfgrass. PlantDisease, 64: 1114-1116
46. Burpee LL, 1992. Biology of Rhizoctonia Species associated with turfgrasses. Plant Disease, 16(2): 112-117
47. Carling DE, Baird EE, Gitaitis RD, et al., 2002. Characterization of AG-13, a newly reported anastomosis group of Rhizoctonia solani. Phytopathology, 92(8): 893-899
48. Chao LJ, Shan XM, Che SC, 2000. Study on causal agent, epidemic lawand integrated control techniques of Turfgrass Brown Patch Disease. Glassland of China. 4: 42-27 (in Chinese)
49. Chet I, Inbar J, Hadar I, 1997 .Fungal antagonists and mycoparasites[M]//Berlin, Wicklow DT, S?derstr?m B. The Mycota IV: Environmentaland microbial relationships: 165-184.
50. Cubeta MA., Echandi E, Abernethy T, et al., 1991. Characterization of anastomosis groups of binucleate Rhizoctonia species using restriction analysis of an amplified ribosomal RNA gene. Phytopathology, 81(11): 1395-1400
51. ELAD Y,2003. Biocontrol of foliar pathogens:mechanisms and application.Commun Agric Appl Biol Sci, 68: 17 - 24.
52. Hayakawa T, Toda T, Ping Q, et al., 2006. A new subgroup of Rhizoctonia solani AG-D, AG-D III, obtained from Japanese zoysia grass exhibiting symptoms of a new disease. Plant Disease. 90: 1389-1394
53. Haygood RA, and Martin SB, 1990. Characterization and pathogenicity of species of Rhizoctonia associated with centipede- grass and St. Augustinegrass in South Carolina. Plant Disease. 74:510-514.
54. Herr LJ, and MM Fulton, 1995. Rhizoctonia solani AG1-IA and AG2-2 IIB cause brown patch on tall fescue and creeping bentgrass in Ohio. Plant Disease. 79:1186.
55. Howell C R, 1998.The role of antibiosis in biocontrol[M] / /HARMAN G E,KUB ICEK C P. Trichoderma and Gliocladium, Vol. 2. Enzymes, biological control and commercial applications. London: Taylor & Francis: 173- 184.
56. Hsiang T, and Dean J D, 2001. DNA Sequencing for anastomosis grouping ofRhizoctonia solani isolates from Poa Annua . International Turfgrass Society.9:674-678
57. Hurd B, and MP Grisham, 1983. Rhizoctonia spp. Associated with brown patch of St. Augustinegrass. Phytopathology 73:1661-1665.
58. Julian MC, Acero J, Salazar O, et al., 1999. Mating typecorrelated molecular markers and demonstration of heterokaryosisin the phytopathogenic fungus Thanatephorus cucumeris (Rhizoctonia solani) AG1-IC by AFLP DNA fingerprinting analysis. Journal of Biotechnology, 67(1): 49-56
59. Kuninaga S, Yokosawa R, 1985. DNA base homology in Rhizoctonia solani Kühn VII Genetic Relatedness between AG-BI and other anastomosis groups. Annual phytopatholoy Society Japan, 51: 133-138
60. Kuninaga S, Yokosawa R, 1983. DNA base homology in Rhizoctonia solani Kühn III genetic relatedness within AG-3, AG-5, AG-7 and AG-BI. Annual phytopatholoy Socity Japan, 49: 648-652
61. Kuninaga S, Natsuaki T, Takeuchi T, 1997. Sequence variation of the rDNA ITS regions within and between anastomosis groups in Rhizoctonia solani. Current Genetics, 32: 237-243
62. Lees AK, Cullen DW, Sullivan L, et al., 2002. Development of conventional and quantitative real-time PCR assays for the detection and identification of Rhizoctonia solani AG-3 in potato and soil. Plant Pathology, 51 (3): 293-302
63. Lilja A, Hietala AM, Karjalainen R, 1996. Idetification of a uninucleate Rhizoctonia spp. By pathogenicity, hyphal anastomosis and RAPD analysis. Plant Pathology, 45: 997-1006
64. Liu ZL, Domier LL, Sinclair JB, 1995. Polymorphism of genes coding for nuclear 18S rRNA intinctiveness of anastomosis group 10 from other groups in the Rhizoctonia solani species complex. Applied and Environmental Microbiology, 61(7): 2659-2664
65. Martin SB, CL Campbell, and LT Lucas,1983. Horizontal distribution and characterization of Rhizoctonia spp. in tall fescue turf. Phytopathology 73:1064-1068.
66. Martin SB, and LT Lucas, 1984. Characterization and pathogenicity of Rhizoctonia spp. and binucleate Rhizoctonia-like fungi from turfgrasses in North Carolina. Phytopathology 74:170-175.
67. Matsumoto M, 2002. Trials of direct detection and identification of Rhizoctonia solani AG 1 and AG 2 subgroups using specifically primed PCR analysis. Mycoscience, 23 (2) :185-189
68. Matsumoto M, Furuya N, Takanami Y, et al., 1997. Rapid detection of Rhizoctonia species, causal agents of rice sheath diseases, by PCR-RFLP analysis using an alkaline DNA extraction method. Mycoscience, 38 (4) :451-454
69. Matsumoto M, Furuya N, Takanami Y, et al., 1996. RFLP analysis of the PCR-amplified 28S rDNA in Rhizoctonia solani. Mycoscience, 37 (3) : 351-356
70. Monte E, 2001.Understanding Trichoderma between biotechnology and micro2 bial ecology. IntMicrobiol, 4: 1 - 4
71. Oniki M, Kobayahsi K, Araki T, et al., 1986. A new disease of turfgrass caused by binucleate Rhizoctonia, AG-Q. Ann. of the Phyto. Socie. of Japan, 52: 850-853
72. Piper CV, and Coe HS, 1919. Rhizoctonia in lawns and pastures. Phytopathology 9:89-95.
73. Salazar O, Julian MC, et al., 2000. Primers based on specific rDNA-ITS sequences for PCR detection of Rhizoctonia solani, R.solani AG2 subgroups and ecological types, and binucleate Rhizoctonia. Mycological Research, 104(3): 281-285
74. Sneh B, Burpee L, Ogoshi A, 1991. Identification of Rhizoctonia species. American Phytopathological Society, St. Paul, Minnesota. 18-78
75. Takamatsu S, Nakano M, Yokota H, et al., 1998. Detection of Rhizoctonia solani AG-2-2Ⅳ, the causal agent of large patch of zovsiagrass, using plasmid DNA as a probe. Annals of the Phytopathological Society of Japan, 64(5): 451-457
76. Takamatsu S, Nakano M, Kunasakdakul K, et al., 1999. Cloning and sequenceanalysis of the plasmid DNA found in Rhizoctonia solani AG-2-2 LP isolate and its potential use for fungal detection. Mycoscience, 20 (1): 29-33
77. Toda T, Hyakumachi M, Suga H, 1999(a). Differentiation of Rhizoctonia AG-D isolates from turfgrass into subgroups I and II based on rDNA and RAPD analyses. European Journal of Plant Pathology 105: 835–846
78. Toda T, Nasu H, Kageyama K, 1998. Hyakumachi-M Genetic identification of web-blight fungus (Rhizoctonia solani AG1) obtained from European pear using RFLP of rDNA-ITS and RAPD analyses. Research Bulletin of the Faculty of Agriculture, Gifu University, 63:1-9
79. Toda T, Hyakumachi M, Arora DK, 1999(b). Genetic relatedness among and within different Rhizoctonia solani anastomosis groups as assessed by RAPD, ERIC and REP-PCR. Microbiological Research, 154(3):247-258
80. Yan W, Xu Y, Li YZ, Chi XZ, 2003. Study on tall fescue brown patch in Shanghai.Journal of Shanghai Jiao Tong University(Agriculture Science), 21: 71-74(in Chinese)
81. Zhang M, and Dernoeden PH, 1995. Facilitating anastomosis grouping of Rhizoctonia solani isolates from cool season turfgrasses. HortScience, 30: 1260-1262
82. Zhang JZ, Shang HS, 2003. Identification of Rhizoctonia species associated with Kentucky bluegrass and Tall fescue. Acta Prataculturae Sinica. 12(4):31-34 (in Chinese)
83. Zhang M, and PH Dernoeden. 1995. Facilitating anastomosis grouping of Rhizoctonia solani isolates from cool season turfgrasses. HortScience 30:1260-1262.