雅安郊区辣椒生长期土壤中木霉种群分布及其对辣椒疫霉病的生防研究
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摘要
对雅安辣椒生长期土壤中木霉的分离、鉴定及室内拮抗试验，筛选到大量的拮抗菌株。利用拮抗作用强的菌株开展了对辣椒疫霉病的生防和促生盆栽试验及木霉在辣椒植株根际定殖模式的研究，为辣椒疫霉病的生防提供理论依据与科学参考。
从雅安郊区辣椒生长期土壤中0-5cm、5-10cm两个深度分别五点取样，共得到木霉58个菌株8个种。它们分别是T.harzianum、T.pseudokoningii T.citrinoviride、T.atroviride、T.longibrachiatum、T.hamatum、T.konigii T.aureoviride。同时土壤5cm深处哈茨木霉、拟康木霉和康氏木霉含量较多，属优势种群，黄绿木霉、桔绿木霉和钩状木霉次之，长枝木霉和深绿木霉没有发现；而土壤10cm处拟康木霉、桔绿木霉和深绿木霉含量较多，属优势种群，康氏木霉、黄绿木霉和长枝木霉次之，钩状木霉和哈茨木霉没有发现。这表明钩状木霉和哈茨木霉习居于土壤表层，长枝木霉和深绿木霉习居于土壤深层，其它木霉种群居于土壤中深度选择性不强。随着辣椒土壤温度升高，土壤含水量的减少以及土壤深度的加深，木霉在辣椒土壤中的种群数量也逐渐减少。
从辣椒土壤分离得到的木霉产生的非挥发性代谢产物对辣椒疫霉病菌的抑制作用表明桔绿木霉、哈次木霉、拟康木霉和康氏木霉对辣椒疫霉菌菌丝有较强的抑制作用，其平均抑制百分率分别为56.4％、69.1％、67.2％、63.6％；而其它几种木霉的非挥发性代谢产物对辣椒疫霉病菌的抑制作用都小于15％，且没有显著性区别。对峙培养表明哈次木霉、拟康木霉和康氏木霉对辣椒疫霉病菌菌丝有溶解、断裂、扭曲、缠绕等作用，最终导致其崩解、死亡。木霉易非发性代产物对辣椒疫霉病菌抑制作不强。
在混施有少量三唑酮和没混施有少量三唑酮的两种辣椒盆栽土中，混施有少量三唑酮的盆栽土中哈次木霉在辣椒植株根际的定殖要比没混施有少量三唑酮的盆栽土中哈次木霉在辣椒植株根际的定殖快。三唑酮可抑制土壤中某些微生物种群，从而加强了木霉在辣椒根际的竞争，以利于引入的木霉在辣椒根际的定殖，达到防治病害的目的。
在使用哈茨木霉生防制剂对辣椒疫霉病进行生物防治的试验中，只施有哈茨木霉
的处理与哈茨木霉并混施有少量三哇酮的两个处理结果表现病情指数为0，对辣椒疫
霉病防治效果达，100%;只施有三哇酮的处理结果表现病情指数为4，对辣椒疫霉病防
治效果达93%.同时有哈茨木霉参加的三个处理中辣椒的株高、茎杆、叶面积和叶片
颜色都有明显的优于其它几个处理。使用哈茨木霉并混施有少量三哇酮比只使用哈
茨木霉处理中的辣椒植株矮、叶片颜色深、茎杆粗，这可能与三哇酮抑制辣椒生长
但能增加辣椒植株的抗逆性有关。
在康氏木霉、拟康木霉与美国康乃尔菌株对辣椒植株的促生作用试验中，辣椒
植株在叶片平均面积、植株平均高度以及茎杆的平均周长都显著优于对照。但是美
国康乃尔菌株与康氏木霉、拟康木霉相比有更显著的促生作用。
Fifty-eight Trichoderma spp. strains were isolated from the capsicum soil in the different depth 5cm and 10cm in ya'an suburb.they were indentified as following species:Tharzianum, T. pseudokoningii, T. citrinoviride, T. atroviride, T. longibrachiatum, T. hamatum, T. koningii,, T. aureoviride.The two Trichoderma isolates were selected and used according to the antagonistic test against Phytophthora capsici .The dominant strains were T. harzianum, T. pseudokoningii, T. koningii.The deeper of the soil located and the higher the soil temperature,the less distribution of Trichoderma spp. were during the capsicum growth.
The antagonistic test showed that T.harzianum, T. pseudokoningii* T. koningii had the better antagonistic effects .The inhabition of metabolites against Phytophthora capsici were 69.1%% 67.2%, and 63.6% repectively.The interaction between T.harzianum and hypha of Phytophthora capsici showed that T.harzianum made hypha of Phytophthora capsici dissolved % twisted and distorted. In the end,it led to collapse of hypha.
The colonization experiment in potted capsicum soil showed that the colonization of Trichoderma spp. in the soil which were mixed with T. harzianum and triadimefon. quicker than that in the soil only applied T.harzianum. The possible reason is some microorganism in soil was inhabited by pesticide to make T. harzianum colonize quickly.
The biological control against Phytophthora capsici with T. harzianum showed that T. harzianum had strongly biological effect on phytophthora capsici .The biological control effect of the treatment of which were mixed with triadimefon and T. harzianum was the same with the treament of which only was used T. harzianum,botii are 100%.But The biological control effect of the treatment in which was only used triadimefon was 93%. At the same time T. harzianum also promoted capsicum growth obvisouly.
T. koningii * T. pseudokoningii and The strains which came from Aemerica kangnaier had obvisouly advantage in capsicum growth.But the biological control effect of kangnaier isolate was superiorer than others two.
从雅安郊区辣椒生长期土壤中0-5cm、5-10cm两个深度分别五点取样，共得到木霉58个菌株8个种。它们分别是T.harzianum、T.pseudokoningii T.citrinoviride、T.atroviride、T.longibrachiatum、T.hamatum、T.konigii T.aureoviride。同时土壤5cm深处哈茨木霉、拟康木霉和康氏木霉含量较多，属优势种群，黄绿木霉、桔绿木霉和钩状木霉次之，长枝木霉和深绿木霉没有发现；而土壤10cm处拟康木霉、桔绿木霉和深绿木霉含量较多，属优势种群，康氏木霉、黄绿木霉和长枝木霉次之，钩状木霉和哈茨木霉没有发现。这表明钩状木霉和哈茨木霉习居于土壤表层，长枝木霉和深绿木霉习居于土壤深层，其它木霉种群居于土壤中深度选择性不强。随着辣椒土壤温度升高，土壤含水量的减少以及土壤深度的加深，木霉在辣椒土壤中的种群数量也逐渐减少。
从辣椒土壤分离得到的木霉产生的非挥发性代谢产物对辣椒疫霉病菌的抑制作用表明桔绿木霉、哈次木霉、拟康木霉和康氏木霉对辣椒疫霉菌菌丝有较强的抑制作用，其平均抑制百分率分别为56.4％、69.1％、67.2％、63.6％；而其它几种木霉的非挥发性代谢产物对辣椒疫霉病菌的抑制作用都小于15％，且没有显著性区别。对峙培养表明哈次木霉、拟康木霉和康氏木霉对辣椒疫霉病菌菌丝有溶解、断裂、扭曲、缠绕等作用，最终导致其崩解、死亡。木霉易非发性代产物对辣椒疫霉病菌抑制作不强。
在混施有少量三唑酮和没混施有少量三唑酮的两种辣椒盆栽土中，混施有少量三唑酮的盆栽土中哈次木霉在辣椒植株根际的定殖要比没混施有少量三唑酮的盆栽土中哈次木霉在辣椒植株根际的定殖快。三唑酮可抑制土壤中某些微生物种群，从而加强了木霉在辣椒根际的竞争，以利于引入的木霉在辣椒根际的定殖，达到防治病害的目的。
在使用哈茨木霉生防制剂对辣椒疫霉病进行生物防治的试验中，只施有哈茨木霉
的处理与哈茨木霉并混施有少量三哇酮的两个处理结果表现病情指数为0，对辣椒疫
霉病防治效果达，100%;只施有三哇酮的处理结果表现病情指数为4，对辣椒疫霉病防
治效果达93%.同时有哈茨木霉参加的三个处理中辣椒的株高、茎杆、叶面积和叶片
颜色都有明显的优于其它几个处理。使用哈茨木霉并混施有少量三哇酮比只使用哈
茨木霉处理中的辣椒植株矮、叶片颜色深、茎杆粗，这可能与三哇酮抑制辣椒生长
但能增加辣椒植株的抗逆性有关。
在康氏木霉、拟康木霉与美国康乃尔菌株对辣椒植株的促生作用试验中，辣椒
植株在叶片平均面积、植株平均高度以及茎杆的平均周长都显著优于对照。但是美
国康乃尔菌株与康氏木霉、拟康木霉相比有更显著的促生作用。
Fifty-eight Trichoderma spp. strains were isolated from the capsicum soil in the different depth 5cm and 10cm in ya'an suburb.they were indentified as following species:Tharzianum, T. pseudokoningii, T. citrinoviride, T. atroviride, T. longibrachiatum, T. hamatum, T. koningii,, T. aureoviride.The two Trichoderma isolates were selected and used according to the antagonistic test against Phytophthora capsici .The dominant strains were T. harzianum, T. pseudokoningii, T. koningii.The deeper of the soil located and the higher the soil temperature,the less distribution of Trichoderma spp. were during the capsicum growth.
The antagonistic test showed that T.harzianum, T. pseudokoningii* T. koningii had the better antagonistic effects .The inhabition of metabolites against Phytophthora capsici were 69.1%% 67.2%, and 63.6% repectively.The interaction between T.harzianum and hypha of Phytophthora capsici showed that T.harzianum made hypha of Phytophthora capsici dissolved % twisted and distorted. In the end,it led to collapse of hypha.
The colonization experiment in potted capsicum soil showed that the colonization of Trichoderma spp. in the soil which were mixed with T. harzianum and triadimefon. quicker than that in the soil only applied T.harzianum. The possible reason is some microorganism in soil was inhabited by pesticide to make T. harzianum colonize quickly.
The biological control against Phytophthora capsici with T. harzianum showed that T. harzianum had strongly biological effect on phytophthora capsici .The biological control effect of the treatment of which were mixed with triadimefon and T. harzianum was the same with the treament of which only was used T. harzianum,botii are 100%.But The biological control effect of the treatment in which was only used triadimefon was 93%. At the same time T. harzianum also promoted capsicum growth obvisouly.
T. koningii * T. pseudokoningii and The strains which came from Aemerica kangnaier had obvisouly advantage in capsicum growth.But the biological control effect of kangnaier isolate was superiorer than others two.
引文
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66 Thomos.E. S. Evaluatin of phenetic species and phylogenetic relationships in the genus Trichoderma by cladistic anlaysis-isozyme polymorphism Mycologia,1989, (89) : 39-40．
67 Zamir, D. etal. Application of enzyme electrophoresis for the identification of the identification of isolation in Trichoderma harzianum can .j. microbio.1985, (31) :578-580．
68 Benhamou N.and chet.J.Hyphal interactions between Tricherma hamatum and Rhizoctonia sonai:Utrastructure and gold cytochemistry of the mycoparasitic process, phytopatology.1993, (83) : 1062-1071．
69 Adams PB.The potential of mycoparasites for biological control of plant diseases [J].Ann. Rev .phytopathol, 1990,28:59-72．
70 杨合同．[博士论文][D]．北京：中国农业大学，1998．
71 Ridout C J Coley-smith J R and J M.Eneyme activity and electrophoretic profile
ofextracellullar protein induced in Trichoderma spp by cell wallsof Rhizoctonia solani[J].J.Gen. Microbiol, 1986,( 132) :2345-23 52．
72 Ivan A .etal.The possible role of competition between Trichoderma harzianum and Fusarium oxysporum on rhizosphere colonization [J]. Phytopathology, 1989,79 (2) : 198-203．
73 angarone B.Royer J C and Nakas J P. Purification and characterization of and endo-l,3-D-glucanase from Trichoderma longibrachintum[J].Apple Environ Microbiol, 1989,(55) :177-184．
74 Lorito M, Harmamn G E and Hays C K etal. Chitinolytic enzymes produced by Trichoderma harzianum: aneifungal activity of purified endo chitinase and chitobiosidase [J]. Phytopathology, 1993, (83) : 302-307．
75 Garcia etal .Cloning and characterization of a chitinase (CHIT42) Cdna From the mycoparasitic fungus Trichoderma harzianum[J].Current Genetics,1994,27(1) :83-89．
76张旭东、刘云龙、张中义等．木霉生防菌对植物生长的影响．云南农业大学学报，2001，16 (4) ：299-302．
77 Changyc,Bakerr,Increased growth of plants in the presence of the biological control agont Trichoderma harzianum [J],Plant Dis ,1986, (70) : 145-148．
78 Wind Hammt, Elady, Bakerr, A mechanism ofr increased plant growth induced by Trichoderma spp. [J], Phto pathology, 1906, (76) : 518-521．
79 Wyssp, Bollert, Wiemeart, Testing the eflect of biological control agents on the formation of vesiclararbs cular mycorrhiza[J]. plant soil, 1992, (147) :159-162．
80 Ahamd JS,Ballerr,Grouth of rhizosphere-competence Mutants of Thrichoderma harzianum on carbon substrates [J]. Can.J. Microbiol, 1998,(34) : 807-814．
81 DeanJ F D.game H R,and Erson J D.The ethylenebiosynthesis-inducing xyganase its induction in Trichoderma viride and certain plant patyoyens [J].Phytopathology, 1989, (79) : 1071-1078．
82 Martin JP use of acid ,rose Bengal and streptomyc in the plate method for estimating soil fungi. Soic science .1950,(143) :215-232．
83 Dinmis, C. R webster, J.Antagonistic prorer ties of species-groups of trichoderma hyphal lateractim Trans.Br.Mycol.Soc. 1991,157:363-369．
84 Dinmis, G, And wbster, J. Antagonistic properitise of species-groups of Trichoderma hyphal Iateractim Trans. Br. Mycoi.Soc.1991,157:363-369.
85 Dinmis, C. webster, J. Antagonistic properties of species-groups of Trichoderma production of Volatile antibiotic Trans: Br. Mycol. soc. 1971,(157):41-52.
86 赵国其，林福呈，陈卫良等．绿色木霉对西瓜枯萎病苗期的控制作用．浙江农业学报，1998，10(4)：206-209．
87 方中达．植病研究方法．
88 常彩涛，王鸣，巩振辉．诱发辣椒疫霉菌大量产孢的最佳方法．西北农业报，1995，23(1)：90-92．
89 方文慧、葛长鹏等．辣椒疫病接种鉴定方法的筛选．天津农业科学，1996，2(3)：12-13．
90 杨雨环．木霉防治辣椒白绢病和青枯病试验研究．贵州农业科学，1996，2：31-34．
91 Bosland P. W, and Lindsey, D.L. A Seeding screen for phytophthora root rot of pepper,capsicum annuelm Plant Disease. 1991,(75):1048-1050.
92 毛爱军，胡恰等．辣椒疫霉病菌接种鉴定技术研究．天津农业科学，1998，16(2)：35-37．
93 AHMAD J S,BAKER R. Rhizosphere competence of Trichoderma harzianum[J]. Phytopathology,1987,77:182-189.
94 AHMAD J S, BAKER R. Rhizosphere competence of benomyl toler-ant mutants of Trichoderma spp.[J].Can. J.Microbiol, 1987, 34:694-696.
95 燕嗣皇，吴石平，陆德清，等．三唑酮对木霉根际竞争定殖的影响[J]．植物病理学报，2000 30：266-270．
96 燕嗣皇，吴石平，陆德清，等．三唑酮对西瓜根际木霉菌与尖孢镰刀菌竞争定殖的影响[J]．植物病理学报，200131：265-270．
97 郭振飞，卢少云，李启明．三唑酮提高水稻幼苗抗旱性的研究[J]，植物学报，199739(6)：541
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64 kim E S etal .Virulence to korea pepper cultivars of isolates of phytophthora capsici from different geographic areas plant disease, 1992,(76) :486-489．
65关于舒，李风云等．我国辣椒疫病的研究现状．辽宁农业科学，1999，(5) ：24-27．
66 Thomos.E. S. Evaluatin of phenetic species and phylogenetic relationships in the genus Trichoderma by cladistic anlaysis-isozyme polymorphism Mycologia,1989, (89) : 39-40．
67 Zamir, D. etal. Application of enzyme electrophoresis for the identification of the identification of isolation in Trichoderma harzianum can .j. microbio.1985, (31) :578-580．
68 Benhamou N.and chet.J.Hyphal interactions between Tricherma hamatum and Rhizoctonia sonai:Utrastructure and gold cytochemistry of the mycoparasitic process, phytopatology.1993, (83) : 1062-1071．
69 Adams PB.The potential of mycoparasites for biological control of plant diseases [J].Ann. Rev .phytopathol, 1990,28:59-72．
70 杨合同．[博士论文][D]．北京：中国农业大学，1998．
71 Ridout C J Coley-smith J R and J M.Eneyme activity and electrophoretic profile
ofextracellullar protein induced in Trichoderma spp by cell wallsof Rhizoctonia solani[J].J.Gen. Microbiol, 1986,( 132) :2345-23 52．
72 Ivan A .etal.The possible role of competition between Trichoderma harzianum and Fusarium oxysporum on rhizosphere colonization [J]. Phytopathology, 1989,79 (2) : 198-203．
73 angarone B.Royer J C and Nakas J P. Purification and characterization of and endo-l,3-D-glucanase from Trichoderma longibrachintum[J].Apple Environ Microbiol, 1989,(55) :177-184．
74 Lorito M, Harmamn G E and Hays C K etal. Chitinolytic enzymes produced by Trichoderma harzianum: aneifungal activity of purified endo chitinase and chitobiosidase [J]. Phytopathology, 1993, (83) : 302-307．
75 Garcia etal .Cloning and characterization of a chitinase (CHIT42) Cdna From the mycoparasitic fungus Trichoderma harzianum[J].Current Genetics,1994,27(1) :83-89．
76张旭东、刘云龙、张中义等．木霉生防菌对植物生长的影响．云南农业大学学报，2001，16 (4) ：299-302．
77 Changyc,Bakerr,Increased growth of plants in the presence of the biological control agont Trichoderma harzianum [J],Plant Dis ,1986, (70) : 145-148．
78 Wind Hammt, Elady, Bakerr, A mechanism ofr increased plant growth induced by Trichoderma spp. [J], Phto pathology, 1906, (76) : 518-521．
79 Wyssp, Bollert, Wiemeart, Testing the eflect of biological control agents on the formation of vesiclararbs cular mycorrhiza[J]. plant soil, 1992, (147) :159-162．
80 Ahamd JS,Ballerr,Grouth of rhizosphere-competence Mutants of Thrichoderma harzianum on carbon substrates [J]. Can.J. Microbiol, 1998,(34) : 807-814．
81 DeanJ F D.game H R,and Erson J D.The ethylenebiosynthesis-inducing xyganase its induction in Trichoderma viride and certain plant patyoyens [J].Phytopathology, 1989, (79) : 1071-1078．
82 Martin JP use of acid ,rose Bengal and streptomyc in the plate method for estimating soil fungi. Soic science .1950,(143) :215-232．
83 Dinmis, C. R webster, J.Antagonistic prorer ties of species-groups of trichoderma hyphal lateractim Trans.Br.Mycol.Soc. 1991,157:363-369．
84 Dinmis, G, And wbster, J. Antagonistic properitise of species-groups of Trichoderma hyphal Iateractim Trans. Br. Mycoi.Soc.1991,157:363-369.
85 Dinmis, C. webster, J. Antagonistic properties of species-groups of Trichoderma production of Volatile antibiotic Trans: Br. Mycol. soc. 1971,(157):41-52.
86 赵国其，林福呈，陈卫良等．绿色木霉对西瓜枯萎病苗期的控制作用．浙江农业学报，1998，10(4)：206-209．
87 方中达．植病研究方法．
88 常彩涛，王鸣，巩振辉．诱发辣椒疫霉菌大量产孢的最佳方法．西北农业报，1995，23(1)：90-92．
89 方文慧、葛长鹏等．辣椒疫病接种鉴定方法的筛选．天津农业科学，1996，2(3)：12-13．
90 杨雨环．木霉防治辣椒白绢病和青枯病试验研究．贵州农业科学，1996，2：31-34．
91 Bosland P. W, and Lindsey, D.L. A Seeding screen for phytophthora root rot of pepper,capsicum annuelm Plant Disease. 1991,(75):1048-1050.
92 毛爱军，胡恰等．辣椒疫霉病菌接种鉴定技术研究．天津农业科学，1998，16(2)：35-37．
93 AHMAD J S,BAKER R. Rhizosphere competence of Trichoderma harzianum[J]. Phytopathology,1987,77:182-189.
94 AHMAD J S, BAKER R. Rhizosphere competence of benomyl toler-ant mutants of Trichoderma spp.[J].Can. J.Microbiol, 1987, 34:694-696.
95 燕嗣皇，吴石平，陆德清，等．三唑酮对木霉根际竞争定殖的影响[J]．植物病理学报，2000 30：266-270．
96 燕嗣皇，吴石平，陆德清，等．三唑酮对西瓜根际木霉菌与尖孢镰刀菌竞争定殖的影响[J]．植物病理学报，200131：265-270．
97 郭振飞，卢少云，李启明．三唑酮提高水稻幼苗抗旱性的研究[J]，植物学报，199739(6)：541
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