绿色粘帚霉对玉米大斑病生物防治研究
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摘要
从雅安郊区玉米农田里,采集玉米大斑病病斑的叶片,经实验室室内分离、纯化、鉴定,确认为玉米大斑病病原菌(Exserohilum turcicum),并在PDA斜面培养基上培养、保存备用,作为本研究的实验菌株。
分别于雅安四川农业大学教学农场和老板山上取土样,从土壤中分离获得绿色粘帚霉(Gliocladium virens)菌株共46个,根据菌株的来源和在PDA培养基上生长的不同特性,选取其中的14个菌株,与实验室保存的16个菌株一起作为本实验的供试菌株。
对玉米大斑病病原菌进行菌丝生长,产孢等生物学特性进行了研。结果表明：大斑病菌菌丝生长的最适条件是全光照、25℃、pH为6、PDA培养基；而菌株产孢最适条件是全黑暗、25℃、pH为6、清水琼脂培养基。
将供试的绿色粘帚霉菌株与大斑病菌做对峙培养：所有绿色粘帚霉菌株对大斑病菌的菌丝生长均有一定的抑制作用,抑制效果最好的为菌株G3,其抑制率达到59.12%。利用绿色粘帚霉难挥发性代谢产物,通过含毒介质法对大斑病菌抑制作用的测定实验中,粘帚霉G3和G8菌株对病原菌菌丝的抑制率最高,分别为64.20%和63.64%；最低的是G37,仅42.50%。在绿色粘帚霉易挥发性代谢产物对大斑病菌抑制作用的实验中,结果表明：抑制率最高的为G8,达到45.00%；最低的为G37,仅21.00%。
对雅安多营农场我校玉米所玉米自交系材料进行大斑病抗病性调查。所调查的60份自交系材料中,病情指数最高的为自交系10ks3-4,达到36.00；最低的为10117002,仅4.75。病情指数达到30.00以上的自交系占调查材料的12.00%；病情指数小于10.00的占调查材料的21.67%。在这些材料中,以10117这类自交系材料发病率及病情指数都较低,而以10ks类的材料发病率及病情指数都偏高。
利用水杨酸、草酸和绿色粘帚霉发酵液对玉米材料进行大斑病抗性诱导实验,通过室内玉米苗期诱导后,酶活性测定实验表明：在受到不同诱导剂处理下,抗病品种的PPO与PAL活性均高于感病品种,材料的抗病性与PPO、PAL活性是呈正相关的。在水杨酸诱导下,各品种植株的PPO和PAL活性达到峰值的幅度变化最大,其次是在绿色粘帚霉发酵液下的处理,再次是草酸的处理,但是这些处理的酶活性达到峰值的变化幅度均大于空白对照实验。在大田苗期诱导实验中,PPO与PAL活性变化与室内诱导实验结果基本一致。
在大斑病盛发期,在大田对经过诱导的玉米苗进行了抗病性调查：福玉1号在水杨酸诱导后,诱抗效果提高到48.89%；而诱抗效果最差的是川单21在草酸中的诱导,为16.04%。通过显著性分析,同一品种在水杨酸中的诱抗效果最佳,其次是在绿色粘帚霉发酵液下的诱导,而且感病品种表现出的诱抗效果比抗病品种差。
The pathogen of Exserohilum turcicum was isolated from the infected leaf of maize in suburb of Ya-an, Sichuan.
46 strains of Gliocladium virens were isolated from different soil in Ya-an. According to the source and strain on the growth in PDA medium with different characteristics,14 strains were selected, used as the tested strains with the 16 strains kept in the lab together.
the experimental results of Exserohilum turcicum hyphal growth, sporulation and other biological characteristics showed that:the optimum hyphal growth conditions were full light,25℃, pH6, PDA medium; while the optimum conditions for sporulation were all dark,25℃, pH 6, water agar medium.
In the dual test of G. virens and the pathogen,the hyphal growth of Exserohilum turcicum was inhibited by G3 with inhibition rate of 59.12%. The non-volatile metabolites of G.virens G3 and G8 had the best inhibition rates,64.20% and 63.64%;respectively, G37 had the lowest inhibition rate-42.50%. The result of the volatile metabolites of G.virens showed that G8 had the inhibition rate which was 45.00%, while G37 had the lowest inhibition rate which was 21.00%.
The result of the disease resistance evaluation indicated that 10ks3-4 had the highest disease index,while 10117002 had the lowest. The disease index of 12% General Materials was more than 30.00, and 10% less than 10.00. Of all these materials,10117's Disease rate and disease index were both low, but 10ks'were opposite.
Useing SA、Oxalic acid and G.virens. fermentation liquor to induce corn northern leaf blight,the enzymatic activity of indoor experiment showed that PPO and PAL enzymatic activity of resistant species was higher than susceptible species.The activity of PPO and PAL were positive correlation with variety resistance. Under the induction of SA、the activity of PPO and PAL of all varieties plants had the greatest changes. Under the induction of G.virens. fermentation liquor was worse than that,but better than Oxalic acid. In short, the changes of the activity by these treatments were greater than confrontation experiment. In the experiment of farmland, the changes of enzymatic activities of PPO and PAL were consistent with the result of indoor'。
The survey of disease resistance of farmland experiment demonstrated that the best effect of induced resistance was FUYU 1-48.89% by the treatments of SA.The worst was CHUANDAN 21-16.04% by the treatments of Oxalic acid. It was obviously that SA had the optimal induced effect under the same sspecies.And the effect of induced resistance expressed by susceptible species was worse than resistant species.
分别于雅安四川农业大学教学农场和老板山上取土样,从土壤中分离获得绿色粘帚霉(Gliocladium virens)菌株共46个,根据菌株的来源和在PDA培养基上生长的不同特性,选取其中的14个菌株,与实验室保存的16个菌株一起作为本实验的供试菌株。
对玉米大斑病病原菌进行菌丝生长,产孢等生物学特性进行了研。结果表明：大斑病菌菌丝生长的最适条件是全光照、25℃、pH为6、PDA培养基；而菌株产孢最适条件是全黑暗、25℃、pH为6、清水琼脂培养基。
将供试的绿色粘帚霉菌株与大斑病菌做对峙培养：所有绿色粘帚霉菌株对大斑病菌的菌丝生长均有一定的抑制作用,抑制效果最好的为菌株G3,其抑制率达到59.12%。利用绿色粘帚霉难挥发性代谢产物,通过含毒介质法对大斑病菌抑制作用的测定实验中,粘帚霉G3和G8菌株对病原菌菌丝的抑制率最高,分别为64.20%和63.64%；最低的是G37,仅42.50%。在绿色粘帚霉易挥发性代谢产物对大斑病菌抑制作用的实验中,结果表明：抑制率最高的为G8,达到45.00%；最低的为G37,仅21.00%。
对雅安多营农场我校玉米所玉米自交系材料进行大斑病抗病性调查。所调查的60份自交系材料中,病情指数最高的为自交系10ks3-4,达到36.00；最低的为10117002,仅4.75。病情指数达到30.00以上的自交系占调查材料的12.00%；病情指数小于10.00的占调查材料的21.67%。在这些材料中,以10117这类自交系材料发病率及病情指数都较低,而以10ks类的材料发病率及病情指数都偏高。
利用水杨酸、草酸和绿色粘帚霉发酵液对玉米材料进行大斑病抗性诱导实验,通过室内玉米苗期诱导后,酶活性测定实验表明：在受到不同诱导剂处理下,抗病品种的PPO与PAL活性均高于感病品种,材料的抗病性与PPO、PAL活性是呈正相关的。在水杨酸诱导下,各品种植株的PPO和PAL活性达到峰值的幅度变化最大,其次是在绿色粘帚霉发酵液下的处理,再次是草酸的处理,但是这些处理的酶活性达到峰值的变化幅度均大于空白对照实验。在大田苗期诱导实验中,PPO与PAL活性变化与室内诱导实验结果基本一致。
在大斑病盛发期,在大田对经过诱导的玉米苗进行了抗病性调查：福玉1号在水杨酸诱导后,诱抗效果提高到48.89%；而诱抗效果最差的是川单21在草酸中的诱导,为16.04%。通过显著性分析,同一品种在水杨酸中的诱抗效果最佳,其次是在绿色粘帚霉发酵液下的诱导,而且感病品种表现出的诱抗效果比抗病品种差。
The pathogen of Exserohilum turcicum was isolated from the infected leaf of maize in suburb of Ya-an, Sichuan.
46 strains of Gliocladium virens were isolated from different soil in Ya-an. According to the source and strain on the growth in PDA medium with different characteristics,14 strains were selected, used as the tested strains with the 16 strains kept in the lab together.
the experimental results of Exserohilum turcicum hyphal growth, sporulation and other biological characteristics showed that:the optimum hyphal growth conditions were full light,25℃, pH6, PDA medium; while the optimum conditions for sporulation were all dark,25℃, pH 6, water agar medium.
In the dual test of G. virens and the pathogen,the hyphal growth of Exserohilum turcicum was inhibited by G3 with inhibition rate of 59.12%. The non-volatile metabolites of G.virens G3 and G8 had the best inhibition rates,64.20% and 63.64%;respectively, G37 had the lowest inhibition rate-42.50%. The result of the volatile metabolites of G.virens showed that G8 had the inhibition rate which was 45.00%, while G37 had the lowest inhibition rate which was 21.00%.
The result of the disease resistance evaluation indicated that 10ks3-4 had the highest disease index,while 10117002 had the lowest. The disease index of 12% General Materials was more than 30.00, and 10% less than 10.00. Of all these materials,10117's Disease rate and disease index were both low, but 10ks'were opposite.
Useing SA、Oxalic acid and G.virens. fermentation liquor to induce corn northern leaf blight,the enzymatic activity of indoor experiment showed that PPO and PAL enzymatic activity of resistant species was higher than susceptible species.The activity of PPO and PAL were positive correlation with variety resistance. Under the induction of SA、the activity of PPO and PAL of all varieties plants had the greatest changes. Under the induction of G.virens. fermentation liquor was worse than that,but better than Oxalic acid. In short, the changes of the activity by these treatments were greater than confrontation experiment. In the experiment of farmland, the changes of enzymatic activities of PPO and PAL were consistent with the result of indoor'。
The survey of disease resistance of farmland experiment demonstrated that the best effect of induced resistance was FUYU 1-48.89% by the treatments of SA.The worst was CHUANDAN 21-16.04% by the treatments of Oxalic acid. It was obviously that SA had the optimal induced effect under the same sspecies.And the effect of induced resistance expressed by susceptible species was worse than resistant species.
引文
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