利用秸秆和肥料增殖木霉及对玉米茎基腐病的生物防治
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摘要
以玉米秸秆为基质,筛选对木霉增殖有利的营养物质,对生防制剂的田间应用提供依据。利用Plack-ett-Burman试验设计筛选影响木霉产孢的主要营养物,采用最陡爬坡试验测定3种营养物的最适浓度范围;通过Box-Benhnken试验设计,对3种营养物浓度进行优化;盆栽试验检测木霉在土壤中的存活情况防治玉米茎基腐病。结果表明,筛选对木霉产孢影响较大的3个营养物质(氯化铵、硫酸钾、七水硫酸亚铁),最佳配方为氯化铵0.97 g/L,硫酸钾0.92 g/L,七水硫酸亚铁0.97 g/L,在此配方条件下孢子产量为2.61×10~9个/g。盆栽试验表明,利用木霉结合秸秆还田和肥料施用,可增强其存活能力和降低玉米茎基腐病的发生率和病情指数。
Experiments were designed to screen the main nutrient factors influencing the sporulation of Trichoderma pseudokoningii strains St-zn-9 by Plackett-Burman method using Minitab software Further their optimal con-centrations of these nutrient ingredients were determined by the steepest ascent design and optimized through theBox-Benhnken experiment. Pot experiments were conducted to test the viability of Trichoderma in soil for biologicallycontrolling corn stalk rot. The results showed that the main influencing nutrient factors related with T. pseudokoningii sporulation were NH_4 Cl, K_2SO_4, and Fe SO_4·7 H_2 O, and the optimized formula was of NH_4Cl at 0.97 g/L, K_2 SO_4 at 0.92 g/L,9 and Fe SO_4·7 H_2O at 0.97 g/L. In this optimized formula, T. pseudokoningii strains St-zn-9 might reach 2.61×10 spores/g. Pot experiments showed that straw returning combined with chemical substances supplied couldgreatly enhance Trichoderma viability and efficiently reduce the incidence and disease index of corn stalk rot.
Experiments were designed to screen the main nutrient factors influencing the sporulation of Trichoderma pseudokoningii strains St-zn-9 by Plackett-Burman method using Minitab software Further their optimal con-centrations of these nutrient ingredients were determined by the steepest ascent design and optimized through theBox-Benhnken experiment. Pot experiments were conducted to test the viability of Trichoderma in soil for biologicallycontrolling corn stalk rot. The results showed that the main influencing nutrient factors related with T. pseudokoningii sporulation were NH_4 Cl, K_2SO_4, and Fe SO_4·7 H_2 O, and the optimized formula was of NH_4Cl at 0.97 g/L, K_2 SO_4 at 0.92 g/L,9 and Fe SO_4·7 H_2O at 0.97 g/L. In this optimized formula, T. pseudokoningii strains St-zn-9 might reach 2.61×10 spores/g. Pot experiments showed that straw returning combined with chemical substances supplied couldgreatly enhance Trichoderma viability and efficiently reduce the incidence and disease index of corn stalk rot.
引文
[1]李世忠,冯海东,解倩,等.宁夏引黄灌区土壤物理性状对玉米秸秆还田的响应[J].农业科学研究,2017,38(2):19-22.Li S Z,Feng H D,Xie Q,et al.Effect of maize straw returning on soil physical properties in Ningxia Yellow River irrigation area[J].Journal of Agricultural Sciences,2017,38(2):19-22.(in Chinese)
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[3]闫贵龙,曹春梅,鲁琳,等.玉米秸秆不同部位主要化学成分和活体外消化率比较[J].中国农业大学学报,2006,11(3):70-74.Yan G L,Cao C M,Lu L,et al.Comparison of main chemical composition and in vitro digestibility in various sections of corn stalks[J].Journal of China Agricultural University,2006,11(3):70-74.(in Chinese)
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[5]Howell C.Cotton seedling preemergence damping-off incited by Rhizopus oryzae and Pythium spp.and its biological control with Trichoderma spp[J].Phytopathology,2002(92):177-180.
[6]郎剑锋,孔凡彬,石明旺,等.哈茨木霉对7种植物病原菌的生防机制研究[J].河南科技学院学报(自然科学版),2013,41(5):32-35.Lang J F,Kong F B,Shi M W,et al.Studies on biocontrol mechanisms of Trichoderma harzianum against seven plant pathogenic fungi[J].Journal of Henan Institute of Science and Technology,2013,41(5):32-35.(in Chinese)
[7]Sawant I S,Sawant S D.A simple method for achieving high CFU of Trichoderma harzianum on organic wastes for field applications[J].Indian Phytopathology,1996,49(2):185-187.
[8]王永东,蒋立科,岳永德,等.生防菌株哈茨木霉H-13固体发酵条件的研究[J].浙江大学学报(农业与生命科学版),2006,32(6):645-650.Wang Y D,Jiang L K,Yue Y D,et al.Studies on the solid fermentation conditions for the biocontrol agents of Trichoderma harzianum strain H-13[J].Journal of Zhejiang University(Agriculture&Life Sciences),2006,32(6):645-650.(in Chinese)
[9]曾庆才,肖荣凤,刘波,等.生防菌哈茨木霉FJAT-9040固体发酵条件的响应面优化[J].福建农业学报,2015,30(2):192-197.Zeng Q C,Xiao R F,Liu B,et al.Optimization of solid-state fermentation condition for the biocontrol strain of Trichoderma harzianum FJAT-9040 by response surface methodology[J].Fujian Journal of Agricultural Sciences,2015,30(2):192-197.(in Chinese)
[10]刘时轮,李勇,傅俊范,等.绿色木霉菌株Tv04-2固体发酵条件研究[J].华北农学报,2008,23(Z2):244-247.Liu S L,Li Y,Fu J F,et al.Study on fermentation conditions of antagonistic Trichoderma viride Tv04-2 strain[J].Acta Agriculturae Boreali-Sinica,2008,23(Z2):244-247.(in Chinese)
[11]田连生,李书生,史延茂,等.利用玉米秸秆制备生物杀菌剂的研究[J].中国生态农业学报,2005,13(2):59-61.Tian L S,Li S S,Shi Y M,et al.Study on production of bio-fungicide by using maize stock[J].Chinese Journal of Eco-Agriculture,2005,13(2):59-61.(in Chinese)
[12]李慧君,李波,吴琳.绿色木霉F6降解玉米秸秆的固体发酵条件优化[J].安徽农业科学,2012,40(24):12204-12207.Li H J,Li B,Wu L.Optimization of the solid state fermentation conditions of Trichoderma viride F6 for corn straw degradation[J].Journal of Anhui Agri.Sci.,2012,40(24):12204-12207.(in Chinese)
[13]梁昌聪,刘磊,张建华,等.响应面法优化绿色木霉H06产孢发酵培养基[J].热带作物学报,2014,35(2):333-338.Liang C C,Liu L,Zhang J H,et al.Optimization of medium components for sporulation quantity by Trichoderma viride strain H06 using response surface methodology[J].Chinese Journal of Tropical Crops,2014,35(2):333-338.(in Chinese)
[14]郎剑锋,孔凡彬,田雪亮,等.不同营养水平下化肥对哈茨木霉生长的影响[J].核农学报,2015,29(10):2020-2025.Lang J F,Kong F B,Tian X L,et al.Effects of five fertilizers under different nutrition levels on mycelial growth and spore production of Trichoderma harzianum[J].Journal of Nuclear Agricultural Sciences,2015,29(10):2020-2025.(in Chinese)
[15]张亚丽,张娟,沈其荣,等.秸秆生物有机肥的施用对土壤供氮能力的影响[J].应用生态学报,2002,13(12):1575-1578.Zhang Y L,Zhang J,Shen Q R,et al.Effect of combined application of bioorganic manure and inorganic nitrogen fertilizer on soil nitrogen supplying characteristics[J].Chinese Journal of Applied Ecology,2002,13(12):1575-1578.(in Chinese)
[16]冯程龙,王晓婷,康文晶,等.利用小麦秸秆生产木霉分生孢子及其生物有机肥对黄瓜的促生效果[J].植物营养与肥料学报,2017,23(5):1286-1295.Feng C L,Wang X T,Kang W J,et al.Trichoderma conidia production using wheat straw and growth promoting effect of its bio-organic fertilizer on cucumber[J].Journal of Plant Nutrition and Fertilizer,2017,23(5):1286-1295.(in Chinese)
[17]方中达.植病研究方法[M].北京:中国农业出版社,1998.
[18]敖新宇,程立君,陈玉惠.生防木霉SS003菌株(Trichoderma atroviride)的固体发酵工艺研究[J].江西农业大学学报,2012,34(6):1256-1261.Ao X Y,Cheng L J,Chen Y H.The solid fermentation technology for bio-control strain SS003 of Trichoderma atroviride[J].Acta Agriculturae Universitatis Jiangxiensis,2012,34(6):1256-1261.(in Chinese)
[19]湛方栋,陈建军,秦丽,等.镉铅污染的玉米秸秆还田对蚕豆生长、养分和镉铅含量的影响[J].农业环境科学学报,2016,35(4):661-668.Zhan F D,Chen J J,Qin L,et al.Effects of applying Cd/Pb contaminated maize stalks on growth and nutrient and Cd and Pb content of faba bean[J].Journal of Agro-Environment Science,2016,35(4):661-668.(in Chinese)
[20]李洪连,徐敬友.农业植物病理学实验实习指导[M].北京:中国农业出版社,2001.
[21]吴晓青,赵忠娟,李哲,等.施加木霉可湿性粉剂对冬小麦田间生长的影响[J].山东科学,2015,28(6):35-42.Wu X Q,Zhao Z J,Li Z,et al.Impact of Trichoderma wettable powder application on winter wheat field growth[J].Shandong Science,2015,28(6):35-42.(in Chinese)
[22]Sarangi S K,Lama T D.Straw composting using earthworm(Eudrilus eugeniae)and fungal inoculant(Trichoderma viridae)and its utilization in rice(Oryza sativa)-groundnut(Arachis hypogaea)cropping system[J].Indian Journal of Agricultural Sciences,2013,83(4):420-425.
[23]任轶,李瑞霞,艾昊,等.减施肥条件下木霉SQR-T037微生物肥对黄瓜产量、品质及养分利用效率的影响[J].江苏农业科学,2014,42(2):143-146.Ren Y,Li R X,Ai H,et al.Effects of Trichoderma microbial fertilizer SQR-T037 on yield,quality and nutrient use efficiency of cucumber under reduced usage of chemical fertilizer[J].Jiangsu Agricultural Sciences,2014,42(2):143-146.(in Chinese)
[24]Danielson R M,Davey C B.Carbon and nitrogen nutrition of Trichoderma[J].Soil Biology&Biochemistry,1973,5(5):505-515.
[25]Mandels M,Aandreotti R E.Problems and challenges in the cellulose to cellulase fermentation[J].Proc.Biochem,1978(13):6-13.
[26]李晓,李亚鑫,张娟霞,等.玉米秸秆还田对冬小麦产量和不同生育期土壤硝态氮累积量的影响[J].干旱地区农业研究,2016,34(6):156-162.Li X,Li Y X,Zhang J X,et al.Effects of maize straw return on yield and soil nitrate accumulation in different growth stages of winter wheat[J].Agricultural Research in the Arid Areas,2016,34(6):156-162.(in Chinese)
[27]Jackson A M,Whipps J M,Lynch J M.Nutritional studies of four fungi with disease biocontrol potential[J].Enzyme Microb.Technol,1991,13(7):456-461.
[2]张素瑜,王和洲,杨明达,等.不同水分条件下玉米秸秆还田对小麦群体发育和干物质积累及产量的影响[J].麦类作物学报,2016,36(9):1183-1190.Zhang S Y,Wang H Z,Yang M D,et al.Influence of returning corn stalks to field under different soil water contents on population development,dry matter accumulation and grain yield in wheat[J].Journal of Triticeae Crops,2016,36(9):1183-1190.(in Chinese)
[3]闫贵龙,曹春梅,鲁琳,等.玉米秸秆不同部位主要化学成分和活体外消化率比较[J].中国农业大学学报,2006,11(3):70-74.Yan G L,Cao C M,Lu L,et al.Comparison of main chemical composition and in vitro digestibility in various sections of corn stalks[J].Journal of China Agricultural University,2006,11(3):70-74.(in Chinese)
[4]乔玉强,曹承富,赵竹,等.秸秆还田与施氮量对小麦产量和品质及赤霉病发生的影响[J].麦类作物学报,2013,33(4):727-731.Qiao Y Q,Cao C F,Zhao Z,et al.Effects of straw-returning and N-fertilizer application on yield,quality and occurrence of Fusarium head blight of wheat[J].Journal of Triticeae Crops,2013,33(4):727-731.(in Chinese)
[5]Howell C.Cotton seedling preemergence damping-off incited by Rhizopus oryzae and Pythium spp.and its biological control with Trichoderma spp[J].Phytopathology,2002(92):177-180.
[6]郎剑锋,孔凡彬,石明旺,等.哈茨木霉对7种植物病原菌的生防机制研究[J].河南科技学院学报(自然科学版),2013,41(5):32-35.Lang J F,Kong F B,Shi M W,et al.Studies on biocontrol mechanisms of Trichoderma harzianum against seven plant pathogenic fungi[J].Journal of Henan Institute of Science and Technology,2013,41(5):32-35.(in Chinese)
[7]Sawant I S,Sawant S D.A simple method for achieving high CFU of Trichoderma harzianum on organic wastes for field applications[J].Indian Phytopathology,1996,49(2):185-187.
[8]王永东,蒋立科,岳永德,等.生防菌株哈茨木霉H-13固体发酵条件的研究[J].浙江大学学报(农业与生命科学版),2006,32(6):645-650.Wang Y D,Jiang L K,Yue Y D,et al.Studies on the solid fermentation conditions for the biocontrol agents of Trichoderma harzianum strain H-13[J].Journal of Zhejiang University(Agriculture&Life Sciences),2006,32(6):645-650.(in Chinese)
[9]曾庆才,肖荣凤,刘波,等.生防菌哈茨木霉FJAT-9040固体发酵条件的响应面优化[J].福建农业学报,2015,30(2):192-197.Zeng Q C,Xiao R F,Liu B,et al.Optimization of solid-state fermentation condition for the biocontrol strain of Trichoderma harzianum FJAT-9040 by response surface methodology[J].Fujian Journal of Agricultural Sciences,2015,30(2):192-197.(in Chinese)
[10]刘时轮,李勇,傅俊范,等.绿色木霉菌株Tv04-2固体发酵条件研究[J].华北农学报,2008,23(Z2):244-247.Liu S L,Li Y,Fu J F,et al.Study on fermentation conditions of antagonistic Trichoderma viride Tv04-2 strain[J].Acta Agriculturae Boreali-Sinica,2008,23(Z2):244-247.(in Chinese)
[11]田连生,李书生,史延茂,等.利用玉米秸秆制备生物杀菌剂的研究[J].中国生态农业学报,2005,13(2):59-61.Tian L S,Li S S,Shi Y M,et al.Study on production of bio-fungicide by using maize stock[J].Chinese Journal of Eco-Agriculture,2005,13(2):59-61.(in Chinese)
[12]李慧君,李波,吴琳.绿色木霉F6降解玉米秸秆的固体发酵条件优化[J].安徽农业科学,2012,40(24):12204-12207.Li H J,Li B,Wu L.Optimization of the solid state fermentation conditions of Trichoderma viride F6 for corn straw degradation[J].Journal of Anhui Agri.Sci.,2012,40(24):12204-12207.(in Chinese)
[13]梁昌聪,刘磊,张建华,等.响应面法优化绿色木霉H06产孢发酵培养基[J].热带作物学报,2014,35(2):333-338.Liang C C,Liu L,Zhang J H,et al.Optimization of medium components for sporulation quantity by Trichoderma viride strain H06 using response surface methodology[J].Chinese Journal of Tropical Crops,2014,35(2):333-338.(in Chinese)
[14]郎剑锋,孔凡彬,田雪亮,等.不同营养水平下化肥对哈茨木霉生长的影响[J].核农学报,2015,29(10):2020-2025.Lang J F,Kong F B,Tian X L,et al.Effects of five fertilizers under different nutrition levels on mycelial growth and spore production of Trichoderma harzianum[J].Journal of Nuclear Agricultural Sciences,2015,29(10):2020-2025.(in Chinese)
[15]张亚丽,张娟,沈其荣,等.秸秆生物有机肥的施用对土壤供氮能力的影响[J].应用生态学报,2002,13(12):1575-1578.Zhang Y L,Zhang J,Shen Q R,et al.Effect of combined application of bioorganic manure and inorganic nitrogen fertilizer on soil nitrogen supplying characteristics[J].Chinese Journal of Applied Ecology,2002,13(12):1575-1578.(in Chinese)
[16]冯程龙,王晓婷,康文晶,等.利用小麦秸秆生产木霉分生孢子及其生物有机肥对黄瓜的促生效果[J].植物营养与肥料学报,2017,23(5):1286-1295.Feng C L,Wang X T,Kang W J,et al.Trichoderma conidia production using wheat straw and growth promoting effect of its bio-organic fertilizer on cucumber[J].Journal of Plant Nutrition and Fertilizer,2017,23(5):1286-1295.(in Chinese)
[17]方中达.植病研究方法[M].北京:中国农业出版社,1998.
[18]敖新宇,程立君,陈玉惠.生防木霉SS003菌株(Trichoderma atroviride)的固体发酵工艺研究[J].江西农业大学学报,2012,34(6):1256-1261.Ao X Y,Cheng L J,Chen Y H.The solid fermentation technology for bio-control strain SS003 of Trichoderma atroviride[J].Acta Agriculturae Universitatis Jiangxiensis,2012,34(6):1256-1261.(in Chinese)
[19]湛方栋,陈建军,秦丽,等.镉铅污染的玉米秸秆还田对蚕豆生长、养分和镉铅含量的影响[J].农业环境科学学报,2016,35(4):661-668.Zhan F D,Chen J J,Qin L,et al.Effects of applying Cd/Pb contaminated maize stalks on growth and nutrient and Cd and Pb content of faba bean[J].Journal of Agro-Environment Science,2016,35(4):661-668.(in Chinese)
[20]李洪连,徐敬友.农业植物病理学实验实习指导[M].北京:中国农业出版社,2001.
[21]吴晓青,赵忠娟,李哲,等.施加木霉可湿性粉剂对冬小麦田间生长的影响[J].山东科学,2015,28(6):35-42.Wu X Q,Zhao Z J,Li Z,et al.Impact of Trichoderma wettable powder application on winter wheat field growth[J].Shandong Science,2015,28(6):35-42.(in Chinese)
[22]Sarangi S K,Lama T D.Straw composting using earthworm(Eudrilus eugeniae)and fungal inoculant(Trichoderma viridae)and its utilization in rice(Oryza sativa)-groundnut(Arachis hypogaea)cropping system[J].Indian Journal of Agricultural Sciences,2013,83(4):420-425.
[23]任轶,李瑞霞,艾昊,等.减施肥条件下木霉SQR-T037微生物肥对黄瓜产量、品质及养分利用效率的影响[J].江苏农业科学,2014,42(2):143-146.Ren Y,Li R X,Ai H,et al.Effects of Trichoderma microbial fertilizer SQR-T037 on yield,quality and nutrient use efficiency of cucumber under reduced usage of chemical fertilizer[J].Jiangsu Agricultural Sciences,2014,42(2):143-146.(in Chinese)
[24]Danielson R M,Davey C B.Carbon and nitrogen nutrition of Trichoderma[J].Soil Biology&Biochemistry,1973,5(5):505-515.
[25]Mandels M,Aandreotti R E.Problems and challenges in the cellulose to cellulase fermentation[J].Proc.Biochem,1978(13):6-13.
[26]李晓,李亚鑫,张娟霞,等.玉米秸秆还田对冬小麦产量和不同生育期土壤硝态氮累积量的影响[J].干旱地区农业研究,2016,34(6):156-162.Li X,Li Y X,Zhang J X,et al.Effects of maize straw return on yield and soil nitrate accumulation in different growth stages of winter wheat[J].Agricultural Research in the Arid Areas,2016,34(6):156-162.(in Chinese)
[27]Jackson A M,Whipps J M,Lynch J M.Nutritional studies of four fungi with disease biocontrol potential[J].Enzyme Microb.Technol,1991,13(7):456-461.
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