复合微生物菌剂对苹果再植病害的生防效果测定及拮抗菌株鉴定
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摘要
为探究复合微生物菌剂对苹果再植病害的生防效果,以苹果砧木实生海棠幼苗为试材,通过盆栽试验测试了木美土里复合微生物菌剂对苹果再植病害的防治效果及对再植海棠的促生作用,然后对该菌剂的可分离微生物进行了分离纯化,并以尖孢镰刀菌Fusarium oxysporum菌株HS2为靶标菌,测试分离获得的10株菌株的拮抗效果,并对拮抗效果显著的1株真菌菌株和2株细菌菌株进行种类鉴定。结果表明,木美土里复合微生物菌剂对苹果再植病害的防治效果达87.78%;经该菌剂处理后,再植海棠苗的株高、茎粗、鲜重、干重及叶面积分别是对照处理的1.51、2.00、5.74、5.21和2.83倍,表明该菌剂有显著的促生作用。木美土里复合微生物菌剂对菌株HS2的抑制率达到68.59%,并从中分离到7株真菌菌株,3株细菌菌株。其中,拮抗效果高于80.00%的真菌菌株有1株,细菌菌株有2株。经鉴定,真菌菌株MMTL-1为哈茨木霉Trichoderma harzianum;细菌菌株BM-01和BM-03均为解淀粉芽胞杆菌Bacillus amyloliquefaciens。
To study the bio-control effect of compound microbial against apple replant disease, pot experiments were applied to evaluate the control effect of Kimidori microbial manure(KMM) against the apple replant diseases(ARD) and whether promoting plant growth of Malus seedlings. Ten microorganism strains isolated from KMM were tested in vitro for their antagonistic activity against Fusarium oxysporum, one of them was the inducing factor of ARD. The species with better inhibition rate were identified through morphology and molecular biology methods. The results showed that the control effect of KMM against ARD could reach 87.78%. Seedling growth was significantly promoted by KMM treatment. The plant height, stem diameter, fresh weight, dry weight and leaf area of KMM treatment increased 1.51, 2.00, 5.74, 5.21 and 2.83 times compare with that of the replant control. The inhibition rate of KMM against F. oxysporum HS2 was 68.59%. Seven fungal isolates and three bacterial strains were isolated from KMM. Among these isolates, one fungal isolate MMTL-1 with the inhibition rate over 80.00% was identified as Trichoderma harzianum. Both the two bacterial isolates BM-01 and BM-03 with the inhibition rate over 80.00% were identified as Bacillus amyloliquefaciens.
To study the bio-control effect of compound microbial against apple replant disease, pot experiments were applied to evaluate the control effect of Kimidori microbial manure(KMM) against the apple replant diseases(ARD) and whether promoting plant growth of Malus seedlings. Ten microorganism strains isolated from KMM were tested in vitro for their antagonistic activity against Fusarium oxysporum, one of them was the inducing factor of ARD. The species with better inhibition rate were identified through morphology and molecular biology methods. The results showed that the control effect of KMM against ARD could reach 87.78%. Seedling growth was significantly promoted by KMM treatment. The plant height, stem diameter, fresh weight, dry weight and leaf area of KMM treatment increased 1.51, 2.00, 5.74, 5.21 and 2.83 times compare with that of the replant control. The inhibition rate of KMM against F. oxysporum HS2 was 68.59%. Seven fungal isolates and three bacterial strains were isolated from KMM. Among these isolates, one fungal isolate MMTL-1 with the inhibition rate over 80.00% was identified as Trichoderma harzianum. Both the two bacterial isolates BM-01 and BM-03 with the inhibition rate over 80.00% were identified as Bacillus amyloliquefaciens.
引文
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Catska V,Hudska G.1993.Use of Agrobacterium radiobacter for biological control of apple replant disease.Acta Horticulturae,324:67-72
Cheng LJ,Xue QH.2012.Laboratory manual of microbiology(2nd edition).Beijing:Science Press(in Chinese)[程丽娟,薛泉宏.2012.微生物学实验技术(第2版).北京:科学出版社]
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Forge T,Neilsen G,Neilsen D.2016.Organically acceptable practices to improve replant success of temperate tree-fruit crops.Scientia Horticulturae,200:205-214
Hu QY,Liu LW,Liu X,Dang JM,Fan JY,Wang ST,Cao KQ.2015.Evaluation of the effect of Kimidori bioorganic fertilizer on the control of apple canker disease.China Fruits,(4):52-55(in Chinese)[胡清玉,刘力伟,刘欣,党建美,范军印,王树桐,曹克强.2015.木美土里生物菌肥对苹果树腐烂病的防治作用评价.中国果树,(4):52-55]
Jiang YM,Ge SF.2016.A coup for apple orchard to“rejuvenate”by improving the soil and reducing the fertilizer.Farmers Daily,10-20(7)(in Chinese)[姜远茂,葛顺峰.2016.苹果园“返老还童”改土减肥有妙招.农民日报,10-20(7)]
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Krumholz MR,Klein RI,McKee CF,Offner SSR,Cunningham AJ.2009.The formation of massive star systems by accretion.Science,323(5915):754-757
Li BZ,Wang YJ,Wang XL.2013.The effect of Kimidori fertilizer on the control of apple scar skin and canker disease.Yantai Fruits,(3):10-12(in Chinese)[李丙智,王洋娟,王晓琳.2013.木美土里防控苹果花脸及腐烂病的效果试验.烟台果树,(3):10-12]
Liu ET,Wang GS,Li YY,Shen X,Chen XS,Song FH,Wu SJ,Chen Q,Mao ZQ.2014.Replanting affects the tree growth and fruit quality of Gala apple.Journal of Integrative Agriculture,13(8):1699-1706
Liu J.2009.The international competition ability research of China apple industry.Master Thesis.Yangling:Northwest A&F University(in Chinese)[刘婧.2009.中国苹果产业国际竞争力研究.硕士学位论文.杨凌:西北农林科技大学]
Liu LW,Liu LY,Hu TL,Wang YN,Wang ST,Cao KQ.2016.Biocontrol effect of Pseudomonas fluorescens strain SS101 against apple replant disease.Journal of Plant Protection,43(5):812-818(in Chinese)[刘力伟,刘丽媛,胡同乐,王亚南,王树桐,曹克强.2016.荧光假单胞菌SS101对苹果再植病害的生防效果.植物保护学报,43(5):812-818]
Liu X,Shao XH,Mao XY,Shan JM.2010.Effects of EM bio-organic fertilizer on growth and soil characteristics of sweet corn.Jiangsu agricultural sciences,(1):301-302(in Chinese)[刘旭,邵孝侯,毛欣宇,单建明.2010.EM生物有机肥对甜玉米生长发育及土壤特性的影响.江苏农业科学,(1):301-302]
Manici LM,Kelderer M,Caputo F,Mazzola M.2015.Auxin-mediated relationships between apple plants and root inhabiting fungi:impact on root pathogens and potentialities of growth-promoting populations.Plant Pathology,64(4):843-851
Manici LM,Kelderer M,Franke-Whittle IH,Ruhmer T,Baab G,Nicoletti F,Caputo F,Topp A,Insam H,Naef A.2013.Relationship between root-endophytic microbial communities and replant disease in specialized apple growing areas in Europe.Applied Soil Ecology,72:207-214
Mazzola M.1998.Elucidation of the microbial complex having a causal role in the development of apple replant disease in Washington.Phytopathology,88(9):930-938
Mei XL,Zhao QY,Tan SY,Xu YC,Shen B,Shen QR.2010.Screening,identification,and biocontrol effect of antagonistic bacteria against Phytophthora capsici.Chinese Journal of Applied Ecology,21(10):2652-2658(in Chinese)[梅新兰,赵青云,谭石勇,徐阳春,沈标,沈其荣.2010.辣椒疫病拮抗菌株筛选、鉴定及其防效.应用生态学报,21(10):2652-2658]
Sun Z,Yang LM.2015.Screening and identification of antagonistic bacteria against Panax ginseng pathogeny fungus.Journal of Plant Protection,42(1):79-86(in Chinese)[孙卓,杨利民,2015.人参病原菌拮抗细菌的分离筛选与鉴定.植物保护学报,42(1):79-86]
Zhang Y.2007.Control effect of bio-product Kangdi 3 on replant diseases of strawberry.Master Thesis.Baoding:Agricultural University of Hebei(in Chinese)[张瑜.2007.生物制剂康地3号对草莓再植病害的防治作用研究.硕士学位论文.保定:河北农业大学]
Zhao YS.2014.A brief analysis of apple industry development in China.Scientific Planting,(9):5-7(in Chinese)[赵玉山.2014.我国苹果产业发展浅析.科学种养,(9):5-7]
Zhou DB,Jing T,Zhang XY,Qi DF,Chen YF,Wang F.2016.Screening and antibacterial activity of antagonistic bacteria against banana Fusarium wilt disease.Journal of Plant Protection,43(6):913-921(in Chinese)[周登博,井涛,张锡炎,起登凤,陈宇丰,王飞.2016.香蕉枯萎病拮抗菌筛选及其抑菌活性.植物保护学报,43(6):913-921]
Zhuang JH,Gao ZG,Chen J,Huang YQ,Xu S.2005.Determination of virulence of 5 fungicides against Trichoderma and Fusarium oxysporum.Plant Protection,31(3):84-86(in Chinese)[庄敬华,高增贵,陈捷,黄艳清,徐韶.2005.5种杀菌剂对木霉菌及尖孢镰刀菌的毒力测定.植物保护,31(3):84-86]
Zou QJ,Wang ST,Liang KJ,Wang YN,Hu TL,Han ZQ,Cao KQ.2014.Suspected pathogenic Fusarium spp.isolated from apple orchard soils in Hebei Province.Mycosystema,33(5):976-983(in Chinese)[邹庆甲,王树桐,梁魁景,王亚南,胡同乐,韩之琪,曹克强.2014.河北省苹果园根际上壤中疑似致病镰抱菌种类.菌物学报,33(5):976-983]
Braun PG.1995.Effects of Cylindrocarpon and Pythium species on apple seedlings and potential role in apple replant disease.Canadian Journal of Plant Pathology,17(4):336-341
Catska V,Hudska G.1993.Use of Agrobacterium radiobacter for biological control of apple replant disease.Acta Horticulturae,324:67-72
Cheng LJ,Xue QH.2012.Laboratory manual of microbiology(2nd edition).Beijing:Science Press(in Chinese)[程丽娟,薛泉宏.2012.微生物学实验技术(第2版).北京:科学出版社]
Dong XZ,Cai MY.2001.Handbook of systematic identification of common bacteria.Beijing:Science Press,pp.349-388(in Chinese)[东秀珠,蔡妙英.2001.常见细菌系统鉴定手册.北京:科学出版社,pp.349-388]
Forge T,Neilsen G,Neilsen D.2016.Organically acceptable practices to improve replant success of temperate tree-fruit crops.Scientia Horticulturae,200:205-214
Hu QY,Liu LW,Liu X,Dang JM,Fan JY,Wang ST,Cao KQ.2015.Evaluation of the effect of Kimidori bioorganic fertilizer on the control of apple canker disease.China Fruits,(4):52-55(in Chinese)[胡清玉,刘力伟,刘欣,党建美,范军印,王树桐,曹克强.2015.木美土里生物菌肥对苹果树腐烂病的防治作用评价.中国果树,(4):52-55]
Jiang YM,Ge SF.2016.A coup for apple orchard to“rejuvenate”by improving the soil and reducing the fertilizer.Farmers Daily,10-20(7)(in Chinese)[姜远茂,葛顺峰.2016.苹果园“返老还童”改土减肥有妙招.农民日报,10-20(7)]
Kandula DRW,Jones EE,Horner IJ,Stewart A.2010.The effect of Trichoderma bio-inoculants on specific apple replant disease(SARD)symptoms in apple rootstocks in New Zealand.Australasian Plant Pathology,39(4):312-318
Krumholz MR,Klein RI,McKee CF,Offner SSR,Cunningham AJ.2009.The formation of massive star systems by accretion.Science,323(5915):754-757
Li BZ,Wang YJ,Wang XL.2013.The effect of Kimidori fertilizer on the control of apple scar skin and canker disease.Yantai Fruits,(3):10-12(in Chinese)[李丙智,王洋娟,王晓琳.2013.木美土里防控苹果花脸及腐烂病的效果试验.烟台果树,(3):10-12]
Liu ET,Wang GS,Li YY,Shen X,Chen XS,Song FH,Wu SJ,Chen Q,Mao ZQ.2014.Replanting affects the tree growth and fruit quality of Gala apple.Journal of Integrative Agriculture,13(8):1699-1706
Liu J.2009.The international competition ability research of China apple industry.Master Thesis.Yangling:Northwest A&F University(in Chinese)[刘婧.2009.中国苹果产业国际竞争力研究.硕士学位论文.杨凌:西北农林科技大学]
Liu LW,Liu LY,Hu TL,Wang YN,Wang ST,Cao KQ.2016.Biocontrol effect of Pseudomonas fluorescens strain SS101 against apple replant disease.Journal of Plant Protection,43(5):812-818(in Chinese)[刘力伟,刘丽媛,胡同乐,王亚南,王树桐,曹克强.2016.荧光假单胞菌SS101对苹果再植病害的生防效果.植物保护学报,43(5):812-818]
Liu X,Shao XH,Mao XY,Shan JM.2010.Effects of EM bio-organic fertilizer on growth and soil characteristics of sweet corn.Jiangsu agricultural sciences,(1):301-302(in Chinese)[刘旭,邵孝侯,毛欣宇,单建明.2010.EM生物有机肥对甜玉米生长发育及土壤特性的影响.江苏农业科学,(1):301-302]
Manici LM,Kelderer M,Caputo F,Mazzola M.2015.Auxin-mediated relationships between apple plants and root inhabiting fungi:impact on root pathogens and potentialities of growth-promoting populations.Plant Pathology,64(4):843-851
Manici LM,Kelderer M,Franke-Whittle IH,Ruhmer T,Baab G,Nicoletti F,Caputo F,Topp A,Insam H,Naef A.2013.Relationship between root-endophytic microbial communities and replant disease in specialized apple growing areas in Europe.Applied Soil Ecology,72:207-214
Mazzola M.1998.Elucidation of the microbial complex having a causal role in the development of apple replant disease in Washington.Phytopathology,88(9):930-938
Mei XL,Zhao QY,Tan SY,Xu YC,Shen B,Shen QR.2010.Screening,identification,and biocontrol effect of antagonistic bacteria against Phytophthora capsici.Chinese Journal of Applied Ecology,21(10):2652-2658(in Chinese)[梅新兰,赵青云,谭石勇,徐阳春,沈标,沈其荣.2010.辣椒疫病拮抗菌株筛选、鉴定及其防效.应用生态学报,21(10):2652-2658]
Sun Z,Yang LM.2015.Screening and identification of antagonistic bacteria against Panax ginseng pathogeny fungus.Journal of Plant Protection,42(1):79-86(in Chinese)[孙卓,杨利民,2015.人参病原菌拮抗细菌的分离筛选与鉴定.植物保护学报,42(1):79-86]
Zhang Y.2007.Control effect of bio-product Kangdi 3 on replant diseases of strawberry.Master Thesis.Baoding:Agricultural University of Hebei(in Chinese)[张瑜.2007.生物制剂康地3号对草莓再植病害的防治作用研究.硕士学位论文.保定:河北农业大学]
Zhao YS.2014.A brief analysis of apple industry development in China.Scientific Planting,(9):5-7(in Chinese)[赵玉山.2014.我国苹果产业发展浅析.科学种养,(9):5-7]
Zhou DB,Jing T,Zhang XY,Qi DF,Chen YF,Wang F.2016.Screening and antibacterial activity of antagonistic bacteria against banana Fusarium wilt disease.Journal of Plant Protection,43(6):913-921(in Chinese)[周登博,井涛,张锡炎,起登凤,陈宇丰,王飞.2016.香蕉枯萎病拮抗菌筛选及其抑菌活性.植物保护学报,43(6):913-921]
Zhuang JH,Gao ZG,Chen J,Huang YQ,Xu S.2005.Determination of virulence of 5 fungicides against Trichoderma and Fusarium oxysporum.Plant Protection,31(3):84-86(in Chinese)[庄敬华,高增贵,陈捷,黄艳清,徐韶.2005.5种杀菌剂对木霉菌及尖孢镰刀菌的毒力测定.植物保护,31(3):84-86]
Zou QJ,Wang ST,Liang KJ,Wang YN,Hu TL,Han ZQ,Cao KQ.2014.Suspected pathogenic Fusarium spp.isolated from apple orchard soils in Hebei Province.Mycosystema,33(5):976-983(in Chinese)[邹庆甲,王树桐,梁魁景,王亚南,胡同乐,韩之琪,曹克强.2014.河北省苹果园根际上壤中疑似致病镰抱菌种类.菌物学报,33(5):976-983]