香蕉-甘蔗轮作模式防控香蕉枯萎病的持续效果与土壤微生态机理(Ⅰ)
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摘要
近年来,珠三角部分蕉农采用香蕉和甘蔗轮作防控香蕉枯萎病,并取得一定成效,我国古籍《广东新语》也有香蕉-甘蔗轮作的记载,但是关于该轮作模式的作用、效果及控病机理尚不明确。本文研究了连作香蕉地轮作不同年限甘蔗对香蕉枯萎病的控制作用,并从土壤微生态角度探讨其作用机理。试验设置4个处理:连作香蕉11年(CK)、香蕉连作地—甘蔗1年—香蕉1年(GZ1)、香蕉连作地—甘蔗2年—香蕉1年(GZ2)和香蕉连作地—甘蔗3年—香蕉1年(GZ3)。研究轮作甘蔗不同年限对后茬香蕉枯萎病发病率的影响,同时通过测定土壤可培养微生物数量,并利用高通量测序技术分析土壤微生物结构,揭示轮作甘蔗影响后茬香蕉枯萎病发生的土壤微生态机制。结果表明,连作蕉地香蕉枯萎病发病率为49.15%,而轮作甘蔗1年后茬香蕉枯萎病发病率降至17.86%,轮作2年和3年甘蔗后茬香蕉枯萎病发病率仅为1.79%。土壤可培养细菌总数随着轮作甘蔗年限的增加而显著增加,可培养放线菌总数和尖孢镰刀菌则呈逐渐减少的趋势,真菌总数无显著变化。土壤细菌多样性指数随轮作甘蔗年限的增加而降低。假单胞菌目(Pseudomonadales)、浮霉菌目(Planctomycetales)、酸微菌目(Acidimicrobiales)和土壤红杆菌目(Solirubrobacterales)的细菌丰度呈随甘蔗种植年限增加而逐年增加的趋势,并与香蕉枯萎病发病率呈显著负相关关系。酸杆菌目(Acidobacteriales)、红螺菌目(Rhodospirillales)和军团菌目(Legionellales)的细菌丰度则呈随甘蔗种植年限逐年降低趋势,并与香蕉枯萎病发病率呈显著正相关。连作香蕉地轮作甘蔗,能够显著降低后茬香蕉枯萎病的发病率,并且随着甘蔗轮作时间年限的增加,发病率逐年降低。轮作甘蔗通过改变土壤微生物群落结构而间接影响尖孢镰刀菌的生存环境,从而降低香蕉枯萎病发病率,轮作2年甘蔗,即可达到显著控病效果。此结果与《广东新语》中关于香蕉-甘蔗轮作年限的记载一致。
Banana wilt caused by Fusarium oxysporum f. sp. cubense is one of the most widespread and severe diseases in the world.In recent years, banana-sugarcane rotation plantation has been used to prevent and control Fusarium wilt of banana in some farmlands in the Pearl River Delta with good effects. Banana-sugarcane rotation had been recorded in ancient Chinese book Guangdong New Sight, but the role, control effect and mechanism of the rotation system was unclear. Therefore, the control effect of banana Fusarium wilt after different years of sugarcane rotation was studied and soil micro-ecological mechanism discussed in this paper.The complete random design method with 4 treatments of medium-resistance ‘Nongke No. 1' banana and ‘Guangdong Yellow Peel'sugarcane were used in the experiment. The farmland with 11 years of continuous cropping of banana(CK) was the control. Other three treatments were continuous cropping of banana ration with sugarcane for one year(GZ1), two years(GZ2) and three years(GZ3) before replanting banana for 1 year. The banana Fusarium wilt disease incidence after various years of sugarcane rotation was investigated, the numbers of soil culturable micro-organisms measured, and soil microbial composition and structure analyzed using the high-throughput DNA sequencing technique. The results showed that that banana Fusarium wilt incidence in banana continuously planting field was 49.15%. After one year of sugarcane rotation, the incidence dropped to 17.86% and the effect of the relative control was 63.66%. After two or three years of sugarcane rotation, the incidence was only 1.79% and the effect of the relative control was above 95%. The amount of cultivable bacteria increased significantly along with increased year of sugarcane rotation, but the amounts of Fusarium oxysporum and actinomycetes significantly decreased. The amount of fungi did not significantly change. Sequence analysis showed that soil bacterial diversity index reduced with the number of years of sugarcane rotation. The relative abundance of bacteria of Pseudomonadales, Planctomycetales, Acidimicrobiales and Solirubrobacterales apparently increased with increasing number of sugarcane rotation years and had significantly negative correlation with the incidence of banana Fusarium wilt.Meanwhile, bacteria of Acidobacteriales, Rhodospirillales, Legionellales and Enterobacteriales apparently decreased and had significantly positive correlation with the incidence of banana Fusarium wilt. In summary, banana-sugarcane rotation significantly reduced banana wilt incidence, compared with continuous banana cropping system. The inhibiting effect apparently increased with increasing number of years of sugarcane rotation. In two years of sugarcane rotation of banana field, the incidence of banana wilt significantly reduced and the yield of banana significantly increased; which was consistent with the results recorded in the Guangdong New Sight.The changes in bacterial community structure in the soil played an important role in the control of banana Fusarium wilt.
Banana wilt caused by Fusarium oxysporum f. sp. cubense is one of the most widespread and severe diseases in the world.In recent years, banana-sugarcane rotation plantation has been used to prevent and control Fusarium wilt of banana in some farmlands in the Pearl River Delta with good effects. Banana-sugarcane rotation had been recorded in ancient Chinese book Guangdong New Sight, but the role, control effect and mechanism of the rotation system was unclear. Therefore, the control effect of banana Fusarium wilt after different years of sugarcane rotation was studied and soil micro-ecological mechanism discussed in this paper.The complete random design method with 4 treatments of medium-resistance ‘Nongke No. 1' banana and ‘Guangdong Yellow Peel'sugarcane were used in the experiment. The farmland with 11 years of continuous cropping of banana(CK) was the control. Other three treatments were continuous cropping of banana ration with sugarcane for one year(GZ1), two years(GZ2) and three years(GZ3) before replanting banana for 1 year. The banana Fusarium wilt disease incidence after various years of sugarcane rotation was investigated, the numbers of soil culturable micro-organisms measured, and soil microbial composition and structure analyzed using the high-throughput DNA sequencing technique. The results showed that that banana Fusarium wilt incidence in banana continuously planting field was 49.15%. After one year of sugarcane rotation, the incidence dropped to 17.86% and the effect of the relative control was 63.66%. After two or three years of sugarcane rotation, the incidence was only 1.79% and the effect of the relative control was above 95%. The amount of cultivable bacteria increased significantly along with increased year of sugarcane rotation, but the amounts of Fusarium oxysporum and actinomycetes significantly decreased. The amount of fungi did not significantly change. Sequence analysis showed that soil bacterial diversity index reduced with the number of years of sugarcane rotation. The relative abundance of bacteria of Pseudomonadales, Planctomycetales, Acidimicrobiales and Solirubrobacterales apparently increased with increasing number of sugarcane rotation years and had significantly negative correlation with the incidence of banana Fusarium wilt.Meanwhile, bacteria of Acidobacteriales, Rhodospirillales, Legionellales and Enterobacteriales apparently decreased and had significantly positive correlation with the incidence of banana Fusarium wilt. In summary, banana-sugarcane rotation significantly reduced banana wilt incidence, compared with continuous banana cropping system. The inhibiting effect apparently increased with increasing number of years of sugarcane rotation. In two years of sugarcane rotation of banana field, the incidence of banana wilt significantly reduced and the yield of banana significantly increased; which was consistent with the results recorded in the Guangdong New Sight.The changes in bacterial community structure in the soil played an important role in the control of banana Fusarium wilt.
引文
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[8]柳红娟,黄洁,刘子凡,等.木薯轮作年限对枯萎病高发蕉园土壤抑病性的影响[J].西南农业学报,2016,29(2):255-259LIU H J,HUANG J,LIU Z F,et al.Effects of different years of cassava rotation on soil disease suppression in high incidence banana plantations[J].Southwest China Journal of Agricultural Sciences,2016,29(2):255-259
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[10]赵飞,倪根金,章家恩.广东增城市农业遗产的调查研究[J].古今农业,2012,(3):52-62ZHAO F,NI G J,ZHANG J E.Investigation on agricultural heritage in Zengcheng City of Guangdong[J].Ancient and Modern Agriculture,2012,(3):52-62
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[14]ZHAO J,NI T,LI Y,et al.Responses of bacterial communities in arable soils in a rice-wheat cropping system to different fertilizer regimes and sampling times[J].PLoS One,2014,9(1):e85301
[15]赵娜.三种茄科蔬菜轮作对高发枯萎病蕉园土壤微生物的调控效应[D].海口:海南大学,2014:40-41ZHAO N.Effecting of crop rotation with three Solanaceae on the soil microbial in banana orchard with serious Fusarium wilt disease[D].Haikou:Hainan University,2014:40-41
[16]ZHANG H,MALLIK A,ZENG R S.Control of Panama disease of banana by rotating and intercropping with Chinese chive(Allium tuberosum Rottler):Role of plant volatiles[J].Journal of Chemical Ecology,2013,39(2):243-252
[17]柳红娟.木薯对香蕉枯萎病的防控效果及其机理研究[D].海口:海南大学,2015:16-17LIU H J.The effect and mechanism of cassava on the control of banana Fusarium wilt[D].Haikou:Hainan University,2015:16-17
[18]辛侃.水稻-香蕉轮作并向土壤中添加有机物料防控香蕉枯萎病的研究[D].海口:海南大学,2014:25-28XIN K.Control of Fusarium oxysporum f.sp.cubense in banana by rice-banana rotation combine with application of organic materials[D].Haikou:Hainan University,2014:25-28
[19]欧阳娴,阮小蕾,吴超,等.香蕉轮作和连作土壤细菌主要类群[J].应用生态学报,2011,22(6):1573-1578OUYANG X,RUAN X L,WU C,et al.Main bacterial groups in banana soil under rotated and continuous cropping[J].Chinese Journal of Applied Ecology,2011,22(6):1573-1578
[20]VAN PEER R,NIEMANN G J,SCHIPPERS B.Induced resistance and phytoalexin accumulation in biological control of Fusarium wilt of carnation by Pseudomonas sp.strain WCS417r[J].Phytopathology,1991,81(7):728-734
[21]AHN I P,LEE S W,SUH S C.Rhizobacteria-induced priming in Arabidopsis is dependent on ethylene,jasmonic acid,and NPR1[J].Molecular Plant-Microbe Interactions,2007,20(7):759-768
[22]SHORESH M,YEDIDIA I,CHET I.Involvement of jasmonic acid/ethylene signaling pathway in the systemic resistance induced in cucumber by Trichoderma asperellum T203[J].Phytopathology,2005,95(1):76-84
[23]STEIN E,MOLITOR A,KOGEL K H,et al.Systemic resistance in Arabidopsis conferred by the mycorrhizal fungus Piriformospora indica requires jasmonic acid signaling and the cytoplasmic function of NPR1[J].Plant and Cell Physiology,2008,49(11):1747-1751
[24]BERENDSEN R L,PIETERSE C M J,BAKKER P A H M.The rhizosphere microbiome and plant health[J].Trends in Plant Science,2012,17(8):478-486
[25]FRIDLENDER M,INBAR J,CHET I.Biological control of soilborne plant pathogens by aβ-1,3 glucanase-producing Pseudomonas cepacia[J].Soil Biology and Biochemistry,1993,25(9):1211-1221
[26]GARBEVA P,VAN VEEN J A,VAN ELSAS J D.Assessment of the diversity,and antagonism towards Rhizoctonia solani AG3,of Pseudomonas species in soil from different agricultural regimes[J].FEMS Microbiology Ecology,2004,47(1):51-64
[27]COUILLEROT O,PRIGENT-COMBARET C,CABALLERO-MELLADO J,et al.Pseudomonas fluorescens and closelyrelated fluorescent pseudomonads as biocontrol agents of soil-borne phytopathogens[J].Letters in Applied Microbiology,2009,48(5):505-512
[28]SANGUIN H,SARNIGUET A,GAZENGEL K,et al.Rhizosphere bacterial communities associated with disease suppressiveness stages of take-all decline in wheat monoculture[J].New Phytologist,2009,184(3):694-707
[29]ZHOU X G,WU F Z.p-Coumaric acid influenced cucumber rhizosphere soil microbial communities and the growth of Fusarium oxysporum f.sp.cucumerinum Owen[J].PLoS One,2012,7(10):e48288
[30]WARD N L,CHALLACOMBE J F,JANSSEN P H,et al.Three genomes from the phylum Acidobacteria provide insight into the lifestyles of these microorganisms in soils[J].Applied and Environmental Microbiology,2009,75(7):2046-2056
[31]RAWAT S R,M?NNIST?M K,BROMBERG Y,et al.Comparative genomic and physiological analysis provides insights into the role of Acidobacteria in organic carbon utilization in Arctic tundra soils[J].FEMS Microbiology Ecology,2012,82(2):341-355
[32]ZOLLA G,BADRI D V,BAKKER M G,et al.Soil microbiomes vary in their ability to confer drought tolerance to Arabidopsis[J].Applied Soil Ecology,2013,68:1-9
[33]CAMPOS S B,LISBOA B B,CAMARGO F A O,et al.Soil suppressiveness and its relations with the microbial community in a Brazilian subtropical agroecosystem under different management systems[J].Soil Biology and Biochemistry,2016,96:191-197
[2]VAN EERD L L,CONGREVES K A,HAYES A,et al.Long-term tillage and crop rotation effects on soil quality,organic carbon,and total nitrogen[J].Canadian Journal of Soil Science,2014,94(3):303-315
[3]XUAN D T,GUONG V T,ROSLING A,et al.Different crop rotation systems as drivers of change in soil bacterial community structure and yield of rice,Oryza sativa[J].Biology and Fertility of Soils,2012,48(2):217-225
[4]NAVARRO-NOYA Y E,GóMEZ-ACATA S,MONTOYA-CIRIACO N,et al.Relative impacts of tillage,residue management and crop-rotation on soil bacterial communities in a semi-arid agroecosystem[J].Soil Biology and Biochemistry,2013,65:86-95
[5]WANG B B,LI R,RUAN Y Z,et al.Pineapple-banana rotation reduced the amount of Fusarium oxysporum more than maize-banana rotation mainly through modulating fungal communities[J].Soil Biology and Biochemistry,2015,86:77-86
[6]XIONG W,ZHAO Q Y,XUE C,et al.Comparison of fungal community in black pepper-vanilla and vanilla monoculture systems associated with vanilla Fusarium wilt disease[J].Frontiers in Microbiology,2016,7:117
[7]辛侃,赵娜,邓小垦,等.香蕉-水稻轮作联合添加有机物料防控香蕉枯萎病研究[J].植物保护,2014,40(6):36-41XIN K,ZHAO N,DENG X K,et al.Effects of rice rotation incorporated with organic materials on suppression of banana Fusarium wilt disease[J].Plant Protection,2014,40(6):36-41
[8]柳红娟,黄洁,刘子凡,等.木薯轮作年限对枯萎病高发蕉园土壤抑病性的影响[J].西南农业学报,2016,29(2):255-259LIU H J,HUANG J,LIU Z F,et al.Effects of different years of cassava rotation on soil disease suppression in high incidence banana plantations[J].Southwest China Journal of Agricultural Sciences,2016,29(2):255-259
[9]HUANG Y H,WANG R C,LI C H,et al.Control of Fusarium wilt in banana with Chinese leek[J].European Journal of Plant Pathology,2012,134(1):87-95
[10]赵飞,倪根金,章家恩.广东增城市农业遗产的调查研究[J].古今农业,2012,(3):52-62ZHAO F,NI G J,ZHANG J E.Investigation on agricultural heritage in Zengcheng City of Guangdong[J].Ancient and Modern Agriculture,2012,(3):52-62
[11]林先贵.土壤微生物研究原理与方法[M].北京:高等教育出版社,2010LIN X G.Principles and Methods of Soil Microbiology Research[M].Beijing:Higher Education Press,2010
[12]KOMADA H.Development of a selective medium for quantitative isolation of Fusarium oxysporum from natural soil[J].Review of Plant Protection Research,1975,8:114-124
[13]SMITH L J,SMITH M K,TREE D,et al.Development of a small-plant bioassay to assess banana grown from tissue culture for consistent infection by Fusarium oxysporum f.sp.cubense[J].Australasian Plant Pathology,2008,37(2):171-179
[14]ZHAO J,NI T,LI Y,et al.Responses of bacterial communities in arable soils in a rice-wheat cropping system to different fertilizer regimes and sampling times[J].PLoS One,2014,9(1):e85301
[15]赵娜.三种茄科蔬菜轮作对高发枯萎病蕉园土壤微生物的调控效应[D].海口:海南大学,2014:40-41ZHAO N.Effecting of crop rotation with three Solanaceae on the soil microbial in banana orchard with serious Fusarium wilt disease[D].Haikou:Hainan University,2014:40-41
[16]ZHANG H,MALLIK A,ZENG R S.Control of Panama disease of banana by rotating and intercropping with Chinese chive(Allium tuberosum Rottler):Role of plant volatiles[J].Journal of Chemical Ecology,2013,39(2):243-252
[17]柳红娟.木薯对香蕉枯萎病的防控效果及其机理研究[D].海口:海南大学,2015:16-17LIU H J.The effect and mechanism of cassava on the control of banana Fusarium wilt[D].Haikou:Hainan University,2015:16-17
[18]辛侃.水稻-香蕉轮作并向土壤中添加有机物料防控香蕉枯萎病的研究[D].海口:海南大学,2014:25-28XIN K.Control of Fusarium oxysporum f.sp.cubense in banana by rice-banana rotation combine with application of organic materials[D].Haikou:Hainan University,2014:25-28
[19]欧阳娴,阮小蕾,吴超,等.香蕉轮作和连作土壤细菌主要类群[J].应用生态学报,2011,22(6):1573-1578OUYANG X,RUAN X L,WU C,et al.Main bacterial groups in banana soil under rotated and continuous cropping[J].Chinese Journal of Applied Ecology,2011,22(6):1573-1578
[20]VAN PEER R,NIEMANN G J,SCHIPPERS B.Induced resistance and phytoalexin accumulation in biological control of Fusarium wilt of carnation by Pseudomonas sp.strain WCS417r[J].Phytopathology,1991,81(7):728-734
[21]AHN I P,LEE S W,SUH S C.Rhizobacteria-induced priming in Arabidopsis is dependent on ethylene,jasmonic acid,and NPR1[J].Molecular Plant-Microbe Interactions,2007,20(7):759-768
[22]SHORESH M,YEDIDIA I,CHET I.Involvement of jasmonic acid/ethylene signaling pathway in the systemic resistance induced in cucumber by Trichoderma asperellum T203[J].Phytopathology,2005,95(1):76-84
[23]STEIN E,MOLITOR A,KOGEL K H,et al.Systemic resistance in Arabidopsis conferred by the mycorrhizal fungus Piriformospora indica requires jasmonic acid signaling and the cytoplasmic function of NPR1[J].Plant and Cell Physiology,2008,49(11):1747-1751
[24]BERENDSEN R L,PIETERSE C M J,BAKKER P A H M.The rhizosphere microbiome and plant health[J].Trends in Plant Science,2012,17(8):478-486
[25]FRIDLENDER M,INBAR J,CHET I.Biological control of soilborne plant pathogens by aβ-1,3 glucanase-producing Pseudomonas cepacia[J].Soil Biology and Biochemistry,1993,25(9):1211-1221
[26]GARBEVA P,VAN VEEN J A,VAN ELSAS J D.Assessment of the diversity,and antagonism towards Rhizoctonia solani AG3,of Pseudomonas species in soil from different agricultural regimes[J].FEMS Microbiology Ecology,2004,47(1):51-64
[27]COUILLEROT O,PRIGENT-COMBARET C,CABALLERO-MELLADO J,et al.Pseudomonas fluorescens and closelyrelated fluorescent pseudomonads as biocontrol agents of soil-borne phytopathogens[J].Letters in Applied Microbiology,2009,48(5):505-512
[28]SANGUIN H,SARNIGUET A,GAZENGEL K,et al.Rhizosphere bacterial communities associated with disease suppressiveness stages of take-all decline in wheat monoculture[J].New Phytologist,2009,184(3):694-707
[29]ZHOU X G,WU F Z.p-Coumaric acid influenced cucumber rhizosphere soil microbial communities and the growth of Fusarium oxysporum f.sp.cucumerinum Owen[J].PLoS One,2012,7(10):e48288
[30]WARD N L,CHALLACOMBE J F,JANSSEN P H,et al.Three genomes from the phylum Acidobacteria provide insight into the lifestyles of these microorganisms in soils[J].Applied and Environmental Microbiology,2009,75(7):2046-2056
[31]RAWAT S R,M?NNIST?M K,BROMBERG Y,et al.Comparative genomic and physiological analysis provides insights into the role of Acidobacteria in organic carbon utilization in Arctic tundra soils[J].FEMS Microbiology Ecology,2012,82(2):341-355
[32]ZOLLA G,BADRI D V,BAKKER M G,et al.Soil microbiomes vary in their ability to confer drought tolerance to Arabidopsis[J].Applied Soil Ecology,2013,68:1-9
[33]CAMPOS S B,LISBOA B B,CAMARGO F A O,et al.Soil suppressiveness and its relations with the microbial community in a Brazilian subtropical agroecosystem under different management systems[J].Soil Biology and Biochemistry,2016,96:191-197