甲壳素对连作条件下平邑甜茶幼苗生长及土壤环境的影响
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摘要
研究在苹果连作土壤中添加甲壳素对苹果幼苗生长、土壤酶及土壤真菌群落结构的影响,探讨甲壳素缓解苹果连作障碍的可能性,为防控苹果连作障碍提供依据。盆栽条件下,以平邑甜茶幼苗为试材,在苹果连作土壤中分别添加0,0.5,1.0和2.5g/kg的甲壳素,测定了连作土壤中添加不同量的甲壳素后,幼苗生物量、根系保护酶活性、土壤主要酶(蔗糖酶、脲酶、磷酸酶等)活性以及土壤中真菌群落结构的变化。9月份结果表明,与对照相比,1.0 g/kg的甲壳素处理连作土,可显著提高平邑甜茶幼苗株高和干鲜重,分别比对照增加了36.8%、82.1%和100.8%;甲壳素处理能增加幼苗根系保护酶活性,其中1.0 g/kg甲壳素处理SOD、POD和CAT活性最高,其次为0.5 g/kg,而2.5 g/kg甲壳素处理显著抑制了幼苗根系保护酶活性。1.0 g/kg甲壳素处理可提高土壤中细菌/真菌值,并且提高了土壤中蔗糖酶、脲酶、蛋白酶、磷酸酶、过氧化氢酶和多酚氧化酶活性,分别比对照提高了8.6%、40.5%、81.1%、15.3%、18.7%和49.8%,2.5 g/kg甲壳素处理则降低土壤酶活性或者使土壤酶活性与对照相当。根据T-RFLP的图谱中OUT的数量、种类及丰度,分别计算了不同处理土壤的真菌多样性,发现1.0 g/kg甲壳素处理的连作土具有最高的多样性、均匀度和丰富度指数,分别比对照增加了52.2%、8.0%和87.1%。主成分分析(PCA)结果显示,不同剂量甲壳素处理的连作土壤中真菌被PC2分成了两部分,其中0.5 g/kg和1.0 g/kg的甲壳素添加量分布在PC2的负方向上,而CK和2.5g/kg的甲壳素处理分布在PC2的正方向上,这说明添加不同量的甲壳素对连作土壤真菌群落多样性有显著影响,添加量太多或者太少均会造成土壤真菌多样性下降,只有适量的甲壳素可提高真菌群落结构多样性。实验结果表明1.0 g/kg的甲壳素可提高连作平邑甜茶幼苗生物量,改善连作土壤环境,有效缓解平邑甜茶的连作障碍。
The effects of different concentrations of chitin on the activity of enzymes and fungal communities existing in replant soils were explored,for enrichment of soils to alleviate apple replant disease( ARD). In this study,Malus hupehensis Rehd. seedlings potted in replant soil were treated with four concentrations of chitin( 0,0.5,1.0,and 2.5 g/kg). The growth of seedlings was monitored though plant height,fresh and dry weight,antioxidant enzyme activities,such as superoxide dismutase( SOD), peroxidase( POD), and catalase( CAT), and lipid peroxidation in roots.Simultaneously,soil enzyme activities,soil microbial load,and fungal community structure,which was measured by terminal-restriction fragment length polymorphism( T-RFLP) profiles,were determined in soil samples subjected to the four chitin treatments. Compared with the control,seedlings treated with 1.0 g/kg chitin showed 36.8%,and 82.1% and 100.8% increase in plant height,and fresh and dry weight,respectively. The optimal dose of chitin could increase antioxidant enzyme activity in the root system of the seedlings,whereas high doses inhibited enzyme activity. The effectiveness of chitin treatment for inhibiting the activity of SOD,POD,and CAT in the root system of M. hupehensis seedlings decreased in the following order: 1.0 g/kg > 0.5 g/kg > 0 g/kg > 2. 5 g/kg. Chitin treatment at 1. 0 g/kg enhanced the bacterial- fungal ratio,and resulted in a "bacteria- rich soil "; consequently,sucrase,urease,protease,phosphatase,catalase,and polyphenol oxidase activities increased by 8.6%,40.5%,81.1%,15.3%,18.7%,and 49.8%,respectively. Remarkable differences in T-RFLP profiles were observed among control,0.5 g/kg,1.0 g/kg,and 2.5 g/kg treatments. A significant difference in fungal community structure was observed with the application of different doses of chitin. The Shannon diversity,evenness,and richness indexes were the highest in replanted soil treated with 1.0 g/kg chitin and the least in soil treated with 2.5 g/kg chitin. Principal component analysis indicated that fungal community structure of soil treated with 1.0g/kg chitin was considerably different from that of the control soil and soil treated with 2.5 g/kg chitin. The present study showed that low concentrations of chitin could alleviate ARD stress in M. hupehensis Rehd. seedlings,whereas high concentrations could aggravate ARD stress. Collectively,these findings suggest that chitin applied to replant soil at 1.0 g/kg obviously enhanced plant height,fresh and dry weight,improved the soil environment,and effectively alleviate ARD stress in M. hupehensis Rehd. seedlings,therefore,a chitin dose of 1.0 g/kg was considered optimal for replant soil.
The effects of different concentrations of chitin on the activity of enzymes and fungal communities existing in replant soils were explored,for enrichment of soils to alleviate apple replant disease( ARD). In this study,Malus hupehensis Rehd. seedlings potted in replant soil were treated with four concentrations of chitin( 0,0.5,1.0,and 2.5 g/kg). The growth of seedlings was monitored though plant height,fresh and dry weight,antioxidant enzyme activities,such as superoxide dismutase( SOD), peroxidase( POD), and catalase( CAT), and lipid peroxidation in roots.Simultaneously,soil enzyme activities,soil microbial load,and fungal community structure,which was measured by terminal-restriction fragment length polymorphism( T-RFLP) profiles,were determined in soil samples subjected to the four chitin treatments. Compared with the control,seedlings treated with 1.0 g/kg chitin showed 36.8%,and 82.1% and 100.8% increase in plant height,and fresh and dry weight,respectively. The optimal dose of chitin could increase antioxidant enzyme activity in the root system of the seedlings,whereas high doses inhibited enzyme activity. The effectiveness of chitin treatment for inhibiting the activity of SOD,POD,and CAT in the root system of M. hupehensis seedlings decreased in the following order: 1.0 g/kg > 0.5 g/kg > 0 g/kg > 2. 5 g/kg. Chitin treatment at 1. 0 g/kg enhanced the bacterial- fungal ratio,and resulted in a "bacteria- rich soil "; consequently,sucrase,urease,protease,phosphatase,catalase,and polyphenol oxidase activities increased by 8.6%,40.5%,81.1%,15.3%,18.7%,and 49.8%,respectively. Remarkable differences in T-RFLP profiles were observed among control,0.5 g/kg,1.0 g/kg,and 2.5 g/kg treatments. A significant difference in fungal community structure was observed with the application of different doses of chitin. The Shannon diversity,evenness,and richness indexes were the highest in replanted soil treated with 1.0 g/kg chitin and the least in soil treated with 2.5 g/kg chitin. Principal component analysis indicated that fungal community structure of soil treated with 1.0g/kg chitin was considerably different from that of the control soil and soil treated with 2.5 g/kg chitin. The present study showed that low concentrations of chitin could alleviate ARD stress in M. hupehensis Rehd. seedlings,whereas high concentrations could aggravate ARD stress. Collectively,these findings suggest that chitin applied to replant soil at 1.0 g/kg obviously enhanced plant height,fresh and dry weight,improved the soil environment,and effectively alleviate ARD stress in M. hupehensis Rehd. seedlings,therefore,a chitin dose of 1.0 g/kg was considered optimal for replant soil.
引文
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[2]Tewoldemedhin Y T,Mazzola M,Botha W J,Spies C F J,Mc Leod A.Characterization of fungi(Fusarium and Rhizoctonia)and oomycetes(Phytophthora and Pythium)associated with apple orchards in South Africa.European Journal of Plant Pathology,2011,130(2):215-229.
[3]Tewoldemedhin Y T,Mazzola M,Labuschagne I,Mc Leod A.A multi-phasic approach reveals that apple replant disease is caused by multiple biological agents,with some agents acting synergistically.Soil Biology and Biochemistry,2011,43(9):1917-1927.
[4]刘素慧,刘世琦,张自坤,尉辉,齐建建,段吉锋.大蒜连作对其根际土壤微生物和酶活性的影响.中国农业科学,2010,43(5):1000-1006.
[5]刘星,邱慧珍,王蒂,张俊莲,沈其荣.甘肃省中部沿黄灌区轮作和连作马铃薯根际土壤真菌群落的结构性差异评估.生态学报,2015,35(12):3938-3948.
[6]樊琳,柴如山,刘立娟,黄利东,章永松.稻草和猪粪发酵残渣配施菌剂对大棚连作土壤的改良作用.植物营养与肥料学报,2013,19(2):437-444.
[7]张国斌,郁继华,冯致,马彦霞,吕剑.NO和ABA对辣椒幼苗自毒作用缓解的生理生化机制.园艺学报,2013,40(3):458-466.
[8]Mazzola M,Hewavitharana S S,Strauss S L.Brassica seed meal soil amendments transform the rhizosphere microbiome and improve apple production through resistance to pathogen reinfestation.Phytopathology,2014,105(4):460-469.
[9]孙海兵,胡艳丽,陈学森,毛志泉.发酵时间对有机物料发酵流体成分含量变化及对连作苹果生物量的影响.植物营养与肥料学报,2012,18(6):1469-1477.
[10]袁玉娟,胡江,凌宁,仇美华,沈其荣,杨兴明.施用不同生物有机肥对连作黄瓜枯萎病防治效果及其机理初探.植物营养与肥料学报,2014,20(2):372-379.
[11]蒋小姝,莫海涛,苏海佳,张小勇.甲壳素及壳聚糖在农业领域方面的应用.中国农学通报,2013,29(6):170-174.
[12]孙世利,骆耀平.壳聚糖对茶树抗性酶调节作用的研究.浙江大学学报:农业与生命科学版,2009,35(1):84-88.
[13]崔健,刘怀锋.甲壳素对苹果幼苗抗旱生理效应的影响.北方园艺,2007,(11):19-22.
[14]Wang Y F,Pan F B,Wang G S,Zhang G D,Wang Y L,Chen X S,Mao Z Q.Effects of biochar on photosynthesis and antioxidative system of Malus hupehensis Rehd.seedlings under replant conditions.Scientia Horticulturae,2014,175:9-15.
[15]赵世杰.植物生理学实验指导.北京:中国农业科技出版社,1998.
[16]关松荫.土壤酶及其研究法.北京:农业出版社,1986.
[17]沈萍,陈向东.微生物学实验(第4版).北京:高等教育出版社,2007.
[18]Hogberg M N,Yarwood S A,Myrold D D.Fungal but not bacterial soil communities recover after termination of decadal nitrogen additions to boreal forest.Soil Biology and Biochemistry,2014,72:35-43.
[19]Tipayno S,Kim C-G,Sa T.T-RFLP analysis of structural changes in soil bacterial communities in response to metal and metalloid contamination and initial phytoremediation.Applied Soil Ecology,2012,61:137-146.
[20]李志鹏,姜娜,刘晗璐,崔学哲,荆祎,杨福合,李光玉.基于DGGE和T-RFLP分析采食不同粗饲料梅花鹿瘤胃细菌区系.中国农业科学,2014,47(4):759-768.
[21]Mazzola M.Replant disease control and soil system resilience to pathogen infestation in response to Brassicaceae seed meal amendment.Phytopathology,2012,102:S4-S4.
[22]Jaiswal A K,Elad Y,Graber E R,Frenkel O.Rhizoctonia solani suppression and plant growth promotion in cucumber as affected by biochar pyrolysis temperature,feedstock and concentration.Soil Biology and Biochemistry,2014,69:110-118.
[23]马彦霞,郁继华,张国斌,曹刚.壳聚糖对水分胁迫下辣椒幼苗氧化损伤的保护作用.中国农业科学,2012,45(10):1964-1971.
[24]程红玉,肖占文,秦嘉海,王瑞.连作对玉米制种田土壤养分和土壤酶活性的影响.土壤,2013,45(4):623-627.
[25]谷岩,邱强,王振民,陈喜凤,吴春胜.连作大豆根际微生物群落结构及土壤酶活性.中国农业科学,2012,45(19):3955-3964.
[26]陈慧,郝慧荣,熊君,齐晓辉,张重义,林文雄.地黄连作对根际微生物区系及土壤酶活性的影响.应用生态学报,2008,18(12):2755-2759.
[27]尹承苗,陈学森,沈向,张兆波,孙海兵,毛志泉.不同浓度有机物料发酵液对连作苹果幼树生物量及土壤环境的影响.植物营养与肥料学报,2013,19(6):1450-1458.
[28]Hallmann J,Rodrguez-Kábana R,Kloepper J W.Chitin-mediated changes in bacterial communities of the soil,rhizosphere and within roots of cotton in relation to nematode control.Soil Biology and Biochemistry,1999,31(4):551-560.
[29]孟玲,王雷.壳聚糖对多种植物病原菌的抑菌效果概述.农药,2009,48(11):781-783.
[30]吕毅,宋富海,李园园,沈向,陈学森,吴树敬,毛志泉.轮作不同作物对苹果园连作土壤环境及平邑甜茶幼苗生理指标的影响.中国农业科学,2014,47(14):2830-2839.
[2]Tewoldemedhin Y T,Mazzola M,Botha W J,Spies C F J,Mc Leod A.Characterization of fungi(Fusarium and Rhizoctonia)and oomycetes(Phytophthora and Pythium)associated with apple orchards in South Africa.European Journal of Plant Pathology,2011,130(2):215-229.
[3]Tewoldemedhin Y T,Mazzola M,Labuschagne I,Mc Leod A.A multi-phasic approach reveals that apple replant disease is caused by multiple biological agents,with some agents acting synergistically.Soil Biology and Biochemistry,2011,43(9):1917-1927.
[4]刘素慧,刘世琦,张自坤,尉辉,齐建建,段吉锋.大蒜连作对其根际土壤微生物和酶活性的影响.中国农业科学,2010,43(5):1000-1006.
[5]刘星,邱慧珍,王蒂,张俊莲,沈其荣.甘肃省中部沿黄灌区轮作和连作马铃薯根际土壤真菌群落的结构性差异评估.生态学报,2015,35(12):3938-3948.
[6]樊琳,柴如山,刘立娟,黄利东,章永松.稻草和猪粪发酵残渣配施菌剂对大棚连作土壤的改良作用.植物营养与肥料学报,2013,19(2):437-444.
[7]张国斌,郁继华,冯致,马彦霞,吕剑.NO和ABA对辣椒幼苗自毒作用缓解的生理生化机制.园艺学报,2013,40(3):458-466.
[8]Mazzola M,Hewavitharana S S,Strauss S L.Brassica seed meal soil amendments transform the rhizosphere microbiome and improve apple production through resistance to pathogen reinfestation.Phytopathology,2014,105(4):460-469.
[9]孙海兵,胡艳丽,陈学森,毛志泉.发酵时间对有机物料发酵流体成分含量变化及对连作苹果生物量的影响.植物营养与肥料学报,2012,18(6):1469-1477.
[10]袁玉娟,胡江,凌宁,仇美华,沈其荣,杨兴明.施用不同生物有机肥对连作黄瓜枯萎病防治效果及其机理初探.植物营养与肥料学报,2014,20(2):372-379.
[11]蒋小姝,莫海涛,苏海佳,张小勇.甲壳素及壳聚糖在农业领域方面的应用.中国农学通报,2013,29(6):170-174.
[12]孙世利,骆耀平.壳聚糖对茶树抗性酶调节作用的研究.浙江大学学报:农业与生命科学版,2009,35(1):84-88.
[13]崔健,刘怀锋.甲壳素对苹果幼苗抗旱生理效应的影响.北方园艺,2007,(11):19-22.
[14]Wang Y F,Pan F B,Wang G S,Zhang G D,Wang Y L,Chen X S,Mao Z Q.Effects of biochar on photosynthesis and antioxidative system of Malus hupehensis Rehd.seedlings under replant conditions.Scientia Horticulturae,2014,175:9-15.
[15]赵世杰.植物生理学实验指导.北京:中国农业科技出版社,1998.
[16]关松荫.土壤酶及其研究法.北京:农业出版社,1986.
[17]沈萍,陈向东.微生物学实验(第4版).北京:高等教育出版社,2007.
[18]Hogberg M N,Yarwood S A,Myrold D D.Fungal but not bacterial soil communities recover after termination of decadal nitrogen additions to boreal forest.Soil Biology and Biochemistry,2014,72:35-43.
[19]Tipayno S,Kim C-G,Sa T.T-RFLP analysis of structural changes in soil bacterial communities in response to metal and metalloid contamination and initial phytoremediation.Applied Soil Ecology,2012,61:137-146.
[20]李志鹏,姜娜,刘晗璐,崔学哲,荆祎,杨福合,李光玉.基于DGGE和T-RFLP分析采食不同粗饲料梅花鹿瘤胃细菌区系.中国农业科学,2014,47(4):759-768.
[21]Mazzola M.Replant disease control and soil system resilience to pathogen infestation in response to Brassicaceae seed meal amendment.Phytopathology,2012,102:S4-S4.
[22]Jaiswal A K,Elad Y,Graber E R,Frenkel O.Rhizoctonia solani suppression and plant growth promotion in cucumber as affected by biochar pyrolysis temperature,feedstock and concentration.Soil Biology and Biochemistry,2014,69:110-118.
[23]马彦霞,郁继华,张国斌,曹刚.壳聚糖对水分胁迫下辣椒幼苗氧化损伤的保护作用.中国农业科学,2012,45(10):1964-1971.
[24]程红玉,肖占文,秦嘉海,王瑞.连作对玉米制种田土壤养分和土壤酶活性的影响.土壤,2013,45(4):623-627.
[25]谷岩,邱强,王振民,陈喜凤,吴春胜.连作大豆根际微生物群落结构及土壤酶活性.中国农业科学,2012,45(19):3955-3964.
[26]陈慧,郝慧荣,熊君,齐晓辉,张重义,林文雄.地黄连作对根际微生物区系及土壤酶活性的影响.应用生态学报,2008,18(12):2755-2759.
[27]尹承苗,陈学森,沈向,张兆波,孙海兵,毛志泉.不同浓度有机物料发酵液对连作苹果幼树生物量及土壤环境的影响.植物营养与肥料学报,2013,19(6):1450-1458.
[28]Hallmann J,Rodrguez-Kábana R,Kloepper J W.Chitin-mediated changes in bacterial communities of the soil,rhizosphere and within roots of cotton in relation to nematode control.Soil Biology and Biochemistry,1999,31(4):551-560.
[29]孟玲,王雷.壳聚糖对多种植物病原菌的抑菌效果概述.农药,2009,48(11):781-783.
[30]吕毅,宋富海,李园园,沈向,陈学森,吴树敬,毛志泉.轮作不同作物对苹果园连作土壤环境及平邑甜茶幼苗生理指标的影响.中国农业科学,2014,47(14):2830-2839.