甘蔗宿根矮化病感病与非感病株根际土壤生物学性状及细菌群落结构特征
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摘要
【目的】比较甘蔗宿根矮化病(ratoon stunting disease,RSD)感病植株与非感病植株根际土壤的生物学性状及细菌群落结构特征,旨在为构建甘蔗健康的根际微环境,筛选高效RSD生防细菌提供参考。【方法】通过田间调查和实验室鉴定,以甘蔗RSD感病植株为试材,非感病植株为对照,采集甘蔗RSD感病植株和非感病植株的根际土壤,并基于传统和现代高通量测序技术,分析了甘蔗RSD感病植株和非感病植株根际土壤的生物学性状和细菌群落结构特征。【结果】与甘蔗RSD非感病植株相比,感病植株根际土壤中指示土壤肥力与健康状况的生物学性状指标β-葡糖苷酶、磷酸酶和氨肽酶活性,以及微生物生物量碳、氮、磷显著降低;同时,指示细菌丰富度的Chao1指数和指示细菌多样性的Shannon指数显著下降。门分类水平与非感病甘蔗植株相比,RSD感病植株根际土壤中Proteobacteria (变形杆菌门)、Actinobacteria (放线菌门)、Gemmatimonadetes (芽单胞菌门)和Nitrospirae (硝化螺旋菌门)等优势门类细菌占比呈倍级降低,但Chloroflexi (绿弯菌门)、Acidobacteria (酸杆菌门)、Firmicutes (厚壁菌门)、Cyanobacteria (蓝细菌门)、Planctomycetes (浮霉菌门)、Bacteroidetes (拟杆菌门)等优势门类细菌占比呈倍级增加;在属分类水平,与非感病甘蔗植株相比,RSD感病植株根际土壤中Xanthobacteraceae (黄色杆菌属)、Acidothermus、Gaiellales、Roseiflexus (玫瑰菌属)、Micromonosporaceae (小单孢菌属)和Nitrospira (硝化螺旋菌属)细菌占比呈倍级降低,但Acidobacteria (嗜酸菌属)细菌及部分未知菌属却呈倍级提高。【结论】甘蔗RSD感病植株根际微环境中指示土壤肥力的生物学指标显著降低,细菌丰富度和多样性显著下降,部分优势细菌门属占比发生剧变可能是导致甘蔗RSD发生的重要原因。
【Objectives】The biological properties and bacterial communities in the rhizosphere of canes infected and non-infected with ratoon stunting disease(RSD) were compared for effective bio-control of the RSD in sugarcanes.【Methods】The cane plant and their rhizosphere soil were identified for infection of RSD firstly in lab. The cane plant and rhizosphere soil samples infected and non-infected by RSD were collected, respectively.The soil biological properties and bacterial community were determined by traditional and high throughput sequencing techniques.【Results】Compared with healthy soil, the activities of β-glucosidase, phosphatase and aminopeptidase in RSD infected soils were significantly inhibited, and the contents of soil microbial biomass carbon, nitrogen and phosphorus were significantly decreased, and the indexes of bacterial diversity and richness including Chao1, Shannon were significantly decreased as well. As for individual bacteria community on phylum level, the ratios of Proteobacteria, Actinobacteria, Gemmatimonadetes and Nitrospirae were all significantly decreased, while those of Chloroflexi, Acidobacteria, Firmicutes, Cyanobacteria, Planctomycetes and Bacteroidetes were significantly increased in rhizospheres of RSD infected canes. On genus level, the ratios of Xanthobacteraceae, Acidothermus, Gaiellales, Roseiflexus, Micromonosporaceae and Nitrospira were significantly decreased, while those of Acidobacteria and unclassified bacteria were significantly increased in rhizospheres of RSD infected canes.【Conclusions】In rhizosphere soils of RSD infected canes, the activities of important soil enzymes and microbial mass carbon, nitrogen and phosphorous contents are low, the diversity and the richness bacterial community are significantly decreased. On the phylum and genus levels, great variations in the abundance of the dominant bacteria have been confirmed.
【Objectives】The biological properties and bacterial communities in the rhizosphere of canes infected and non-infected with ratoon stunting disease(RSD) were compared for effective bio-control of the RSD in sugarcanes.【Methods】The cane plant and their rhizosphere soil were identified for infection of RSD firstly in lab. The cane plant and rhizosphere soil samples infected and non-infected by RSD were collected, respectively.The soil biological properties and bacterial community were determined by traditional and high throughput sequencing techniques.【Results】Compared with healthy soil, the activities of β-glucosidase, phosphatase and aminopeptidase in RSD infected soils were significantly inhibited, and the contents of soil microbial biomass carbon, nitrogen and phosphorus were significantly decreased, and the indexes of bacterial diversity and richness including Chao1, Shannon were significantly decreased as well. As for individual bacteria community on phylum level, the ratios of Proteobacteria, Actinobacteria, Gemmatimonadetes and Nitrospirae were all significantly decreased, while those of Chloroflexi, Acidobacteria, Firmicutes, Cyanobacteria, Planctomycetes and Bacteroidetes were significantly increased in rhizospheres of RSD infected canes. On genus level, the ratios of Xanthobacteraceae, Acidothermus, Gaiellales, Roseiflexus, Micromonosporaceae and Nitrospira were significantly decreased, while those of Acidobacteria and unclassified bacteria were significantly increased in rhizospheres of RSD infected canes.【Conclusions】In rhizosphere soils of RSD infected canes, the activities of important soil enzymes and microbial mass carbon, nitrogen and phosphorous contents are low, the diversity and the richness bacterial community are significantly decreased. On the phylum and genus levels, great variations in the abundance of the dominant bacteria have been confirmed.
引文
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[9]韦金菊,宋修鹏,黄伟华,等.广西主要蔗区甘蔗宿根矮化病调查[J].南方农业学报,2017,48(7):1216-1219.Wei J J,Song X P,Huang W H,et al.Investigation on sugarcane ratoon stuning disease in main sugarcane area of Guangxi[J].Journal of Southern Agriculture,2017,48(7):1216-1219.
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[11]赵久成.嫁接对西瓜根际土壤微生物多样性及生物学性状的影响[D].南宁:广西大学硕士学位论文,2014.Zhao J C.Effects of grafting on soil microbial diversity and biological properties in rhizosphere of watermelon[D].Nanning:MSThesis of Guangxi University,2014.
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[17]吴金水,肖和艾,陈桂秋,等.旱地土壤微生物磷测定方法研究[J].土壤学报,2003,40(1):70-78.Wu J S,Xiao H A,Chen G Q,et al.Measurement of microbial biomass-P in upland soils in China[J].Acta Pedologica Sinica,2003,40(1):70-78.
[18]Grice E A,Kong H H,Conlan S,et al.Topographical and temporal diversity of the human skin microbiome[J].Science,2009,324(5931):1190-1192.
[19]Bainard L D,Koch A M,Gordon A M,Klironomos J N.Growth response of crops to soil microbial communities from conventional monocropping and tree-based intercropping systems[J].Plant and Soil,2013,363(1-2):345-356.
[20]杨尚东,赵久成,郭伊娟,等.番茄青枯病感病植株和健康植株根际土壤细菌群落结构的初步分析[J].中国蔬菜,2014,(8):25-29.Yang S D,Zhao J C,Guo Y J,et al.Characterization of soil bacterial community structures in rhizosphere of tomatoes infected with bacterial wilt and its non-infected plants[J].Chinese Vegetables,2014,(8):25-29.
[21]朱丽霞,章家恩,刘文高.根系分泌物与根际微生物相互作用研究综述[J].生态环境,2003,(1):102-105.Zhu L X,Zhang J E,Liu W G.Review of interactions between root exudates and rhizosphere microorganisms[J].Ecological Environment,2003,(1):102-105.
[22]钟文辉,蔡祖聪.土壤管理措施及环境因素对土壤微生物多样性影响研究进展[J].生物多样性,2004,12(4):456-465.Zhong W H,Cai Z C.Effect of soil management practices and environmental factors on soil microbial diversity:a review[J].Biodiversity Science,2004,12(4):456-465.
[23]杨宁,杨满元,雷玉兰,等.衡阳紫色土丘陵坡地土壤酶活性对植被恢复的响应[J].生态环境学报,2014,23(4):575-580.Yang N,Yang M Y,Lei Y L,et al.Response of soil enzyme activities to re-vegetation on sloping-land with purple soils in Hengyang of Hunan province[J].Ecology and Environmental Sciences,2014,23(4):575-580.
[24]Sowerb Y A,Emmet T B,Beie R C,et al.Microbial community changes in health and soil communities along a geographical gradient:Interaction with climate change manipulations[J].Soil Biology and Biochemistry,2005,37:1805-1813.
[25]Ahamadou B,Huang Q,Chen W,et al.Microcalorimetric assessment of microbial activity in long-term fertilization experimental soils of Southern China[J].Fems Microbiology Ecology,2009,70(2):186-195.
[26]Powlson D S,Brookes P C,Christensen B T.Measurement of soil microbial biomass provides an early indication of changes in total soil organic matter due to straw incorporation[J].Soil Biology&Biochemistry,1987,19(2):159-164.
[2]李文凤.甘蔗种苗传播病害病原检测与分子鉴定[A].中国植物病理学会.2016年学术年会论文集[C].中国植物病理学会,2016.Li W F.Pathogen detection and molecular identification of sugarcane seedling transmission diseases[A].Chinese Plant Pathology Society,Proceedings of 2016 Annual Academic Conference[C].Chinese Plant Pathology Society,2016.
[3]郭莺,汪文华,刘黎卿,等.甘蔗宿根矮化病多克隆抗体和免疫磁珠的制备[J].生物技术通报,2018,34(6):79-83.Guo Y,Wang W H,Liu L Q,et al.Preparation of polyclonal antibody against ratoon stunting disease and its immunomagnetic beads[J].Biotechnology Bulletin,2018,34(6):79-83.
[4]陈明辉,杨丽涛,谢晓娜,等.甘蔗宿根矮化病研究进展[J].南方农业学报,2011,42(3):280-283.Chen M H,Yang L T,Xie X N,et al.Research advances in ratoon stunting disease of sugarcane[J].Southern Journal of Agricultural Sciences,2011,42(3):280-283.
[5]Davis M J,Gillaspie A G,Harris R W,et al.Ratoon stunting disease of sugarcane:Isolation of the causal bacterium[J].Science,1980,210(4476):1365-1367.
[6]张小秋.宿根矮化病病原菌特性及其侵染后的甘蔗生理和基因差异表达[D].南宁:广西大学博士学位论文,2017.Zhang X Q.Characteristics of leifsonia xyli subsp.xyli and influences of its inoculation on physiology and gene expressions in sugarcane[D].Nanning:PhD Dissertation of Guangxi University,2017.
[7]Rao G P,Singh M,Singh H N.Alternative hosts of sugarcane diseases[J].Sugar Cane,1990,2(2):77-81.
[8]张荣跃,李文凤,黄应昆,等.PCR检测甘蔗宿根矮化病研究简述[J].中国糖料,2014,(3):69-70.Zhang R Y,Li W F,Huang Y K,et al.Review of PCR detection for ratoon stunting disease in sugarcane[J].Chinese Sugar,2014,(3):69-70.
[9]韦金菊,宋修鹏,黄伟华,等.广西主要蔗区甘蔗宿根矮化病调查[J].南方农业学报,2017,48(7):1216-1219.Wei J J,Song X P,Huang W H,et al.Investigation on sugarcane ratoon stuning disease in main sugarcane area of Guangxi[J].Journal of Southern Agriculture,2017,48(7):1216-1219.
[10]James G.A review of ratoon stunting disease[J].Sugar Cane,1997,(4):9-14.
[11]赵久成.嫁接对西瓜根际土壤微生物多样性及生物学性状的影响[D].南宁:广西大学硕士学位论文,2014.Zhao J C.Effects of grafting on soil microbial diversity and biological properties in rhizosphere of watermelon[D].Nanning:MSThesis of Guangxi University,2014.
[12]Hayano K A method for the determination ofβ-glucosidase activity in soil[J].Soil Science&Plant Nutrition,1973,19(2):103-108.
[13]Ladd J N.Properties of proteolytic enzymes extracted from soil[J].Soil Biology&Biochemistry,1972,4(2):227-237.
[14]Tabatabai M A,Bremner J M.Use of P-nitrophenyl phosphate for assay of soil phosphatase activity[J].Soil Biology&Biochemistry,1969,1(4):301-307.
[15]Vance E D,Brookes P C,Jenkinson D S.An extraction method for measuring soil microbial biomass C[J].Soil Biology&Biochemistry,1987,19(6):703-707.
[16]Joergensen R G,Brookes P C.Ninhydrin-reactive nitrogen measurements of microbial biomass in 0.5 M K2SO4 soil extracts[J].Soil Biology&Biochemistry,1990,22:1023-1027.
[17]吴金水,肖和艾,陈桂秋,等.旱地土壤微生物磷测定方法研究[J].土壤学报,2003,40(1):70-78.Wu J S,Xiao H A,Chen G Q,et al.Measurement of microbial biomass-P in upland soils in China[J].Acta Pedologica Sinica,2003,40(1):70-78.
[18]Grice E A,Kong H H,Conlan S,et al.Topographical and temporal diversity of the human skin microbiome[J].Science,2009,324(5931):1190-1192.
[19]Bainard L D,Koch A M,Gordon A M,Klironomos J N.Growth response of crops to soil microbial communities from conventional monocropping and tree-based intercropping systems[J].Plant and Soil,2013,363(1-2):345-356.
[20]杨尚东,赵久成,郭伊娟,等.番茄青枯病感病植株和健康植株根际土壤细菌群落结构的初步分析[J].中国蔬菜,2014,(8):25-29.Yang S D,Zhao J C,Guo Y J,et al.Characterization of soil bacterial community structures in rhizosphere of tomatoes infected with bacterial wilt and its non-infected plants[J].Chinese Vegetables,2014,(8):25-29.
[21]朱丽霞,章家恩,刘文高.根系分泌物与根际微生物相互作用研究综述[J].生态环境,2003,(1):102-105.Zhu L X,Zhang J E,Liu W G.Review of interactions between root exudates and rhizosphere microorganisms[J].Ecological Environment,2003,(1):102-105.
[22]钟文辉,蔡祖聪.土壤管理措施及环境因素对土壤微生物多样性影响研究进展[J].生物多样性,2004,12(4):456-465.Zhong W H,Cai Z C.Effect of soil management practices and environmental factors on soil microbial diversity:a review[J].Biodiversity Science,2004,12(4):456-465.
[23]杨宁,杨满元,雷玉兰,等.衡阳紫色土丘陵坡地土壤酶活性对植被恢复的响应[J].生态环境学报,2014,23(4):575-580.Yang N,Yang M Y,Lei Y L,et al.Response of soil enzyme activities to re-vegetation on sloping-land with purple soils in Hengyang of Hunan province[J].Ecology and Environmental Sciences,2014,23(4):575-580.
[24]Sowerb Y A,Emmet T B,Beie R C,et al.Microbial community changes in health and soil communities along a geographical gradient:Interaction with climate change manipulations[J].Soil Biology and Biochemistry,2005,37:1805-1813.
[25]Ahamadou B,Huang Q,Chen W,et al.Microcalorimetric assessment of microbial activity in long-term fertilization experimental soils of Southern China[J].Fems Microbiology Ecology,2009,70(2):186-195.
[26]Powlson D S,Brookes P C,Christensen B T.Measurement of soil microbial biomass provides an early indication of changes in total soil organic matter due to straw incorporation[J].Soil Biology&Biochemistry,1987,19(2):159-164.