两种配方废弃菌棒堆肥中微生物多样性研究
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摘要
以废弃菌棒为主要原料,添加牛粪和草炭,进行堆肥发酵,取腐熟完成后的2种肥料样品DNA,采用16SrDNA V3+V4区和ITS2高通量测序方法,研究不同配方粗制肥料中微生物多样性和群落结构的差异。结果表明,"牛粪+菌棒"制得的有机肥生物多样性高于"草炭+菌棒",且真菌差异大于细菌;"牛粪+菌棒"有机肥相对"草炭+菌棒"有机肥具有10个优势细菌属和6个真菌属,"草炭+菌棒"有机肥相对"牛粪+菌棒"有机肥具有14个优势细菌属和4个真菌属。两种配方堆肥制得的有机肥微生物群落结构无显著性差异。研究发现了多种具有农业价值的功能菌,可为功能性微生物的开发利用提供科学依据。此外,也发现一些潜在危害因子,如黄曲霉和杂色曲霉亚种在"牛粪+菌棒"有机肥中丰度较高,产生的黄曲霉毒素、杂曲霉毒素易造成农作物污染。
Based on the high-throughput DNA sequencing of Illumina,the microbial community composition of the compost ofmushroom residue added with cow dung and turf as the nitrogen source was entirely clarified,respectively. The results showed that the biodiversity of organic manure produced by "cow dung + mushroom residue" was higher than that of "turf + mushroom residue" ,and the fungal difference was greater than that of bacteria; compared with the compost of "turf + mushroom residue" ,the compost of "cow dung + mushroom residue" had 10 dominant bacteria and six dominant fungi at genus level,at the same time,the compost of "turf + mushroom residue" had 14 dominant genus and four dominant fungal genus relative to "cattle manure + fungus" . Therefore,the microbial communities had no significant difference between two formula composts. At the same time,it was found that a variety of functional bacteria with agricultural value can provide a scientific basis for the development and utilization of functional microorganisms in the future,such as Chaetomium. In addition,some potential hazard factors,such as Aspergillus_ flavus and Aspergillus_subversicolor were found to be highly abundant in the "cow dung + mushroom residue" organic fertilizer,which produced the toxin was likely to cause crop pollution. Agricultural producers and related management departments should pay attention to the risks.
Based on the high-throughput DNA sequencing of Illumina,the microbial community composition of the compost ofmushroom residue added with cow dung and turf as the nitrogen source was entirely clarified,respectively. The results showed that the biodiversity of organic manure produced by "cow dung + mushroom residue" was higher than that of "turf + mushroom residue" ,and the fungal difference was greater than that of bacteria; compared with the compost of "turf + mushroom residue" ,the compost of "cow dung + mushroom residue" had 10 dominant bacteria and six dominant fungi at genus level,at the same time,the compost of "turf + mushroom residue" had 14 dominant genus and four dominant fungal genus relative to "cattle manure + fungus" . Therefore,the microbial communities had no significant difference between two formula composts. At the same time,it was found that a variety of functional bacteria with agricultural value can provide a scientific basis for the development and utilization of functional microorganisms in the future,such as Chaetomium. In addition,some potential hazard factors,such as Aspergillus_ flavus and Aspergillus_subversicolor were found to be highly abundant in the "cow dung + mushroom residue" organic fertilizer,which produced the toxin was likely to cause crop pollution. Agricultural producers and related management departments should pay attention to the risks.
引文
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[14] TAN S Y,JIANG Y,SONG S,et al. Two Bacillus Myloliquefaciensstrains isolated using the competitive tomato root enrichment method and their effects on suppressing ralstonia solanacearum and promoting tomato plant growth[J]. Crop Prot,2013,43:134-140.
[15] OLUWASEYI S O. Streptomyces:implications and interactions in plant growth promotion[J]. Applied Microbiology and Biotechnology,2019,103(3):1179-1188.
[16] WANG J,LIANG J,GAO S. Biodegradation of lignin monomers vanillic,p-Coumaric,and Syringic Acid by the bacterial strain,Sphingobacterium sp. HY-H[J]. Current Microbiology,2018,75(9):1156-1164.
[17] STORY S P,KLINE E L,HUGHES T A,et al. Degradation of aromatic hydrocarbons by Sphingomonas paucimobilis strain EPA505[J]. Arch. Environ. Contam. Toxicol.,2004,47:168-176.
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[19] RYCKEBOER J R,MERGAERT J,VAES K,et al. A survey of bacteria and fungi occurring during composting and selfheating processes[J]. Annals of Microbiology,2003,53(4):349-410.
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[21] SHAUKAT S,BAJWA M A,WAQAR M A,et al. Production,optimization,purification and characterization of glucomalyase from Arachniotus sp[J]. Chem. Soc. Pak.,2006,28:368-373.
[22]张初署.中国四个生态区花生土壤中黄曲霉菌分布、产毒特征及遗传多样性研究[D].北京:中国农业科学院,2013.ZHANG Chushu. Study on the distribution,aflatoxin production and genetic diversity of Aspergillus flavus in soils of peanut fields in four agroecological zones of China[D]. Beijing:Chinese Academy of Agricultural Sciences,2013.(in Chinese)
[23] JURJEVIE Z,PETERSON S W,SOLFRIZZO M,et al. Sterigmatocystin production by nine newly described Aspergillus species in section Versicolores grown on two different media[J]. Mycotoxin Research,2013,29(3):141-145.
[24] YUE Haimei,WANG Mu,GONG Wenfeng,et al. The screening and identification of the biological control fungi Chaetomium spp against wheat common root rot[J]. FEMS Microbiol. Letter,2018,365(22). http:∥doi. org/10. 1093/femsle/fny242.
[2]吕雄华,王芳.我国食用菌出口结构对其贸易竞争力指数的灰色动态模型分析[J].广东农业科学,2017,44(4):159-162.Lü Xionghua,WANG Fang. Analysis of China's mushroom export structure versus its trade competitiveness index with grey dynamic models[J]. Guangdong Agricultural Science,2017,44(4):159-162.(in Chinese)
[3]张平,郑志安,赵祖松颖.我国食用菌产业发展变化及对策分析[J].北方园艺,2017(22):167-174.ZHANG Ping,ZHENG Zhian,ZHAO Zusongying,et al. Development and countermeasures of edible fungi industry in China[J].Norther Horticulture,2017(22):167-174.(in Chinese)
[4]邓媛方.蘑菇废弃菌棒和猪粪混合厌氧发酵特性研究[D].杨凌:西北农林科技大学,2012.DENG Yuanfang. Research on anaerobic co-digestion of mushroom cultivation wastes and pig manure[D]. Yangling:Northwest A&F University,2012.(in Chinese)
[5]熊小兴,王飞,李小毛.菇渣发酵有机肥在小白菜上的应用试验[J].江西农业学报,2009,21(7):100-101.XIONG Xiaoxing,WANG Fei,LI Xiaomao. Application of fermented mushroom dregsin pakchoi[J]. Acta Agriculturae Jiangxi,2009,21(7):100-101.(in Chinese)
[6]徐剑波,胡庆存.香菇废弃菌棒作葡萄园基肥施用效果研究[J].现代农业科技,2014(6):236-237.XU Jianbo,HU Qingcun. Study on the effect of applying mushroom waste stick to the base fertilizer of vineyard[J]. Modern Agricultural Technology,2014(6):236-237.(in Chinese)
[7]朱琳,曾椿淋,高凤,等.水稻秸秆堆肥制备粗制肥料中微生物多样性研究[J/OL].农业机械学报,2018,49(7):228-234.ZHU Lin,ZENG Chunlin,GAO Feng,et al. Characteristic analysis of microbial diversity in crud fertilizer form compost of rice straw[J/OL]. Transactions of the Chinese Society for Agricultural Machinery,2018,49(7):228-234. http:∥www. j-csam.org/jcsam/ch/reader/view_abstract. aspx? file_no=20180727&flag=1. DOI:10. 6041/j. issn. 1000-1298. 2018. 07. 027.(in Chinese)
[8]陈竞,代金平,冯蕾,等.几种配方制备有机肥菌群分析及其育苗效果评价[J].新疆农业科学,2017,54(6):1099-1107.CHEN Jing,DAI Jinping,FENG Lei,et al. Preliminary analysis of several kinds of formulated organic fertilizer flora and their effects evaluation[J]. Xinjiang Agricultural Sciences,2017,54(6):1099-1107.(in Chinese)
[9] LIU Ling,WANG Shuqi,GUO Xiaoping,et al. Succession and diversity of microorganisms and their association with physicochemical properties during green waste thermophilic composting[J]. Waste Management,2018,73:101-112.
[10] SHOKRALLA S,SPALL J L,GIBSON J F,et al. Next-generation sequencing technologies for environmental DNA research[J]. Molecular Ecology,2012,21(8):1794-1805.
[11]丁建莉,姜昕,关大伟,等.东北黑土微生物群落对长期施肥及作物的响应[J].中国农业科学,2016,49(22):4408-4418.DING Jianli,JIANG Xin,GUAN Dawei,et al. Responses of micropopulation in black soil of northeast China to long-term fertilization and crops[J]. Scientia Agricultura Sinica,2016,49(22):4408-4418.(in Chinese)
[12] JOICE D C,DEBORA C C,GILDA A F. Subgingival microbiota dysbiosis in systemic lupus erythematosus:association with periodontal status[J]. Microbiome,2017,5(1):34.
[13]林先贵,冯有智.潮土农田微生物研究进展[J].中国生态农业学报,2016,24(4):416-434.LIN Xiangui,FENG Youzhi. Research progresses of farmland microorganisms in fluvo-aquic soil of China[J]. Chinese Journal of Eco-Agriculture,2016,24(4):416-434.(in Chinese)
[14] TAN S Y,JIANG Y,SONG S,et al. Two Bacillus Myloliquefaciensstrains isolated using the competitive tomato root enrichment method and their effects on suppressing ralstonia solanacearum and promoting tomato plant growth[J]. Crop Prot,2013,43:134-140.
[15] OLUWASEYI S O. Streptomyces:implications and interactions in plant growth promotion[J]. Applied Microbiology and Biotechnology,2019,103(3):1179-1188.
[16] WANG J,LIANG J,GAO S. Biodegradation of lignin monomers vanillic,p-Coumaric,and Syringic Acid by the bacterial strain,Sphingobacterium sp. HY-H[J]. Current Microbiology,2018,75(9):1156-1164.
[17] STORY S P,KLINE E L,HUGHES T A,et al. Degradation of aromatic hydrocarbons by Sphingomonas paucimobilis strain EPA505[J]. Arch. Environ. Contam. Toxicol.,2004,47:168-176.
[18] HARTMANN M,FREY B,MAYER J,et al. Distinct soil microbial diversity under long-term organic and conventional farming[J]. ISME Journal,2015,9(5):1177-1194.
[19] RYCKEBOER J R,MERGAERT J,VAES K,et al. A survey of bacteria and fungi occurring during composting and selfheating processes[J]. Annals of Microbiology,2003,53(4):349-410.
[20] WATANABE K,NAGAO N,YAMAMOTO S,et al. Thermobacillus composti sp. nov,a moderately thermophilic bacterium isolated from a composting reactor[J]. Int. J. Syst. Evol. Microbiol.,2007,57(7):1473-1477.
[21] SHAUKAT S,BAJWA M A,WAQAR M A,et al. Production,optimization,purification and characterization of glucomalyase from Arachniotus sp[J]. Chem. Soc. Pak.,2006,28:368-373.
[22]张初署.中国四个生态区花生土壤中黄曲霉菌分布、产毒特征及遗传多样性研究[D].北京:中国农业科学院,2013.ZHANG Chushu. Study on the distribution,aflatoxin production and genetic diversity of Aspergillus flavus in soils of peanut fields in four agroecological zones of China[D]. Beijing:Chinese Academy of Agricultural Sciences,2013.(in Chinese)
[23] JURJEVIE Z,PETERSON S W,SOLFRIZZO M,et al. Sterigmatocystin production by nine newly described Aspergillus species in section Versicolores grown on two different media[J]. Mycotoxin Research,2013,29(3):141-145.
[24] YUE Haimei,WANG Mu,GONG Wenfeng,et al. The screening and identification of the biological control fungi Chaetomium spp against wheat common root rot[J]. FEMS Microbiol. Letter,2018,365(22). http:∥doi. org/10. 1093/femsle/fny242.
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