添加菌剂制作堆肥的肥效及菌群多样性分析

doi:10.11924/j.issn.1000-6850.casb14100027

摘要/Abstract

摘要： 为了加速低温地区农牧废弃物转化为有机肥以用于培肥地力，筛选一组能够在14天内转化稻杆重量36%的菌剂。利用气质联用检测挥发性产物，发现其中含有L-乳酸、甲酸、乙酸和丙酸等多种能够培肥地力的有机质。在东北低温条件下，添加该菌剂制作堆肥与不添加菌剂的堆肥相比，添加菌剂堆温最高可达63.0℃，55.0℃以上的高温期为7天，比未添加菌剂的堆肥腐熟时间缩短17天。发酵57天后发现添加菌剂与不添加菌剂堆肥样品中可溶性糖含量分别为11.80和8.92 mg/L，C/N分别为12.65和13.69。添加菌剂培育的番茄种子和番茄幼苗长势也明显优于不添加菌剂的对照组。454高通量技术检测原菌剂与添加该菌剂制作的堆肥样品的菌群多样性，发现Bacteroides sp.、Firmicutes sp.和Clostridium sp.在2个菌群中都属于优势种，但添加菌剂制作的堆肥，其菌群多样性更加丰富。试验证明，添加菌剂制作堆肥不仅可缩短有机肥堆置时间还可以增加肥效。

关键词: 纤维亚麻, 纤维亚麻, 盐胁迫, 半定量RT-PCR, NHX基因

Abstract: In order to accelerate agriculture and animal waste transform into organic fertilizer to increase soil fertility, a stable cellulose decomposing bacterial community was screened out which could decompose 36% of rice straw within 14 days. Many kinds of volatile products (L-lactic acid, formic acid, acetic acid and propionic acid) were detected by GCMS, most of them were advantageous to increase soil organic matter. In the northeast of China, two composts were piled up and the bacteria agent was added to only one of the composts. We found that the highest temperature was 63.0℃, temperature over 55.0℃ maintained for 7 days in compost added with bacteria agent. In addition, time of thoroughly decomposition of the compost added with bacteria agent was 17 days shorter than the compost without bacteria agent. After piled for 57 days, in compost with and without bacteria agent, the soluble sugar was 11.80 and 8.92 mg/L, respectively, and C/N was 12.65 and 13.69, respectively. The 454 pyrosequencing technology was used to survey the whole bacterial community. The predominant strains belonged to three genera: Bacteroides sp., Firmicutes sp. and Clostridium sp.. But the bacterial diversity was much abundant of the added with bacteria agent. The resulted showed that the compost added with bacteria agent can not only cut down organic fertilizer’s stacking time but also increase its fertilizer efficiency.

刘长莉,李娜,王宝鑫,李春玲,齐英杰. 添加菌剂制作堆肥的肥效及菌群多样性分析[J]. 中国农学通报, 2015, 31(8): 145-153.

Liu Changli,Li Na,Wang Baoxin,Li Chunling and Qi Yingjie. Analysis on Fertilizer Efficiency and Microbial Diversity of Compost with Bacterium Agent[J]. Chinese Agricultural Science Bulletin, 2015, 31(8): 145-153.

参考文献

[1] Vargas-gare M D C, Rez-estrella F F S, Pemjose L, et al. Influence of microbialinoeulation and co-composting material on the evolution of humic-like substanees during composting of horticultural wastes[J].Process Biochemistry,2006,41(2):1438-1443.
[2] 张陇利,刘青,徐智,等.复合微生物菌剂对污泥堆肥的作用效果研究[J].环境工程学报,2008,2(2):266-269.
[3] 籍宝霞,张丽萍,程辉彩,等.低温发酵剂在鸡粪冬季处理中的作用[J].江苏农业科学,2009(6):418-420.
[4] Yu H. Microbial community succession and lignocellulose degradation during agricultural waste composting[J].2007,18(6):793-802.
[5] 王伟东,王小芬,刘长莉,等.木质纤维素分解菌复合系WSC-6分解稻秆过程中的产物及pH动态[J].环境科学,2008,29(1):219-224.
[6] 鲁如坤.土壤农业化学分析方法[M].北京:中国农业科学出版社,2000:146-166,289-295.
[7] Galan M, Guivier E, Caraux G, et al. A 454 multiplex sequencing method for rapid and reliable genotyping of highly polymorphic genes in large-scale studies [J].BMC Genomics,2010,11(1):296.
[8] Jutta G, Carsten N. Isolation and characterisation of new microsatellite markers for the stonefly Brachyptera braueri comparing a traditional approach with high throughput 454 sequencing [J].Conservation Genetics Resources,2013,5(2):413-416.
[9] 高洁,汤烈贵.高分子化学.纤维素科学[M].北京:北京科学出版社,1996:238-240.
[10] 刘稳,高培基.半纤维素酶的分子生物学[J].纤维素科学与技术,1998,6( 1):9-15.
[11] Johnston T M. Effect of fatty acid mixtures on the rice styles nematode (Tylenchorhynchus martini Fielding)[J].Nature,1959,183· (4672):1392.
[12] Jatala P. Biological control of plant-parasitic nematodes[J].Annual Review of Phytophath,1986,24:453-489.
[13] Wang J J, Tang Z H. The Regulation of Soluble Sugars in the Growth and Development of Plants[J].Botanical Research,2014,DOI:10.12677/BR.2014.33011.
[14] Plaza C, Hernndez D, Garca , et al. Microbial activity in pig slurry amended soils under semiarid conditions[J].Soil Biology and Biochemistry,2004,36(10):1577-1585.
[15] Kautz T, Wirth S, Ellmer F. Microbial activity in a sandy arable soil is governed by the fertilization regime[J].European Journal of Soil Biology,2004,40(2):87-94 .
[16] 徐阳春,沈其荣,冉伟.长期免耕与施用有机肥对土壤微生物生物量碳、氮、磷的影响[J].土壤学报,2002,39(1):89-96.
[17] Günther S, Holger K. Bulk soil C to N ratio as a simple measure of net N mineralization from stabilized soil organic matter in sandy arable soils[J].Soil Biology and Biochemistry,2003,35(4):629-632.
[18] 胡启武,欧阳华,刘贤德.祁连山北坡垂直带土壤碳氮分布特征[J].山地学报,2006,24(6):654-661.
[19] 葛晓光,新编蔬菜育苗大全[M].北京:中国农业出版社,2004:88-100.
[20] Pandey S, Singh S, Yadav A N, et al. Phylogenetic diversity and characterization of novel and efficient cellulase producing bacterial isolates from various extreme environments[J].Bioscience,biotechnology,and biochemistry,2013,77(7):1474-1480.
[21] Zverlov V, Mahr S, Riedel K, et al. Properties and gene structure of a bifunctional cellulolytic enzyme (CelA) from the extreme thermophile ‘Anaerocellum thermophilum’ with separate glycosyl hydrolase family 9 and 48 catalytic domains[J].Microbiology,1998,144(2):457-465.
[22] Riedel K, Bronnenmeier K. Intramolecular synergism in an engineered exo-endo-1, 4-β-glucanase fusion protein[J].Molecular Microbiology,1998,28(4):767-775.
[23] Zverlov V V, Kellermann J, Schwarz W H. Functional subgenomics of Clostridium thermocellum cellulosomal genes:identification of the major catalytic components in the extracellular complex and detection of three new enzymes[J].PROTEOMICS,2005,5(14):3646-3653.
[24] Gold N D, Martin V J. Global view of the Clostridium thermocellum cellulosome revealed by quantitative proteomic analysis[J].Journal of Bacteriology,2007,189(19):6787-6795.
[25] Blanc M, Marilley L, Beffa T, et al. Thermophilic bacterial communities in hot composts as revealed by most probable number counts and molecular (16S rDNA) methods[J].FEMS Microbiology Ecology,1999,28(2):141-149.
[26] Pantazaki A, Pritsa A, Kyriakidis D. Biotechnologically relevant enzymes from Thermus thermophilus[J].Applied Microbiology and Biotechnology,2002,58(1):1-12.
[27] Tang J, Kanamori T, Inoue Y, et al. Changes in the microbial community structure during thermophilic composting of manure as detected by the quinone profile method[J].Process Biochemistry,2004,39(12):1999-2006.
[28] Watanabe A, Katoh K, Kimura M. Effect of Rice StrawApplicationon CH4 Emission from Paddy Field[J].Soil Sci PlantNutr.1993,39(4):707-712.
[29] Yabe S, Aiba Y, Sakai Y, et al. Sphingobacterium thermophilum sp. nov.,within the phylum Bacteroidetes isolated from compost[J].International Journal of Systematic and Evolutionary Microbiology,2012,50:20-24.
[30] Yang D C, Im W T, Kim M K, et al. Pseudoxanthomonas koreensis sp. nov. and Pseudoxanthomonas daejeonensis sp. nov.[J].International Journal of Systematic and Evolutionary Microbiology,2005,55(2):787-791.
[31] Suihko M L, Eija S. Characterisation of aerobically grown non-spore-forming bacteria from paper mill pulps containing recycled fibres[J].Journal of Industrial Microbiology & Biotechnology,2009,36(1):53-64.
[32] 乔江涛,郭荣波,袁宪正,等.玉米秸秆厌氧降解复合菌系的微生物群落结构[J].环境科学,2013,04:1531-1539.
[33] 赵银瓶.厌氧纤维素降解复合菌系群落结构及功能微生物研究[D].北京:中国农业科学院,2012:30-34.
[34] Murashima K A, Kosugi D R H. Synergistic effects of cellulosomal xylanase and cellulases from Clostridium cellulovorans on plant cell wall degradation[J].Journal of Bacteriology,2003,185(5):1518-1524.
[35] Koukiekolo R, Cho H Y, Kosugi A, et al. Degradation of corn fiber by Clostridium cellulovorans cellulases and hemicellulases and contribution of scaffolding protein CbpA[J].Applied and Environmental Microbiology,2005,71(7):3504-3511.
[36] Kato S, Haruta S, Cui Z J, et al. Effective cellulose degradation by a mixed-culture system composed of a cellulolytic Clostridium and aerobic non-cellulolytic bacteria[J].FEMS Microbiology Ecology,2004,51(1):133-142.
[37] Hatsumi S, Hironori I, Shohei A, et al. Isolation and characterization of a new Clostridium sp. that performs effective cellulosic waste digestion in a thermophilic methanogenic bioreactor[J].Applied and Environmental Microbiology,2006,72(5):3702-3709.