摘要
【目的】探究番茄秸秆与玉米秸秆共堆肥发酵过程中的关键微生物及其功能,以进一步加速其发酵进程。【方法】以番茄秸秆和玉米秸秆为原料,二者按质量比3∶1混合进行条垛式堆肥,每周采集发酵堆20~50 cm深度样品检测木质纤维素酶活力的变化,以酶活力最高时期的代表性样品为研究对象,对其进行高通量焦磷酸测序和Orbitrap宏蛋白质组学研究。【结果】多数真菌隶属于子囊菌门Ascomycota,其中嗜热丝孢菌Thermomyces的相对丰度最高,约占真菌全部序列的70.5%,该属真菌主要分泌内切-1,4-β-木聚糖酶,是主要的半纤维素降解真菌。主要细菌为放线菌门Actinobacteria、变形菌门Proteobacteria和厚壁菌门Firmicutes,三者共占细菌总序列数的87.0%。其中,嗜热裂孢菌属Thermobifida和糖单孢菌属Saccharomonospora是主要的放线菌属,其相对丰度分别占全部细菌序列的16.5%和1.36%。Thermobifida是唯一的纤维素降解菌,鉴定到4种内切纤维素酶组分和3种外切纤维素酶组分;同时产生一种果胶裂解酶和一种β-木聚糖酶。Saccharomonospora分别产生一种β-木聚糖酶、两种丝氨酸蛋白酶和两种胰蛋白酶,在半纤维素的降解和蛋白质降解方面发挥重要作用。Idiomarina是变形菌门下的主要细菌属,平均相对丰度15.6%,主要参与蛋白质的降解。而厚壁菌门下的清野氏菌属Planifilum虽基因丰度不高(仅为1.23%),但却在半纤维素的降解过程中发挥重要作用。【结论】将高通量焦磷酸测序和Orbitrap宏蛋白质组学结合的整合宏组学方法在探明复杂生境中关键微生物的群落结构及功能方面具有可行性。
【Objective】The integrated meta-omics was applied to reveal the functional microorganisms in the tomato stalks/maize straws compost.【Method】Sampling from the pile of a compost of tomato stalks and maize straws in the ratio of3:1(m:m)at the height of 20–50 cm was carried out once every week for lignocellulosic enzyme assay.When a peak enzymatic activity appeared,specimens were collected and the integrated meta-omic analysis that combined high-throughput pyrosequencing and Orbitrap studies was conducted.【Result】The greatest population of fungi in the compost belonged to the phylum Ascomycota,in which,Thermomyces was the most abundant accounting for 70.5%.Thermomyces is known to secret endo-1,4-β-xylanase that degrades hemicellulose in biomass.Of the total bacteria population,Actinobacteria,Proteobacteria,and Firmicutes phyla made up 87.0%,and the two genera of Actinobacteria,Thermobifida and Saccharomonospora,accounted for16.5%and 1.36%,respectively.Thermobifida is capable of degrading cellulose with its 4 endoglucanases and 3 cellobiohydrolases.It also secrets enzymes associated with hemicellulose and pectate degradation.Saccharomonospora produces aβ-xylanase,2 serine proteases and 2 trypsins involved in the decomposition of hemicelluloses and proteins.Idiomarina was the major genus of Proteobacteria found in the compost with a relative abundance of 15.6%.It involves in protein degradation.Although low in abundance(merely 1.23%),Planifilum of Firmicutes phylum plays an import role in degrading hemicelluloses.【Conclusion】The integrated meta-omics that combined pyrosequencing and Orbitrap analysis provided a new tool for studying the community structure and functions of the microorganisms in a complex habitat such as a compost of different bio-materials.With the information obtained,ways to accelerate the composting process could be explored.
作者
朱屹
李俊良
焦博
朱倩倩
张小梅
ZHU Yi;LI Junliang;JIAO Bo;ZHU Qianqian;ZHANG Xiaomei(College of Resource and Environment,Qingdao Agricultural University,Qingdao,Shandong266109,China)
出处
《福建农业学报》
CAS
CSCD
北大核心
2020年第7期764-772,共9页
Fujian Journal of Agricultural Sciences
基金
国家重点研发计划项目(2017YFD0800403)
国家自然科学基金项目(31600084)
山东省重大科技创新工程项目(2019JZZY010721)
关键词
整合宏组学
番茄秸秆
功能微生物
嗜热丝孢菌
Integrated meta-omics
tomato stalk
functional microorganism
Thermomyces
