利用高通量测序技术对水稻秸秆中、低温降解菌系的比较分析

采用Illumina MiSeq高通量测序技术,对中、低温(中温25℃、低温10℃)富集驯化所得水稻秸秆降解菌系进行测序;采用生物信息学方法对中、低温秸秆降解菌系群落生物多样性进行分析。结果表明,测序质控后获得601489条16S rDNA序列,平均长度273 bp,经Silval 132数据库比对,中温降解菌系包括18个门,172个属,302个OTU,其中优势菌属为弓形杆菌属(Arcobacter)、拟杆菌属(Bacteroides)、屠场杆菌属(Macellibacteroides)、假单胞菌属(Pseudomonas)和梭形杆菌属(Lysinibacillus);低温降解菌系包括16个门,169个属,280个OTU,其中优势菌属为弓形杆菌属、屠场杆菌属、拟杆菌属、丛毛单胞属(Comamonas)和假单胞菌属。温度对降解菌系中优势菌门的相对丰度无显著影响,中、低温秸秆降解菌系主要菌属相对丰度差异显著。

Soil Microbial Characteristics Under Long-Term Heavy Metal Stress:A Case Study in Zhangshi Wastewater Irrigation Area,Shenyang

Soil samples were collected from Zhangshi Wastewater Irrigation Area in the suburb of Shenyang City,China,an area with a 30-year irrigation history with heavy metal-containing wastewater.The chemical properties and microbial characteristics of the soils were examined to evaluate the present situation of heavy metal pollution and to assess the soil microbial characteristics under long-term heavy metal stress.In light of the National Environmental Quality Standards of China,the soil in the test area was heavily polluted by Cd and to a lesser degree by Zn and Cu,even though wastewater irrigation ceased in 1993.Soil metabolic quotient (qCO_2) had a significant positive correlation,while soil microbial quotient (qM) had a negative correlation with content of soil heavy metals.Soil microbial biomass carbon (MBC) had significantly negative correlation with Cd,but soil substrate-induced respiration (SIR),dehydrogenase activity (DHA),cellulase activity, and culturable microbial populations had no persistent correlations with soil heavy metal content.Soil nutrients,except for phosphorous,showed positive effects on soil microbial characteristics,which to a certain degree obscured the adverse effects of soil heavy metals.Soil Cd contributed more to the soil microbial characteristics,but qM and qCO_2 were more sensitive and showed persistent responses to heavy metals stress.It could be concluded that qM and qCO_2 can be used as bioindicators of heavy metal pollution in soils.

Toxic effects of acetochlor, methamidophos and their combination on nifH gene in soil

Toxic effects of two agrochemicals on nifH gene in agricultural black soil were investigated using denaturing gradient gel electrophoresis （DGGE） and sequencing approaches in a microcosm experiment. Changes of soil nifH gene diversity and composition were examined following the application of acetochlor, methamidophos and their combination. Acetochlor reduced the nifH gene diversity （both in gene richness and diversity index values） and caused changes in the nifH gene composition. The effects of acetochlor on nifH gene were strengthened as the concentration of acetochlor increased. Cluster analysis of DGGE banding patterns showed that nifH gene composition which had been affected by low concentration of acetochlor （50 mg/kg） recovered firstly. Methamidophos reduced nifH gene richness that except at 4 weeks. The medium concentration of methamidophos （150 mg/kg） caused the most apparent changes in nifH gene diversity at the first week while the high concentration of methamidophos （250 mg/kg） produced prominent effects on nifH gene diversity in the following weeks. Cluster analysis showed that minimal changes of nifH gene composition were found at 1 week and maximal changes at 4 weeks. Toxic effects of acetochlor and methamidophos combination on nifH gene were also apparent. Different nifH genes （bands） responded differently to the impact of agrochemicals： four individual bands were eliminated by the application of the agrochemicals, five bands became predominant by the stimulation of the agrochemicals, and four bands showed strong resistance to the influence of the agrochemicals. Fifteen prominent bands were partially sequenced, yielding 15 different nifH sequences, which were used for phylogenetic reconstructions. All sequences were affiliated with the alpha- and beta-proteobacteria, showing higher similarity to eight different diazotrophic genera.

Effects of Acetochlor and Methamidophos on Fungal Communities in Black Soils

Using plate counting and ergosterol assay, single and joint effects of acetochlor and methamidophos on the dynamicsof soil fungal population and total fungal biomass in the black soil zone of Northeast China were investigated. The resultsdemonstrated that acetochlor at high concentration levels (150 and 250 mg kg-1) had an acute and mostly chronic toxicityon both the soil fungal population and total fungal biomass, but at a low concentration (50 mg kg-1) generally had astimulating effect that was stronger with total fungal biomass than with the soil fungal population. Methamidophos ata high concentration level (250 mg kg-1) alone and almost all of its combinations with various dosages of acetochlorincreased the soil fungal population, whereas at most sampling dates with 250 mg methamidophos kg-1 soil, total fungalbiomass increased, but in combination with acetochlor it was decreased in the early period of incubation and then increased28 days after incubation. Thus, through measuring the number of colony forming unit of the soil fungal population alongwith the total fungal biomass, a better understanding on effects of agrochemicals on soil fungi could be made.

降解菌系和助腐剂对不同还田方式下水稻秸秆降解特征的影响

基于田间小区试验,设置两种秸秆还田处理(以秸秆混土处理模拟旋耕还田,以秸秆不混土处理模拟秸秆沟埋还田),采用尼龙网袋法,通过测定秸秆降解率及秸秆半纤维素、纤维素和木质素的含量,研究两种还田方式下施用降解菌系和助腐剂后水稻秸秆的降解规律。结果表明:在试验前5个月,不混土处理的秸秆降解率始终高于混土处理4.5%~11.6%;至第12个月,混土处理秸秆降解率显著高于不混土处理4.9%~13.0%。在前2个月,各处理水稻秸秆降解较快,秸秆降解率均超过50%,此后秸秆降解进入缓慢期。混土处理中,降解菌系和助腐剂仅能在第1个月提高秸秆的降解效率,此时秸秆降解率分别比对照组高11.0%和10.2%(P<0.01);不混土处理中,降解菌系在1~5个月内显著提高秸秆降解效率4.3%~9.7%,助腐剂在1~12个月显著提高秸秆降解效率7.3%~14.4%。试验至第5个月时,喷施助腐剂的秸秆半纤维素、纤维素和木质素的降解率分别达79.1%、80.2%和34.7%,且与喷施菌系处理差异不显著。综上所述,秸秆降解菌系和秸秆助腐剂均能提高水稻秸秆的降解效率,且不混土处理的施用效果好于混土处理。