溴甲烷熏蒸对土壤反硝化作用及nosZ型反硝化微生物群落结构的影响

Effects of methyl bromide fumigation on community structure of denitrifying bacteria with nitrousoxide reductase gene (nosZ) in soil

【目的】探究溴甲烷熏蒸对土壤反硝化作用的影响及机制。【方法】本研究采用nos Z-PCR-RFLP(Restriction fragment length polymorphism)方法、nos Z-MPN-PCR(Most-Probable-Number-PCR)计数法和土壤硝酸根的消灭率方法研究溴甲烷熏蒸对土壤nos Z型反硝化细菌群落结构、数量及反硝化活性的影响。【结果】实验结果说明溴甲烷熏蒸剂熏蒸土壤100 d土壤的反硝化作用与对照无显著差异(P>0.05)。溴甲烷熏蒸土壤和对照土样nos Z型反硝化细菌群落的Margalef指数,Shannon-Wiener和Evenness指数无显著差异(P>0.05)。溴甲烷熏蒸土壤和对照土壤中存在Uncultured bacterium partial rhodopsendomonas、pseudomonas fluorescens、Herbacspirillum、Mesorhizobium和Bradyrhizobium,并且此6种微生物均是试验土样和对照土样的优势种群;对照土壤中检测到Azospirillum、Rhizobium melibei和Nitrosospira multiformis,在溴甲烷熏蒸土壤中未检测到;而溴甲烷熏蒸土壤中检测到Uncultured Azospirillum,Mesorhizobium在对照土壤中未检测到。通过nos Z-MPN-PCR计数法得出反硝化细菌数量比对照低1.4倍。【结论】说明溴甲烷熏蒸100d土壤的nos Z型反硝化微生物群落组成和反硝化细菌数量发生变化,而土壤的反硝化作用与对照无显著差异。

[ Objective ] We studied the effect of methyl bromide fumigation on soil edaphic denitrification. [ Methods ] We adopted nosZ-PCR-RFLP （restriction fragment length polymorphism） method, nosZ-MPN-PCR （Most-Probable-Number- PCR） counting method and soil nitrate elimination rate method, to explore the effect of methyl bromide fumigation on community structure, quantity and activity of denitrifying bacteria in soil. [ Result] After methyl bromide fumigating soil for 100 d, soil denitrification did not change obviously （P 〉 0. 05）. Margalef index, Shannon-wiener index and Evenness index had no significant difference （P 〉 0. 05） in nosZ denitrifying bacterial communities between fumigated soil and the control. There were Rhodopsendomonas, Pseudomonas fluorescens, Herbacspirillum, uncultured bacterium partial in both of them. However, Azospirillum, Rhizobium melibei, Nitrosospira multiformis were exclusively found in the control, and Uncultured Azospirillum sp, Mesorhizobium sp were in fumigated one. Moreover, the number of denitrifying bacteria in the control resolved by nosZ-MPN-PCR （Most-Probable-Number-PCR） was 1.4 times higher than that of the fumigated one. [ Conclusion] After 100 d fumigating soil, the composition of nosZ denitrifying microbial community and the population of denitrifying bacteria changed. Furthermore, there was no difference in denitrification between the fumigated soil and the control.