生物炭不同施用量对烟草内生细菌多样性的影响

Effect of Different Biochar Application Rates on Diversity of Tobacco Endophytic Bacteria

【目的】为了对施用生物炭后的烟叶细菌的种类变化进行全面而准确的调查。【方法】以烟叶为研究对象,设置4个处理:处1理为施用生物炭50 g/株(Y4.1);处理2为施用生物炭100 g/株(Y5.1);处理3为施生物炭150 g/株(Y6.1)和处理4不施用生物炭作为对照(Y1.1)。应用Illumina测序平台的Mi Seq高通量测序仪对施用生物炭后烟叶内生细菌的16S r DNA-V4区扩增子进行测序。并应用Qiime和Mothur等软件整理和统计样品序列数目和操作分类单元(OTUs)数量,分析物种的丰度和Alpha多样性。【结果】在97%的序列相似性水平上把所测得的序列划分为1177、1429、1228、1182操作分类单元(OTUs)。通过OTU丰度聚类分析表明:施用生物炭处理后样品的OTUs数低于不施用生物炭处理,说明施用生物炭能提高烟叶细菌群落丰度;施用生物炭50g/株处理后样品的OTUs数高于施用生物炭100 g/株处理,施用生物炭100 g/株处理后样品的OTUs数高于施用生物炭150 g/株处理,说明随着施炭量增加到100、150 g/株细菌群落丰度又开始减少。【结论】通过属的水平分析表明:鞘氨醇单胞菌属(Sphingomonas)在施用生物炭处理的样品中丰度比不施用生物炭处理的高,说明施用生物炭有利于增加该菌属的丰度。芽球菌属(Blastococcus)、Solirubrobacter属、Gaiella属、节细菌属(Arthrobacter)这4个属在施用生物炭50 g/株处理的样品中丰度大于不施用生物炭处理,在不施用生物炭处理的样品中丰度大于施用生物炭100、150 g/株处理,说明施用生物炭超过50 g会降低这4个菌属的丰度。

【Objective】The study aims to conduct a comprehensive and accurate investigation on the species change of bacteria in tobacco leaves after the application of biochar. 【Method】Tobacco was chosen as the object of this study,and 4 treatments were set including treatment 1,the application of biochar 50 g each plant（ Y4. 1）; Treatment 2,the application of biochar 100 g each plant（ Y5. 1）; Treatment3,the application of biochar 150 g each plant（ Y6. 1）; And treatment 4,as control group without biochar（ Y1. 1）. Amplicons in 16 S r DNA-V4 region of endophytic bacteria from biochar-applied tobacco were sequenced by the use of Mi Seq high-throughput sequencer from Illumina sequencing platform. And Qiime and Mothur were adopted to sort out and calculate the number of sample sequences and operational taxonomic units（ OTUs） and analyze abundance and Alpha diversity of species. 【Result】The measured sequences were divided into 1177,1429,1228 and 1182 OTUs at 97 % of sequence similarity level. OTU abundance clustering analysis showed that the number of OTUs with the application of biochar was higher than that of non-biochar treatment,which indicated that the application of biochar could increase the abundance of tobacco bacteria groups; The number of OTUs with the treatment of 50 g each plant biochar was higher than that of100 g each plant biochar and the later was higher than that of150 g each plant,which indicated that as the weight of biochar increased to 100 g each plant and 150 g each plant,the abundance of tobacco bacteria groups began to decrease. 【Conclusion】The analysis on genus level showed that the abundance of Sphingomonas in the samples treated with biochar was higherthan that without biochar treatment,which demonstrated that the application of biochar was beneficial to the abundance of this genus. The abundances of Blastococcus,Solirubrobacter,Gaiella and Arthrobacter in samples treated with 50 g each plant biochar were higher than those without biochar treatment,and the later were greater than those with 100 g each plant and 150 g each plant treatments,indicating that the application of biochar over 50 g each plant would inhibit the growth of these 4 genera.