Wheat roots and soil DNA extraction under two CO2 levels Raw sequence reads

During plant growth, from seed germination until maturity, roots release specificcompounds to the surrounding soil. The composition of root deposits depends on planttype and age, leading to altering structure and function of root microbiome reacting tothese compounds. Atmospheric CO2 levels had been increasing for the last 150 years,stimulating growth and photosynthesis in C3 plants (e.g. wheat), increasing rootdeposition.In the current study, effect of elevated CO2 on wheat-root microbiome composition andplant development was studied, with emphasis on denitrifying communities.Wheat plants were grown in greenhouse equipped with two CO2 chambers: 1) ambientCO2 of 400 ppm (aCO2); 2) elevated CO2 of 850 ppm (eCO2). Soil, roots and leaveswere sampled during six weeks of growth and chemical and microbial parameters weremeasured. Total bacterial community was quantified using qPCR with universal 16SRNA gene primers, while denitrifying genes (e.g. nirK, nirS, nosZ) were measured usingspecific primers constructed based on metagenome sequencing. In addition, total andN 2 O reducing bacterial community in soil and root were sequenced.

在植物生长周期中，从种子萌发至成熟阶段，植物根系会向周围土壤释放特定化合物。根系沉积物质的组成因植物种类与株龄而异，会改变响应此类化合物的根系微生物组的结构与功能。近150年来，大气二氧化碳（CO₂）浓度持续攀升，可促进C3植物（例如小麦）的生长与光合作用，进而增加根系沉积物质的释放量。本研究探讨了高浓度CO₂对小麦根系微生物组组成及植株生长发育的影响，重点关注反硝化微生物群落。实验在配备两个CO₂培养舱的温室中开展：1）本底CO₂浓度组（aCO₂）：400 ppm；2）高CO₂浓度组（eCO₂）：850 ppm。在小麦生长的六周周期内，分别采集土壤、根系与叶片样本，并测定相关化学与微生物学参数。采用通用16S核糖体RNA（16S rRNA）基因引物，通过实时荧光定量PCR（qPCR）对总细菌群落进行定量；反硝化功能基因（如nirK、nirS、nosZ）则使用基于宏基因组测序设计的特异性引物进行检测。此外，还对土壤与根系中总细菌群落及氧化亚氮（N₂O）还原细菌群落进行了测序。