Table 1 Primer Sequences, Peptide Templates and Positions

Supplementary materials

Table S1 Information of the selected protein sequences of nitrate and nirtrous oxide reductases (coded by narG and nosZ) from GenBank, including the accession numbers and the names of strains

a The accession numbers were locus of the protein sequences of nitrate and nirtrous oxide reductases encoded by narG and nosZ from these bacteria.

Table S2 Bacterial strains used in this study to test the specificity of the narG andnosZprimers

+, PCR product of the right size obtained; －, no PCR product obtained; ＋ PCR product was cloned and sequenced and the sequence was deposited in the data banks. Pseudomonas stutzeri A1501 were provided by prof. Min Lin, The Chinese Academy of Agricultural Sciences (CAAS).

Table S3 Information of standard curves of the denitrifying genes

Table S4 The clonesand sequences information of the denitrifying genes

a Only the total OTUs from the four treatments of each gene were shown here as the OTUs from the four treatments overlapped.

Fig S1 Relative abundances of narG(a), qnorB (b) and nosZ(c)T-RFs from triplicate plots of long-term fertilization treatments. The relative abundance of T-RFs is given as a percentage of the total peak height. Fragment sizes within the graph indicate the sizes (bp) of the experimental T-RFs by T-RFLP.

Fig S2 Standard curves for 16S rRNA (a), narG(b), nirK(c), nirS(d), qnorB(e) andnosZ(f) assays obtained by plotting the concentration of plasmid DNA versus the threshold cycle. Red multiple sign present the amplification and quantity of soil samples.

Fig S3 Melt curves for 16S rRNA (a), narG(b), nirK(c), nirS(d), qnorB(e) andnosZ(f) genes. The Y-axis is the derivative of the slope of the signal intensity as temperature increases.

Fig S1a narG

Fig S1bqnorB

Fig S1cnosZ

Fig S2 a: 16S rRNA, b: narG, c: nirK, d: nirS, e: qnorB, f: nosZ

Fig S3 a: 16S rRNA, b: narG, c: nirK, d: nirS, e: qnorB, f: nosZ