Table 4 Primers used in this study.

Internal gene fragment

nifR

F

5'- GCAAGCTT CCCAATCCGCCCGTCCCTC - 3'

R

5'- CAGAGCTC GAAATGGTCGCCAGCCGC - 3'

ppx1 (AGRO_2774)

F

5'- CTAAGCTT GCAGCGAGGTCCAGTGGGTG - 3'

R

5'- CAGAGCTC CAGTTCCGGGTCGTCGATG - 3'

ppx2 (AGRO_2552)

F

5'- CTAAGCTT GCTCCAATCCGCCAGTTCGC - 3'

R

5'- CAGAGCTC GGTCGGTGGTACATGGCGAA - 3'

rpoN

F

5'- GCAAGCTT CATCGAATCGTAACCCGCG - 3'

R

5'- GCGAGCTC AGCGACCACCTCAATCAG - 3'

rrpP

F

5'- GTAAGCTT GATCAGCGCCGTCACGACAAG - 3'

R

5'- CAGAGCTC CCGCGCAACCCTGCTACCAT - 3'

relA/spoT (AGRO_1497)

F

5'- GTAAGCTT GTATGCTCCAGGAACTCTTC - 3'

R

5'- GAGAGCTC CGTCAAGGGACGTCAGAAAA - 3'

AGRO_0033

F

5'- CGTGACAAGCTT TTCATAGATGCAGGCAACGTC - 3'

R

5'- GAACGTGAGCTC CTGGTGGGCGGATGATTGT - 3'

AGRO_0636

F

5'- CCTAGAAAGCTT CACAATAGTCGCTGCCAAGT - 3'

R

5'- CCTAGAGAGCTC GCTTGTCTCGTGACGCTCATT - 3'

AGRO_1729

F

5'- CGTGACAAGCTT CGGGATGGCAATGGTCTCGT - 3'

R

5'- CTTGTAGAGCTC CTGGCGTTGATACCCATGCT - 3'

AGRO_3967

F

5'- GAGTGTAAGCTT TCCAGCACATAATAGGGCGA - 3'

R

5'- CTTGGAGAGCTC GACACTTGCGGTTGTCATCG - 3'

Gene knockout confirmation

GM^R

F

5'- GATGCCCATACTTGAGCCACCTAAC - 3'

nifR

R

5'- AAGATCATCATTTGCCTCTTCCGGAG - 3'

ppx1 (AGRO_2774)

R

5'- GCGAAGCGATCCCGTCGTGGCAAGA - 3'

ppx2 (AGRO_2552)

R

5'- GACTCGATCAGAAGCACAGGGGC - 3'

rpoN

R

5'- ATGACGCATTTCGAGCTGACGCAGTT - 3'

rrpP

R

5'- ATGCGGTGTCCTGTCCGTGGTTTA - 3'

relA/spoT (AGRO_1497)

R

5'- CGCTTTCTTTTTGTGGAATATCGCA - 3'

AGRO_0033

R

5'- GTCGTGTTTCAAGTTTCAGGGTCCG - 3'

AGRO_0636

R

5'- GATGTTGGGTCTTTGGAAAAAACCG - 3'

AGRO_1729

R

5'- ATGAATGGTGAAAAGCGGGGGAGC - 3'

AGRO_3967

R

5'- GGTGAATCGCAGATGATGGAATTGAT - 3'

Quantitative RT-PCR

crdS

F

5'- ATCCAATTCAGCACAATCTCG -3'

R

5'- ACATATCCCCTTTCCATCAG -3'

rpoD

F

5'- GGAAATCCAGAACCTCTCCAC -3'

R

5'- GAACTTGTAACCACGGCGATA -3'