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Molecular detection of nifH gene‐containing Paenibacillus in the rhizosphere of sorghum ( Sorghum bicolor ) sown in Cerrado soil
Author(s) -
Coelho M.R.R.,
Carneiro N.P.,
Marriel I.E.,
Seldin L.
Publication year - 2009
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.2009.02578.x
Subject(s) - rhizosphere , sorghum , sorghum bicolor , biology , sweet sorghum , foxtail , agronomy , poaceae , botany , paenibacillus , bacteria , 16s ribosomal rna , genetics
Aims: To develop a polymerase chain reaction (PCR)‐based approach for the detection of nifH gene‐containing Paenibacillus in environmental samples. Methods and Results: The primers, nifHPAENf and nifHPAENr, were designed and tested with DNA from: (i) strains of different nitrogen‐fixing Paenibacillus species, (ii) strains of other nitrogen‐fixing genera and (iii) rhizosphere of sorghum sown in Cerrado soil amended with either 12 or 120 kg ha −1 of nitrogen fertilizer. All nitrogen‐fixing Paenibacillus strains tested and the DNA samples from rhizosphere soil were amplified when these primers were used, generating a 280 bp fragment. When the PCR products obtained from both sorghum rhizospheres were cloned and sequenced, the majority of the clones analysed could be identified as Paenibacillus durus . Moreover, a greater diversity in the nifH sequences could be observed in the rhizosphere treated with a high amount of nitrogen fertilizer. Conclusions: Nitrogen fertilization slightly influenced the structure of the nifH gene‐containing Paenibacillus community in sorghum rhizospheres cultivated in Cerrado soil. Significance and Impact of the Study: The PCR detection method developed is adequate to assess the presence of nifH gene‐containing Paenibacillus in the environment and can be used in future to determine the ecological role of this group of micro‐organisms for the nitrogen input to the plants.