Journal
EUROPEAN JOURNAL OF SOIL SCIENCE
Volume 74, Issue 2, Pages -Publisher
WILEY
DOI: 10.1111/ejss.13365
Keywords
interference; plant residue; qPCR; sequencing; soil bacterial community
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Plant residues used as soil amendments in laboratory experiments may release compounds interfering with soil DNA extraction and molecular biological analyses. However, our study found no significant difference in microbial community composition or abundance between soil samples with and without added plant residue through DNA extraction, 16S rRNA gene sequencing, and quantitative PCR analysis.
Plant residues are often used as soil amendments in laboratory experiments, but they can reportedly release compounds interfering with soil DNA extraction and subsequent molecular biological analyses. Theoretically, for accurate comparison of microbial community composition in soils with and without added plant residues after a period of incubation, no significant difference at the beginning of the experiment is required between the amended and unamended control soils. We mixed plant residue into soil and immediately (within 10 min) commenced DNA extraction, and then performed 16S rRNA gene sequencing and quantitative PCR (qPCR) to determine bacterial community composition and abundance. Soil without plant residue addition served as a control. Five commonly used DNA extraction kits, 16S rRNA gene primer pairs, and soils, and two types (rice straw and alfalfa shoots) and three addition rates (2%, 4%, and 6%; w/w) of plant residue, were tested. In all cases, we found no significant difference in measured bacterial community composition or abundance between the treatments with and without added plant residue.
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