Abstract
Weed resistance to herbicides constitutes a serious problem to world crop production. One of the weeds that are significantly threatening the crops’ yield and quality is Apera spica-venti. The target-site resistance (TSR) mechanism of A. spica-venti has been widely studied, though, little is known about its non-target-site resistance (NTSR) mechanisms at the molecular level. Molecular examination of NTSR is, to a great extent, based on the expression profiles of selected genes, e.g. those participating in detoxification. However, to obtain reliable results of gene expression analysis, the use of a normalizer is required. The aim of this study was to select the best reference genes in A. spica-venti plants of both populations, susceptible and resistant to ALS inhibitor, under treatment with herbicide. Eleven housekeeping genes were chosen for their expression stability assessment. The efficiency correction of raw quantification cycles (Cq) was included in the gene expression stability analyses, which resulted in indicating the TATA-box binding protein (TBP), glyceraldehyde-3-phosphate dehydrogenase, cytosolic (GAPC), and peptidyl-prolyl cis–trans isomerase CYP28 (CYP28) genes as the most stably expressed reference genes. The obtained results are of vital importance for future studies on the expression of genes associated with the non-target-site resistance mechanisms in the A. spica-venti populations susceptible and resistant to herbicides.
Highlights
Weed resistance to herbicides constitutes a serious problem to world crop production
The elevated levels of cytochrome P450 monooxygenases (CYP450s), GT, and glutathione S-transferases (GST) typically contribute to herbicide detoxification, which is divided in three phases
In order to choose A. spica-venti populations to be subjected to HKGs stability analysis, plants from the populations resistant and susceptible to pyroxsulam were examined for the absence of mutations in the acetolactate synthase (ALS) sequence
Summary
Weed resistance to herbicides constitutes a serious problem to world crop production. The aim of this study was to select the best reference genes in A. spica-venti plants of both populations, susceptible and resistant to ALS inhibitor, under treatment with herbicide. The obtained results are of vital importance for future studies on the expression of genes associated with the non-target-site resistance mechanisms in the A. spica-venti populations susceptible and resistant to herbicides. RT-qPCR is the most popular and the most commonly used method for the assessment of genes expression level It is frequently used for the validation of results obtained in high-throughput analyses. The studies on reference gene expression stability in weeds belonging to the Poaceae family, Alopecurus myosuroides and Eleusine indica, exposed to herbicide stress have revealed that in the plants subjected to the same stress, different genes exhibited the most stable expressions. TUB was identified to show unstable expression in E. indica
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