Abstract
Background: The spread of herbicide-resistance Ambrosia artemisiifolia threatens not only the production of agricultural crops, but also the composition of weed communities. The reduction of their spread would positively affect the biodiversity and beneficial weed communities in the arable habitats. Detection of resistant populations would help to reduce herbicide exposure which may contribute to the development of sustainable agroecosystems. Methods: This study focuses on the application of target-site resistance (TSR) diagnostic of A. artemisiifolia caused by different herbicides. We used targeted amplicon sequencing (TAS) on Illumina Miseq platform to detect amino acid changes in herbicide target enzymes of resistant and wild-type plants. Results: 16 mutation points of four enzymes targeted by four herbicide groups, such as Photosystem II (PSII), Acetohydroxyacid synthase (AHAS), 5-enolpyruvylshikimate 3-phosphate synthase (EPSPS) and protoporphyrinogen IX oxidase (PPO) inhibitors have been identified in common ragweed populations, so far. All the 16 mutation points were analyzed and identified. Out of these, two mutations were detected in resistant biotypes. Conclusions: The applied next-generation sequencing-targeted amplicon sequencing (NGS-TAS) method on A. artemisiifolia resistant and wild-type populations enable TSR detection of large sample numbers in a single reaction. The NGS-TAS provides information about the evolved herbicide resistance that supports the integrated weed control through the reduction of herbicide exposure which may preserve ecological properties in agroecosystems.
Highlights
This article is an open access articleExtensive weed management in agricultural fields is a major threat to agroecosystem biodiversity as weed communities play a key role in maintaining heterogeneity
To design correct Next generation sequencing (NGS)-targeted amplicon sequencing (TAS) primer pairs, it is essential to know at least the sequences at the intron-exon boundaries close to the mutation points
Gene sequences and point mutations of the herbicide target enzymes were determined in sensitive genotypes by using in silico NGS transcriptome analysis (NGS-TA), SS Sanger sequencing, and next-generation sequencing-targeted amplicon sequencing (NGS-TAS) (Figure 1)
Summary
Extensive weed management in agricultural fields is a major threat to agroecosystem biodiversity as weed communities play a key role in maintaining heterogeneity. The total number of weed species in agricultural fields has increased since the 1950s. This has resulted in a more diverse composition of agricultural weeds, it has negatively affected landscape structure and land-use intensity. Crop intensification methods, such distributed under the terms and conditions of the Creative Commons. As herbicide treatment, have become critical to control weed communities to improve agricultural crop production [1]. It is important not to endanger beneficial weed species while using weed control. To define the correct management of invasive species, local environmental drivers have to be identified due to their severe effects on individual performance [2]
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