Background: We have previously demonstrated that an aldo‐keto reductase (AKR) from Echinochloa colona (EcAKR4‐1) can metabolize glyphosate and confers glyphosate resistance. This study aims to investigate if the EcAKR4‐1 orthologs from Lolium rigidum also play a role in glyphosate resistance in non‐target‐site based, glyphosate‐resistant (R) L. rigidum populations from Western Australia. Results: The full‐length L. rigidum AKR gene ( LrAKR4C10 ) orthologous to EcAKR4‐1 , together with a distinct LrAKR1 , were cloned from plants of a glyphosate‐susceptible (S) (VLR1) and three glyphosate R L. rigidum populations (WALR50, WALR60 and WALR70). Reverse transcription quantitative polymerase chain reaction (RT‐qPCR) results showed that basal expression levels of the two LrAKR genes did not differ between the R and S populations, but their expression was significantly induced by glyphosate (up to 4.3‐fold) or 2,4‐D treatment (up to 3.4‐fold) in R populations. Escherichia coli cells transformed respectively with LrAKR4C10 and LrAKR1 were more tolerant to glyphosate. Rice ( Oryza sativa ) seedlings overexpressing each of the two LrAKR gene survived glyphosate rates that were lethal to the green fluorescence protein (GFP) control plants. Structural modeling predicts a similar way of glyphosate binding and detoxification by LrAKR4C10 and EcAKR4‐1, but an alternative way of glyphosate binding by LrAKR1. Relatively lower capacity of the two LrAKRs in conferring glyphosate resistance than the known EcAKR4‐1 was discussed in relation to structural interaction. Conclusion: Glyphosate‐induced higher expression of the two LrAKR genes in L. rigidum populations contributes to a moderate level of glyphosate resistance likely through enhanced glyphosate metabolism. The herbicide 2,4‐D can also induce the LrAKR expression, indicating the potential antagonistic effect of 2,4‐D to glyphosate. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

The evolution of herbicide resistance in weedy plants leads to various adaptation traits including flowering time and seed germination. In our previous studies, we found an association of the early flowering phenotype with the ACCase inhibitor herbicide resistance genotype in a population of Polypogon fugax. MADS-box transcription factors are known to play pivotal roles in regulating plant flowering time. In this study, a SHORT VEGETATIVE PHASE (SVP)-like gene, belonging to the StMADS11 subfamily in the MADS-box family, was cloned from the early flowering P. fugax population (referred to as PfMADS16) and resistant to the herbicide clodinafop- propargyl. Overexpression of the SVP-like gene PfMADS16 in Arabidopsis thaliana resulted in early flowering and seed abortion. This is consistent with the phenotypic characters of resistant P. fugax plants, but contrary to the conventional role of SVP-like genes that usually suppress flowering. In addition, down regulation of the seed formation gene AtKTN1 in flowers of PfMADS16 transgenic Arabidopsis plants indicates that PfMADS16 may be indirectly associated with seed viability. Furthermore, one protein (PfMADS2) from the APETALA1 (AP1) subfamily interacting with PfMADS16 in P. fugax was identified with relevance to flowering time regulation. These results suggest that the PfMADS16 gene is an early flowering regulation gene associated with seed formation and viability in resistant P. fugax population. Our study provides potential application of PfMADS16 for integrated weed management (such as genetic-based weed control strategies) aiming to reduce the soil weed seedbank.

Asia minor bluegrass (Polypogon fugax, P. fugax), a weed that is both distributed across China and associated with winter crops, has evolved resistance to acetyl-CoA carboxylase (ACCase) herbicides, but the resistance mechanism remains unclear. The goal of this study was to analyze the transcriptome between resistant and sensitive populations of P. fugax at the flowering stage. Populations resistant and susceptible to clodinafop-propargyl showed distinct transcriptome profiles. A total of 206,041 unigenes were identified; 165,901 unique sequences were annotated using BLASTX alignment databases. Among them, 5904 unigenes were classified into 58 transcription factor families. Nine families were related to the regulation of plant growth and development and to stress responses. Twelve unigenes were differentially expressed between the clodinafop-propargyl-sensitive and clodinafop-propargyl-resistant populations at the early flowering stage; among those unigenes, three belonged to the ABI3VP1, BHLH, and GRAS families, while the remaining nine belonged to the MADS family. Compared with the clodinafop-propargyl-sensitive plants, the resistant plants exhibited different expression pattern of these 12 unigenes. This study identified differentially expressed unigenes related to ACCase-resistant P. fugax and thus provides a genomic resource for understanding the molecular basis of early flowering.

The transition from puddled-transplanted rice (Oryza sativa L.) (PTR) to direct-seeded rice (DSR) is gaining popularity in central China. In contrast, the PTR system is the most common practice in southwest China. Weeds are a major problem in the paddy fields of the DSR systems, and herbicides are widely used for weed control. However, the increased frequency and rate of herbicide use leads to the rapid evolution of resistance. Smallflower umbrella sedge (Cyperus difformis L.) is a troublesome weed species in rice fields of China and is usually controlled by the acetolactate synthase (ALS)-inhibiting herbicide bensulfuron-methyl. Here, we collected 32 C. difformis populations from DSR systems (Hunan Province) and PTR systems (Guangxi Province) and investigated their resistance to bensulfuron-methyl. Results revealed 80% (8 out of 10) populations from Hunan Province and 14% (3 out of 22) populations from Guangxi Province had evolved resistance to bensulfuron-methyl. Five populations from Hunan Province (HN-2, HN-3, HN-5, HN-9, HN-10) possessing the Trp-574-Leu mutation had high-level resistance (ranging from 169- to >1,309-fold) based on GR₅₀ ratios. The resistant populations from Guangxi Province had a lower level of resistance to bensulfuron-methyl due to a Pro-197-Ser mutation. The Asp-376-Glu mutation was only identified in the HN-4 population. In addition, the GX-3 population from the PTR systems was resistant to bensulfuron-methyl without ALS gene mutations, indicating non–target site resistance (NTSR). Although some resistant populations of both regions exhibited cross-resistance to multiple ALS-inhibiting herbicides, including pyrazosulfuron-ethyl, bispyribac-sodium, penoxsulam, and imazapic, sensitivity was also detected to the auxin herbicide MCPA and the photosystem II–inhibiting herbicides bentazone and propanil. These results indicate that cultivation practices affect resistance evolution in C. difformis. DSR systems exert high selection pressure by selecting the Trp-574-Leu mutation, resulting in high-level resistance. In contrast, a mutation at Pro-197 plus NTSR likely plays a significant role in ALS resistance in the PTR systems.

In our previous study with a wild radish (Raphanus raphanistrum) population resistant to HPPD-inhibiting herbicides we indicated that two additional candidate P450 genes, CYP71A28 and CYP72A13-like, may also contribute to the resistance. This study investigates the role of these two P450 genes in mesotrione resistance in the wild radish population. The full-length R. raphanistrum P450 genes (thereafter named as RrCYP71A28 and RrCYP72A15) were cloned from mesotrione-resistant (R) and -susceptible (S) wild radish plants. RT-qPCR results showed that basal expression levels of the two P450 genes are significantly higher (up to 3-fold) in the R than the S plants. Escherichia coli cells transformed respectively with RrCYP71A28 and RrCYP72A15 were more tolerant to mesotrione. Transgenic Arabidopsis plants expressing RrCYP72A15 showed a modest level of resistance to mesotrione. UPLC-MS/MS analysis demonstrated that tissue mesotrione levels in RrCYP72A15 transgenic Arabidopsis plants were significantly lower (up to 2-fold) than that in the wild type. Structural modelling predicts CYP72A15 can bind to RrCYP72A15 and metabolize mesotrione likely through formation of 4-OH-mesotrione. Although the RrCYP72A15 gene confers a modest level of resistance, overexpression of the multiple herbicide-metabolizing genes could contribute to the low level of mesotrione resistance observed in the R wild radish population.

Roegneria kamoji Ohwi (Poaceae), a wild relative plant of wheat which is widely distributed across China, has become a dominant and problematic weed in wheat fields in some regions. We have previously confirmed that R. kamoji is highly tolerant to foliar-applied acetyl-CoA carboxylase (ACCase) and acetolactate synthase (ALS) inhibitors in wheat (Triticum aestivum L.). The sensitivity of R. kamoji to pre-emergence (PRE) herbicides and the basis of fenclorim increase selectivity to butachlor between wheat and R. kamoji were evaluated in this study. Screenhouse bioassay showed that R. kamoji exhibited similar sensitivity to wheat to PRE herbicides at their recommended field doses (RFD); it also showed that buatchlor provides the highest relative control for R. kamoji (53.4% emergence and 81.5% fresh weight reduction, respectively), while it had no impact on seedling emergence of wheat among the six PRE herbicides. When butachlor was applied at four-fold RFD, no R. kamoji seedlings emerged; however, it significantly reduced the above-ground biomass of wheat compared with the non-treated control. Pre-treatment with herbicide safener fenclorim by seed soaking increased the ED₁₀ value of butachlor to wheat from 221.8 to 1600.1 g a.i. ha⁻¹, thus increasing the selectivity index from 9.6 to 68.9 between wheat and R. kamoji. The activities of α-amylase activity and protein content during germination, and glutathione-S-transferase (GST) and β-ketoacyl-CoA synthase (KCS) in the seedlings, could be induced by butachlor in both wheat seeds with or without fenclorim pre-soaking. These results suggested that butachlor provides the highest control for R. kamoji and did not affect germination and emergence in wheat. The basis of fenclorim-increased selectivity to butachlor was associated with the induced GST and KCS-mediated enhanced herbicide metabolism in wheat.