Segregation of non‐target‐site resistance to herbicides in multiple‐resistant Alopecurus myosuroides plants

Non‐target‐site resistance (NTSR) comprises a set of mechanisms conferring resistance to multiple modes of action. Investigation of the number of loci involved in NTSR will aid in the understanding of these resistance mechanisms. Therefore, six different multiple herbicide‐resistant Alopecurus myosuroides plants with different herbicide history were crossed in two generations with a susceptible wild type. Seeds from the backcrossing generation were studied for their segregation rate for resistance to five herbicides with four different modes of action (HRAC groups C₂, A, B and K₃). Taking into account that NTSR is a set of quantitative traits, the numbers of loci controlling NTSR were estimated using a normal mixture model fitted by the NLMIXED procedure of SAS. Each herbicide was controlled by a different number of loci comparing the six plants. In most of the cases, chlorotoluron resistance was controlled by one locus, whereas resistance to fenoxaprop‐P‐ethyl needed one or two loci. Resistance to pinoxaden was in all plants conferred by two loci. Cross‐resistance of fenoxaprop‐P‐ethyl and pinoxaden was found in all backcrossings, indicating that at least one of the two loci is responsible for both resistances. Resistance to mesosulfuron + iodosulfuron was conferred by a minimum of two loci. Results indicated that a minimum of five different loci can be involved in a multiple NTSR plant. Furthermore, the plant‐specific accumulation of NTSR loci was demonstrated. Such behaviour should be taken into account when evaluating the development and further spread of herbicide resistance.     

Herbicide‐resistant weed management: focus on glyphosate

This review focuses on proactive and reactive management of glyphosate‐resistant (GR) weeds. Glyphosate resistance in weeds has evolved under recurrent glyphosate usage, with little or no diversity in weed management practices. The main herbicide strategy for proactively or reactively managing GR weeds is to supplement glyphosate with herbicides of alternative modes of action and with soil‐residual activity. These herbicides can be applied in sequences or mixtures. Proactive or reactive GR weed management can be aided by crop cultivars with alternative single or stacked herbicide‐resistance traits, which will become increasingly available to growers in the future. Many growers with GR weeds continue to use glyphosate because of its economical broad‐spectrum weed control. Government farm policies, pesticide regulatory policies and industry actions should encourage growers to adopt a more proactive approach to GR weed management by providing the best information and training on management practices, information on the benefits of proactive management and voluntary incentives, as appropriate. Results from recent surveys in the United States indicate that such a change in grower attitudes may be occurring because of enhanced awareness of the benefits of proactive management and the relative cost of the reactive management of GR weeds. Copyright © 2011 Society of Chemical Industry     

Our top 10 herbicide‐resistant weed management practices

Although proactive or reactive herbicide‐resistant weed management (HRWM) practices have been recommended to growers in different agroecoregions globally, there is a need to identify and prioritise those having the most impact in mitigating or managing herbicide selection pressure in the northern Great Plains of North America. Our perspective on this issue is based on collaborative research, extension activities and dialogue with growers or farming experience (cereal, oilseed and pulse crop production) during the past 30 years. We list our top 10 HRWM practices, concluding with the number 1 practice which is the foundation of the other nine practices: crop diversity. Although our top 10 HRWM practices have broad applicability across agroecoregions, their ranking may vary widely. © 2017 Society of Chemical Industry     

Differentially expressed genes in dicamba‐resistant and dicamba‐susceptible biotypes

In an ongoing effort to investigate the mechanism of auxinic herbicide resistance in Kochia scoparia (kochia), polymerase chain reaction‐based cDNA suppression subtractive hybridization was used to identify genes that are differentially expressed between dicamba‐resistant (HRd) and dicamba‐susceptible (S1) kochia biotypes in response to herbicide treatment. Both the HRd and S1 adaptor‐ligated cDNAs were used in separate hybridizations in order to generate biotype‐specific clones. A total of 710 cDNAs, representing putative differentially expressed mRNAs, were isolated and subjected to further screening. The false‐positive cDNAs were removed by conducting two colony hybridizations and at least one Northern hybridization. Differential or biotype‐specific expression was confirmed for six clones each from the HRd and S1 plants. The S1‐related genes were constitutively expressed at higher levels than in the HRd plants, but none had significant sequence similarity to known genes. Among the HRd‐related genes, HRd‐88 had 42% amino acid sequence identity to a conserved domain within thiol peptidases, which might be involved in auxin‐regulated gene expression. The constitutively expressed and inducible (by the dicamba treatment) HRd‐39 had 40% identity and 60% similarity to a domain from the Fe(II)/α‐ketoglutarate‐dependent hydroxylase superfamily. The HRd‐39 gene product had the characteristics of an enzyme that is able to detoxify dicamba via oxidative hydroxylation and thus its overexpression might confer the dicamba resistance phenotype.     

Cross‐resistance study and biochemical mechanisms of thiamethoxam resistance in B‐biotype Bemisia tabaci (Hemiptera: Aleyrodidae)

BACKGROUND: B‐biotype Bemisia tabaci (Gennadius) has invaded China over the past two decades. To understand the risks and to determine possible mechanisms of resistance to thiamethoxam in B. tabaci, a resistant strain was selected in the laboratory. Cross‐resistance and the biochemical mechanisms of thiamethoxam resistance were investigated in the present study. RESULTS: A 66.3‐fold thiamethoxam‐resistant B. tabaci strain (TH‐R) was established after selection for 36 generations. Compared with the susceptible strain (TH‐S), the selected TH‐R strain showed obvious cross‐resistance to imidacloprid (47.3‐fold), acetamiprid (35.8‐fold), nitenpyram (9.99‐fold), abamectin (5.33‐fold) and carbosulfan (4.43‐fold). No cross‐resistance to fipronil, chlorpyrifos or deltamethrin was seen. Piperonyl butoxide (PBO) and triphenyl phosphate (TPP) exhibited significant synergism on thiamethoxam effects in the TH‐R strain (3.14‐ and 2.37‐fold respectively). However, diethyl maleate (DEM) did not act synergistically with thiamethoxam. Biochemical assays showed that cytochrome P450 monooxygenase activities increased 1.21‐ and 1.68‐fold respectively, and carboxylesterase activity increased 2.96‐fold in the TH‐R strain. However, no difference was observed for glutathione S‐transferase between the two strains. CONCLUSION: B‐biotype B. tabaci develops resistance to thiamethoxam. Cytochrome P450 monooxygenase and carboxylesterase appear to be responsible for the resistance. Reasonable resistance management that avoids the use of cross‐resistance insecticides may delay the development of resistance to thiamethoxam in this species. Copyright © 2009 Society of Chemical Industry     

Seedbank persistence,germination and early growth of glyphosate‐resistant Kochia scoparia

This study describes the seedbank persistence of glyphosate‐resistant (GR) Kochia scoparia at two sites in western Canada and examines if GRK. scoparia from western Canada and mid‐western United States (USA) differ from their susceptible counterparts in seed germination and early growth characteristics at low‐temperature regimes. Site or depth of seed burial (surface, 2.5 cm, 10 cm) did not affect seed viability over time and time to 50% and 90% loss of viability averaged 210 and 232 days respectively. Glyphosate‐resistant K. scoparia generally germinated later and had lower cumulative germination than glyphosate‐susceptible (GS) K. scoparia from Saskatchewan, Canada; and Kansas, USA; but not Colorado, USA. Similarly, time to 10% first leaf of GSK. scoparia from Saskatchewan and Kansas tended to be sooner than that of GRK. scoparia, with a greater percentage of GS vs. GR seedlings of populations from all regions having attained first leaf by the end of the experiment. The short seedbank longevity and delayed and reduced germination and time to first leaf of GRK. scoparia may potentially be exploited to maximise management efficacy through delayed preseeding weed control or alternatively by early seeding date to enhance crop competitiveness.     

Simple sequence repeat analysis of genetic diversity among Acetyl‐CoA carboxylase inhibitor‐resistant and ‐susceptible Echinochloa crus‐galli and E. oryzicola populations in Korea

Echinochloa species are amongst the most problematic weeds in rice fields of Korea. The steady reliance on the Acetyl‐CoA carboxylase (ACCase) and acetolactate synthase inhibiting herbicides for control of these weeds has led to resistance to these herbicides. The objective of this study was to assess the genetic diversity among populations of ACCase inhibitor‐resistant and ‐susceptible Echinochloa crus‐galli and E. oryzicola in Korea, to better understand their population structure and possible origins of resistance. Seven simple sequence repeat markers were applied to assess the genetic diversity between resistant and susceptible E. crus‐galli and E. oryzicola from 12 populations in Korea. Genetic diversity was slightly higher in the resistant group. The Unweighted Pair Group Method using Arithmetic algorithm (UPGMA) dendrogram generated two distinct clades. One clade consisted of Echinochloa spp. from three populations, i.e. Anmyeondo, Gimje 4 and Gongju, which are resistant and differentiated from the susceptible populations, and the other clade contained the rest of the populations. Structure modelling supported two clades of UPGMA clustering. Based on these data, we can infer that some resistant populations are greatly differentiated, whereas other resistant biotypes are still building up resistance in rice fields in Korea. Resistance traits will be fixed and continue to spread over time without proper control measures.     

Identification and expression pattern of a glutathione S‐transferase in Echinochloa crus‐galli

Plant glutathione S‐transferase (GST) forms a major part of the herbicide detoxification enzyme network in plants. A GST cDNA was isolated from Echinochloa crus‐galli and characterised. The gene, designated EcGST1 (E. crus‐galli GeneBank no: JX518596 ), has a 684 bp open reading frame predicted to encode a 25 kD protein. Sequence alignment showed that EcGST1 is a GST homologue. Its expression in response to quinclorac treatment was monitored in seedlings (leaves and roots) and adult plants (leaves, roots, stems and seeds) of quinclorac‐resistant (R) and susceptible (S) biotypes of E. crus‐galli. EcGST1 expression was 1.5–3 times greater in the R plants than in the S plants. However, after exposure to quinclorac, the difference in the expression levels of EcGST1 in R plants, compared with S plants, increased to a ratio of 6–10. Enhanced EcGST1 levels should enable greater quinclorac detoxification following quinclorac stimulation in R plants. GST‐based metabolism may be partially responsible for resistance to quinclorac in E. crus‐galli. The results suggest a new resistance mechanism for this R biotype in Chinese rice fields.     

Distribution of herbicide‐resistant acetyl‐coenzyme A carboxylase alleles in Lolium rigidum across grain cropping areas of South Australia

Resistance to the acetyl‐coenzyme A carboxylase (ACCase)‐inhibiting herbicides in Lolium rigidum is widespread in grain cropping areas of South Australia. To better understand the occurrence and spread of resistance to these herbicides and how it has changed with time, the carboxyl transferase (CT) domain of the ACCase gene from resistant L. rigidum plants, collected from both random surveys of the mid‐north of Southern Australia over 10 years as well as stratified surveys in individual fields, was sequenced and target site mutations characterised. Amino acid substitutions occurring as a consequence of these target site mutations, at seven positions in the ACCase gene previously correlated with herbicide resistance, were identified in c. 80% of resistant individuals, indicating target site mutation is a common mechanism of resistance in L. rigidum to this herbicide mode of action. Individuals containing multiple amino acid substitutions (two, and in two cases, three substitutions) were also found. Substitutions at position 2041 occurred at the highest frequency in all years of the large area survey, while substitutions at position 2078 were most common in the single farm analysis. This study has shown that target site mutations leading to amino acid substitutions in ACCase of L. rigidum are widespread across South Australia and that these mutations have likely evolved independently in different locations. The results indicate that seed movement, both within and between fields, may contribute to the spread of resistance in a single field. However, over a large area, the independent appearance and selection of target site mutations conferring resistance through herbicide use is the most important factor.     

Development of a high‐throughput real‐time PCR assay for the detection of the R81T mutation in the nicotinic acetylcholine receptor of neonicotinoid‐resistant Myzus persicae

BACKGROUND: Myzus persicae is a globally important aphid pest that is mainly controlled through the application of chemical insecticides. Recently, a clone of M. persicae exhibiting control‐compromising levels of resistance to neonicotinoid insecticides was described. The resistance of this clone was associated with reduced affinity of imidacloprid for the target site (the nicotinic acetylcholine receptor) as a result of mutation of a key amino acid residue (R81T) in the loop D region of a nAChR β1 subunit. The potent levels of resistance conferred by this mechanism are cause for considerable concern, and the frequency and distribution of the mutation in worldwide populations of M. persicae require careful monitoring. In this study, a high‐throughput assay has been developed that allows detection of the mutation in individual aphids. RESULTS: A real‐time TaqMan assay to detect the R81T substitution was developed that proved to be sensitive and specific in tests of analytical sensitivity and in a blind genotyping trial of DNA extracted from individual aphids comprising the three possible genotypes. The assay was then used to examine the frequency of the R81T mutation in aphids collected and stored in ethanol from peach orchards in southern France. The R81T frequency varied from 33 to 100% in seven populations from the department of Gard, France. CONCLUSIONS: This study describes a rapid and sensitive assay that very effectively detects the R81T mutation in individual aphids. The results also have practical significance for the control of M. persicae in southern France and provide contemporary data to inform current resistance management strategies. Copyright © 2012 Society of Chemical Industry     

Occurrence of sulfonylurea resistance in Sagittaria trifolia,a basal monocot species,based on target‐site and non‐target‐site resistance

Sagittaria trifolia L. is one of the most serious weeds in paddy fields in Japan. Since the late 1990s, severe infestations of S. trifolia have occurred following applications of sulfonylurea herbicides in Akita prefecture. In this study, two accessions of S. trifolia, R1 and R2, were collected from paddy fields with severe infestations and their resistance profiles were determined in comparison to a susceptible accession, S1. R1 and R2 were highly resistant to bensulfuron‐methyl. R1 was also highly resistant to pyrazosulfuron‐ethyl, but R2 was susceptible. Relative to S1, R1 had an amino acid substitution at the Pro₁₉₇ residue of acetolactate synthase (ALS), a well‐known mutation that confers sulfonylurea resistance, suggesting that R1 has a target‐site‐based resistance (TSR) mechanism. The sequence of the ALS gene in R2 was identical to that in S1. A Southern blot analysis indicated that there was only one copy of the ALS gene in S1 and R2. These results suggest that R2 has a non‐target‐site‐based resistance (NTSR) mechanism. R2 was moderately resistant to imazosulfuron but susceptible to thifensulfuron‐methyl. R2 and S1 were susceptible to pretilachlor, benfuresate, MCPA‐ethyl and bentazon. The results reveal the occurrence of two sulfonylurea‐resistant biotypes of S. trifolia that show different mechanisms of cross‐resistance to sulfonylureas related to TSR in R1 and NTSR in R2.