Cross-resistance Patterns Research Articles

Herbicide resistance is complex: a global review of cross-resistance in weeds within herbicide groups

Abstract Herbicides have been placed in global Herbicide Resistance Action Committee (HRAC) herbicide groups based on their sites of action (e.g., acetolactate synthase–inhibiting herbicides are grouped in HRAC Group 2). A major driving force for this classification system is that growers have been encouraged to rotate or mix herbicides from different HRAC groups to delay the evolution of herbicide-resistant weeds, because in theory, all active ingredients within a herbicide group physiologically affect weeds similarly. Although herbicide resistance in weeds has been studied for decades, recent research on the biochemical and molecular basis for resistance has demonstrated that patterns of cross-resistance are usually quite complicated and much more complex than merely stating, for example, a certain weed population is Group 2-resistant. The objective of this review article is to highlight and describe the intricacies associated with the magnitude of herbicide resistance and cross-resistance patterns that have resulted from myriad target-site and non–target site resistance mechanisms in weeds, as well as environmental and application timing influences. Our hope is this review will provide opportunities for students, growers, agronomists, ag retailers, regulatory personnel, and research scientists to better understand and realize that herbicide resistance in weeds is far more complicated than previously considered when based solely on HRAC groups. Furthermore, a comprehensive understanding of cross-resistance patterns among weed species and populations may assist in managing herbicide-resistant biotypes in the short term by providing growers with previously unconsidered effective control options. This knowledge may also inform agrochemical company efforts aimed at developing new resistance-breaking chemistries and herbicide mixtures. However, in the long term, nonchemical management strategies, including cultural, mechanical, and biological weed management tactics, must also be implemented to prevent or delay increasingly problematic issues with weed resistance to current and future herbicides.

Unveiling environmental transmission risks: comparative analysis of azole resistance in Aspergillus fumigatus clinical and environmental isolates from Yunnan, China.

Azole resistance in Aspergillus fumigatus poses a significant clinical challenge globally. Our previous epidemiological analysis revealed a remarkably high frequency (~80%) of azole-resistant A. fumigatus in Yunnan's greenhouse environments, prompting increased local and regional research for targeted control strategies. In this study, we analyzed 94 clinical A. fumigatus isolates from Yunnan, comparing their susceptibility profiles and genotypic characteristics with environmental strains previously isolated. While the overall frequency of azole resistance in clinical isolates was lower than that in environmental samples, a significant prevalence of cross-resistance, with varying resistance patterns based on minimum inhibitory concentration (MIC) levels was observed, which exceeded rates in other regions of China. Specific mutation combinations in the cyp51A gene were linked to elevated MIC values in clinical and/or environmental samples, while some resistant strains with wild-type cyp51A remain unexplained, indicating a need for further investigation into their resistance mechanisms. The differences in unique genetic elements and the distinct genetic differentiation observed between clinical and environmental isolates can be attributed to Yunnan's unique geomorphology and potential genotype importation from other provinces and abroad. Extensive allele exchanges and sharing contributed to the selection of azole-resistant clinical isolates, suggesting a common environmental origin, and the transmission routes of local drug-resistant strains cannot be excluded. These findings emphasize the imperative for regional and targeted surveillance to monitor resistance trends and guide effective antifungal therapy, and management strategies to mitigate invasive aspergillosis risk in this region.IMPORTANCEAzole resistance in Aspergillus fumigatus is a major global health concern, with particularly high rates (~80%) observed in Yunnan's greenhouse environments. This study compares azole resistance in 94 clinical isolates from Yunnan with environmental strains, revealing lower clinical resistance but significant cross-resistance and distinct resistance patterns. Specific mutations in the cyp51A gene were associated with elevated minimum inhibitory concentration values, though some resistant strains had wild-type cyp51A, highlighting the need for further research. The unique genetic profiles and potential external genotype influences in Yunnan emphasize the need for targeted regional surveillance. Effective monitoring and control strategies are essential to manage and mitigate the risk of invasive aspergillosis.

PfGSTF2 endows resistance to quizalofop-p-ethyl in Polypogon fugax by GSH conjugation.

Populations of Polypogon fugax have developed resistance to many acetyl-CoA carboxylase (ACCase)-inhibiting herbicides. This resistance threats the effectiveness and sustainability of herbicide use. In our previous research, a field P. fugax population exhibited GST-based metabolic resistance to the widely used ACCase-inhibiting herbicide quizalofop-p-ethyl. Here, in this current study, we identified and characterized two GST genes (named as PfGSTF2 and PfGSTF58) that showed higher expression levels in the resistant than the susceptible population. Transgenic rice calli overexpressing PfGSTF2, but not PfGSTF58, became resistant to quizalofop-p-ethyl and haloxyfop-R-methyl. This reflects similar cross-resistance pattern to what was observed in the resistant P. fugax population. Transgenic rice seedlings overexpressing PfGSTF2 also exhibited resistance to quizalofop-p-ethyl. In contrast, CRISPR/Cas9 knockout of the orthologue gene in rice seedlings increased their sensitivity to quizalofop-p-ethyl. LC-MS analysis of in vitro herbicide metabolism by Escherichia coli-expressed recombinant PfGSTF2 revealed that quizalofop (but not haloxyfop) was detoxified at the ether bond, generating the GSH-quizalofop conjugate and a propanoic acid derivative with greatly reduced herbicidal activity. Equally, these two metabolites accumulated at higher levels in the resistant population than the susceptible population. In addition, both recombinant PfGSTF2 and PfGSTF58 can attenuate cytotoxicity by reactive oxygen species (ROS), suggesting a role in plant defence against ROS generated by herbicides. Furthermore, the GST inhibitor (NBD-Cl) reversed resistance in the resistant population, and PfGSTF2 (but not PfGSTF58) responded to NBD-Cl inhibition. All these suggest that PfGSTF2 plays a significant role in the evolution of quizalofop resistance through enhanced herbicide metabolism in P. fugax.

Genetic basis of resistance to Cry1Ac in Rachiplusia nu (Lepidoptera: Noctuidae): inheritance mode, cross-resistance patterns and fitness cost.

Rachiplusia nu (Guenée) was historically a secondary soybean pest in Brazil, but a key soybean pest in Argentina. From 2021 onwards, injury caused by R. nu has been reported in soybean that expresses the Cry1Ac toxin from Bacillus thuringiensis (Berliner) in both countries. In this study, we selected resistant and susceptible strains of R. nu to Cry1Ac using Cry1Ac-containing leaf tissue and characterized the inheritance of resistance, cross-resistance patterns and fitness cost. Neonates of the Cry1Ac-resistant strain of R. nu were able to develop on Cry1Ac soybean leaves and emerge as fertile adults, while neonates from the susceptible and heterozygous strains did not survive beyond 10 days. The resistance ratio to Cry1Ac estimated in diet-overlay bioassays in the resistant strain was > 736.92-fold. The inheritance pattern of Cry1Ac resistance in R. nu was characterized as autosomal recessive and monogenic. The Cry1Ac-resistant strain of R. nu also exhibited high resistance to Cry1A.105 (resistance ratio > 159.87-fold), but negligible resistance to Cry2Ab2 (resistance ratio = 1.25-fold). Life history data showed that the resistance to Cry1Ac in R. nu is not associated with a substantial fitness cost. The inheritance pattern of Cry1Ac resistance in R. nu is autosomal recessive, monogenic and not associated with obvious fitness costs. Cross-resistance occurred between Cry1Ac and Cry1A.105 in R. nu but not between Cry1Ac and Cry2Ab2, indicating that Cry1A.105/Cry2Ab2/Cry1Ac soybean is a valuable tool to manage Cry1Ac resistance in R. nu. This is the first study reporting the genetic basis of Cry1Ac resistance in R. nu. © 2024 Society of Chemical Industry. Published by John Wiley & Sons Ltd.

Downy brome (Bromus tectorum) management and herbicide resistance in dryland wheat production across northeastern Oregon

Abstract Downy brome (Bromus tectorum L.) is a difficult species to control in the dryland wheat (Triticum aestivum L.) production areas of northeastern Oregon. The selection of herbicide-resistant B. tectorum populations has further complicated B. tectorum management. A survey of wheat growers was conducted in 2021 and 2022 to understand B. tectorum management practices. The survey included four questions based on the growing seasons from 2017 to 2022 related to crop rotation, tillage versus no-tillage, irrigation versus dryland, and herbicide programs. To determine the extent of herbicide resistance, seeds were collected from 49 B. tectorum populations in wheat fields in northeastern Oregon and tested for resistance. Herbicides tested were clethodim, glyphosate, imazamox, mesosulfuron, metribuzin, propoxycarbazone, pyroxasulfone, pyroxsulam, quizalofop, and sulfosulfuron. Winter wheat–summer fallow rotation was the most predominant cropping system in the region. Most of the fields were in no-tillage systems, and none were irrigated. Pyroxasulfone applied preemergence and acetolactate synthase (ALS) inhibitors applied postemergence were the most often used herbicides for B. tectorum control in winter wheat. Glyphosate was the most frequently used herbicide for B. tectorum control in summer fallow. Resistance screenings confirmed that 46 of the 49 B. tectorum populations were resistant to ALS inhibitors with different cross-resistance patterns. Two populations were resistant to metribuzin and exhibited multiple resistance to ALS inhibitors. All populations were susceptible to clethodim, glyphosate, pyroxasulfone, and quizalofop. The widespread occurrence of ALS inhibitor–resistant B. tectorum populations limits effective postemergence herbicide options in winter wheat.

Cross-resistance patterns in field strains of the sheep blowfly following laboratory-based selection pressure with dicyclanil or imidacloprid

Control of the sheep blowfly relies largely on the use of insecticides applied prophylactically in advance of expected fly activity. However, the blowfly has shown an ability to develop resistance to some of these insecticides. Recent reports of the co-occurrence of resistance to both dicyclanil and imidacloprid in in vitro bioassays with field-collected fly strains has raised the possibility that the two resistances may represent cross-resistance linked by a common mechanism. We investigated this by imposing insecticide selection pressure on larvae of two insecticide-resistant field strains over a number of generations using either dicyclanil or imidacloprid and then measuring changes in sensitivity to both the selecting chemical and the alternate chemical. Larvae selected over six generations with each chemical showed significant increases in resistance to the selecting chemical: resistance ratios at the IC50 5.5 - 8.1-fold higher for dicyclanil, and 3.1 - 3.8-fold for imidacloprid. The larvae also showed significant increases in levels of resistance towards the alternate chemical: resistance ratios 2.6 - 3.1-fold higher towards dicyclanil following selection with imidacloprid, and 2.2 - 3.2-fold higher towards imidacloprid following selection with dicyclanil. The increases in resistance to both chemicals after exposure to either suggests a common mechanism of resistance, at least in our laboratory-selected populations. Assays with the cytochrome P450 inhibitor aminobenzotriazole showed that this synergist was able to remove the increased resistance to both compounds in strains selected with either compound, suggesting that cytochrome P450 is responsible for the resistance observed to both chemicals. Our results confirm that cross-resistance exists between dicyclanil and imidacloprid in the sheep blowfly, and hence the two compounds should be considered as related entities in insecticide rotation strategies for flystrike control.

Metabolism of 2,4-D in plants: comparative analysis of metabolic detoxification pathways in tolerant crops and resistant weeds.

The commercialization of 2,4-D (2,4-dichlorophenoxyacetic acid) latifolicide in 1945 marked the beginning of the selective herbicide market, with this active ingredient playing a pivotal role among commercial herbicides due to the natural tolerance of monocots compared with dicots. Due to its intricate mode of action, involving interactions within endogenous auxin signaling networks, 2,4-D was initially considered a low-risk herbicide to evolve weed resistance. However, the intensification of 2,4-D use has contributed to the emergence of 2,4-D-resistant broadleaf weeds, challenging earlier beliefs. This review explores 2,4-D tolerance in crops and evolved resistance in weeds, emphasizing an in-depth understanding of 2,4-D metabolic detoxification. Nine confirmed 2,4-D-resistant weed species, driven by rapid metabolism, highlight cytochrome P450 monooxygenases in Phase I and glycosyltransferases in Phase II as key enzymes. Resistance to 2,4-D may also involve impaired translocation associated with mutations in auxin/indole-3-acetic acid (Aux/IAA) co-receptor genes. Moreover, temperature variations affect 2,4-D efficacy, with high temperatures increasing herbicide metabolism rates and reducing weed control, while drought stress did not affect 2,4-D efficacy. Research on 2,4-D resistance has primarily focused on non-target-site resistance (NTSR) mechanisms, including 2,4-D metabolic detoxification, with limited exploration of the inheritance and genetic basis underlying these traits. Resistance to 2,4-D in weeds is typically governed by a single gene, either dominant or incompletely dominant, raising questions about gain-of-function or loss-of-function mutations that confer resistance. Future research should unravel the physiological and molecular-genetic basis of 2,4-D NTSR, exploring potential cross-resistance patterns and assessing fitness costs that may affect future evolution of auxin-resistant weeds. © 2024 Society of Chemical Industry.

Occurrence and Mechanisms Conferring Multiple Resistance to ALS-Inhibiting and Auxins Mimics Herbicides in Papaver rhoeas from Tunisia

Herbicide-resistant corn poppy (Papaver rhoeas L.) is one of the most important broadleaved weeds and the number of resistant cases is still growing. The aims of this study were to confirm the resistance of P. rhoeas from Tunisia to ALS inhibitors and auxin mimics and investigate the mechanisms of Target-Site Resistance (TSR) and Non-Target Site Resistance (NTSR) involved. Dose–response trials to determine cross-resistance patterns for ALS inhibitors and auxin mimics were conducted in a greenhouse. In this study, multiple resistance to tribenuron-methyl and dicamba but not to 2,4-D was found in P. rhoeas populations. Cross-resistance to imazamox was confirmed as well. Sequence analysis of the ALS gene detected target-site mutations in codon 197 of the ALS gene, namely, Pro197His, Pro197Thr, Pro197Leu, and Pro197Asn. In this study, the metabolism experiments with malathion (a cytochrome P450 inhibitor) showed that malathion reduced resistance to imazamox, indicating that P450 is involved in the resistance. TSR and NTSR mechanisms to ALS inhibitors likely coexist. The findings of this study revealed a significant synergistic interaction between malathion and dicamba in particular populations, suggesting that the resistance to auxin mimics can be conferred by enhanced metabolism.

Cross-Resistance to Imidacloprid in Brown Planthopper <i>Nilaparvata lugens</i> (Stal)

Cross-resistance pattern of imidacloprid resistance in brown planthopper, Nilaparvata lugens (Stål) to three insecticides viz., dinotefuran, pymetrozine and triflumezopyrim was evaluated with laboratory developed imidacloprid resistant (IMI-R) and susceptible populations (IMI-S). IMI-R population had a 6-fold resistance compared to IMI-S population. The LC50 values for dinotefuran in IMI-R and IMI-S populations were 466.6 and 129.2 mg/ L with a resistance ratio of 3.6-fold. Whereas, LC50 values for triflumezopyrim and pymetrozine in IMI-R and IMI-S populations were 0.27 and 0.17 mg/L; 49.07 and 31.5 mg/L with resistant ratios of 1.58 and 1.55, respectively. These findings clearly demonstrated cross-resistance between imidacloprid and dinotefuran but no obvious cross-resistance to triflumezopyrim and pymetrozine.

Resistance patterns and molecular basis to ACCase-inhibiting herbicides

Abstract Digitaria ciliaris var. chrysoblephara (Fig. & De Not.) R.R. Stewart is an annual xeromorphic weed that severely infests direct-seeded rice fields in China. Herbicide resistance is emerging in D. ciliaris var. chrysoblephara owing to extensive and recurrent use of the acetyl-CoA carboxylase (ACCase)-inhibiting herbicide metamifop. In this study, a total of 53 D. ciliaris var. chrysoblephara populations randomly sampled from direct-seeded rice fields across Jiangsu Province were investigated for metamifop resistance and potential resistance-endowing mutations. Single-dose assays revealed that 17 (32.1%) populations evolved resistance to metamifop and 5 (9.4%) populations were in the process of developing resistance. The resistance index (RI) of metamifop-resistant populations ranged from 2.7 to 32.1. Amino acid substitutions (Ile-1781-Leu, Trp-2027-Cys/Ser, and Ile-2041-Asn) in ACCase genes were detected in resistant D. ciliaris var. chrysoblephara plants and caused various cross-resistance patterns to ACCase-inhibiting herbicides. All of four resistant populations (YC07, YZ09, SQ03, and HA06), with different ACCase mutations, exhibited cross-resistance to the aryloxyphenoxypropionate (APP) herbicides cyhalofop-butyl (RI values: 10.0 to 19.9), fenoxaprop-P-ethyl (RI values: 53.7 to 132.8), and haloxyfop-P-methyl (RI values: 6.2 to 62.6), and the phenylpyrazoline (DEN) pinoxaden (RI values: 2.3 to 5.4), but responded differently to the cyclohexanedione (CHD) herbicides clethodim and sethoxydim. It is noteworthy that four postemergence herbicides used for rice cropping, including bispyribac-sodium, pyraclonil, quinclorac, and anilofos, showed poor control effect against D. ciliaris var. chrysoblephara, suggesting few alternations for managing this weed in rice fields except ACCase inhibitors. In conclusion, this work demonstrated that the D. ciliaris var. chrysoblephara had developed resistance to ACCase-inhibiting herbicides in rice cultivation of China, and target-site amino acid substitutions in ACCase were primarily responsible for metamifop resistance.

Abstract PS08-03: Sequencing Antibody-Drug Conjugate after Antibody-Drug Conjugate in Metastatic Breast Cancer (A3 study): Multi-Institution Experience and Biomarker Analysis

Abstract Background: Antibody-drug conjugates (ADCs) improve survival in patients with metastatic breast cancer (MBC) and offer the potential for targeted delivery of highly potent therapy. Many patients are now candidates for multiple ADCs, but optimal strategies for sequencing are unknown. We previously reported on a single institution experience of patients receiving multiple ADCs for MBC (Abelman, ASCO 2023). Here we report a multi-institution update with biomarker analysis. Methods: We included all patients treated at three academic medical institutions who received multiple ADCs for MBC. Patients were included if they had hormone receptor positive, HER2-negative (HR+/HER2-) breast cancer or triple-negative breast cancer (TNBC); patients with HER2+ metastatic breast cancer were excluded. Clinical information was determined by chart review. The metric of “cross-resistance” to the second ADC was defined as patients with progressive disease on first restaging assessment or progression within 60 days of treatment initiation. Every subsequent ADC beyond the first was compared against the prior ADC for presence of identical “antibody target” and “payload”. Comparisons across ADCs were performed using Fisher’s exact test. Significance was determined to be a type I error less than 0.05. A subset of patients had available whole exome tissue sequencing through commercially available sequencing platforms (BostonGene and Caris) performed around the time of receipt of ADC. All sequencing reports were examined for presence of pathogenic variants, variants of uncertain significance, and currently undefined variants and fusions as defined by each sequencing platform. Results: 68 patients were identified who received two or more ADCs for metastatic HR+/HER2- breast cancer or TNBC from August 2014-June 2023. 30 patients (44.1%) had HR+/HER2- disease and 38 patients (55.9%) had TNBC; 50 patients (73.5%) had HER2-low disease. Median age at time of second ADC was 59.6 (range 29.9-88.6). Patients had received a median of 4 lines of treatment in the metastatic setting prior to initiation of the second ADC. At time of first restaging, cross-resistance was present in 38/64 evaluable cases (59.4%). When the antibody target of the latter ADC was the same as the prior, cross-resistance was present in 11/14 cases (78.5%) compared to 26/49 cases (53.1%) when the later ADC targeted a different tumor-associated antigen. Relatively similar patterns of cross-resistance were observed regardless of whether the later ADC contained an identical payload to prior (6/10 cases, 60%) versus a different payload (22/42, 52.4%). Sequencing information was available for 20 patients who received multiple ADCs with 15 unique reports performed at the time of resistance to ADC1, prior to initiation of ADC2, or after ADC2 if presence of cross-resistance. Variants in topoisomerase-I associated genes (TOP1, TOP2A, TOP3A, TOP3B) were identified in a subset of patients mediating cross-resistance to the second ADC with a topoisomerase-I inhibitor payload. Conclusions: In this multi-institution study, cross-resistance to the second ADC appears to be driven by the antibody target in some patients versus the payload in others, highlighting the heterogeneity of mechanisms related to ADC resistance. Tumor sequencing revealed candidate resistance mutations that may guide optimal sequencing for patients with MBC. Citation Format: Rachel Occhiogrosso Abelman, Laura Spring, Geoffrey Fell, Andrew Davis, Whitney Hensing, Phoebe Ryan, Neelima Vidula, Seth Wander, Arielle Medford, Jennifer Shin, Elizabeth Abraham, Steven Isakoff, Beverly Moy, Leif Ellisen, Aditya Bardia. Sequencing Antibody-Drug Conjugate after Antibody-Drug Conjugate in Metastatic Breast Cancer (A3 study): Multi-Institution Experience and Biomarker Analysis [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PS08-03.

Distribution and Mechanism of Japanese Brome (Bromus japonicus) Resistance to ALS-Inhibiting Herbicides in China.

Bromus japonicus is a common monocot weed that occurs in major winter wheat fields in the Huang-Huai-Hai region of China. Pyroxsulam is a highly efficient and safe acetolactate synthase (ALS)-inhibiting herbicide that is widely used to control common weeds in wheat fields. However, B. japonicus populations in China have evolved resistance to pyroxsulam by different mutations in the ALS gene. To understand the resistance distribution, target-site resistance mechanisms, and cross-resistance patterns, 208 B. japonicus populations were collected from eight provinces. In the resistant population screening experiment, 59 populations from six provinces showed different resistance levels to pyroxsulam compared with the susceptible population, of which 17 B. japonicus populations with moderate or high levels of resistance to pyroxsulam were mainly from the Hebei (4), Shandong (4) and Shanxi (9) Provinces. Some resistant populations were selected to investigate the target site-resistance mechanism to the ALS-inhibiting herbicide pyroxsulam. Three pairs of primers were designed to amplify the ALS sequence, which was assembled into the complete ALS sequence with a length of 1932 bp. DNA sequencing of ALS revealed that four different ALS mutations (Pro-197-Ser, Pro-197-Thr, Pro-197-Phe and Asp-376-Glu) were found in 17 moderately or highly resistant populations. Subsequently, five resistant populations, QM21-41 with Pro-197-Ser, QM20-8 with Pro-197-Thr and Pro-197-Phe, and QM21-72, QM21-76 and QM21-79 with Asp-376-Glu mutations in ALS genes, were selected to characterize their cross-resistance patterns to ALS inhibitors. The QM21-41, QM20-8, QM21-72, QM21-76 and QM21-79 populations showed broad-spectrum cross-resistance to pyroxsulam, mesosulfuron-methyl and flucarbazone-sodium. This study is the first to report evolving cross-resistance to ALS-inhibiting herbicides due to Pro-197-Phe mutations in B. japonicus.

Sensitivity of Lithuanian Zymoseptoria tritici to Quinone Outside Inhibitor and Succinate Dehydrogenase Inhibitor Fungicides

Septoria tritici blotch (STB) ais one of the most damaging winter wheat diseases worldwide, presenting a significant threat to its yields. The causal STB agent, Zymoseptoria tritici, also presents a challenge to control due to its rapid adaptation to fungicides. This requires researchers to continuously monitor the pathogen and investigate and explore strategies to manage the spread of the disease and the development of resistance in the pathogen. Therefore, this study presents the current situation and describes changes in the sensitivity of Z. tritici isolates from Lithuania to quinone outside inhibitors (QoIs) and succinate dehydrogenase inhibitors (SDHIs) for the years 2019–2022. The isolates were tested at five different concentrations of two QoI fungicides (azoxystrobin and pyraclostrobin) and three SDHI fungicides (fluxapyroxad, benzovindiflupyr, and bixafen). During the test period, the EC50 values of the tested QoIs increased, while no clear changes were observed in the SDHIs. The most pronounced shift was observed for the active QoI substance pyraclostrobin. The distribution of the EC50 values of the SDHI fungicides showcased one isolate with an outstandingly high EC50 value of 2.6 mg L−1. The results of this study did not reveal any strong patterns of cross-resistance between the fungicides tested. However, a significant positive, moderate correlation (r = 0.55) was found between fluxapyroxad and benzovindiflupyr. Overall, the results of this study contribute to the understanding of the fungicide-resistance situation of Z. tritici in Lithuania and may complement management strategies for the pathogen and its fungicide resistance.

The Pro-197-Thr mutation in the ALS gene confers novel resistance patterns to ALS-inhibiting herbicides in Bromus japonicus in China.

Bromus japonicus is one of the most notorious agricultural weeds in China. The long-term use of ALS-inhibiting herbicides has led to rapid evolution of herbicide resistance in B. japonicus. B. japonicus population (BJ-R) surviving mesosulfuron-methyl treatment was collected from wheatland. Here, we aimed to confirm the resistance mechanisms in this putative resistant population. The dose-reponse tests were used to test the resistance level of the B. japonicus to ALS-inhibiting herbicides. Pretreatment with P450 and GST inhibitors and GST activity assays were used to determine whether P450 or GST was involved in the resistance of the BJ-R population. Sanger sequencing was used to analyse the ALS mutation of the BJ-R population. RT-qPCR was used to confirm the the expression levels of the ALS gene in mesosulfuron-methyl -resistant (BJ-R) and-susceptible (BJ-S) B. japonicus. An in vitro ALS activity assay was used to determine the ALS activity of the BJ-R and BJ-S populations. Homology modelling and docking were used to determine the binding energy of the BJ-R and BJ-S populations with ALS-inhibiting herbicides. B. japonicus population (BJ-R) was confirmed to be 454- and 2.7-fold resistant to the SU herbicides mesosulfuron-methyl and nicosulfuron, and 7.3-, 2.3-, 1.1- and 10.8-fold resistant to the IMI herbicide imazamox, the TP herbicide penoxsulam, the PTB herbicide pyribenzoxim and the SCT herbicide flucarbazone-sodium, respectively, compared with its susceptible counterpart (BJ-S). Neither a P450 inhibitor nor a GST inhibitor could reverse the level of resistance to mesosulfuron-methyl in BJ-R. In addition, no significant differences in GST activity were found between the BJ-R and BJ-S. ALS gene sequencing revealed a Pro-197-Thr mutation in BJ-R, and the gene expression had no significant differences between the BJ-R and BJ-S. The ALS activity of BJ-R was 106-fold more tolerant to mesosulfuron-methyl than that of BJ-S. Molecular docking showed that the binding energy of the ALS active site and mesosulfuron-methyl was changed from -6.67 to -4.57 kcal mol-1 due to the mutation at position 197. These results suggested that the Pro-197-Thr mutation was the main reason for the high resistance level of BJ-R to mesosulfuron-methyl. Unlike previous reports of the cross-resistance pattern conferred by this mutation, we firstly documented that the Pro-197-Thr mutation confers broad cross-resistance spectrums to ALS-inhibiting herbicides in B. japonicus.

Identification of candidate genes involved with dicamba resistance in waterhemp (Amaranthus tuberculatus) via transcriptomics analyses

AbstractWaterhemp [Amaranthus tuberculatus (Moq.) Sauer] is one of the most troublesome weeds in the United States. An A. tuberculatus population (CHR) was identified in Illinois, USA, as resistant to herbicides from six different site-of-action groups. Recently, the same population was also recognized as dicamba resistant. This study aimed to identify key resistance genes and the putative dicamba resistance mechanism in A. tuberculatus via transcriptomics analysis. Multiple differentially expressed (DE) genes and co-expression gene modules were identified as associated with dicamba resistance. Specifically, genes encoding glutathione S-transferases (GSTs), ATP-binding cassette transporters, peroxidases, and uridine diphosphate (UDP)-glycosyltransferases (UGTs) were identified. Results indicated enhanced oxidative stress tolerance as the primary mechanism for reducing dicamba toxicity. Results also point to potential glycosylation via UGTs and conjugation via GSTs of dicamba and its by-products. This is the first transcriptomics characterization of dicamba resistance in A. tuberculatus. Multiple non-target-site resistance genes were identified, indicating a cross-resistance pattern in the CHR population leading to a putative-enhanced oxidative stress response. Regions of multiple DE genes (i.e., genomic hot spots) across the A. tuberculatus genome corroborate previous results and potentially add to the complexity of non-target-site resistance traits.

In Doxorubicin-Adapted Hodgkin Lymphoma Cells, Acquiring Multidrug Resistance and Improved Immunosuppressive Abilities, Doxorubicin Activity Was Enhanced by Chloroquine and GW4869.

Classical Hodgkin lymphoma (cHL) is a highly curable disease (70-80%), even though long-term toxicities, drug resistance, and predicting clinical responses to therapy are major challenges in cHL treatment. To solve these problems, we characterized two cHL cell lines with acquired resistance to doxorubicin, KM-H2dx and HDLM-2dx (HRSdx), generated from KM-H2 and HDLM-2 cells, respectively. HRSdx cells developed cross-resistance to vinblastine, bendamustin, cisplatin, dacarbazine, gemcitabine, brentuximab vedotin (BV), and γ-radiation. Both HDLM-2 and HDLM-2dx cells had intrinsic resistance to BV but not to the drug MMAE. HDLM-2dx acquired cross-resistance to caelyx. HRSdx cells had in common decreased CD71, CD80, CD54, cyt-ROS, HLA-DR, DDR1, and CD44; increased Bcl-2, CD58, COX2, CD26, CCR5, and invasive capability; increased CCL5, TARC, PGE2, and TGF-β; and the capability of hijacking monocytes. In HRSdx cells less sensitive to DNA damage and oxidative stress, the efflux drug transporters MDR1 and MRP1 were not up-regulated, and doxorubicin accumulated in the cytoplasm rather than in the nucleus. Both the autophagy inhibitor chloroquine and extracellular vesicle (EV) release inhibitor GW4869 enhanced doxorubicin activity and counteracted doxorubicin resistance. In conclusion, this study identifies common modulated antigens in HRSdx cells, the associated cross-resistance patterns, and new potential therapeutic options to enhance doxorubicin activity and overcome resistance.

Helicoverpa armigera (Hubner) Associated Cross Resistance Patterns in South Indian Crop Ecosystem

Aim: To study the cross resistance patterns associated with Mahaboobnagar, Raichur, Nagpur populations of Helicoverpa armigera (Hubner).
 Study design: Bioassay
 Place and Duration of Study: The experiment was carried out from February 2010 to May 2011 at Department of Entomology, College of Agriculture, Rajendranagar, Hyderabad, Telangana.
 Methodology: Helicoverpa armigera was selected for indoxacarb in F1 and F2 continuously then the population subjected to different selected insecticides to know the cross resistance patterns associated.
 Results: Mahaboobnagar population recorded 1.109 and 0.816 fold resistance at LD50 and LD90, respectively, while Raichur population has developed still higher levels of relative resistance by 1.591 and 0.846 fold when compared with the Nagpur population at LD50 and LD90, respectively. Similarly, the Raichur population has developed 1.435 and 1.037 folds relative resistance at LD50­ and LD90, respectively as compared with the Mahaboobnagar population.
 The Mahaboobnagar population resistant to indoxacarb at F3, when subjected to selected insecticides like cypermethrin, methomyl, spinosad showed a negative cross resistance ratio of 0.665, 0.830, 0.916 to cypermethrin, methomyl, spinosad respectively, and a positive cross resistance ratio of 1.019 to indoxacarb, while similar trend was displayed by Raichur population showing a negative cross resistance ratio of 0.932, 0.565, 0.803 to cypermethrin, methomyl, spinosad respectively and positive cross resistance of 1.036 indoxacarb further, same trend was shown by Nagpur population by displaying a negative cross resistance ratio of 0.610, 0.735, 0.519 to cypermethrin, methomyl, spinosad and positive cross resistance ratio of 1.026 to indoxacarb.
 Conclusion: Continuous application of single insecticide belonging to a specific group across the generations increases the resistance from F1 to F3. Alternating the new chemistries with old conventional chemicals resulted in no cross resistance development as it was observed in all test populations.

First report of target-site resistance to ACCase-inhibiting herbicides in Bromus tectorum L.

The prevalent and repeated use of acetyl-coenzyme A carboxylase (ACCase)-inhibiting herbicides for Bromus tectorum L. control in fine fescue (Festuca L. spp) grown for seed has selected ACCase-resistant B. tectorum populations. The objectives of this study were to (1) evaluate the response of nine B. tectorum populations to the ACCase inhibitors clethodim, sethoxydim, fluazifop-P-butyl, and quizalofop-P-ethyl and the acetolactate synthase (ALS) inhibitor sulfosulfuron and (2) characterize the resistance mechanisms. Bromus tectorum populations were confirmed to be resistant to the ACCase-inhibiting herbicides tested. The levels of resistance varied among the populations for clethodim (resistance ratio, RR = 5.1-14.5), sethoxydim (RR = 18.7-44.7), fluazifop-P-butyl (RR = 3.1-40.3), and quizalofop-P-ethyl (RR = 14.5-36). Molecular investigations revealed that the mutations Ile2041Thr and Gly2096Ala were the molecular basis of resistance to the ACCase-inhibiting herbicides. The Gly2096Ala mutation resulted in cross-resistance to the aryloxyphenoxypropionate (APP) herbicides fluazifop-P-butyl and quizalofop-P-ethyl, and the cyclohexanedione (CHD) herbicides clethodim, and sethoxydim, whereas Ile2041Thr mutation resulted in resistance only to the two APP herbicides. All B. tectorum populations were susceptible to sulfosulfuron (RR = 0.3-1.7). This is the first report of target-site mutations conferring resistance to ACCase-inhibiting herbicides in B. tectorum. The results of this study suggest multiple evolutionary origins of resistance and contribute to understanding the patterns of cross-resistance to ACCase inhibitors associated with different mutations in B. tectorum. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Characterization of Two Novel Inhibitors of the Mycobacterium tuberculosis Cytochrome bc1 Complex.

Drug-resistant tuberculosis is a global health care threat calling for novel effective treatment options. Here, we report on two novel cytochrome bc1 inhibitors (MJ-22 and B6) targeting the Mycobacterium tuberculosis respiratory chain with excellent intracellular activities in human macrophages. Both hit compounds revealed very low mutation frequencies and distinct cross-resistance patterns with other advanced cytochrome bc1 inhibitors.

Application of ALS inhibitors at pre-emergence is effective in controlling resistant barnyardgrass biotypes depending on the mechanism of resistance

Echinochloa species are one of the most troublesome weeds in rice crops, and their control is hampered due to herbicide resistance. The aim of this study was to identify the cross-resistance pattern and the differential resistance level for pre- and post-emergence applications of imazethapyr and penoxsulam in populations of barnyardgrass [Echinochloa crus-galli (L.) Beauv.] with different ALS gene mutations. Of 26 biotypes, 23 were imazethapyr-resistant, and 10 were cross-resistant to penoxsulam. The resistance index (RI) to imazethapyr was 5.7–19.5 for Ser653Asn, 26.7–68.3 for Ala122Thr and Ala205Asn, and 70.9–252.9 for Trp574Leu. Only Trp574Leu also resulted in resistance to penoxsulam. The double ALS mutation Ala122Asn + Trp574Leu resulted in a RI for imazethapyr and penoxsulam higher than 2800. The ED50 for penoxsulam applied at pre-emergence was three and six times lower than at post-emergence for the susceptible and resistant biotypes, respectively. The application of imazethapyr at pre-emergence was more effective than at post-emergence only for the biotypes with low resistance level mutation Ser653Asn. The efficacy of the herbicide quinclorac was similar for the application at pre- and post-emergence for susceptible and resistant biotypes. The ALS mutations Ala205Asn and Ala122Asn + Trp574Leu are reported for the first time in barnyardgrass. The use of ALS inhibitors at pre-emergence should be prioritized over post-emergence in fields with resistant barnyardgrass and in need of using these products to control other non-resistant weeds.