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The redlegged earth mite, Halotydeus destructor, is an economically important pest of winter field crops and pastures in Australia, and has evolved field resistance to pyrethroid chemicals through a polymorphism in the voltage-gated parasodium channel leading to knockdown resistance (kdr). In this study, we quantified the rate of reversion to susceptibility of partially resistant H. destructor populations under field conditions in the absence of pesticide exposure. This was conducted over multiple years at two geographically distant locations with mite populations known to possess pyrethroid resistance. Fitness costs associated with pyrethroid resistance were identified through reductions in the frequency of kdr resistance alleles in the absence of pesticides. This was assessed using an amplicon sequencing approach targeting known resistance alleles. We also found that resistance can increase rapidly in frequency after only a single pyrethroid application in the field. Our results highlight that, once established in H. destructor populations, pyrethroid resistance will not easily be lost even after several years, emphasizing the importance of limiting the evolution of resistance in the first place. This helps to explain why pyrethroid resistance in H. destructor continues to persist at very high frequencies in the field and continues to expand within Australia despite the existence of fitness costs. Understanding field fitness costs associated with pesticide resistance is important when devising resistance management strategies for this pest. [ABSTRACT FROM AUTHOR]

ABSTRACT Multi-fungicide resistance (MFR) is a serious environmental problem, which results in the excessive use of fungicides. Fitness penalty, as a common phenomenon in MFR, can partially counteract the issue of resistance due to the weakened vigor of MFR pathogens. Their underlying mechanism and relationship remain unexplained. By Oxford Nanopore Technologies sequencing and dot blot, we found that N6-methyloxyadenine (6mA) modification, the dominate epigenetic marker in Phytophthora capsici, was significantly altered after MFR emerged. Among the differently methylated genes, PcGSTZ1 could efficiently detoxify SYP-14288, a novel uncoupler, through complexing the fungicide with glutathione and induce MFR. Interestingly, PcGSTZ1 overexpression was induced by elevated 6mA levels and chromatin accessibility to its genomic loci. Moreover, the overexpression led to reactive oxygen species burst and ferroptosis in SYP-14288-resistant mutants, which enhanced the resistance and induced fitness penalty in P. capsici through triggering low energy shock adaptive response. Furthermore, this study revealed that the 6mA-PcGSTZ1-ferroptosis axis could mediate intergenerational resistance memory transmission and enabled adaptive advantage to P. capsici. In conclusion, the findings provide new insights into the biological role of 6mA as well as the mechanisms underlying the trade-off between MFR and fitness. These could also benefit disease control through the blockade of the epigenetic axis to resensitize resistant isolates.IMPORTANCEN6-methyloxyadenine (6mA) modification on DNA is correlated with tolerance under different stress in prokaryotes. However, the role of 6mA in eukaryotes remains poorly understood. Our current study reveals that DNA adenine methyltransferase 1 (DAMT1)-mediated 6mA modification at the upstream region of GST zeta 1 (GSTZ1) is elevated in the resistant strain. This elevation promotes the detoxification uncoupler and induces multifungicide resistance (MFR). Moreover, the overexpression led to reactive oxygen species burst and ferroptosis in SYP-14288-resistant mutants, which enhanced the resistance and induced fitness penalty in Phytophthora capsici through triggering low energy shock adaptive response. Furthermore, this study revealed that the 6mA-PcGSTZ1-ferroptosis axis could mediate intergenerational resistance memory transmission and enabled adaptive advantage to P. capsici. Overall, our findings uncover an innovative mechanism underlying 6mA modification in regulating PcGSTZ1 transcription and the ferroptosis pathway in P. capsici.

BACKGROUND: Cucumber fruit rot (CFR) caused by Fusarium incarnatum is a devastating fungal disease in cucumber. In recent years, CFR has occurred frequently, resulting in serious yield and quality losses in China. Phenamacril exhibits a specific antifungal activity against Fusarium species. However, no data for phenamacril against F. incarnatum is available. RESULTS: The sensitivity of 80 F. incarnatum strains to phenamacril was determined. The half maximal effective concentration (EC50) values ranged from 0.1134 to 0.3261 μg mL−1 with a mean EC50 value of 0.2170 ± 0.0496 μg mL−1. A total of seven resistant mutants were obtained from 450 mycelial plugs by phenamacril‐taming on potato dextrose agar (PDA) plates with 10 μg mL−1 of phenamacril, and the resistant frequency was 1.56%. Phenamacril‐resistant mutants showed decreased mycelial growth, conidiation and virulence as compared with the corresponding wild‐type strains, indicating that phenamacril resistance suffered a fitness penalty in F. incarnatum. In addition, using sequence analysis, the point mutations of S217P or I424S were discovered in Fimyosin‐5 (the target of phenamacril). The site‐directed mutagenesis of the S217P, P217S, I424S and S424I substitutions were constructed to reveal the relationship between the point mutations and phenamacril resistance. The results strongly demonstrated that the mutations of S217P and I424S in Fimyosin‐5 conferred phenamacril‐resistance in F. incarnatum. CONCLUSION: Phenamacril‐resistant mutants were easily induced and their resistance level was high. The S217P or I424S substitutions in Fimyosin‐5 conferring phenamacril resistance were detected and futherly verified by transformation assay with site‐directed mutagenesis. Thus, we proposed that the resistance development of F. incarnatum to phenamacril is high risk. © 2022 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

BACKGROUND: Resistance to protoporphyrinogen oxidase (PPO)‐inhibiting herbicides is endowed primarily by target‐site mutations at the PPX2 gene that compromise binding of the herbicide to the catalytic domain. In Amaranthus spp. PPX2, the most prevalent target mutations are deletion of the G210 codon, and the R128G and G339A substitutions. These mutations strongly affect the dynamic of the PPO2 binding pocket, resulting in reduced affinity with the ligand. Here we investigated the likelihood of co‐occurrence of the most widespread target site mutations in the same PPX2 allele. RESULTS: Plants carrying R128G+/+ ΔG210+/−, where + indicates presence of the mutation, were crossed with each other. The PPX2 of the offspring was subjected to pyrosequencing and E. coli‐based Sanger sequencing to determine mutation frequencies and allele co‐occurrence. The data show that R128G ΔG210 can occur in one allele only; the second allele carries only one mutation. Double mutation in both alleles is less likely because of significant loss of enzyme activity. The segregation of offspring populations derived from a cross between heterozygous plants carrying ΔG210 G399A also showed no co‐occurrence in the same allele. The offspring exhibited the expected mutation distribution patterns with few exceptions. CONCLUSIONS: Homozygous double‐mutants are not physiologically viable. Double‐mutant plants can only exist in a heterozygous state. Alternatively, if two mutations are detected in one plant, each mutation would occur in a separate allele. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Abstract Climate change and pesticide resistance are two of the most imminent challenges human society is facing today. Knowledge of how the evolution of pesticide resistance may be affected by climate change such as increasing air temperature on the planet is important for agricultural production and ecological sustainability in the future but is lack in scientific literatures reported from empirical research. Here, we used the azoxystrobin‐Phytophthora infestans interaction in agricultural systems to investigate the contributions of environmental temperature to the evolution of pesticide resistance and infer the impacts of global warming on pesticide efficacy and future agricultural production and ecological sustainability. We achieved this by comparing azoxystrobin sensitivity of 180 P. infestans isolates sampled from nine geographic locations in China under five temperature schemes ranging from 13 to 25°C. We found that local air temperature contributed greatly to the difference of azoxystrobin tolerance among geographic populations of the pathogen. Both among‐population and within‐population variations in azoxystrobin tolerance increased as experimental temperatures increased. We also found that isolates with higher azoxystrobin tolerance adapted to a broader thermal niche. These results suggest that global warming may enhance the risk of developing pesticide resistance in plant pathogens and highlight the increased challenges of administering pesticides for effective management of plant diseases to support agricultural production and ecological sustainability under future thermal conditions.

Resistance to insecticides used to control pests is an issue of increasing concern for agriculture. The grain aphid, Sitobion avenae, is a pest of cereals and grasses worldwide, and one of growing concern due to the evolution of resistance to certain insecticides. Resistance confers benefits to insects by enabling them to survive exposure to insecticide compounds; however, the mutations conferring resistance may also penalise the insect in pesticide‐free environments due to fitness costs associated with the new phenotype. Here we tested the hypothesis of a reproductive penalty linked to the knockdown resistance mutation (kdr) to pyrethroid insecticides. The mutation occurs predominantly in a single SA3 clone. To date, only heterozygous‐resistant forms (kdr‐SR) have been detected in populations in Ireland and the UK, and this suggests that a fitness penalty may preclude the formation of both male and female heterozygous‐resistant sexual forms. By designing an experiment which included a resistant and a non‐resistant clone, we were able to simulate reduced daylight and temperature conditions which, in nature, trigger sexual reproduction and therefore study the responses of each clone. This allowed us to detect the switch from asexual females to sexual females and males and report on the conditions associated with the production of sexual forms. The results showed that both aphid clones were able to produce sexual forms with no difference in the onset of sexual reproduction, although reproductive strategies differed between clones. The later onset of male forms in the SA3 clone may decrease the likelihood of mating interactions to create fully resistant (kdr‐RR) genotypes and this may constitute a fitness penalty due to pyrethroid resistance. [ABSTRACT FROM AUTHOR]

Abstract Background Cross-resistance, a phenomenon that a pathogen resists to one antimicrobial compound also resists to one or several other compounds, is one of major threats to human health and sustainable food production. It usually occurs among antimicrobial compounds sharing the mode of action. In this study, we determined the sensitivity profiles of Alternaria alternata, a fungal pathogen which can cause diseases in many crops to two fungicides (mancozeb and difenoconazole) with different mode of action using a large number of isolates (234) collected from seven potato fields across China. Results We found that pathogens could also develop cross resistance to fungicides with different modes of action as indicated by a strong positive correlation between mancozeb and difenoconazole tolerances to A. alternata. We also found a positive association between mancozeb tolerance and aggressiveness of A. alternata, suggesting no fitness penalty of developing mancozeb resistance in the pathogen and hypothesize that mechanisms such as antimicrobial compound efflux and detoxification that limit intercellular accumulation of natural/synthetic chemicals in pathogens might account for the cross-resistance and the positive association between pathogen aggressiveness and mancozeb tolerance. Conclusions The detection of cross-resistance among different classes of fungicides suggests that the mode of action alone may not be an adequate sole criterion to determine what components to use in the mixture and/or rotation of fungicides in agricultural and medical sects. Similarly, the observation of a positive association between the pathogen’s aggressiveness and tolerance to mancozeb suggests that intensive application of site non-specific fungicides might simultaneously lead to reduced fungicide resistance and enhanced ability to cause diseases in pathogen populations, thereby posing a greater threat to agricultural production and human health. In this case, the use of evolutionary principles in closely monitoring populations and the use of appropriate fungicide applications are important for effective use of the fungicides and durable infectious disease management.

Climate change and pesticide resistance are two of the most imminent challenges human society is facing today. Knowledge of how the evolution of pesticide resistance may be affected by climate change such as increasing air temperature on the planet is important for agricultural production and ecological sustainability in the future but is lack in scientific literatures reported from empirical research. Here, we used the azoxystrobin‐Phytophthora infestans interaction in agricultural systems to investigate the contributions of environmental temperature to the evolution of pesticide resistance and infer the impacts of global warming on pesticide efficacy and future agricultural production and ecological sustainability. We achieved this by comparing azoxystrobin sensitivity of 180 P. infestans isolates sampled from nine geographic locations in China under five temperature schemes ranging from 13 to 25°C. We found that local air temperature contributed greatly to the difference of azoxystrobin tolerance among geographic populations of the pathogen. Both among‐population and within‐population variations in azoxystrobin tolerance increased as experimental temperatures increased. We also found that isolates with higher azoxystrobin tolerance adapted to a broader thermal niche. These results suggest that global warming may enhance the risk of developing pesticide resistance in plant pathogens and highlight the increased challenges of administering pesticides for effective management of plant diseases to support agricultural production and ecological sustainability under future thermal conditions. [ABSTRACT FROM AUTHOR]

Multi-fungicide resistance (MFR) is a serious environmental problem, which results in the excessive use of fungicides. Fitness penalty, as a common phenomenon in MFR, can partially counteract the issue of resistance due to the weakened vigor of MFR pathogens. Their underlying mechanism and relationship remain unexplained. By Oxford Nanopore Technologies sequencing and dot blot, we found that N6-methyloxyadenine (6mA) modification, the dominate epigenetic marker in Phytophthora capsici , was significantly altered after MFR emerged. Among the differently methylated genes, PcGSTZ1 could efficiently detoxify SYP-14288, a novel uncoupler, through complexing the fungicide with glutathione and induce MFR. Interestingly, PcGSTZ1 overexpression was induced by elevated 6mA levels and chromatin accessibility to its genomic loci. Moreover, the overexpression led to reactive oxygen species burst and ferroptosis in SYP-14288-resistant mutants, which enhanced the resistance and induced fitness penalty in P. capsici through triggering low energy shock adaptive response. Furthermore, this study revealed that the 6mA-PcGSTZ1-ferroptosis axis could mediate intergenerational resistance memory transmission and enabled adaptive advantage to P. capsici . In conclusion, the findings provide new insights into the biological role of 6mA as well as the mechanisms underlying the trade-off between MFR and fitness. These could also benefit disease control through the blockade of the epigenetic axis to resensitize resistant isolates.IMPORTANCEN6-methyloxyadenine (6mA) modification on DNA is correlated with tolerance under different stress in prokaryotes. However, the role of 6mA in eukaryotes remains poorly understood. Our current study reveals that DNA adenine methyltransferase 1 (DAMT1)-mediated 6mA modification at the upstream region of GST zeta 1 (GSTZ1) is elevated in the resistant strain. This elevation promotes the detoxification uncoupler and induces multifungicide resistance (MFR). Moreover, the overexpression led to reactive oxygen species burst and ferroptosis in SYP-14288-resistant mutants, which enhanced the resistance and induced fitness penalty in Phytophthora capsici through triggering low energy shock adaptive response. Furthermore, this study revealed that the 6mA-PcGSTZ1-ferroptosis axis could mediate intergenerational resistance memory transmission and enabled adaptive advantage to P. capsici . Overall, our findings uncover an innovative mechanism underlying 6mA modification in regulating PcGSTZ1 transcription and the ferroptosis pathway in P. capsici ., Competing Interests: The authors declare no conflict of interest.

Despite considerable differences in cropping systems around the globe, chemical weed control is a key tool in conventional agroecosystems, which has led to an increase in herbicide resistance. Although mutations causing resistance are thought to have an adaptation cost in resistant plants compared to the susceptible ones under herbicide-free conditions, such cost may not always express or will express under certain ecological conditions. To ensure that herbicides will keep going as viable instruments in agricultural production, strategies to minimize resistance are needed. Proactive or reactive strategies for weed control should utilize an overall integrated weed management approach by combining as many weed management practices as possible. The term 'superweed' was used initially to describe the phenomenon in which genetically engineered crops would become troublesome weeds and that the genes of interest would spread into related weeds, rendering them problematic, or into wild species, turning them into troublesome weeds. Contrary to the above definition, the use of this term in the literature has often been linked with herbicide resistance, mostly related to the cultivation of genetically engineered crops and the related increase in the use of glyphosate, which rapidly selected resistant weed populations. From a scientific point of view, weeds are better survivors than non-weedy species and cause crop problems because they have several unique traits, e.g., they are aggressive, adapt easily to different environments, produce many seeds, compete strongly with crops, disperse easily, are difficult to control, traits which occur whether weeds are herbicide-resistant or not. We propose that the term 'superweed' should be referred to weeds with resistant populations to several herbicides with diverse modes of action (MOAs). • Chemical weed control is a key tool in conventional agroecosystems, which has led to an increase in herbicide resistance. • Resistant weeds may have an adaptation cost under herbicide-free conditions, but such cost may not always express. • Proactive or reactive strategies for weed control should utilize an overall integrated weed management. • The term 'superweed' has been linked with herbicide resistance, related to the cultivation of genetically engineered crops. • Weeds have several unique traits, which occur whether plants are resistant or not. [ABSTRACT FROM AUTHOR]

Quantitative resistance (QR) to crop diseases has usually been much more durable than major-gene, effector-triggered resistance. It has been observed that the effectiveness of some QR has eroded as pathogens adapt to it, especially when deployment is extensive and epidemics occur regularly, but it generally declines more slowly than effector-triggered resistance. Changes in aggressiveness and specificity of diverse pathogens on cultivars with QR have been recorded, along with experimental data on fitness costs of pathogen adaptation to QR, but there is little information about molecular mechanisms of adaptation. Some QR has correlated or antagonistic effects on multiple diseases. Longitudinal data on cultivars' disease ratings in trials over several years can be used to assess the significance of QR for durable resistance in crops. It is argued that published data likely underreport the durability of QR, owing to publication bias. The implications of research on QR for plant breeding are discussed. [ABSTRACT FROM AUTHOR]

Summary: Plant defence traits, such as herbicide resistance mutations, may incur a fitness cost to plants that become evident when the trait is not needed. However, individuals with multiple herbicide resistance traits may decrease fitness beyond that of plants with a single herbicide resistance mutation. Multiple herbicide‐resistant (MHR) Amaranthus tuberculatus populations are becoming more prevalent in Midwest United States agroecosystems. The objective was to determine whether selected MHR A. tuberculatus populations express differential development when grown in a herbicide‐free environment. The hypothesis was that MHR A. tuberculatus populations become increasingly less fit when additional herbicide resistances evolve. Multiple herbicide‐resistant and herbicide‐susceptible A. tuberculatus populations were grown in a herbicide‐free field for 20 weeks for two seasons. Differences (P < 0.001) in apical growth were detected 5 and 7 weeks after transplanting for all populations in 2016 and 2017 respectively. Gender and population influenced (P < 0.001) flowering date, with males flowering up to 1.5 weeks earlier than females, but did not cause pollination asynchrony. Shoot biomass was not different (P = 0.84) across A. tuberculatus populations, but there were differences (P < 0.001) for gender and year. Seed production was different amongst A. tuberculatus populations (P = 0.001), but was not influenced by the number of MHR traits. Conversely, a negative quadratic relationship between seed mass and the number of MHR traits was observed (r2 = 0.32; P < 0.001). The experiment results demonstrate that MHR in A. tuberculatus populations is not incurring a fitness penalty that will remove the populations in the immediate future. [ABSTRACT FROM AUTHOR]

Annual bluegrass (Poa annua L.) is an annual weed that is particularly troublesome in managed turfgrass. It has been controlled conventionally with herbicides, including acetolactate synthase (ALS) inhibitors. However, resistance to ALS inhibitors has been documented throughout the southeastern United States since 2012. A rate-response trial was conducted to confirm and determine the resistance level of suspected resistant P. annua biotypes from Mississippi (Reunion), followed by DNA sequencing to determine whether the mechanism of resistance is a target-site mutatio n. In addition, a fitness assay was conducted together with a susceptible biotype to determine whether resistance to ALS inhibitors is associated with decreased fitness. Reunion was at least 45 times more resistant to foramsulfuron than the standard susceptible biotype based on I50 estimates [I50 is the rate of herbicide giving a 50% response (50% visual necrosis)], requiring a predicted 331 g a.i./ha foramsulfuron for 50% control. DNA sequencing results identified a Trp574-to-Leu mutation in the ALS gene of the Reunion biotype, which has been shown by other studies to confer resistance to ALS inhibitors. Measurement of fitness parameters among the Reunion and susceptible biotypes demonstrated reduced seed yield, tillering, and flowering time in the resistant Reunion biotype, suggesting that ALS inhibitor resistance is possibly correlated to decreased fitness in P. annua. Alternative methods to control P. annua need to be considered as a result of the evolution of herbicide-resistant biotypes. An integrated management strategy to control P. annua weeds will help prevent further evolution of resistance. Because this study evaluated only the target-site mechanism of resistance, it is also necessary to determine whether the resistant biotype has reduced uptake, translocation, or enhanced metabolism as additional mechanisms of resistance. Consequently, a fitness study encompassing a more comprehensive list of plant parameters will provide conclusions of the fitness costs associated with ALS inhibitor resistance in P. annua. Chemical names: Foramsulfuron {1-(4,6-dimethoxypyrimidin-2-yl)-3-[2-(dimethylcarbamoyl)-5-formamidophenylsulfonyl] urea}. [ABSTRACT FROM AUTHOR]

Echinochloa colona L. (Link) (awnless barnyard grass) is one of the top three most problematic weeds of summer crops in Australia. This weed has evolved resistance to glyphosate. A study was conducted to evaluate the effect of environmental factors on the germination and seedling emergence of a glyphosate-resistant (GR) and a glyphosate-susceptible (GS) biotype of E. colona. The two biotypes had similar germination and emergence responses to light and temperature conditions, water stress, solution pH, sorghum residue cover and seed burial depth. Light stimulated germination more than dark conditions, and seeds germinated at a wide range of alternating day/night temperatures, from 20°C/10°C to 35°C/25°C, whereas no seeds germinated at 15°C/5°C. These results suggest that E. colona can emerge in spring, summer and autumn in Queensland. The sodium chloride concentration required to inhibit 50% germination was greater for the GR biotype (209 mm) than the GS biotype (174 mm). Seed germination was not affected by pH in the range 4–10. Water stress reduced germination by 50% at an osmotic potential of –0.44 MPa. In a shade-house study, retention of sorghum residue cover on the soil surface reduced the seedling emergence of E. colona. Emergence was 70% in the absence of crop residue, whereas a residue amount of 8 t ha–1 reduced emergence to 47%. Emergence was greatest for seeds placed on the soil surface and declined linearly with increasing burial depth; no seedlings emerged from 8 cm depth. The GR biotype had higher germination than the GS biotype under high sodium chloride concentrations; therefore, this biotype may be highly competitive with crops under highly saline conditions. Because germination was high on the soil surface and was stimulated by light, this weed will remain problematic under no-till farming systems in Australia. Awnless barnyard grass seeds germinated at alternating day/night temperatures ranging from 20/10°C to 35/25°C. Seedling emergence was highest on the soil surface, decreased with increased burial depth, but no seedlings emerged from 8 cm. Emergence reduced by 47% with the sorghum residue load of 8 t ha–1. [ABSTRACT FROM AUTHOR]

Conyza bonariensis (L.) Cronquist is one of the main glyphosate-resistant weeds in no-till fields in Brazil. Here, the seed quality of glyphosate-resistant (R) and -susceptible (S) C. bonariensis biotypes, collected from different sites, was evaluated under stressful conditions by different seed tests. Glyphosate resistance was confirmed by dose-response and shikimate accumulation assays. The resistance factors were 6.9 (R1/S1), 4.5 (R2/S2), and 5.8 (R3/S3). Biotypes S1, S2, and S3 accumulated 2.7, 2.4, and 2.8 times more shikimic acid than biotypes R1, R2, and R3, respectively. Stress-free seed viability and germination potential ranged from 39% to 57% and from 37% to 57%, respectively, with no difference between R and S biotypes within each collection site. Seed incubation at 8 °C over 7 days (cold test) promoted greater germination in S biotypes (54% to 79%) compared to R ones (28% to 39%). In the accelerated aging tests (incubation at 42 °C over 48 hours), the germination decreased in both S (11% to 27%) and R (6% to 16%) biotypes. In the high-temperature stress tests, there were no differences in germination within biotypes at 35 and 45 °C; however, at 60 °C, the germination of the S1, R1, S2, R2, S3, and R3 biotypes was reduced by approximately 51%, 54%, 63%, 59%, 40%, and 30%, respectively. Under non-stressful conditions, germination potential and seed viability were similar in R and S biotypes; however, under cold or heat stress conditions, R biotypes reduced their germination rates, revealing that glyphosate resistance causes a fitness penalty in C. bonariensis at the seed level. However, because seed viability was not determined after experiments, it cannot be stated that such reduction in germination was due to the death or only a dormant phase of the seeds.

Wild poinsettia (Euphorbia heterophylla L.) is a difficult-to-control weed in soybean production in Brazil that has developed resistance to herbicides, including acetolactate synthase inhibitors. We investigated the potential fitness cost associated to the Ser-653-Asn mutation that confers imazamox resistance in this weed. Plant height, leaf and stem dry weight, leaf area and seed production per plant as well as the growth indices of specific leaf area, leaf area ratio, relative growth rate and net assimilation in F2 homozygous resistant (R) and susceptible (S) wild poinsettia progenies were pairwise compared. S plants were superior in most of the traits studied. Plant heights for S and R biotypes, recorded at 95 days after planting (DAP), were 137 and 120 cm, respectively. Leaf areas were 742 and 1048 cm2 in the R and S biotypes, respectively. The dry weights of leaves and stems in the S plants were 30 and 35%, respectively, higher than in the R plants. In both biotypes, the leaves had a greater share in dry weight at early development stages, but from 50 DAP, the stem became the main contributor to the dry weight of the shoots. The R biotype produced 110 ± 4 seed plant−1, i.e., 12 ± 3% less seeds per plant than that of the S one (125 ± 7 seed plant−1). The growth indices leaf area ratio and specific leaf area were generally higher in the S biotype or similar between both biotypes; while the relative growth rate and net assimilation rate were punctually superior in the R biotype. These results demonstrate that the Ser-653-Asn mutation imposed a fitness cost in imazamox R wild poinsettia.

After four years of cold storage, dimethachlon resistance of two laboratory-induced resistant Sclerotinia sclerotiorum isolates SCG7 and LA50 declined by 99.5% and 98.9%, respectively, and cross resistance to iprodione and procymidone also declined dramatically. Along with the decline of fungicide resistance, osmotic sensitivity to sodium chloride and glucose decreased tremendously; mycelial growth rate, sclerotia number and weight per potato dextrose agar (PDA) plate increased on average by 118.6%, 85. 5% and 64.5%, respectively; and virulence to detached leaves of oilseed rape increased by 72.7% on average. Significant negative correlations were detected between dimethachlon resistance levels and mycelial growth rate on PDA (r = −0.980, P = 0.021), and between resistance levels and lesion diameters on detached leaves of oilseed rape plants (r = −0.997, P = 0.002). These results have profound implications for assessing the potential risk for resistance development to dicarboximide fungicides in S. sclerotiorum.

Hickory trunk canker (HTC), primarily caused by Botryosphaeria dothidea , is an aggravating disease that threatens an important regional economic tree species of Chinese hickory and few information is available in the control of this disease. Here, the sensitivity of 93 isolates to fludioxonil and the resistance risk were investigated. All the isolates tested were sensitive to fludioxonil and the EC 50 ranged from 0.0028 to 0.0569 μg/mL. The tamed fludioxonil-resistant mutants remained highly resistant to fludioxonil even after 10 consecutive transfers to fludioxonil-free PDA plates. As for fitness penalty, the fludioxonil-resistant mutants demonstrated a reduction in conidia production and virulence as well as increased sensitivity to high osmotic stress. While, variations in mycelial growth and responses to SDS and H 2 O 2 were not detected in all the resistant mutants. In addition, the resistant mutants demonstrated positive cross-resistance to iprodione but not to fungicides of other modes of action. Sequential analysis of BdNik1 showed that premature stop codon occurred in all the resistant mutants despite of point mutation (BD16–22R9 and BD16–22R20) or frameshift mutation (BD16–22R8, BD16–22R11 and BD16–22R18). Our study suggested that fludioxonil exhibited excellent inhibition activity on mycelial growth of B. dothidea in vitro , the resistance risk of B. dothidea to fludioxonil should be low to moderate and fludioxonil would be a nice candidate in controlling HTC caused by B. dothidea. [Display omitted] • The population of B. dothidea causing hickory trunk canker is sensitive to fludioxonil. • All the obtained resistant mutants were high resistant to fludioxonil and the high resistance was stable. • The resistant mutants showed fitness penalty and positive cross-resistance to iprodione. • The resistance to fludioxonil was resulted from the occurrence of premature stop codon in BdNik1. [ABSTRACT FROM AUTHOR]

Attention should be devoted to weeds evolving herbicide resistance with non-target-site resistance (NTSR) mechanism due to their unpredictable resistance patterns. Quantification of fitness cost can be used in NTSR management strategies to determine the long-term fate of resistant plants in weed populations. To our knowledge, this is the first report evaluating potential fecundity and vegetative fitness of a NTSR black-grass (Alopecurus myosuroides Huds), the most important herbicide resistant weed in Europe, with controlled genetic background. The susceptible (S) and NTSR sub-populations were identified and isolated from a fenoxaprop-P-ethyl resistant population by a plant cloning technique. Using a target-neighborhood design, competitive responses of S and NTSR black-grass sub-populations to increasing density of winter wheat were quantified for 2 years in greenhouse and 1 year in field. Fitness traits including potential seed production, vegetative biomass and tiller number of both sub-populations significantly decreased with increasing density of winter wheat. More importantly, no statistically significant differences were found in fitness traits between S and NTSR sub-populations either grown alone (no competition) or in competition with winter wheat. According to the results, the NTSR black-grass is probably to persist in field even in the cessation of fenoxaprop-P-ethyl. So, effective herbicide resistant management strategies are strongly suggested to prevent and stop the spread of the NTSR black-grass, otherwise NTSR loci conferring resistance to a range of herbicides in black-grass will persist in the gene pool even in the absence of herbicide application. Consequently, herbicide as an effective tool for control of black-grass will gradually be lost in fields infested by NTSR black-grass.

The alleles responsible for herbicide resistance in weeds can result in a fitness cost within affected plants. Over 200 cases of resistance to triazine herbicides have been confirmed in a wide range of weed species globally. In New Zealand, Chenopodium album L. was the first species reported as resistant to triazines. Several studies have already shown that triazine resistance in weeds is associated with fitness costs. Our current study provides further information about fitness penalties caused by triazine resistance during the vegetative growth phase of C. album. Triazine-resistant phenotypes produced less biomass and were shorter than susceptible ones prior to the onset of flowering. At an early stage of growth, triazine-resistant plants had lower photosynthetic efficacy and growth rates than susceptible plants, indicated by lower net assimilation rate (NAR) and relative growth rate (RGR), respectively. However, at a later stage of growth, the resistant plants had greater RGR values than susceptible phenotypes, though there were no significant differences in NAR between triazine-resistant and susceptible plants at this later stage. The triazine-resistant plants had less capacity for vegetative growth than susceptible plants during competition with wheat, indicating less ability to capture resources by triazine-resistant plants under competition. Overall, this study has revealed that the triazine resistance allele caused a substantial fitness cost to C. album only at the early phase of vegetative growth stage; thus, the use of crop competition to try managing triazine-resistant C. album plants should occur during this early phase.

Attention should be devoted to weeds evolving herbicide resistance with non-target-site resistance (NTSR) mechanism due to their unpredictable resistance patterns. Quantification of fitness cost can be used in NTSR management strategies to determine the long-term fate of resistant plants in weed populations. To our knowledge, this is the first report evaluating potential fecundity and vegetative fitness of a NTSR black-grass (Alopecurus myosuroides Huds), the most important herbicide resistant weed in Europe, with controlled genetic background. The susceptible (S) and NTSR sub-populations were identified and isolated from a fenoxaprop-P-ethyl resistant population by a plant cloning technique. Using a target-neighborhood design, competitive responses of S and NTSR black-grass sub-populations to increasing density of winter wheat were quantified for 2 years in greenhouse and 1 year in field. Fitness traits including potential seed production, vegetative biomass and tiller number of both sub-populations significantly decreased with increasing density of winter wheat. More importantly, no statistically significant differences were found in fitness traits between S and NTSR sub-populations either grown alone (no competition) or in competition with winter wheat. According to the results, the NTSR black-grass is probably to persist in field even in the cessation of fenoxaprop-P-ethyl. So, effective herbicide resistant management strategies are strongly suggested to prevent and stop the spread of the NTSR black-grass, otherwise NTSR loci conferring resistance to a range of herbicides in black-grass will persist in the gene pool even in the absence of herbicide application. Consequently, herbicide as an effective tool for control of black-grass will gradually be lost in fields infested by NTSR black-grass. [ABSTRACT FROM AUTHOR]

'Gene-for-gene' theory predicts that gain of virulence by an avirulent pathogen on plants expressing resistance ( R) genes is associated with fitness loss in susceptible hosts. However, the validity of this prediction has been studied in only a few plant viral pathosystems. In this study, the Soybean mosaic virus (SMV)- Rsv4 pathosystem was exploited to test this prediction. In Rsv4-genotype soybeans, P3 of avirulent SMV strains provokes an as yet uncharacterized resistance mechanism that restricts the invading virus to the inoculated leaves. A single amino acid substitution in P3 functionally converts an avirulent to a virulent strain, suggesting that the genetic composition of P3 plays a crucial role in virulence on Rsv4-genotype soybeans. In this study, we examined the impact of gain of virulence mutation(s) on the fitness of virulent variants derived from three avirulent SMV strains in a soybean genotype lacking the Rsv4 gene. Our data demonstrate that gain of virulence mutation(s) by all avirulent viruses on Rsv4-genotype soybean is associated with a relative fitness loss in a susceptible host. The implications of this finding on the durable deployment of the Rsv4 gene in soybean are discussed. [ABSTRACT FROM AUTHOR]

BACKGROUND: Fenhexamid, a sterol biosynthesis inhibitor effective against Botrytis, inhibits the 3-ketoreductase (Erg27) involved in C-4 demethylation. Several fenhexamid-resistant phenotypes have been detected in Botrytis cinerea populations from French vineyards. The field isolates with the highest resistance levels display amino acid changes in Erg27 (F412S, F412I or F412V). RESULTS: Fenhexamid-resistant mutants were generated by site-directed mutagenesis of the erg27 gene in a sensitive recipient strain to overcome the impact of different genetic backgrounds. The wild-type erg27 allele was replaced by the three mutated alleles ( erg27 F412 S/ I/ V) by homologous recombination. These isogenic strains were shown to be fenhexamid-resistant and were used to quantify the impact of F412 mutations on fungal fitness. Several parameters, including radial growth, the production of sclerotia and conidia, freezing resistance and aggressiveness, were quantified in laboratory conditions. Analysis of variance demonstrated significant differences between the mutant and parental strains for some characters. In particular, the mutants grew more slowly than the wild-type strain and displayed variations in the production of sclerotia and conidia with temperature and susceptibility to freezing. CONCLUSIONS: The results highlight a moderate but significant impact of F412 mutations on the survival capacity of B. cinerea strains displaying high levels of resistance to fenhexamid in laboratory conditions, potentially limiting their dispersal and persistence, particularly in terms of overwintering, in field conditions. Copyright © 2011 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

The advantage of the resistance conferred by a mutation can sometimes be offset by a high fitness-cost penalty. This balance will affect possible fate of the resistance allele. Few studies have explored the impact of the genetic background on the expression of the resistance fitness cost and none has attempted to measure the variation in fitness-cost dominance. However, both the fitness penalty and its dominance may modify evolutionary trajectory and outcome. Here the impact of Arabidopsis thaliana intraspecific genetic diversity on fitness cost and its associated dominance was investigated by analysing 12 quantitative traits in crosses between a mutant conferring resistance to the herbicide 2,4-D and nine different natural genetic backgrounds. Fitness cost values were found to be more affected by intraspecific genetic diversity than fitness cost dominance, even though this effect depends on the quantitative trait measured. This observation has implications for the choice of the best strategy for preventing herbicide resistance development. In addition, our results pinpoint a potential compensatory improvement of the resistance fitness cost and its associated dominance by the genetic diversity locally present within a species.Heredity (2008) 101, 499–506; doi:10.1038/hdy.2008.92; published online 3 September 2008 [ABSTRACT FROM AUTHOR]

Two greenhouse trials were conducted over a 2 year period (2004 and 2005) to compare the relative growth potential and fitness of triazine-susceptible (TS) and triazine-resistant (TR) waterhemp biotypes. Waterhemp plants from each biotype were grown in 6 L pots in the greenhouse in a completely randomized design. There was no difference in leaf number, plant height, plant biomass, time to bud or time to flower between the two biotypes. The TR female waterhemp plants produced 30% less reproductive biomass and 39% less seed than the TS biotype. The fitness penalty associated with triazine resistance was related to reproductive capability, not to vegetative growth. [ABSTRACT FROM AUTHOR]

Wild poinsettia (Euphorbia heterophylla L.) is a difficult-to-control weed in soybean production in Brazil that has developed resistance to herbicides, including acetolactate synthase inhibitors. We investigated the potential fitness cost associated to the Ser-653-Asn mutation that confers imazamox resistance in this weed. Plant height, leaf and stem dry weight, leaf area and seed production per plant as well as the growth indices of specific leaf area, leaf area ratio, relative growth rate and net assimilation in F2 homozygous resistant (R) and susceptible (S) wild poinsettia progenies were pairwise compared. S plants were superior in most of the traits studied. Plant heights for S and R biotypes, recorded at 95 days after planting (DAP), were 137 and 120 cm, respectively. Leaf areas were 742 and 1048 cm2 in the R and S biotypes, respectively. The dry weights of leaves and stems in the S plants were 30 and 35%, respectively, higher than in the R plants. In both biotypes, the leaves had a greater share in dry weight at early development stages, but from 50 DAP, the stem became the main contributor to the dry weight of the shoots. The R biotype produced 110 ± 4 seed plant−1, i.e., 12 ± 3% less seeds per plant than that of the S one (125 ± 7 seed plant−1). The growth indices leaf area ratio and specific leaf area were generally higher in the S biotype or similar between both biotypes; while the relative growth rate and net assimilation rate were punctually superior in the R biotype. These results demonstrate that the Ser-653-Asn mutation imposed a fitness cost in imazamox R wild poinsettia. [ABSTRACT FROM AUTHOR]

Conyza bonariensis (L.) Cronquist is one of the main glyphosate-resistant weeds in no-till fields in Brazil. Here, the seed quality of glyphosate-resistant (R) and -susceptible (S) C. bonariensis biotypes, collected from different sites, was evaluated under stressful conditions by different seed tests. Glyphosate resistance was confirmed by dose-response and shikimate accumulation assays. The resistance factors were 6.9 (R1/S1), 4.5 (R2/S2), and 5.8 (R3/S3). Biotypes S1, S2, and S3 accumulated 2.7, 2.4, and 2.8 times more shikimic acid than biotypes R1, R2, and R3, respectively. Stress-free seed viability and germination potential ranged from 39% to 57% and from 37% to 57%, respectively, with no difference between R and S biotypes within each collection site. Seed incubation at 8 °C over 7 days (cold test) promoted greater germination in S biotypes (54% to 79%) compared to R ones (28% to 39%). In the accelerated aging tests (incubation at 42 °C over 48 hours), the germination decreased in both S (11% to 27%) and R (6% to 16%) biotypes. In the high-temperature stress tests, there were no differences in germination within biotypes at 35 and 45 °C; however, at 60 °C, the germination of the S1, R1, S2, R2, S3, and R3 biotypes was reduced by approximately 51%, 54%, 63%, 59%, 40%, and 30%, respectively. Under non-stressful conditions, germination potential and seed viability were similar in R and S biotypes; however, under cold or heat stress conditions, R biotypes reduced their germination rates, revealing that glyphosate resistance causes a fitness penalty in C. bonariensis at the seed level. However, because seed viability was not determined after experiments, it cannot be stated that such reduction in germination was due to the death or only a dormant phase of the seeds. [ABSTRACT FROM AUTHOR]

The alleles responsible for herbicide resistance in weeds can result in a fitness cost within affected plants. Over 200 cases of resistance to triazine herbicides have been confirmed in a wide range of weed species globally. In New Zealand, Chenopodium album L. was the first species reported as resistant to triazines. Several studies have already shown that triazine resistance in weeds is associated with fitness costs. Our current study provides further information about fitness penalties caused by triazine resistance during the vegetative growth phase of C. album. Triazine-resistant phenotypes produced less biomass and were shorter than susceptible ones prior to the onset of flowering. At an early stage of growth, triazine-resistant plants had lower photosynthetic efficacy and growth rates than susceptible plants, indicated by lower net assimilation rate (NAR) and relative growth rate (RGR), respectively. However, at a later stage of growth, the resistant plants had greater RGR values than susceptible phenotypes, though there were no significant differences in NAR between triazine-resistant and susceptible plants at this later stage. The triazine-resistant plants had less capacity for vegetative growth than susceptible plants during competition with wheat, indicating less ability to capture resources by triazine-resistant plants under competition. Overall, this study has revealed that the triazine resistance allele caused a substantial fitness cost to C. album only at the early phase of vegetative growth stage; thus, the use of crop competition to try managing triazine-resistant C. album plants should occur during this early phase. [ABSTRACT FROM AUTHOR]

Sensitivity monitoring of Venturia effusa , cause of pecan scab, has revealed insensitivity to fentin hydroxide and tebuconazole, but recent research indicates that the insensitivity to fentin hydroxide is not stable. A study was undertaken to determine if a fitness cost may be responsible for this instability. In this study, experiments were conducted to evaluate fitness components and phenotypic stability of insensitivity of V. effusa to fentin hydroxide and tebuconazole. Conidial production, conidial germination, microcolony growth, sensitivity to osmotic stress, and sensitivity to oxidative stress in the absence of fungicide were compared for isolates with differing sensitivities to both fungicides. Percent conidial germination decreased linearly with increasing fentin hydroxide insensitivity, and microcolony growth on 1.0 mM H 2 O 2 decreased linearly with increasing tebuconazole insensitivity. Stability of resistance was assessed on concentrations of 1.0, 3.0, and 10 µg/ml of both fungicides prior to and after five transfers on non-fungicide-amended medium. Tebuconazole insensitivity was stable after transfers, but fentin hydroxide insensitivity on 1.0 and 3.0 µg/ml decreased significantly after transfers, indicating instability. Here we provide evidence that in V. effusa tebuconazole insensitivity is stable and fentin hydroxide insensitivity is not. These results suggest that fentin-hydroxide-resistant V. effusa isolates have reduced conidial viability compared with sensitive isolates, which may allow the population to regain sensitivity in the absence of this frequently used fungicide.

Background: The potential for emergence of antiviral drug resistance during influenza pandemics has raised great concern for public health. Widespread use of antiviral drugs is a significant factor in producing resistant strains. Recent studies show that some influenza viruses may gain antiviral drug resistance without a fitness penalty. This creates the possibility of strategic interaction between populations considering antiviral drug use strategies., Methods: To explain why, we develop and analyze a classical 2-player game theoretical model where each player chooses from a range of possible rates of antiviral drug use, and payoffs are derived as a function of final size of epidemic with the regular and mutant strain. Final sizes are derived from a stochastic compartmental epidemic model that captures transmission within each population and between populations, and the stochastic emergence of antiviral drug resistance. High treatment levels not only increase the spread of the resistant strain in the subject population but also affect the other population by increasing the density of the resistant strain infectious individuals due to travel between populations., Results: We found two Nash equilibria where both populations treat at a high rate, or both treat at a low rate. Hence the game theoretical analysis predicts that populations will not choose different treatment strategies than other populations, under these assumptions. The populations may choose to cooperate by maintaining a low treatment rate that does not increase the incidence of mutant strain infections or cause case importations to the other population. Alternatively, if one population is treating at a high rate, this will generate a large number of mutant infections that spread to the other population, in turn incentivizing that population to also treat at a high rate. The prediction of two separate Nash equilibria is robust to the mutation rate and the effectiveness of the drug in preventing transmission, but it is sensitive to the volume of travel between the two populations., Conclusions: Model-based evaluations of antiviral influenza drug use during a pandemic usually consider populations in isolation from one another, but our results show that strategic interactions could strongly influence a population's choice of antiviral drug use policy. Furthermore, the high treatment rate Nash equilibrium has the potential to become socially suboptimal (i.e. non-Pareto optimal) under model assumptions that might apply under other conditions. Because of the need for players to coordinate their actions, we conclude that communication and coordination between jurisdictions during influenza pandemics is a priority, especially for influenza strains that do not evolve a fitness penalty under antiviral drug resistance.

After four years of cold storage, dimethachlon resistance of two laboratory-induced resistant Sclerotinia sclerotiorum isolates SCG7 and LA50 declined by 99.5% and 98.9%, respectively, and cross resistance to iprodione and procymidone also declined dramatically. Along with the decline of fungicide resistance, osmotic sensitivity to sodium chloride and glucose decreased tremendously; mycelial growth rate, sclerotia number and weight per potato dextrose agar (PDA) plate increased on average by 118.6%, 85. 5% and 64.5%, respectively; and virulence to detached leaves of oilseed rape increased by 72.7% on average. Significant negative correlations were detected between dimethachlon resistance levels and mycelial growth rate on PDA (r = -0.980, P = 0.021), and between resistance levels and lesion diameters on detached leaves of oilseed rape plants (r = -0.997, P = 0.002). These results have profound implications for assessing the potential risk for resistance development to dicarboximide fungicides in S. sclerotiorum.