Inheritance and mechanism of glyphosate resistance in annual bluegrass (Poa annua L.).

BACKGROUND: In initial screening, glyphosate was ineffective in controlling five Poa annua populations. These populations were tested for resistance, and studies undertaken to determine resistance mechanisms and inheritance pattern. RESULTS: Dose-response studies conducted at 16/12°C and 27/20°C on the five putative resistant populations showed low-level resistance (1.4- to 2.5-fold) to glyphosate. Shikimic acid accumulation in response to glyphosate confirmed differences among the populations, with greater shikimic acid accumulation in the susceptible population. The EPSPS gene copy number was 0.5- to 5.2-fold greater in one resistant population (HT) than in the susceptible (S) population, but not in the others. EPSPS gene expression was five- to tenfold higher in HT compared with the susceptible population. Target site mutations, differences in glyphosate absorption or translocation or altered expression of aldo-keto reductase (AKR) were not identified in any of the resistant populations. Crosses were successful between one resistant population and the susceptible population (P262-16♂ ✕ S♀) and inheritance of glyphosate resistance appears to be controlled by a single, nuclear dominant gene in this population. CONCLUSION: Our study identified EPSPS gene amplification in a South Australian glyphosate-resistant P. annua population (HT). This mechanism of resistance was not identified in the other four glyphosate-resistant populations, and other common mechanisms were excluded. Although the resistance mechanism in some P. annua populations remains unknown, inheritance studies with one population suggest the involvement of a single dominant gene. © 2021 Society of Chemical Industry.

Increasing the value and efficiency of herbicide resistance surveys.

The scale of herbicide resistance within a cropping region can be estimated and monitored using surveys of weed populations. The current approach to herbicide resistance surveys is time-consuming, logistically challenging and costly. Here we review past and current approaches used in herbicide resistance surveys with the aims of (i) defining effective survey methodologies, (ii) highlighting opportunities for improving efficiencies through the use of new technologies and (iii) identifying the value of repeated region-wide herbicide resistance surveys. One of the most extensively surveyed areas of the world's cropping regions is the Australian grain production region, with >2900 fields randomly surveyed in each of three surveys conducted over the past 15 years. Consequently, recommended methodologies are based on what has been learned from the Australian experience. Traditional seedling-based herbicide screening assays remain the most reliable and widely applicable method for characterizing resistance in weed populations. The use of satellite or aerial imagery to plan collections and image analysis to rapidly quantify screening results could complement traditional resistance assays by increasing survey efficiency and sampling accuracy. Global management of herbicide-resistant weeds would benefit from repeated and standardized surveys that track herbicide resistance evolution within and across cropping regions. © 2021 Society of Chemical Industry.

Loss of trifluralin metabolic resistance in Lolium rigidum plants exposed to prosulfocarb recurrent selection.

BACKGROUND: Resistance to the dinitroaniline herbicide trifluralin in Lolium rigidum (annual ryegrass) often is mediated by the enhanced capacity to metabolize the herbicide to less toxic polar conjugates and/or by functionally recessive target-site mutations in α-tubulin. RESULTS: In two L. rigidum populations possessing enhanced trifluralin metabolism, resistance was largely reversed by recurrent selection with the thiocarbamate herbicide prosulfocarb (i.e. plant survival was two- to >20-fold lower). Their ability to metabolize trifluralin was significantly decreased (by ≈2.3-fold) following recurrent prosulfocarb selection, to levels comparable to those observed in susceptible plants or when trifluralin metabolism was inhibited by treatment with the insecticide phorate. CONCLUSIONS: This study provides evidence that trait(s) enabling efficient trifluralin metabolism in L. rigidum are purged from the population under prosulfocarb recurrent selection. The level of trifluralin metabolism in vitro and its inhibition caused by phorate action on trifluralin-metabolizing enzyme(s) is equivalent to the effect produced by prosulfocarb selection. The hypothetical link between the two phenomena is that the putative monooxygenase(s) conferring trifluralin metabolic resistance also mediate the activation of prosulfocarb to its toxic sulfoxide. Thus, we speculate that survival to prosulfocarb via a lack of metabolic herbicide activation, and survival to trifluralin conferred by enhanced herbicide metabolism, are mutually exclusive. These findings not only open up a new research direction in terms of the interaction between different herbicide resistance mechanisms in L. rigidum, but also offer strategies for immediate management of the population dynamics of metabolism-based resistance in the field. © 2020 Society of Chemical Industry.

Diversity and extent of mutations endowing resistance to the acetolactate synthase (AHAS)-inhibiting herbicides in Indian hedge mustard (Sisymbrium orientale) populations in Australia.

Indian hedge mustard (Sisymbrium orientale) (IHM) is an important broadleaf weed across southern Australia. Resistance to sulfonylurea (SU) herbicides that inhibit acetohydroxyacid synthase (AHAS) is extensive in Australia, but resistance to imidazolinone (IMI) herbicides has only been reported recently. The AHAS-mutation profile of 65 IHM populations collected randomly from cropped fields was investigated to better understand the extent and types of resistance present. Resistance to SU herbicides was present in 40% of the populations and resistance to IMI herbicides in 11%. Mutations were identified in SoAHAS by sequence analysis, and included previously reported amino-acid substitutions at Pro197 and Trp574, but also new substitutions at Pro197 and Asp376 for this species. One population with possible non-target-site resistance was identified. Germination studies with fresh seed found no significant effect by mutations in SoAHAS on germination; however, population factors had a large effect on germination in S. orientale. Resistance to AHAS-inhibiting herbicides in populations of S. orientale is endowed by mutations in SoAHAS in all but one population examined. Mutations at Pro197 conferring resistance to SU herbicides were most common, while mutations at Trp574 that provide resistance to IMI herbicides are also present.

The mechanism of diflufenican resistance and its inheritance in oriental mustard (Sisymbrium orientale L.) from Australia.

BACKGROUND: An oriental mustard population (P3) collected near Quambatook, Victoria was identified as being resistant to diflufenican by screening with the field rate (200 g a.i. ha-1 ) of the herbicide. The mechanism(s) of diflufenican resistance and its inheritance in this population were therefore investigated. RESULTS: Dose-response experiments confirmed that population P3 was 140-fold more resistant to diflufenican than susceptible populations, as determined by the comparison of 50% lethal (LD50 ) values. The phytoene desaturase (PDS) gene from five individuals each of the S1 [susceptible (S)] and P3 [resistant (R)] populations was sequenced, and a substitution of valine for leucine at position 526 (Leu-526-Val) was detected in all five individuals of P3, but not in the S1 population. Inheritance studies showed that diflufenican resistance is encoded in the nuclear genome and is dominant, as the response to diflufenican at 200 g a.i. ha-1 of F1 families was equivalent to that of the resistant biotype. The segregation of F2 phenotypes fitted a 3:1 inheritance model. Segregation of 42 F2 individuals by genotype sequencing fitted a 1:2:1 (ss:Rs:RR) ratio. CONCLUSION: Resistance to diflufenican in oriental mustard is conferred by the Leu-526-Val mutation in the PDS gene. Inheritance of resistance is managed by a single gene with high levels of dominance. © 2018 Society of Chemical Industry.

EPSPS gene amplification conferring resistance to glyphosate in windmill grass (Chloris truncata) in Australia.

BACKGROUND: Five glyphosate-resistant populations of Chloris truncata originally collected from New South Wales were compared with one susceptible (S) population from South Australia to confirm glyphosate resistance and elucidate possible mechanisms of resistance. RESULTS: Based on the amounts of glyphosate required to kill 50% of treated plants (LD50 ), glyphosate resistance (GR) was confirmed in five populations of C. truncata (A536, A528, T27, A534 and A535.1). GR plants were 2.4-8.7-fold more resistant and accumulated less shikimate after glyphosate treatment than S plants. There was no difference in glyphosate absorption and translocation between GR and S plants. The EPSPS gene did not contain any point mutation that had previously been associated with resistance to glyphosate. The resistant plants (A528 and A536) contained up to 32-48 more copies of the EPSPS gene than the susceptible plants. CONCLUSION: This study has identified EPSPS gene amplification contributing to glyphosate resistance in C. truncata. In addition, a Glu-91-Ala mutation within EPSPS was identified that may contribute to glyphosate resistance in this species. © 2017 Society of Chemical Industry.

Target-site mutations conferring resistance to glyphosate in feathertop Rhodes grass (Chloris virgata) populations in Australia.

BACKGROUND: Chloris virgata is a warm-season, C4 , annual grass weed affecting field crops in northern Australia that has become an emerging weed in southern Australia. Four populations with suspected resistance to glyphosate were collected in South Australia, Queensland and New South Wales, Australia, and compared with one susceptible (S) population to confirm glyphosate resistance and elucidate possible mechanisms of resistance. RESULTS: Based on the rate of glyphosate required to kill 50% of treated plants (LD50 ), glyphosate resistance (GR) was confirmed in four populations of C. virgata (V12, V14.2, V14.16 and V15). GR plants were 2-9.7-fold more resistant and accumulated less shikimate after glyphosate treatment than S plants. GR and S plants did not differ in glyphosate absorption and translocation. Target-site EPSPS mutations corresponding to Pro-106-Leu (V14.2) and Pro-106-Ser (V15, V14.16 and V12) substitutions were found in GR populations. The population with Pro-106-Leu substitution was 2.9-4.9-fold more resistant than the three other populations with Pro-106-Ser substitution. CONCLUSION: This report confirms glyphosate resistance in C. virgata and shows that target-site EPSPS mutations confer resistance to glyphosate in this species. The evolution of glyphosate resistance in C. virgata highlights the need to identify alternative control tactics. © 2016 Society of Chemical Industry.

Reduced translocation in 2,4-D-resistant oriental mustard populations (Sisymbrium orientale L.) from Australia.

BACKGROUND: Two oriental mustard populations (P2 and P13) collected from Port Broughton, South Australia were identified as resistant to 2,4-D. The level of resistance, mechanism and the mode of inheritance for 2,4-D resistance in these populations were investigated. RESULTS: Populations P2 and P13 were confirmed to be resistant to 2,4-D at the field rate (600 g a.e. ha-1 ). P2 and P13 were 81- and 67-fold more resistant than the susceptible populations (S1 and S2) at the dose required for 50% mortality (LD50 ), respectively. No predicted amino acid modification was detected in sequences of potential target-site genes (ABP, TIR1 and AFB5). Resistant populations had reduced 2,4-D translocation compared with the susceptible populations, with 77% of [14 C]2,4-D retained in the treated leaf versus 32% at 72 h after treatment. Resistance to 2,4-D is encoded on the nuclear genome and is dominant, as the response to 2,4-D of all F2 individuals were similar to the resistant biotypes. The segregation of F2 phenotypes fitted a 3: 1 (R: S) inheritance model. CONCLUSION: Resistance to 2,4-D in oriental mustard is likely due to reduced translocation of 2,4-D out of the treated leaf. Inheritance of 2,4-D resistance is conferred by a single gene with a high level of dominance. © 2017 Society of Chemical Industry.

Basis of ACCase and ALS inhibitor resistance in Hordeum glaucum Steud.

BACKGROUND: Acetyl coenzyme-A carboxylase (ACCase) and/or acetolactate synthase (ALS) inhibitor resistance has been identified by herbicide resistance screening in eight populations obtained from cropping regions of South Australia. This study aimed to quantify the level of resistance and characterise the molecular basis of resistance to ACCase and ALS inhibitors in these H. glaucum populations. RESULTS: H. glaucum populations from the Upper-North region were highly resistant (resistance index RI > 12) to the aryloxyphenoxypropionate (APP) herbicides quizalofop and haloxyfop and less resistant (RI = 2-12) to cyclohexanedione (CHD) herbicide clethodim, and some Mid-North populations had a low level of resistance (RI = 2-6) to the sulfonylurea (SU) herbicide mesosulfuron. Gene sequencing confirmed the presence of Ile-1781-Leu, Ile-2041-Asn and Gly-2096-Ala mutations in the ACCase gene, with no mutation found in the ALS gene. The use of the known metabolic inhibitor malathion in combination with mesosulfuron enhanced the activity of this herbicide. These populations were also susceptible to SU herbicide sulfometuron. CONCLUSION: This study has documented APP-to-CHD herbicide cross-resistance, the first case of ACCase inhibitor resistance due to Ile-2041-Asn mutation, and characterised the resistance to ALS inhibitors in H. glaucum. Resistance to ACCase inhibitors is due to a target-site mutation. The reversal of SU resistance by malathion and susceptibility to sulfometuron suggests that non-target-site mechanisms confer resistance to ALS inhibitors. © 2016 Society of Chemical Industry.

EPSPS gene amplification in glyphosate-resistant Bromus diandrus.

BACKGROUND: Glyphosate is the most widely used herbicide in the world and has been intensively used to control B. diandrus, a problematic weed of crops and pastures in southern Australia. RESULTS: Resistance to glyphosate was identified in two populations of B. diandrus that were nearly fivefold more resistant to glyphosate than wild-type plants. Both populations contained EPSPS gene amplification, with resistant plants having an average of around 20-fold the number of copies of EPSPS compared with susceptible plants. EPSPS expression was also increased in resistant plants of both populations; however, expression levels were not correlated with the number of EPSPS copies. Amplification of only one of the four EPSPS genes present in B. diandus was detected. Investigation into the inheritance of glyphosate resistance found no segregation in the F2 generation. Every individual in the F2 populations contained between three and 30 copies of EPSPS; however, on average they contained fewer copies compared with the parent resistant population. CONCLUSIONS: Glyphosate resistance in B. diandrus is due to EPSPS gene amplification. Resistance is heritable but complex.

Temperature influences the level of glyphosate resistance in barnyardgrass (Echinochloa colona).

BACKGROUND: Echinochloa colona is an important summer-growing weed species in cropping regions of northern Australia that has evolved resistance to glyphosate owing to intensive use of this herbicide in summer fallow. RESULTS: Pot trials conducted at 20 and 30 °C on six E. colona populations showed a significant increase in the level of glyphosate resistance in resistant populations at 30 °C compared with 20 °C. However, there was no influence of growth temperature on glyphosate susceptibility of the sensitive population. Sequencing of the target-site gene (EPSPS) of the six populations identified a mutation at position 106 leading to a change from proline to serine in the most resistant population A533.1 only. EPSPS gene amplification was not detected in any of the resistant populations examined. Examining (14) C-glyphosate uptake on two resistant and one susceptible population showed a twofold increase at 20 °C; however, few differences in glyphosate translocation occurred from the treated leaf to other plant parts between populations or temperatures. CONCLUSION: There is reduced efficacy of glyphosate at high temperatures on resistant E. colona populations, making these populations harder to control in summer.