No abstract

Lolium rigidum (annual or rigid ryegrass) is a widespread annual weed in cropping systems of southern Australia, and herbicide resistance in L. rigidum is a common problem in this region. In 2010, a random survey was conducted across the grain belt of Western Australia to determine the frequency of herbicide-resistant L. rigidum populations and to compare this with the results of previous surveys in 1998 and 2003. During the survey, 466 cropping fields were visited, with a total of 362 L. rigidum populations collected. Screening of these populations with the herbicides commonly used for control of L. rigidum revealed that resistance to the ACCase-and ALS-inhibiting herbicides was common, with 96% of populations having plants resistant to the ACCase herbicide diclofop-methyl and 98% having plants resistant to the ALS herbicide sulfometuron. Resistance to another ACCase herbicide, clethodim, is increasing, with 65% of populations now containing resistant plants. Resistance to other herbicide modes of action was significantly lower, with 27% of populations containing plants with resistance to the pre-emergent herbicide trifluralin, and glyphosate, atrazine and paraquat providing good control of most of the populations screened in this survey. Ninety five per cent of L. rigidum populations contained plants with resistance to at least two herbicide modes of action. These results demonstrate that resistance levels have increased dramatically for the ACCase-and ALS-inhibiting herbicides since the last survey in 2003 (>95% vs. 70-90%); therefore, the use of a wide range of integrated weed management options are required to sustain these cropping systems in the future.

In 2003, a random survey was conducted across the Western Australian wheatbelt to establish the frequency and distribution of herbicide resistance in ryegrass populations infesting crop fields. Five hundred cropping fields were visited at crop maturity, and ryegrass seed was collected in 452 of these fields. Subsequently, each crop field population was screened with herbicides of various modes of action that are commonly used for ryegrass control in Australian cropping systems. Most of these ryegrass populations were found to be resistant to the ACCase-inhibitor herbicide diclofop-methyl (68%) and the ALS-inhibitor herbicide sulfometuron (88%). A comparison of resistance levels in the same agronomic zones surveyed 5 years earlier determined that there had been an increase of 20 percentage points in the frequency of resistance over this 5-year period. This survey also determined that the majority (64%) of populations were found to be multiple resistant to both diclofop-methyl and sulfometuron. The distribution patterns of the collected populations indicated that there were higher frequencies of resistant and developing resistance populations occurring in the intensively cropped regions of the wheatbelt, which had greater herbicide selection pressure. Of concern is that 24% and 8% of populations were found to be developing resistance to trifluralin and clethodim, respectively. Currently these herbicides are heavily relied upon for control of ACCase and ALS herbicide resistant ryegrass. Nearly all populations remain susceptible to glyphosate. Ryegrass across the WA wheatbelt now exhibits multiple resistance across many but not all herbicides, posing severe management and sustainability challenges.

This study investigates mechanisms of multiple resistance to glyphosate, acetyl-coenzyme A carboxylase (ACCase) and acetolactate synthase (ALS)-inhibiting herbicides in two Lolium rigidum populations from Australia. When treated with glyphosate, susceptible (S) plants accumulated 4- to 6-fold more shikimic acid than resistant (R) plants. The resistant plants did not have the known glyphosate resistance endowing mutation of 5-enolpyruvylshikimate-3 phosphate synthase (EPSPS) at Pro-106, nor was there over-expression of EPSPS in either of the R populations. However, [(14)C]-glyphosate translocation experiments showed that the R plants in both populations have altered glyphosate translocation patterns compared to the S plants. The R plants showed much less glyphosate translocation to untreated young leaves, but more to the treated leaf tip, than did the S plants. Sequencing of the carboxyl transferase domain of the plastidic ACCase gene revealed no resistance endowing amino acid substitutions in the two R populations, and the ALS in vitro inhibition assay demonstrated herbicide-sensitive ALS in the ALS R population (WALR70). By using the cytochrome P450 inhibitor malathion and amitrole with ALS and ACCase herbicides, respectively, we showed that malathion reverses chlorsulfuron resistance and amitrole reverses diclofop resistance in the R population examined. Therefore, we conclude that multiple glyphosate, ACCase and ALS herbicide resistance in the two R populations is due to the presence of distinct non-target site based resistance mechanisms for each herbicide. Glyphosate resistance is due to reduced rates of glyphosate translocation, and resistance to ACCase and ALS herbicides is likely due to enhanced herbicide metabolism involving different cytochrome P450 enzymes.

This work demonstrates that metabolic herbicide resistance is commonly occurring in L. rigidum, and coevolution of both metabolic resistance and target-site resistance is an evolutionary reality. Metabolic herbicide resistance can potentially endow resistance to many herbicides and poses a threat to herbicide sustainability and thus crop production, calling for major research and management efforts.