Herbicide resistance in rigid ryegrass is an escalating problem in grain-cropping fields of southeastern Australia due to increased reliance on herbicides as the main method for weed control. Weed surveys were conducted between 1998 and 2009 to identify the extent of herbicide-resistant rigid ryegrass across this region to dinitroaniline, and acetolactate synthase- and acetyl coenzyme A (CoA) carboxylase-inhibiting herbicides. Rigid ryegrass was collected from cropped fields chosen at random. Outdoor pot studies were conducted during the normal winter growing season for rigid ryegrass with PRE-applied trifluralin and POST-applied diclofop-methyl, chlorsulfuron, tralkoxydim, pinoxaden, and clethodim. Herbicide resistance to trifluralin in rigid ryegrass was identified in one-third of the fields surveyed from South Australia, whereas less than 5% of fields in Victoria exhibited resistance. In contrast, resistance to chlorsulfuron was detected in at least half of the cropped fields across southeastern Australia. Resistance to the cereal-selective aryloxyphenoxypropionate-inhibiting herbicides diclofop-methyl, tralkoxydim, and pinoxaden ranged between 30 and 60% in most regions, whereas in marginal cropping areas less than 12% of fields exhibited resistance. Resistance to clethodim varied between 0 and 61%. Higher levels of resistance to clethodim were identified in the more intensively cropped, higher-rainfall districts where pulse and canola crops are common. These weed surveys demonstrated that a high incidence of resistance to most tested herbicides was present in rigid ryegrass from cropped fields in southeastern Australia, which presents a major challenge for crop producers.
Glyphosate resistance was first discovered in populations of rigid ryegrass in Australia in 1996. Since then, glyphosate resistance has been detected in additional populations of rigid ryegrass and Italian ryegrass in several other countries. Glyphosate-resistant rigid ryegrass and Italian ryegrass have been selected in situations where there is an overreliance on glyphosate to the exclusion of other weed control tactics. Two major mechanisms of glyphosate resistance have been discovered in these two species: a change in the pattern of glyphosate translocation such that glyphosate accumulates in the leaf tips of resistant plants instead of in the shoot meristem; and amino acid substitutions at Pro 106 within the target site, 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS). There are also populations with both mechanisms. In the case of glyphosate resistance, the target site mutations tend to provide a lower level of resistance than does the altered translocation mechanism. Each of these resistance mechanisms is inherited as a single gene trait that is largely dominant. As these ryegrass species are obligate outcrossers, this ensures resistance alleles can move in both pollen and seed. Some glyphosate-resistant rigid ryegrass populations appear to have a significant fitness penalty associated with the resistance allele. Field surveys show that strategies vary in their ability to reduce the frequency of glyphosate resistance in populations and weed population size, with integrated strategies—including alternative weed management and controlling seed set of surviving plants—the most effective.
Three Australian Sisymbrium orientale and one Brassica tournefortii biotypes are resistant to acetolactate synthase (ALS)-inhibiting herbicides due to their possession of an ALS enzyme with decreased sensitivity to these herbicides. Enzyme kinetic studies revealed no interbiotypic differences within species in K m (pyruvate) (the substrate concentration at which the reaction rate is half maximal) but a greater V max (the rate when the enzyme is fully saturated with substrate) for two of the resistant S orientale biotypes over susceptible levels. F 1 hybrids from reciprocal crosses between resistant and susceptible biotypes of S orientale showed an intermediate response to chlorsulfuron compared to the parental plants. ALS herbicide resistance in S orientale segregated in a 3:1 (resistant:susceptible) ratio in F 2 plants with a single rate of chlorsulfuron, indicating that resistance is inherited as a single, incompletely dominant nuclear gene. Two regions of the ALS structural gene known to vary in ALSresistant biotypes were ampli®ed and sequenced. Resistant S orientale biotypes NS01 and SS03 contained a single nucleotide substitution in Domain B, predicting a Trp (in susceptible) to Leu (in resistant) amino acid change. Two adjacent nucleotide substitutions (CCT to ATT) predicting a Pro (in susceptible) to Ile (in resistant) change in the primary amino acid sequence were identi®ed in Domain A of resistant S orientale biotype SS01. Likewise, a single nucleotide substitution at the same site in the resistant B tournefortii biotype predicts a Pro (in susceptible) to Ala (in resistant) substitution. No other interbiotypic nucleotide differences predicted amino acid changes in the sequenced regions, suggesting that the amino acid substitutions reported above are responsible for resistance to ALS-inhibiting herbicides in the respective biotypes.
Glyphosate is widely used for weed control in the grape growing industry in southern Australia. The intensive use of glyphosate in this industry has resulted in the evolution of glyphosate resistance in rigid ryegrass. Two populations of rigid ryegrass from vineyards, SLR80 and SLR88, had 6- to 11-fold resistance to glyphosate in dose-response studies. These resistance levels were higher than two previously well-characterized glyphosate-resistant populations of rigid ryegrass (SLR77 and NLR70), containing a modified target site or reduced translocation, respectively. Populations SLR80 and SLR88 accumulated less glyphosate, 12 and 17% of absorbed glyphosate, in the shoot in the resistant populations compared with 26% in the susceptible population. In addition, a mutation within the target enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) where Pro106had been substituted by either serine or threonine was identified. These two populations are more highly resistant to glyphosate as a consequence of expressing two different resistance mechanisms concurrently.
A diagnostic test (Syngenta Quick-Test, QT) used for testing grass weed survivors to herbicides in the field for resistance was evaluated. Cuttings from grass weeds were transplanted into pots to regenerate new leaves, then treated with herbicide. In greenhouse experiments, resistance of known herbicide-resistant blackgrass biotypes to the aryloxyphenoxypropanoate herbicides CGA 184927 and fenoxaprop-ethyl and to the phenylurea herbicide isoproturon was verified by the QT. The findings were similar to those for seedlings grown from seed. Rigid ryegrass from suspect resistant fields in South Australia was sampled and sent by post to Switzerland for QT analysis. Resistance was confirmed in less than 4 wk, which verified resistance as responsible for the field failures. The added features of the QT over current resistance tests suggest a likely fit for in-season testing of surviving weeds and possible follow-up action.
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