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1.

Background

Annual sowthistle is a weed that is difficult to control in lentil crops in southern Australia due to a lack of herbicide options, widespread herbicide resistance and prolific production of highly mobile seed. This study investigates herbicide resistance in annual sowthistle in the Mid-North (MN) and Yorke Peninsula (YP) regions of South Australia, identifies and characterizes the mechanisms of acetolactate-synthase (ALS)-inhibitor resistance in this amphidiploid species, and combines this with analyses of population structure and gene flow.

Results

ALS-inhibitor-resistant annual sowthistle is widespread across the YP and MN of South Australia and is associated with a variety of Proline-197 mutations of the ALS gene, including leucine, alanine, arginine, serine, threonine and histidine. These mutations were found in different combinations on either of the two copies of the ALS gene. An additional 200 tissue samples were collected from across a single field on the YP and the ALS gene was sequenced for all these individuals. Different ALS-inhibitor resistance profiles were evident between mutation combinations and within mutation combinations, possibly mediated by differing subgenome assortment of the mutations, or altered gene experession of the two ALS homeologs. Population genetics analysis showed evidence of long-distance dispersal, resulting in highly mobile resistance genes, and multiple instances of resistance mutation evolution.

Conclusions

Continuing selection of Sonchus oleraceus populations with ALS-inhibiting herbicides has resulted in the accumulation of additional mutations within the ALS gene. New practices to control herbicide-resistant S. oleraceus should be examined, and control should focus on reducing seed set and dispersal to prevent the spread of emerging cases of resistance. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.  相似文献   

2.
BACKGROUND: Hordeum populations are becoming increasingly difficult to control in cropping fields. Two herbicide‐resistant H. leporinum populations were identified during a random crop survey after herbicides were applied. The study aimed to determine the herbicide resistance profile of these H. leporinum biotypes to a range of herbicides used for their control. RESULTS: Based on dose–response studies, one H. leporinum population was very highly resistant to sulfosulfuron and sulfometuron (both sulfonylurea herbicides) and also displayed low‐level resistance to imazamox (an imidazolinone herbicide). Reduced sensitivity of the ALS enzyme was identified with in vitro activity assays. Gene sequence analysis revealed a proline‐to‐threonine substitution at amino acid position 197 of ALS, which is likely to be the molecular basis for resistance in this population. Herbicide screening also revealed a different H. leporinum population with resistance to the bipyridyl herbicide paraquat. CONCLUSION: This study established the first cases of (1) sulfonylurea‐to‐imidazolinone cross‐resistance and (2) field‐evolved paraquat resistance in a Hordeum species in Western Australia. Copyright © 2012 Society of Chemical Industry  相似文献   

3.
Sulfonylurea-resistant biotypes of Schoenoplectus juncoides were collected from Nakafurano, Shiwa, Matsuyama, and Yurihonjyo in Japan. All of the four biotypes showed resistance to bensulfuron-methyl and thifensulfuron-methyl in whole-plant experiments. The growth of the Nakafurano, Shiwa, and Matsuyama biotypes was inhibited by imazaquin-ammonium and bispyribac-sodium, whereas the Yurihonjyo biotype grew normally after treatment with these herbicides. The herbicide concentration required to inhibit the acetolactate synthase (ALS) enzyme by 50% (I50), obtained using in vivo ALS assays, indicated that the four biotypes were > 10-fold more resistant to thifensulfuron-methyl than a susceptible biotype. The Nakafurano, Shiwa, and Matsuyama biotypes exhibited no or little resistance to imazaquin-ammonium, whereas the Yurihonjyo biotype exhibited 6700-fold resistance to the herbicide. The Nakafurano and Shiwa biotypes exhibited no resistance to bispyribac-sodium, but the Matsuyama biotype exhibited 21-fold resistance and the Yurihonjyo biotype exhibited 260-fold resistance to the herbicide. Two S. juncoides ALS genes (ALS1 and ALS2) were isolated and each was found to contain one intron and to encode an ALS protein of 645 amino acids. Sequencing of the ALS genes revealed an amino acid substitution at Pro197 in either encoded protein (ALS1 or ALS2) in the biotypes from Nakafurano (Pro197 → Ser197), Shiwa (Pro197 → His197), and Matsuyama (Pro197 → Leu197). The ALS2 of the biotype from Yurihonjyo was found to contain a Trp574 → Leu574 substitution. The relationships between the responses to ALS-inhibiting herbicides and the amino acid substitutions, which are consistent with previous reports in other plants, indicate that the substitutions at Pro197 and Trp574 are the basis of the resistance to sulfonylureas in these S. juncoides biotypes.  相似文献   

4.
Schoenoplectiella juncoides is a noxious sedge weed in rice paddy fields that has evolved resistance to sulfonylurea (SU) herbicides. The molecular basis of resistance is amino acid substitutions at Pro197, Trp574 or Asp376 in the acetolactate synthase (ALS) enzyme, which is the target of SUs. Schoenoplectiella juncoides has two ALS genes and resistant plants have point mutations that cause amino acid substitutions in either encoded protein. Single‐nucleotide substitutions at the codon for Pro197 in the ALS genes can cause six types of amino acid substitutions and all of these substitutions have been found in both ALS genes among Japanese SU‐resistant biotypes. Whole‐plant herbicide responses differ among the amino acid substitution types. Furthermore, analyses of ALS activity in plant extracts show that the extracts’ responses to herbicides differ, depending on which ALS gene is mutated. The activity responses of the ALS extracts to the SU, imazosulfuron, showed double‐sigmoid curves with plateaus of ~30% inhibition for Pro197 substitutions in ALS1 and ~70% for Pro197 substitutions in ALS2. This indicates that ALS1 and ALS2 contribute to the responses with a proportion of 7:3. The double‐sigmoid curves can be reconstructed to show the responses of the resistant and susceptible enzymes separately by regression analysis. The resistance levels of the separate ALS1 or ALS2 mutated enzyme are highly correlated with the whole‐plant responses, with a relationship that the former is the square of the latter. This could provide a quantitative insight into the physiological basis of resistance.  相似文献   

5.
Yu Q  Han H  Powles SB 《Pest management science》2008,64(12):1229-1236
BACKGROUND: In the important grass weed Lolium rigidum (Gaud.), resistance to ALS‐inhibiting herbicides has evolved widely in Australia. The authors have previously characterised the biochemical basis of ALS herbicide resistance in a number of L. rigidum biotypes and established that resistance can be due to a resistant ALS and/or enhanced herbicide metabolism. The purpose of this study was to identify specific resistance‐endowing ALS gene mutation(s) in four resistant populations and to develop PCR‐based molecular markers. RESULTS: Six resistance‐conferring ALS mutations were identified: Pro‐197‐Ala, Pro‐197‐Arg, Pro‐197‐Gln, Pro‐197‐Leu, Pro‐197‐Ser and Trp‐574‐Leu. All six mutations were found in one population (WLR1). Each Pro‐197 mutation conferred resistance to the sulfonylurea (SU) herbicide sulfometuron, whereas the Trp‐574‐Leu mutation conferred resistance to both sulfometuron and the imidazolinone (IMS) herbicide imazapyr. A derived cleaved amplified polymorphic sequences (dCAPS) marker was developed for detecting resistance mutations at Pro‐197. Furthermore, cleaved amplified polymorphic sequences (CAPS) markers were developed for detecting each of the six mutant resistant alleles. Using these markers, the authors revealed diverse ALS‐resistant alleles and genotypes in these populations and related them directly to phenotypic resistance to ALS‐inhibiting herbicides. CONCLUSION: This study established the existence of a diversity of ALS gene mutations endowing resistance in L. rigidum populations: 1–6 different mutations were found within single populations. At field herbicide rates, resistance profiles were determined more by the specific mutation than by whether plants were homo‐ or heterozygous for the mutation. Copyright © 2008 Society of Chemical Industry  相似文献   

6.
BACKGROUND: Wild radish, a problem weed worldwide, is a severe dicotyledonous weed in crops. In Australia, sustained reliance on ALS‐inhibiting herbicides to control this species has led to the evolution of many resistant populations endowed by any of several ALS mutations. The molecular basis of ALS‐inhibiting herbicide resistance in a novel resistant population was studied. RESULTS: ALS gene sequencing revealed a previously unreported substitution of Tyr for Ala at amino acid position 122 in resistant individuals of a wild radish population (WARR30). A purified subpopulation individually homozygous for the Ala‐122‐Tyr mutation was generated and characterised in terms of its response to the different chemical classes of ALS‐inhibiting herbicides. Whole‐plant dose‐response studies showed that the purified subpopulation was highly resistant to chlorsulfuron, metosulam and imazamox, with LD50 or GR50 R/S ratio of > 1024, > 512 and > 137 respectively. The resistance to imazypyr was found to be relatively moderate (but still substantial), with LD50 and GR50 R/S ratios of > 16 and > 7.8 respectively. In vitro ALS activity assays showed that Ala‐122‐Tyr ALS was highly resistant to all tested ALS‐inhibiting herbicides. CONCLUSION: The molecular basis of ALS‐inhibiting herbicide resistance in wild radish population WARR30 was identified to be due to an Ala‐122‐Tyr mutation in the ALS gene. This is the first report of an amino acid substitution at Ala‐122 in the plant ALS that confers high‐level and broad‐spectrum resistance to ALS‐inhibiting herbicides, a remarkable contrast to the known mutation Ala‐122‐Thr endowing resistance to imidazolinone herbicide. Copyright © 2012 Society of Chemical Industry  相似文献   

7.
Compared with natural seed dispersal, human‐mediated seed dispersal could spread herbicide resistance genes on a much larger scale. Herbicide‐resistant weed seeds have been reported as contaminants in commercial grain. We investigated the contamination of seeds of Lolium species with target‐site mutations conferring resistance to acetolactate synthase (ALS)‐inhibiting herbicides in wheat imported from the USA, Canada and Australia into Japan. We also investigated the establishment of ALS‐inhibiting herbicide‐resistant Lolium species in 12 seaports in Japan that are major entry points for international commodities. We found herbicide‐resistant Lolium spp. seeds from all classes of wheat samples. Resistant individuals became established at six of eight ports where more than 50 kt of imported wheat is unloaded every year. The establishment of resistant Lolium spp. individuals was common at major grain landing ports. Monitoring over 3 years at one port revealed that the frequency of resistant individuals did not fluctuate between years. Many resistant individuals were distributed in front of the entrance of a fodder company, but a few resistant individuals were found in areas 2 km away from the port. The results indicate that gene flow is rare through pollen or seed movement from resistant plants to peripheral populations. Further extensive and long‐term monitoring is necessary to perform a comprehensive risk assessment of herbicide‐resistant plants entering Japan through major commercial ports.  相似文献   

8.
Resistance to the acetyl‐coenzyme A carboxylase (ACCase)‐inhibiting herbicides in Lolium rigidum is widespread in grain cropping areas of South Australia. To better understand the occurrence and spread of resistance to these herbicides and how it has changed with time, the carboxyl transferase (CT) domain of the ACCase gene from resistant L. rigidum plants, collected from both random surveys of the mid‐north of Southern Australia over 10 years as well as stratified surveys in individual fields, was sequenced and target site mutations characterised. Amino acid substitutions occurring as a consequence of these target site mutations, at seven positions in the ACCase gene previously correlated with herbicide resistance, were identified in c. 80% of resistant individuals, indicating target site mutation is a common mechanism of resistance in L. rigidum to this herbicide mode of action. Individuals containing multiple amino acid substitutions (two, and in two cases, three substitutions) were also found. Substitutions at position 2041 occurred at the highest frequency in all years of the large area survey, while substitutions at position 2078 were most common in the single farm analysis. This study has shown that target site mutations leading to amino acid substitutions in ACCase of L. rigidum are widespread across South Australia and that these mutations have likely evolved independently in different locations. The results indicate that seed movement, both within and between fields, may contribute to the spread of resistance in a single field. However, over a large area, the independent appearance and selection of target site mutations conferring resistance through herbicide use is the most important factor.  相似文献   

9.
BACKGROUND: Papaver rhoeas (L.) has evolved resistance to tribenuron in winter wheat fields in northern Greece owing to multiple Pro197 substitutions. Therefore, the cross‐resistance pattern to other sulfonylurea and non‐sulfonylurea ALS‐inhibiting herbicides of the tribenuron resistant (R) and susceptible (S) corn poppy populations was studied by using whole‐plant trials and in vitro ALS catalytic activity assays. RESULTS: The whole‐plant trials revealed that tribenuron R populations were also cross‐resistant to sulfonylureas mesosulfuron + iodosulfuron, chlorsulfuron and triasulfuron. The whole‐plant resistance factors (RFs) calculated for pyrithiobac, imazamox and florasulam ranged from 12.4 to > 88, from 1.5 to 28.3 and from 5.6 to 25.4, respectively, and were lower than the respective tribenuron RF values (137 to > 2400). The ALS activity assay showed higher resistance of the ALS enzyme to sulfonylurea herbicides (tribenuron > chlorsulfuron) and lower resistance to non‐sulfonylurea ALS‐inhibiting herbicides (pyrithiobac > florasulam ≈ imazamox). CONCLUSION: These findings indicate that Pro197 substitution by Ala, Ser, Arg or Thr in corn poppy results in a less sensitive ALS enzyme to sulfonylurea herbicides than to other ALS‐inhibiting herbicides. The continued use of sulfonylurea herbicides led to cross‐resistance to all ALS‐inhibiting herbicides, making their use impossible in corn poppy resistance management programmes. Copyright © 2011 Society of Chemical Industry  相似文献   

10.
An investigation, using herbicidal pot tests in a greenhouse condition, was conducted to determine the whole‐plant dose–response relationships to several acetolactate synthase (ALS)‐inhibiting herbicides of sulfonylurea (SU)‐resistant Schoenoplectus juncoides with various Pro197 mutations in ALS that was collected from Japanese rice paddy fields. All the tested SU‐resistant accessions with a Pro197 mutation were highly resistant to two commonly used SU herbicides (imazosulfuron and bensulfuron‐methyl), but were much less resistant to another SU herbicide, metsulfuron‐methyl, and were substantially not resistant to imazaquin‐ammonium. These cross‐resistance patterns have been known previously in fragments of S. juncoides and other weed species and were comprehensively confirmed in this study with a whole set of Pro197 mutations. The analyses of resistance levels, based on ED90 values, newly showed that different accessions with a common amino acid substitution in ALS1 showed similar responses to these herbicides (confirmed with four amino acid substitutions), that the rankings of resistance levels that were conferred by various Pro197 mutations in ALS1 differed among the SU herbicides and that the resistance levels of the ALS2‐mutated accessions were higher than, lower than or similar to those of the corresponding ALS1‐mutated accessions, depending on the compared pair, but the deviation patterns were generally similar among the SU herbicides in each compared pair. The final finding might suggest that the abundance of ALS2 is not as stable as that of ALS1. In addition, as a result of these new findings, together with expected further research, a suggested possibility is that substituting amino acids at Pro197 generally could be estimated by plotting each accession's ED90 values of imazosulfuron and bensulfuron‐methyl in a two‐dimensional graph.  相似文献   

11.
Imidazolinone herbicides inhibit the first enzyme of branched-chain amino acid biosynthesis, acetohydroxyacid synthase. The inhibition of the enzyme in vitro by these herbicides increases with incubation time, but is not irreversible, as deduced by reaction progress curves. In contrast to this result is the apparent irreversible inhibition of the enzyme by these imidazolinones that occurs when the herbicide is applied to intact corn plants. Plants treated with imidazolinone herbicide and then extracted showed dramatically reduced enzyme activity. This effect on extractable enzyme level occurred with several different imidazolinone herbicides in either foliar or soil application. The decrease in extractable enzyme activity could be observed within four hours after treatment. Herbicides other than imidazolinones did not reduce the extractable enzyme level. These findings suggest that the enzyme- imidazolinone interaction in vivo may be different from the interaction observed in vitro.  相似文献   

12.
A biotype of Sonchus oleraceus L. and two bio types of Sisymbrium orientate Torn., SSO 3 and NSO 1, are the first dicot weeds in Australia to develop resistance to ALS-inhibiting herbicides. The resistant biotypes had been exposed to va rying periods of selection with sulfonylurea her bicides. All three biotypes are resistant to a range of sulfonylurea and imidazolinone herbicides. The S. orientale biotypes are also resistant to the triazolopyrimidine herbicide, flumetsulam. LD50 ratios of resistant Sonchus oleraceus for sulfony lurea and imidazolinone herbicides are greater than 64-fold and 4.5-fold, respectively. GR50 ratios are greater than 9 for sulfonylureas and 7.4 for imazapyr. The LD50 ratios for both S. orien tale biotypes for chlorsulfuron, sulfometuron methyl, metsulfuron-methyl, flumetsulam and imazethapyr are greater than 110-, 15-, 7-, 24- and 29-fold, respectively. All resistant biotypes are susceptible to MCPA, diuron and diflufenican, herbicides which do not inhibit ALS.  相似文献   

13.
Suspected sulfonylurea (SU)‐resistant Schoenoplectus juncoides plants were collected from rice paddy fields at 24 sites in Japan in order to discover the occurrence pattern of target‐site substitutions on a nationwide scale and at a local field scale. A genetic analysis of the two acetolactate synthase (ALS) genes, ALS1 and ALS2, of the collected plants confirmed that a single‐nucleotide mutation at the Pro197, Asp376 or Trp574 site of either ALS1 or ALS2 existed in each suspected SU‐resistant plant. On a nationwide scale, it was shown that the ALS1 mutations and the ALS2 mutations occurred at a similar frequency, that the P197S and the P197L substitutions were found most frequently among all the substitutions, and that the W574L substitutions (known as global resistance to any ALS‐inhibiting herbicide) were found at a relatively low frequency but in a geographically wide range. In the local field‐scale survey, which was conducted at two sites in Hyogo Prefecture, it was shown that the substitutions were less diverse, compared to on a nationwide scale, probably because the investigation involved a limited number of local fields, and that several substitutions and a susceptible biotype were found in single fields suggesting that a number of collections is required in order to understand the local SU‐resistant status of S. juncoides. In addition, this study reported new findings, that of the P197R, P197T and D376E substitutions in S. juncoides. This set of diverse substitutions in a weed species can be used for further research purposes.  相似文献   

14.
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 Km (pyruvate) (the substrate concentration at which the reaction rate is half maximal) but a greater Vmax (the rate when the enzyme is fully saturated with substrate) for two of the resistant S orientale biotypes over susceptible levels. F1 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 F2 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 ALS-resistant biotypes were amplified 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 (CC T to AT T) predicting a Pro (in susceptible) to Ile (in resistant) change in the primary amino acid sequence were identified 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. © 1999 Society of Chemical Industry  相似文献   

15.
Primisulfuron‐resistant (AR and MR) and ‐susceptible (AS and MS) Bromus tectorum biotypes were collected from a Poa pratensis field at Athena, Oregon, and in research plots at Madras, Oregon. Studies were conducted to characterize the resistance of the B. tectorum biotypes. Whole plant bioassay and acetolactate synthase (ALS) enzyme assay revealed that the AR biotype was highly resistant to the sulfonylurea (SU) herbicides, primisulfuron and sulfosulfuron and to a sulfonylaminocarbonyltriazolinone (SCT) herbicide, propoxycarbazone‐sodium. However, the AR biotype was not resistant to imazamox, an imidazolinone (IMI) herbicide. Results of the whole plant bioassay studies showed that the MR biotype was moderately resistant to all ALS inhibitors tested. However, there were no differences in ALS sensitivities between the MR and MS biotypes. The nucleotide and amino acid sequence analysis of the als gene demonstrated a single‐point mutation from C to T, conferring the exchange of the amino acid proline to serine at position 197 in the AR biotype. However, this mutation was not found in the MR biotype. Results of this research indicate that: the resistance of the AR biotype to SU and SCT herbicides is based on an altered target site due to a single‐point mutation; resistance in the MR biotype is not due to a target site mutation.  相似文献   

16.
A population of Bromus tectorum infesting an olive grove at Córdoba (Spain) survived simazine use rates of 3.0 kg a.i. ha−1 over two consecutive years. Non‐tillage olive monoculture and two annual simazine applications had been carried out for 10 years. The resistant biotype showed a higher ED50 value (7.3 kg a.i. ha−1) than that of the susceptible control (0.1 kg a.i. ha−1), a 73‐fold increase in herbicide tolerance. The use of fluorescence, Hill reaction, absorption, translocation and metabolism assays showed that simazine resistance in this biotype was caused by a modification of the herbicide target site, since chloroplasts from the resistant biotype of B. tectorum were more than 300 times less sensitive to simazine than those from the susceptible biotype. In addition, non‐treated resistant plants of B. tectorum displayed a significant reduction in the QA to QB electron transfer rate when compared with the susceptible biotype, a characteristic that has been linked to several mutations in the protein D1 conferring resistance to PS II inhibiting herbicides. Resistant plants showed cross‐resistance to other groups of triazine herbicides with the hierarchy of resistance level being methoxy‐s‐triazines ≥chloro‐s‐triazines > methylthio‐s‐triazines > cis‐triazines. The results indicate a naturally occurring target‐site point mutation is responsible for conferring resistance to triazine herbicides. This represents the first documented report of target site triazine resistance in this downy brome biotype.  相似文献   

17.
Herbicide resistance in Lolium rigidum is widespread across much of the agricultural land in Australia. As the incidence of herbicide resistance has increased, so has the incidence of multiple herbicide resistance. This reduces the herbicide options available for control of this weed. This study reports on the successful amplification and sequencing of the acetolactate synthase (ALS) gene of L. rigidum using primers designed from sequence information of related taxa. This enables, for the first time, the successful determination of a mutation in the ALS gene of this species that provides resistance to ALS‐inhibiting herbicides. This mutation causes amino acid substitution at Trp574 (numbering standardised to Arabidopsis thaliana) to Leu which had been reported to confer a high level of resistance against all classes of ALS inhibitor herbicides. In addition, multiple resistance to ALS‐inhibiting and acetyl‐coenzyme A carboxylase‐inhibiting herbicides is acquired through the independent accumulation of mutant alleles for the target sites. This may thus explain some of the irregular, mosaic resistance patterns that occur in this predominantly outcrossing species.  相似文献   

18.
19.
Timely detection of herbicide resistance at an early stage of crop cultivation is essential to help farmers find alternative solutions to manage herbicide resistance in their fields. In this study, maximum quantum yield of PS II [Fv/Fm = (FmFo)/Fm] was measured at the 4–5 leaf stage to discriminate between herbicide‐resistant and susceptible biotypes of Echinochloa species. The differences in Fv/Fm between herbicide‐resistant and susceptible Echinochloa spp. were consistent with the whole‐plant assay based on I50 (herbicide doses causing a 50% inhibition of Fv/Fm) and GR50 (herbicide doses causing a 50% reduction in plant fresh weight) values and R/S ratios (herbicide resistance index), regardless of the mode of action of the tested herbicides. A PS II inhibitor caused the fastest inhibition of Fv/Fm, compared with ACCase and ALS inhibitors, after herbicide treatment. The required time for discrimination between herbicide‐resistant and susceptible Echinochloa spp. was 64 h after PS II inhibitor treatment, much shorter than those of ACCase and ALS inhibitor‐treated plants, which required 168 and 192 h respectively. The leaf chlorophyll fluorescence assay provided reliable diagnostics of herbicide resistance in Echinochloa spp. with significant time savings and convenient measurement in field conditions compared with the conventional whole‐plant assay.  相似文献   

20.
BACKGROUND: Horseweed is a weed commonly found in agronomic crops, waste areas and roadsides. Resistance to ALS‐inhibiting herbicides in horseweed was first reported in 1993 in a population from Israel. Resistance to ALS‐inhibiting herbicides in horseweed is now widespread, but, as of now, the resistance mechanism has not been reported. RESULTS: Two of three populations evaluated (P116 and P13) were found to be uniform for resistance (>98% of individuals survived 8.8 g AI ha?1 of cloransulam), whereas a third population, P525, contained about 85% resistant individuals. Cross‐resistance to cloransulam, chlorimuron, imazethapyr and bispyribac was observed in the P116 population. P525 and P13 were both sensitive to imazethapyr but resistant to chlorimuron, imazethapyr and bispyribac. Enzyme activity assays indicated that resistance in P13 was due to an altered target site. Southern blot analysis indicated that the ALS target site is encoded by a single copy gene. Overlapping ALS gene regions were amplified and sequenced from each population. Amino acid substitutions of Ser for Pro at position 197 (P197S) was detected from P13, Ala for Pro (P197A) was identified from P525 and substitution of Glu for Asp (D376E) at position 376 was found in P116. Molecular markers were developed to differentiate between wild‐type and resistant codons at positions 197 and 376 of horseweed ALS. CONCLUSION: Resistance to ALS‐inhibiting herbicides in horseweed is conferred by target‐site mutations that have also been identified in other weed species. Identification of the mutations within horseweed ALS gene sequence enables molecular assays for rapid detection and resistance diagnosis. Copyright © 2011 Society of Chemical Industry  相似文献   

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