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Several UK populations of the grass weed Alopecurus myosuroides were identified where high proportions of individuals showed resistance to the acetolactate synthase (ALS)‐inhibiting herbicides, mesosulfuron‐methyl + iodosulfuron‐methyl sodium mixture and sulfometuron‐methyl. Screening with sulfometuron, followed by DNA sequencing of the ALS gene from resistant and susceptible individuals, led to the identification of eight populations where a single point mutation segregated with resistance to sulfometuron. All highly resistant individuals from seven of eight populations showed a single‐nucleotide polymorphism (SNP) in the first position of the Pro197 codon of an A. myosuroides ALS gene, conferring a predicted proline to threonine target‐site change. One population showed resistant individuals with single‐nucleotide polymorphism in the second position of the Trp574 codon, conferring a predicted tryptophan to leucine substitution. No other mutations segregating with resistance were found. Enzyme assays confirmed that resistance was due to an altered form of ALS enzyme, which was less susceptible to inhibition by sulfonylureas, making this one of the first fully characterised cases of ALS target‐site resistance in a European grass weed. Increased information regarding the nature and distribution of ALS target‐site mutation may help support sustainable management strategies, allowing continued use of mesosulfuron + iodosulfuron against this weed in the UK. 相似文献
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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
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V Kati L Scarabel D Thiery‐Lanfranchi V Kioleoglou S Liberopoulou C Dlye 《Weed Research》2019,59(5):367-376
The issue of cross‐ or multiple resistance to acetolactate synthase (ALS) inhibitors and the auxinic herbicide 2,4‐D was investigated in Papaver rhoeas L., a common and troublesome weed in winter cereals, in a broad‐scale study across four European countries. A combination of herbicide sensitivity bioassays and molecular assays targeting mutations involved in resistance was conducted on 27 populations of P. rhoeas originating from Greece (9), Italy (5), France (10) and Spain (3). Plants resistant to the field rate of 2,4‐D were observed in 25 of the 27 populations assayed, in frequencies ranging from 5% to 85%. Plants resistant to ALS‐inhibiting herbicides (sulfonylureas) were present in 24 of the 27 populations, in frequencies ranging from 4% to 100%. Plants resistant to 2,4‐D co‐occurred with plants resistant to sulfonylureas in 23 populations. In four of these, the probability of presence of plants with cross‐ or multiple resistance to 2,4‐D and sulfonylureas was higher than 0.5. ALS genotyping of plants from the field populations or of their progenies, identified ALS alleles carrying a mutation at codon Pro197 or Trp574 in 2,4‐D‐sensitive and in 2,4‐D‐resistant plants. The latter case confirmed multiple resistance to 2,4‐D and ALS inhibitors at the level of individual plants in all four countries investigated. This study is the first to identify individual plants with multiple resistance in P. rhoeas, an attribute rarely assessed in other weed species, but one with significant implications in designing chemical control strategies. 相似文献
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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. 相似文献
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荠菜对乙酰乳酸合成酶抑制剂类除草剂的抗性水平及其分子机制 总被引：1，自引：1，他引：0
为明确荠菜种群对苯磺隆的抗性水平及其靶标抗性产生的分子机制,采用整株水平测定法测定了荠菜对苯磺隆及其他5种乙酰乳酸合成酶（ALS）抑制剂类除草剂的抗性水平,同时扩增和比对了荠菜抗性和敏感种群之间ALS基因的差异。结果显示:与敏感种群15-ZMD-1相比,抗性种群15-ZMD-5对苯磺隆产生了高水平抗性,抗性倍数为219.6;15-ZMD-5种群不同单株中共存在3种突变方式,分别为ALS基因197位点脯氨酸（CCT）突变为亮氨酸（CTT）、574位点色氨酸（TGG）突变为亮氨酸（TTG）以及单株同时发生上述197和574位点的氨基酸突变。15-ZMD-5抗苯磺隆种群对嘧草硫醚、啶磺草胺和氟唑磺隆均产生了高水平的交互抗性,抗性倍数分别为41.2、79.3和87.8;对双氟磺草胺和咪唑乙烟酸产生了低水平的交互抗性,抗性倍数分别为8.5和5.6。分析表明,荠菜抗性种群ALS基因发生的氨基酸突变可能是导致其对ALS抑制剂类除草剂产生抗性的重要原因之一。 相似文献
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Molecular characterisation of resistance to ALS-inhibiting herbicides in Hordeum leporinum biotypes 总被引：1，自引：0，他引：1
BACKGROUND: The acetolactate synthase (ALS)-inhibiting herbicide sulfosulfuron is registered in Australia for the selective control of Hordeum leporinum Link. in wheat crops. This herbicide failed to control H. leporinum on two farms in Western Australia on its first use. This study aimed to determine the level of resistance of three H. leporinum biotypes, identify the biochemical and molecular basis and develop molecular markers for diagnostic analysis of the resistance. RESULTS: Dose-response studies revealed very high level (>340-fold) resistance to the sulfonylurea herbicides sulfosulfuron and sulfometuron. In vitro ALS assays revealed that resistance was due to reduced sensitivity of the ALS enzyme to herbicide inhibition. This altered ALS sensitivity in the resistant biotypes was found to be due to a mutation in the ALS gene resulting in amino acid proline to serine substitution at position 197. In addition, two- to threefold higher ALS activities were consistently found in the resistant biotypes, compared with the known susceptible biotype. Two cleaved amplified polymorphic sequence (CAPS) markers were developed for diagnostic testing of the resistant populations. CONCLUSION: This study established the first documented case of evolved ALS inhibitor resistance in H. leporinum and revealed that the molecular basis of resistance is due to a Pro to Ser mutation in the ALS gene. 相似文献
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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
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为明确河南省部分地区的多花黑麦草Lolium multiflorum种群对乙酰辅酶A羧化酶（acetylCoA carboxylase,ACCase）和乙酰乳酸合成酶（acetolactate synthase,ALS）抑制剂类除草剂的抗性水平和抗性机理,采用整株生物测定法测定采自新乡市和驻马店市的多花黑麦草种群对ACCase抑制剂类除草剂精噁唑禾草灵、炔草酯、唑啉草酯和ALS抑制剂类除草剂甲基二磺隆、氟唑磺隆、啶磺草胺的抗性水平,并对多花黑麦草ACCase和ALS靶标酶编码基因进行克隆及氨基酸序列比对,分析其靶标抗性机理。结果显示,与多花黑麦草敏感种群HNXX01相比,HNZMD04和HNXX05种群对6种除草剂均产生了抗性,HNZMD04种群对精噁唑禾草灵和啶磺草胺的相对抗性倍数分别为44.65和40.31,对炔草酯和氟唑磺隆的相对抗性倍数分别为11.91和11.93;HNXX05种群对精噁唑禾草灵和氟唑磺隆的相对抗性倍数分别为27.70和25.67。HNZMD04和HNXX05抗性种群的ACCase基因均发生了D2078G突变,2个种群的突变率分别为55%和70%;HNZMD04... 相似文献
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BACKGROUND: Bromus rigidus is a common weed species that has increased in cropping fields owing to limited control options. During a random field survey in Western Australia, six B. rigidus populations that had survived in‐crop weed control programmes were collected. The study aimed to determine the resistance profile of these six populations. RESULTS: Based on dose–response studies, all six B. rigidus populations had a low‐level resistance to sulfosulfuron and sulfometuron (both sulfonylurea herbicides) while remaining susceptible to herbicides with other modes of action. ALS in vitro activity assays revealed no differences in enzyme sensitivity between susceptible and resistant populations, while the use of malathion (a cytochrome P450 inhibitor) in combination with sulfosulfuron caused the resistant populations to behave like the susceptible population. CONCLUSION: This study established that these six B. rigidus populations have a low‐level resistance to the ALS‐inhibiting sulfonylurea herbicides, but are able to be controlled by other herbicide modes of action. The low‐level, malathion‐reversible resistance, together with a sensitive ALS, strongly suggest that a non‐target‐site enhanced metabolism is the mechanism of resistance. Copyright © 2012 Society of Chemical Industry 相似文献
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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
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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
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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
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Zheng D Kruger GR Singh S Davis VM Tranel PJ Weller SC Johnson WG 《Pest management science》2011,67(12):1486-1492
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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Physiological and molecular basis for ALS inhibitor resistance in Bromus tectorum biotypes 总被引：2，自引：0，他引：2
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
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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
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Sulfonylurea resistance in Stellaria media [L.] Vill. 总被引：1，自引：1，他引：1
A sulfonylurea resistant biotype of common chickweed (Stellaria media L. Vill.) was found in a field treated with chlorsulfuron or metsulfuron for eight consecutive years. In pot experiments the biotype was resistant to postemergence treatments with the following acetolactate synthase (ALS) inhibitors: chlorsulfuron, metsulfuron, tribenuron, triasulfuron, rimsulfuron, sulfometuron, flumetsulam and imazapyr. The level of resistance to chlorsulfuron and sulfometuron was higher than to the other sulfonylurea herbicides. Whereas the level of cross resistance to the triazolopyrimidine herbicide, flumetsulam was comparable to that of metsulfuron, that of imazapyr was significantly lower. In contrast to imazapyr the biotype was not resistant to imazethapyr, an other imidazolinone herbicide. ALS in vitro assays revealed that resistance was due to an ALS enzyme that was less sensitive to ALS inhibiting herbicides. Herbicides with different modes of action were equally effective on the susceptible and resistant biotypes. 相似文献
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Ten accessions of sulfonylurea‐resistant Schoenoplectus juncoides were collected from paddy fields in Japan. In order to characterize acetolactate synthase from sulfonylurea‐resistant S. juncoides, acetolactate synthase amino acid substitutions, whole‐plant growth inhibition and acetolactate synthase enzyme inhibition were examined. Schoenoplectus juncoides has two acetolactate synthase genes (ALS1 and ALS2). The sulfonylurea‐resistant accessions harbored amino acid substitutions at Pro197 or Trp574 in either ALS1 or ALS2 (the amino acid number is standardized to the Arabidopsis thaliana sequence). The whole plants of all the sulfonylurea‐resistant accessions showed resistance to imazosulfuron. The resistance level depended on the altered amino acid residues in acetolactate synthase. The acetolactate synthase enzyme that was partially purified from all the sulfonylurea‐resistant accessions was less sensitive to imazosulfuron, compared to the susceptible accession, suggesting that the resistance is related to the altered acetolactate synthase enzyme. In addition, the concentration–response inhibition of acetolactate synthase activity by imazosulfuron in the sulfonylurea‐resistant accessions was remarkably different with the presence of an amino acid substitution in either ALS1 or ALS2. Furthermore, the concentration–response inhibition of acetolactate synthase activity in the sulfonylurea‐resistant accessions with a P197S, P197T or W574L mutation showed a double‐sigmoid curve. The regression analysis of enzyme inhibition suggested that the abundance ratio of ALS1 to ALS2 enzymes was approximately 70:30%, with a range of ±15%. Taken together, these results suggest that the resistance of sulfonylurea‐resistant accessions of S. juncoides is related to altered acetolactate synthase in either ALS1 or ALS2, although the abundance of the altered acetolactate synthase in the plants is different among the sulfonylurea‐resistant accessions. 相似文献
20.
BACKGROUND: Lolium rigidum Gaud. is one of the most common weed species in winter cereals in north‐eastern Spain, with populations that have evolved resistance to herbicides becoming more widespread since the mid‐1990s. Nine trials on commercial fields with herbicide‐resistant L. rigidum were conducted during the cropping seasons 2001–2002 to 2003–2004, testing the efficacy of 20 herbicides and mixtures pre‐ and post‐emergence and as sequential applications. Weed populations chosen had different resistance patterns to chlortoluron, chlorsulfuron, diclofop‐methyl and tralkoxydim, representative of the resistance problems faced by farmers. RESULTS: In pre‐emergence, prosulfocarb mixed with trifluralin, chlortoluron or triasulfuron was effective on six populations. In post‐emergence, iodosulfuron alone or mixed with mesosulfuron gave the best results but did not control three resistant populations. At Ferran 1, none of the herbicide combinations reached 90% efficacy. CONCLUSIONS: The diverse efficacy patterns of the different populations demonstrate the need for detailed knowledge of the populations before using herbicides. Moreover, the unexpected insufficient efficacy of the new herbicide iodosulfuron prior to its field use shows the need to combine herbicides with other non‐chemical weed control methods to control resistant L. rigidum in north‐eastern Spain. Copyright © 2010 Society of Chemical Industry 相似文献