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1.
A laboratory test was developed to assess the sensitivity of field populations of Phytophthora infestans to metalaxyl. Discs of potato leaf tissue were floated upon solutions of the fungicide at different concentrations and inoculated with spores. The extent of symptom development was noted after incubation under standard conditions for 5–6 days. In preliminary experiments growth of isolates of P. infestans obtained from culture collections was severely inhibited in discs treated at 2 μg/ml. By contrast the development of an isolate obtained from a crop in Eire in which blight control with metalaxyl had failed, and known to be markedly less sensitive in vitro, was unaffected in discs treated at 100 μg/ml. During the summer of 1980, 234 samples of P. infestans were obtained from 20 sites in south-west England, 10 of which had received sprays containing metalaxyl and 10 of which had not. All samples were sensitive to metalaxyl applied at 2 μg/ml. In 1981, 35 sites within the same area, 30 of which had received sprays containing either metalaxyl or ofurace (a related fungicide), were similarly surveyed. Most of the 79 samples of P. infestans examined proved sensitive and at all sites the amount of blight was small. However, at three sites, including one not treated with acylalanine fungicides, strains were found which were unaffected by 100 μg/ml metalaxyl in leaf disc tests. These findings are discussed in relation to the development of resistant blight in other areas and to the use of fungicide mixtures.  相似文献   

2.
During the growing seasons between the years 2001 and 2004, 98 isolates of Pseudoperonospora cubensis from nine regions of Czech Republic were collected and screened for tolerance/resistance to the three frequently used fungicides (propamocarb, fosetyl‐Al, metalaxyl). Fungicides were tested in five different concentrations, using a floating disc bioassay. Fungicide effectiveness varied considerably. Propamocarb appeared most effective and all the isolates collected in the years 2001–2003 were found sensitive to all tested concentrations [607–9712 μg active ingredient (a.i.)/ml]. In 2004, some strains with increased resistance to propamocarb were detected. These strains were characterized by tolerance at the lowest concentrations (607 μg a.i./ml, eventually on 1214 μg a.i./ml); however, they were controlled by 2428 μg a.i./ml. Fosetyl‐Al was effective at the recommended concentration of 1600 μg a.i./ml against all isolates. However, the occurrence of isolates (collected in 2001) which sporulated at low concentrations (400 and 800 μg a.i./ml) indicated that the selection for tolerance occurs in the pathogen population. Nevertheless, this phenomenon was not confirmed with the P. cubensis isolates collected between the years 2002 and 2004. Metalaxyl was found ineffective, because 97% of the isolates showed the resistance to the recommended concentration (200 μg a.i./ml), and the other 3% of isolates expressed tolerant response. The majority of the isolates showed profuse and/or limited sporulation at higher concentrations (400 and 800 μg a.i./ml). A substantial shift to highly metalaxyl resistant strains was evident in the Czech P. cubensis populations during 2001–2004.  相似文献   

3.
Leaf blight caused by Phytophthora colocasiae is the most destructive disease affecting taro (Colocasia esculenta) worldwide including India. Fungicides (primarily metalaxyl) remain as an important strategy to manage taro leaf blight in India over decades. It is important to monitor isolate sensitivity to identify build-up of fungicide resistance and thereby modify fungicide usage strategies. P. colocasiae isolates representing four different geographical regions of India were evaluated for their sensitivity to metalaxyl and three other commercially available fungicides viz. Samarth, Biofight and Akoton by poisoned media technique. All the isolates tested were sensitive to metalaxyl, nevertheless there is an increase in the effective concentration compared to the previous reports. Among the other fungicides, Samarth was found to be superior in completely inhibiting mycelial growth at 0.05% followed by Biofight at 1%. Metalaxyl and Akoton® shared a common inhibitory concentration at 2%. The most effective fungicide determined by the in vitro method was evaluated in vivo for studying the pattern of inhibition before and after the disease development in detached taro leaf. The results of the study revealed that build-up on metalaxyl resistance in P. colocasiae is in its course and caution should be taken while administering against taro leaf blight. Fungicide Samarth could be used as an alternative to metalaxyl for management of taro leaf blight.  相似文献   

4.
Over a 3-yr period 261 isolates of 17 species of Pythium were tested for sensitivity to metalaxyl at concentrations of 5, 50 or 100 μ/ml. A wide range of responses was observed, from isolates where growth ceased at 5 μg/ml to those where growth at 100 μg/ml was similar to that of the untreated controls. In further tests isolates of 11 different species had ED50's < 1 μg/ml. A lower sensitivity was detected in isolates of six Pythium spp. where values in the range 1–10 μg/ml were obtained. This lower sensitivity was not related to previous known use of metalaxyl. Three isolates of Pythium dissotocum from sites where the fungicide had been used repeatedly had ED50's > 100 μg/ml and were considered resistant. The resistance was stable over a 2-yr period and isolates were cross-resistant to furalaxyl, benalaxyl, ofurace, cyprofuram and oxadixyl. Increasing concentrations of metalaxyl reduced or prevented the production of zoospores by four species of Pythium, although when zoospores were produced, this was followed by the normal processes of encystment and germination. Culturing P. dissotocum on different sub-lethal concentrations of metalaxyl for 18 wk did not induce a high level of resistance to the fungicide.  相似文献   

5.
Abstract: The effect of timing of application on the incompatibility of three fungicides (metalaxyl, mancozeb, copper oxide) and isolate MK2001 of the entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin was assessed in vitro . Isolate MK 2001 at 1 × 107 conidia/ml concentration was highly pathogenous to adults of Lygus lineolaris (Hemiptera) (84% mortality at day 4 post-treatment). The fungicides at the manufacturer recommended rate (1X) were strongly fungistatic and inhibited the fungal isolate radial growth on Sabouraud dextrose agar (SDA). Mancozeb, metalaxyl and copper oxide exhibited insecticidal activity of 24, 40 and 48% respectively, on L. lineolaris adults. It is clear that application of the fungal isolate MK2001 2–4 days before applying fungicides metalaxyl, mancozeb or copper oxide synergized the insecticidal effect of the isolate. On the contrary, application of metalaxyl, mancozeb, copper oxide 2–4 days before applying isolate MK2001 antagonized the insecticidal effect. The simultaneous use of each fungicide (metalaxyl, mancozeb or copper oxide) and the isolate gave lesser insect mortality. This study revealed that although some fungicides are incompatible for use alongside fungal isolates, the proper evaluation of their time of use could be beneficial in biological control or IPM programmes. Furthermore, the application of B. bassiana isolate MK2001 followed by fungicide application could synergize insecticidal activity.  相似文献   

6.
Biodegradation process and the nature of metabolism of metalaxyl in soil   总被引:1,自引:0,他引:1  
The enhanced biodegradation of metalaxyl was studied in tobacco, citrus, avocado and corn soils. The most rapid degradation of metalaxyl occurred in a tobacco soil in which the half-life (50% degradation) of metalaxyl was 6 days. The main breakdown product of metalaxyl in all soils was the acid metabolite. Ring labelled [14C]metalaxyl incubated for 4 wk in 6 soils demonstrated a low rate of 14CO2 evolution ranging from 2.1% to 11.3% which was unrelated to the biodegradation properties of the soil. A relationship between the concentration of metalaxyl and the subsequent rate of biodegradation was found in the tobacco soils. Higher concentrations of metalaxyl resulted in faster biodegradation rates. A single exposure of tobacco and corn soils to metalaxyl (100 μg/ml or 200 μg/g dry weight of soil) significantly increased their subsequent capacity to degrade the fungicide. Addition of the fungicide thiram or the antibiotics streptomycin and chloramphenicol to an avocado soil resulted in 75% and 51% inhibition of metalaxyl degradation, respectively. A combination of the fungicide and antibiotics resulted in 89% inhibition. The results indicate that enhanced microbial degradation of metalaxyl can occur in a wide range of soils. Under experimental conditions using soil solutions or soil systems, a single application of the fungicide may trigger this event. A wide range of fungi and bacteria appear to take part in degrading metalaxyl.  相似文献   

7.
Trichophyton species susceptibility to green and red propolis from Brazil   总被引:1,自引:0,他引:1  
Aims:  The in vitro antifungal activity of Brazilian green and red propolis was tested against different species of Trichophyton .
Methods and Results:  The antifungal activity of the Brazilian aqueous and alcoholic extracts of the green propolis and the alcoholic extract of red propolis was observed against Trichophyton rubrum , Trichophyton tonsurans and Trichophyton mentagrohytes samples, using as controls itraconazole and terbinafine. The minimal inhibitory concentration was determined following the microdilution method indicated by the 'Clinical and Laboratory Standards Institute'. The minimal fungicide concentration was determined by the absence of growth in liquid sabouraud culture medium. The data obtained showed that the green propolis alcoholic extract's antifungal activity was from 64 to 1024 μg ml−1, whereas the red propolis alcoholic extract was from 8 to 1024 μg ml−1.
Conclusions:  The antifungal activity of the red propolis alcoholic extract was more efficient than the green propolis alcoholic extract for all three species studied. The T. rubrum samples were shown to be more sensitive to the antifungal activity of the alcoholic extracts of the propolis.
Significance and Impact of the Study:  The antifungal potential of the alcoholic extracts of green and red propolis demonstrated suggest an applicable potential as an alternative treatment for dermatophytosis caused by these species.  相似文献   

8.
Isolates of Phytophthora cactorum resistant to the systemic fungicide metalaxyl were obtained by exposing them to sequentially increased concentrations of metalaxyl. A linear relationship was observed between the concentrations of metalaxyl and percentage inhibition of mycelial growth of P. cactorum. The stability of metalaxyl-resistant isolates 150R and 250R was confirmed after six serial transfers on corn meal agar without fungicide. The in vitro metalaxyl-resistant isolate (Ph10) was less aggressive on apple rootstocks compared with the Ph07 isolated from metalaxyl-treated trees and the Ph03 isolated from untreated trees. Metalaxyl-resistant and sensitive isolates remained sensitive to the chemically unrelated fungicide fosetyl-Al at high concentration (600 μg/ml), to mancozeb, and to a mixture of metalaxyl + mancozeb. Significant differences in resistance to metalaxyl existed among P. cactorum field isolates.  相似文献   

9.
Abstract:  Entomopathogenic fungi are a poorly exploited source of insecticidal proteins, which may be used for the development of new natural insecticides and as alternative molecules for transgenic deployment. The crude soluble protein extracts in Adamek's liquid medium of 25 fungal isolates belonging to the fungal species Metarhizum anisopliae , Beauveria bassiana , Beauveria brongniartii and Scopulariopsis brevicaulis were screened by per os toxicity on Spodoptera littoralis larvae. Whilst extracts from two M. anisopliae and two B. bassiana isolates gave significant mortalities when applied either on alfalfa leaf discs or incorporated into artificial diet, the one from M. anisopliae 01/58-Su isolate was the only most toxic that showed promise for S. littoralis control. In leaf disc assays, this extract exhibited strong dose-related toxic and antifeedant activity against the larvae. At 10, 20 and 40  μ g protein/insect, the extract gave 61.3%, 96.6% and 100.0% mortality, respectively, and average survival time of 5.7, 4.3 and 3.1 days respectively. Not only the antifeeding index was dose-related, but it significantly increased over time in a dose-related manner. Longer exposure times led to a dose-related significant increase in larval mortality. The exposure times for 50% mortality were 91.3 h and 62.1 h for 20 and 40  μ g protein/insect respectively. The crude extract when exposed to higher temperature or protease treatment lost toxicity, indicating that toxicity was protein mediated. In addition, the liquid medium composition did not influence its insecticidal activity. The effects of the protein extract on midgut cells of second instar larvae of S. littoralis were investigated by using both light and electron microscopy. A progressive bleeding of the midgut epithelium into the gut lumen was observed along with lysis of the epithelium and deterioration of the microvilli.  相似文献   

10.
The leaf disc agroinoculation system was applied to study tomato yellow leaf curl virus (TYLCV) replication in explants from susceptible and resistant tomato genotypes. This system was also evaluated as a potential selection tool in breeding programmes for TYLCV resistance. Leaf discs were incubated with a head-to-tail dimer of the TYLCV genome cloned into the Ti plasmid ofAgrobacterium tumefaciens. In leaf discs from susceptible cultivars (Lycopersicon esculentum) TYLCV single-stranded genomic DNA and its double-stranded DNA forms appeared within 2–5 days after inoculation. Whiteflies (Bemisia tabaci) efficiently transmitted the TYLCV disease to tomato test plants following acquisition feeding on agroinoculated tomato leaf discs. This indicates that infective viral particles have been produced and have reached the phloem cells of the explant where they can be acquired by the insects. Plants regenerated from agroinfected leaf discs of sensitive tomato cultivars exhibited disease symptoms and contained TYLCV DNA concentrations similar to those present in field-infected tomato plants, indicating that TYLCV can move out from the leaf disc into the regenerating plant. Leaf discs from accessions of the wild tomato species immune to whitefly-mediated inoculation,L. chilense LA1969 andL. hirsutum LA1777, did not support TYLCV DNA replication. Leaf discs from plants tolerant to TYLCV issued from breeding programmes behaved like leaf discs from susceptible cultivars.The Hebrew University of Jerusalem, Faculty of Agriculture, Department of Field and Vegetable Crops  相似文献   

11.
Aims:  The aim of this study was to identify fungal populations in unroasted cocoa beans stored in Spain in order to evaluate the ochratoxin A (OTA)-production ability of certain Aspergillus isolates.
Methods and Results:  Twenty batches of cocoa beans from different origins and with different OTA content were selected for this study. Three Aspergillus carbonarius and 13 Aspergillus niger aggregate strains isolated from these cocoa bean samples were selected to evaluate their OTA synthesis ability, being the only A. carbonarius isolates which are OTA producers [−1 culture medium; LOD = 6  μ g kg−1 culture medium].
Conclusions:  No correspondence was found between the OTA levels in cocoa beans and the presence of OTA-producing fungi. Nonetheless, some samples contained A. carbonarius with a high OTA-producing ability and, consequently, specific fungal controls should be set up during storage to avoid this toxin.
Significance and Impact of the Study:  Toxigenic fungi in cocoa beans are not well understood. This study attempted to identify these fungi and evaluate their OTA-producing ability.  相似文献   

12.
Pepper Phytophthora blight (PPB), caused by Phytophthora capsici, is an important disease of pepper in China. The extensive application of metalaxyl has resulted in widespread resistance to this fungicide in field. This study has evaluated the activities of several fungicides against the mycelial growth and sporangium germination of metalaxyl‐sensitive and metalaxyl‐resistant P. capsici isolates by determination of EC50 values. The results showed that the novel carboxylic acid amide (CAA) fungicide mandipropamid exhibited excellent inhibitory activity against PPB both in vitro and in vivo, with averagely EC50 values of 0.075 and 0.004 μg/ml in mycelial growth and sporangium germination, respectively, and over 88% efficacy in controlling PPB. The other three CAA fungicides also provided over 70% efficacy in controlling PPB. The mycelial growth was less sensitive to quinone outside inhibitor (QoI) fungicides azoxystrobin and trifloxystrobin than that of sporangium germination in P. capsici isolates. However, azoxystrobin and trifloxystrobin provided over 80% efficacy in controlling PPB. It was noted that propamocarb and cymoxanil did not exhibit activity against the mycelial growth or sporangium germination of P. capsici isolates in the in vitro tests, with over 70% efficacy in controlling PPB. The new fungicide mixture 62.5 g/l fluopicolide + 625 g/l propamocarb (trade name infinito, 687.5 g/l suspension concentrate (SC)) produced over 88% efficacy in controlling PPB caused by both metalaxyl‐sensitive and metalaxyl‐resistant isolates. The data of this study also proved that there was obviously no cross‐resistance between metalaxyl and the other tested fungicides. Therefore, these fungicides should be good alternatives to metalaxyl for the control of PPB and management of metalaxyl resistance.  相似文献   

13.
Aims:  Because of the lack of a standard method, the aim of this work is to evaluate the suitability of the broth microdilution method CLSI M38-A in determining the resistance level of some Penicillium expansum isolates to thiabendazole (TBZ). The ability of the isolates to produce patulin (PAT) and citrinin (CIT) has been also assessed.
Methods and Results:  Penicillium expansum isolates (128) were assayed (apples, pears, grapes and five reference strains). It was observed that 69·4% of the strains isolated from apples and pears were resistant to TBZ. Sensitive isolates were inhibited at 0·25–0·5 μg ml−1 whilst resistant isolates still grew at 512 μg ml−1. PAT was produced by all P. expansum isolates. CIT was detected in 98·8% of TBZ-resistant isolates and in 89·1% of the TBZ-sensitive isolates.
Conclusions:  The preliminary screening method combined with the adaptation of the method CLSI M38-A, can be a good strategy to be used in assessing the in vitro activity of TBZ against a large number of isolates.
Significance and Impact of the Study:  The proposed methodology can be a contribution to the standardization of susceptibility tests to fungicides against P. expansum.  相似文献   

14.
Antifeeding activity of limonoids from Khaya senegalensis (Meliaceae)   总被引:1,自引:0,他引:1  
Abstract: Fifteen B,D-secolimonoids of mexicanolide, rearranged phragmalin, methyl angolensate and glycoside types have been isolated from ether and acetone extracts of the stem bark of Khaya senegalensis (Desr.). The antifeedant activity of the isolated compounds was assessed by conventional choice leaf disc method on the third-instar larvae of Spodoptera littoralis (Boisd.). Khayalactol, 1- O -acetylkhayanolide A, 2-hydroxyseneganolide, khayanolide A, khayanolide D and methyl angolensate displayed strong antifeedant activity at 1000  μ g/ml with antifeedant percentages of 83.8, 61.9, 60.1, 59.5, 57.1 and 55.7, respectively. 2-Hydroxyseneganolide and khayanolide D showed high activity at 500, 300 and 200  μ g/ml while 1- O -acetylkhayanolide A was the only compound that revealed antifeedant activity at a concentration as low as 100  μ g/ml. Antifeedant activity was dose-dependent in some of the isolated compounds. Correlation between antifeedant activity of the isolated compounds and chemical structure was discussed.  相似文献   

15.
The fungus Syncephalastrum racemosum (Cohn) Schroeter was found to transform the fungicide metalaxyl [N-(2,6-dimethylphenyl)-N-(methoxyacetyl)-alanine methyl ester] in pure culture. After 21 days of incubation in a basal medium amended with 5 μg of metalaxyl per ml, more than 80% of the compound was transformed by the fungus. The transformation rates decreased as the concentrations of metalaxyl increased from 5 to 100 μg/ml. No transformation was observed when the concentration of metalaxyl was higher than 200 μg/ml. Two isomeric metabolites and a mixture of two other isomeric metabolites were isolated from the organic extract of the growth medium and identified as N-(2-methyl-6-hydroxymethylphenyl)-N- and N-(2-hydroxymethyl-6-methylphenyl)-N-(methoxyacetyl)-alanine methyl ester and N-(3-hydroxy- and N-(5-hydroxy-2,6-dimethyl-phenyl)-N-(methoxyacetyl)-alanine methyl ester according to their mass-spectral and nuclear magnetic resonance-spectral characteristics. Benzylic hydroxylation of the methyl side chains and/or aromatic hydroxylation appeared to be the major reactions involved in the metabolism of metalaxyl.  相似文献   

16.
Three Types of Sensitivity to Metalaxyl in Plasmopara viticola   总被引:2,自引:0,他引:2  
The sensitivity of Plasmopara-isolates to metalaxyl was determined by a leaf disc test. For the first time this permitted the distinction and clear characterization of not only sensitive and resistant isolates but also of isolates belonging to a third type of sensitivity, termed the reduced sensitive type.  相似文献   

17.
1.  1. It has been accepted that aquatic hyphomycetes colonising submerged leaves increase the nutritional value of leaf detritus and suggested that fungal biomass plays a greater role in the growth of shredders than leaf tissue itself. However, it is not clear what proportion of the nutritional needs of shredders is met by fungal biomass.
2.  We fed Pycnopsyche gentilis larvae with tulip poplar ( Liriodendron tulipifera ) leaf discs colonised by the aquatic hyphomycete, Anguillospora filiformis , which had been radiolabelled to quantify the contribution of fungal carbon to the growth of the shredder at different larval developmental stages. Instantaneous growth rates of larvae on this diet were also estimated.
3.  When provided with fungal-colonised leaves (14–16% fungal biomass), the third and the fifth instar larvae of P. gentilis grew at the rates of 0.061 and 0.034 day−1, respectively, but on a diet of sterile leaves, both larval instars lost weight. The incorporation rates of fungal carbon were 31.6 μg C mg−1 AFDM day−1, accounting for 100% of the daily growth rate of the third instar larvae and 8.6 μg C mg−1 AFDM day−1, accounting for 50% of the daily growth rate of the fifth instar larvae.
4.  These results suggest that leaf material colonised by A. filiformis is a high quality food resource for P. gentilis larvae, and that fungal biomass can contribute significantly to the growth of these larvae. Differences in feeding behaviour and digestive physiology may explain the significantly greater assimilation of fungal biomass by the earlier instar than the final instar. To satisfy their nutritional needs the fifth instar larvae would have to assimilate detrital mass that may have been modified by fungal exoenzymes.  相似文献   

18.
This study was undertaken to evaluate the range of 20-hydroxyecdysone (20HE) concentrations which induce cell proliferation and imaginal differentiation in lepidopteran wing discs in vitro . Wing discs were cultured in medium containing various doses of 20HE. During imaginal differentiation in vitro , wing discs were observed histologically and the number of mitosis was counted every day. Wing discs differentiated adult features in medium containing 0.02–0.2 μg/mL 20HE, and these doses also increased the number of mitosis in disc cells. Wing discs developed the same in vitro as they do in vivo . The concentration of 20HE over 0.2 μg/mL inhibited both mitosis and imaginal differentiation. Cell proliferation, cuticle deposition and tissue elongation were successively observed in vitro the same as observed in vivo . These results suggest that a moderate concentration of ecdysteroid can induce cell proliferation followed by imaginal differentiation.  相似文献   

19.
Spinach (Spinacia oleracea L.) leaf discs floating on buffer solution were treated with Kresoxim-methyl (KROM), an inhibitor of respiratory electron transport. In the leaf tissue, actual and maximal nitrate reductase (NR) activities, nitrite content and ATP levels were determined. In darkened leaf discs incubated without KROM (control) actual NR activity decreased to 20% after 6 h in the dark. Treatment with 10 μg ml−1 (corresponding to 32 μM) KROM totally prevented inactivation of NR in the dark and also diminished NR-protein degradation during prolonged darkness. Due to restricted nitrite reduction in darkened leaf tissues, nitrite accumulated in KROM-treated discs. Inhibition of respiration decreased ATP and increased AMP levels in KROM-treated discs. In illuminated leaf discs, NR was highly activated to 65%. Nevertheless, KROM-treatment caused an additional activation of NR (activation state 76%) in the light. Possible side-effects of KROM on nitrite reduction and photosynthesis were also checked in the leaf-disc system. Neither nitrite reduction nor photosynthesis were altered in KROM-treated discs. The extent of KROM-induced activation of NR was dependent on the applied concentration and on the pH of the external medium. The highest activation of NR was achieved at an external pH of 4.8, confirming previous results (Kaiser and Brendle-Behnisch, 1995, Planta 196: 1–6) that cytosolic acidification might play an important role in the modulation of NR activity. Received: 17 June 1998 / Accepted: 2 September 1998  相似文献   

20.
Aims:  To evaluate the potential of Actinoplanes campanulatus , Micromonospora chalcea and Streptomyces spiralis endophytic in cucumber roots, to promote plant growth and to protect seedlings and mature plants of cucumber from diseases caused by Pythium aphanidermatum , under greenhouse conditions.
Methods and Results:  Three endophytic isolates, out of 29, were selected through tests aimed at understanding their mechanisms of action as biocontrol agents and plant growth promoters. When applied individually or in combination, they significantly promoted plant growth and reduced damping-off and crown and root rot of cucumber. The combination of the three isolates resulted in significantly better suppression of diseases and plant growth promotion, than where the plants were exposed to individual strains.
Conclusions:  The three selected actinomycete isolates colonized cucumber roots endophytically for 8 weeks, promoted plant growth and suppressed pathogenic activities of P. aphanidermatum on seedling and mature cucumber plants.
Significance and Impact of the Study:  The results clearly show that the endophytic, glucanase-producing actinomycetes used, especially as a combined treatment, could replace metalaxyl, which is the currently recommended fungicide for Pythium diseases in the United Arab Emirates. These endophytic isolates also have the potential to perform as plant growth promoters, which is a useful attribute for crop production in nutrient impoverished soils.  相似文献   

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