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1.
Groundnut bruchid (Caryedon serratus Olivier) is a major storage insect pest that significantly lowers the quality and market acceptance of the produce. Screening for resistance against groundnut bruchid in field conditions is difficult due to the variation in environmental factors and possible occurrence of biotypes. Hence, identification of tightly linked markers or quantitative trait loci (QTLs) is needed for selection and pyramiding of resistance genes for durable resistance. A population of recombinant inbred lines derived from a cross between VG 9514 (resistant) and TAG 24 (susceptible) was screened for five component traits of bruchid resistance in 2 years. The same population was genotyped with 221 polymorphic marker loci. A genetic linkage map covering 1,796.7 cM map distance was constructed with 190 marker loci in cultivated groundnut. QTL analysis detected thirteen main QTLs for four components of bruchid resistance in nine linkage groups and 31 epistatic QTLs for total developmental period (TDP). Screening in 2 years for bruchid resistance identified two common main QTLs. The common QTL for TDP, qTDP-b08, explained 57–82 % of phenotypic variation, while the other common QTL for adult emergence, qAE2010/11-a02, explained 13–21 % of phenotypic variation. Additionally, three QTLs for TDP, adult emergence and number of holes and one QTL for pod weight loss were identified which explained 14–39 % of phenotypic variation. This is the first report on identification of multiple main and epistatic loci for bruchid resistance in groundnut.  相似文献   

2.
Genetically modified (GM) cowpea seeds expressing αAI-1, an α-amylase inhibitor from the common bean, have been shown to be immune against several bruchid species. Effective control of such pests by growing GM cowpea could promote the spread of bruchid species that are αAI-1 tolerant. Consequently, the sustainability of bruchid pest control could be increased by combining GM seeds and hymenopteran parasitoids. However, there are concerns that αAI-1 could interfere with the biological control provided by parasitoids. Here, we assessed the impact of GM cowpea seeds expressing αAI-1 on the αAI-1-tolerant bruchid Zabrotes subfasciatus and its parasitoid Dinarmus basalis. αAI-1 in cowpea seeds did not increase resistance to Z. subfasciatus or affect the mortality rate of Z. subfasciatus larvae. Parasitism of Z. subfasciatus by D. basalis and fitness of D. basalis offspring were not affected by the presence of αAI-1. Thus, αAI-1-expressing cowpeas and parasitoids should be compatible for the control of bruchid pests.  相似文献   

3.
Bruchid, Callosobruchus chinensis (L.) is an important pest of Vigna radiata during storage. RFLP and PCR based markers identified, linked with bruchid resistance gene in wild accession of greengram (V. radiata var. Sublobata) either collected from Madagaskar or Australia. Whether these markers will be useful for marker assisted introgression of bruchid resistance gene from the Indian accession into the existing cultivars are not known. Here, we employed two STS based markers which were found earlier, to be linked with bruchid resistance gene in Australian accession ACC41. Only one primer pair, STSbr1 showed polymorphism among Indian Sublobata accession (Sub2) and other twelve green gram cultivars. Analysis of 113 segregating lines (F6) of a cross between a popular cultivar of West Bengal, B1 and Sub2 showed a cent percent co-segregation of resistant locus with the polymorphic fragment. STSbr1 behave as a dominant marker among Indian genotypes although it has been shown earlier a co-dominant banding pattern between ACC41 and other Australian Susceptible cultivars. Other STS marker, STSbr2, does not produce any polymorphic fragment among Sub2 and 18 greengram genotypes. STSbr1 employed in screening of 50 green gram accessions and found high efficiency in screening of bruchid resistant genotypes also. So STSbr1 will be useful for marker assisted selection and germplasm screening for development of bruchid resistant greengram.  相似文献   

4.
Bruchid larvae cause major losses in grain legume crops throughout the world. Some bruchid species, such as the cowpea weevil, are pests that damage stored seeds. Plants synthesize a variety of molecules, including proteinaceous proteinase inhibitors, to defend themselves against attack by insects. In this work, a trypsin inhibitor (DMTI-II) isolated from Dimorphandra mollis seeds was tested for anti-insect activity against Callosobruchus maculatus larvae. The inhibitor produced ca. 67% mortality to this bruchid when incorporated into an artificial diet at a level of 1%. The doses necessary to cause 50% mortality (LD50) and to reduce weight by 50% (ED50) for DMTI-II were ca. 0.50% and 0.60%, respectively. The action of DMTI-II on C. maculatus larvae may involve the inhibition of trypsin-like activity of larval midgut extracts, the absence of digestion by midgut preparations or with a mixture of pepsin and papain, and its association with a chitin column and chitinous structure in the midgut of this insect.  相似文献   

5.
1. A globulin fraction prepared from the meal of Callosobruchus maculatus-resistant cowpea (Vigna wiguiculata) seeds was shown to be detrimental to this bruchid when incorporated in artificial seeds.2. The performance of C. maculatus was also shown to be strongly hindered by vicilins from resistant seeds when these storage proteins were incorporated in artificial seeds at the level of 2%.3. The purified vicilins from seeds of both resistant and susceptible cowpea varieties were shown to have the same SDS-PAGE pattern but different mobilities in non-denaturing polyacrylamide gel electrophoresis.4. These results and previous ones obtained by us (Silva and Xavier-Filho, 1991; Sales et al., 1992) strongly suggest that the resistance of cowpea seeds from the cultivar TVu 2027 and from others bred from it is associated with the presence of vicilin molecules which are refractory to digestion by bruchid midgut proteinases.  相似文献   

6.
Bruchid resistance, controlled by a single dominant gene (Br) in a wild mungbean accession (TC1966), has been incorporated into cultivated mungbean (Vigna radiata). The resistance gene simultaneously confers inhibitory activity against the bean bug, Riptortus clavatus Thunberg (Hemiptera: Alydidae). The resultant isogenic line (BC20 generation) was characterized by the presence of a group of novel cyclopeptide alkaloids, called vignatic acids. A linkage map was constructed for Br and the vignatic acid gene (Va) using restriction fragment length polymorphism (RFLP) markers and a segregating BC20F2 population. By screening resistant and susceptible parental lines with 479 primers, eight randomly amplified polymorphic DNA (RAPD) markers linked to Br were identified and cloned for use as RFLP probes. All eight RAPD-based markers, one mungbean, and four common bean genomic clones were effectively integrated around Br within a 3.7-cM interval. Br was mapped to a 0.7-cM segment between a cluster consisting of six markers and a common bean RFLP marker, Bng110. The six markers are closest to the bruchid resistance gene, approximately 0.2?cM away. The vignatic acid gene, Va, cosegregated with bruchid resistance. However, one individual was identified in the BC20F2 population that retained vignatic acids in spite of its bruchid susceptibility. Consequently, Va was mapped to a single locus at the same position as the cluster of markers and 0.2?cM away from Br. These results suggest that the vignatic acids are not the principal factors responsible for bruchid resistance in V. radiata but will facilitate the use of map-based cloning strategies to isolate the Br gene.  相似文献   

7.
Aliquots of powdered dried leaves of Tephrosia vogelii (Leguminosae – Papilionoideae) were extracted with hexane, acetone and ethanol. The extracts were tested for their potential to protect stored legume seeds from damage by the bruchids Acanthoscelides obtectus, Callosobruchus maculatus and Callosobruchus chinensis (Coleoptera – Bruchidae). While the acetone and ethanol extracts were ineffective except for a significant but low effect on A. obtectus, the hexane extract had a relatively high efficacy, providing similar control of all three bruchid species. This effect was manifested in slowly‐reduced adult survival within 7 days and in reduced numbers of eggs laid and of F1 progeny produced, resulting in seed damage averaging 7.1% compared to 99.8% in the untreated controls after one generation. These findings are discussed in the context of previously identified constituents of T. vogelii, the rotenoids rotenone, tephrosin and deguelin.  相似文献   

8.
Bruchid beetles infest various seeds. The seed coat is the first protective barrier against bruchid infestation. Although non-host seed coats often impair the oviposition, eclosion and survival of the bruchid Callosobruchus maculatus larvae, morphological and biochemical aspects of this phenomenon remain unclear. Here we show that Phaseolus vulgaris (non-host) seed coat reduced C. maculatus female oviposition about 48%, increased 83% the seed penetration time, reduced larval mass and survival about 62 % and 40 % respectively. Interestingly, we found no visible effect on the major events of insect embryogenesis, namely the formation of the cellular blastoderm, germ band extension/retraction, embryo segmentation, appendage formation and dorsal closure. Larvae fed on P. vulgaris seed coat have greater FITC fluorescence signal in the midgut than in the feces, as opposed to what is observed in control larvae fed on Vigna unguiculata. Cysteine protease, α-amylase and α-glucosidase activities were reduced in larvae fed on P. vulgaris natural seed coat. Taken together, our results suggest that although P. vulgaris seed coat does not interfere with C. maculatus embryonic development, food digestion was clearly compromised, impacting larval fitness (e.g. body mass and survivability).  相似文献   

9.
Rotenone, a naturally occurring ketone, is widely employed for the management of invasive fish species. The use of rotenone poses serious challenges to conservation practitioners due to its impacts on non-target organisms including amphibians and macroinvertebrates. Using laboratory studies, we investigated the effects of different rotenone concentrations (0, 12.5, 25, 37.5, 50, 100 μg L-1) on selected invertebrate groups; Aeshnidae, Belostomatids, Decapods, Ephemeroptera, Pulmonata and zooplankton over a period of 18 hours. Based on field observations and body size, we hypothesized that Ephemeropterans and zooplankton would be more susceptible to rotenone than Decapods, Belostomatids and snails. Experimental results supported this hypothesis and mortality and behaviour effects varied considerably between taxa, ranging from no effect (crab Potamonuates sidneyi) to 100% mortality (Daphnia pulex and Paradiaptomus lamellatus). Planktonic invertebrates were particularly sensitive to rotenone even at very low concentrations. Future research should investigate the recovery time of invertebrate communities after the application of rotenone and conduct field assessments assessing the longer term effects of rotenone exposure on the population dynamics of those less sensitive organisms.  相似文献   

10.
Rotenone is a widely used pesticide that induces Parkinson’s disease-like symptoms in rats and death of dopaminergic neurons in culture. Although rotenone is a potent inhibitor of complex I of the mitochondrial electron transport chain, it can induce death of dopaminergic neurons independently of complex I inhibition. Here we describe effects of rotenone in the fission yeast, Schizosaccharomyces pombe, which lacks complex I and carries out rotenone-insensitive cellular respiration. We show that rotenone induces generation of reactive oxygen species (ROS) as well as fragmentation of mitochondrial networks in treated S. pombe cells. While rotenone is only modestly inhibitory to growth of wild type S. pombe cells, it is strongly inhibitory to growth of mutants lacking the ERK-type MAP kinase, Pmk1, or protein kinase A (PKA). In contrast, cells lacking the p38 MAP kinase, Spc1, exhibit modest resistance to rotenone. Consistent with these findings, we provide evidence that Pmk1 and PKA, but not Spc1, are required for clearance of ROS in rotenone treated S. pombe cells. Our results demonstrate the usefulness of S. pombe for elucidating complex I-independent molecular targets of rotenone as well as mechanisms conferring resistance to the toxin.  相似文献   

11.
A simple and reproducible method for the treatment of Arabidopsis thaliana leaves with rotenone is presented. Rosette leaves were incubated with rotenone and Triton X-100 for at least 15 h. Treated leaves showed increased expression of COX19 and BCS1a, 2 genes known to be induced in Arabidopsis cell cultures after rotenone treatment. Moreover, rotenone/Triton X-100 incubated leaves presented an inhibition of oxygen uptake. The simplicity of the procedure shows this methodology is useful for studying the effect of the addition of rotenone to a photosynthetic tissue in situ.  相似文献   

12.
The deployment in common beans (Phaseolus vulgaris L.) of arcelin-based bruchid resistance could help reduce post-harvest storage losses to the Mexican bean weevil [(Zabrotes subfasciatus (Boheman)]. Arcelin is a member of the arcelin-phytohemagglutinin-α-amylase inhibitor (APA) family of seed proteins, which has been extensively studied but not widely used in bean breeding programs. The purpose of this study was to evaluate microsatellite markers for genetic analysis of arcelin-based bruchid resistance and to determine the orientation of markers and the rate of recombination around the APA locus. A total of 10 previously developed microsatellites and 22 newly developed markers based on a sequenced BAC from the APA locus were screened for polymorphism and of these 15 were mapped with an F2 population of 157 individuals resulting from a susceptible × resistant cross of SEQ1006 × RAZ106 that segregated for both the arcelin 1 allele and resistance to the bruchid, Z. subfasciatus. Microsatellites derived from APA gene sequences were linked within 0.8 cM of each other and were placed relative to the rest of the b04 linkage group. In a comparison of genetic to physical distance on the BAC sequence, recombination was found to be moderate with a ratio of 125 kb/cM, but repressed within the APA locus itself. Several markers were predicted to be very effective for genetic studies or marker-assisted selection, based on their significant associations with bruchid resistance and on low adult insect emergence and positions flanking the arcelin and phytohemagglutinin genes.  相似文献   

13.

Purpose

Carbon monoxide (CO) is an accepted cytoprotective molecule. The extent and mechanisms of protection in neuronal systems have not been well studied. We hypothesized that delivery of CO via a novel releasing molecule (CORM) would impart neuroprotection in vivo against ischemia-reperfusion injury (IRI)-induced apoptosis of retinal ganglion cells (RGC) and in vitro of neuronal SH-SY5Y-cells via activation of soluble guanylate-cyclase (sGC).

Methods

To mimic ischemic respiratory arrest, SH-SY5Y-cells were incubated with rotenone (100 nmol/L, 4 h) ± CORM ALF186 (10–100 µmol/L) or inactivated ALF186 lacking the potential of releasing CO. Apoptosis and reactive oxygen species (ROS) production were analyzed using flow-cytometry (Annexin V, mitochondrial membrane potential, CM-H2DCFDA) and Western blot (Caspase-3). The impact of ALF186± respiratory arrest on cell signaling was assessed by measuring expression of nitric oxide synthase (NOS) and soluble guanylate-cyclase (sGC) and by analyzing cellular cGMP levels. The effect of ALF186 (10 mg/kg iv) on retinal IRI in Sprague-Dawley rats was assessed by measuring densities of fluorogold-labeled RGC after IRI and by analysis of apoptosis-related genes in retinal tissue.

Results

ALF186 but not inactivated ALF186 inhibited rotenone-induced apoptosis (Annexin V positive cells: 25±2% rotenone vs. 14±1% ALF186+rotenone, p<0.001; relative mitochondrial membrane potential: 17±4% rotenone vs. 55±3% ALF186+rotenone, p<0.05). ALF186 increased cellular cGMP levels (33±5 nmol/L vs. 23±3 nmol/L; p<0.05) and sGC expression. sGC-inhibition attenuated ALF186-mediated protection (relative mitochondrial membrane potential: 55±3% ALF186+rotenone vs. 20±1% ODQ+ALF186+rotenone, p<0.05). ALF186 protected RGC in vivo (IRI 1255±327 RGC/mm2 vs. ALF186+IRI 2036±83; p<0.05) while sGC inhibition abolished the protective effects of ALF186 (ALF186+IRI 2036±83 RGC/mm2 vs. NS-2028+ALF186+IRI 1263±170, p<0.05).

Conclusions

The CORM ALF186 inhibits IRI-induced neuronal cell death via activation of sGC and may be a useful treatment option for acute ischemic insults to the retina and the brain.  相似文献   

14.
Vicilins are seed proteins, and they constitute 70–80% of the total protein in leguminous seeds; with amolecular mass between 150 and 190 kDa, they are composed of subunits without disulfide bridges, with high affinity for chitin-binding. They are also associated with seed defense against insect pests. The chitin-binding vicilin from Anadenanthera colubrina seeds was purified by ammonium sulfate, followed by affinity chromatography on a chitin column, molecular exclusion on Superdex 75 Tricorn in FPLC system and Phenomenex C8 chromatography in HPLC system. The A. colubrina vicilin, named AcV, is a tetrameric glycoprotein composed of 1.55% carbohydrates and molecular weight determined by SDS-PAGE, consisting of 70, 73, 43 and 41 kDa. The AcV homogeneity was confirmed in native PAGE, where it was observed to be a unique band with slow mobility in this gel, with approximately 230 kDa. AcV added to the Callosobruchus maculatus diet in the bioassays resulted in a strong effect on adult emergence (ED50 of 0.096%), and in larvae caused a marked reduction in mass (WD50 of 0.32%) and lethality (LD50 of 0.33%) (w:w). The digestibility of AcV was evaluated in vitro with the digestive enzymes of larvae of C. maculatus of fourth instar, showing major fragments of 10 and 30 kDa. AcV showed reactivity against the anti-EvV antibody from Erythrina velutina vicilin. The deleterious effects of AcV are likely to be associated with the chitin-binding fragments generated by proteolysis in the bruchid gut, similarly to that found for vicilins from other leguminous plant species, Enterolobium contortisiliquum and Vigna unguiculata. AcV might be a candidate protein for a possible bioinsecticidal control of the bruchid weevil, C. maculatus.  相似文献   

15.
An impact assessment study was undertaken to determine seed predation rates by two bruchid beetles, Bruchidus sahlbergi Schilsky and Caryedon serratus Olivier, on the invasive shrub Acacia nilotica (L.) Willd. ex Del. The former bruchid was released as a biological control agent for A. nilotica, whereas the latter is naturalized in Australia. We attempted to determine the dynamics and magnitude of bruchid predation, following a number of differing reports of their effectiveness. To investigate the importance of seed availability on bruchid numbers, we compared seed death in high- and low-seed-density habitats, both before and after pods had dropped from tree branches. Bruchid predation was initially low (<2%) in pods on tree branches in both habitats, but increased abruptly to 12% in riparian and 32% in nonriparian exclosures by the third collection date. Bruchid numbers then crashed to below 2% in both habitats, when pods dropped to the ground. B. sahlbergi predation later increased markedly (up to 65%) again at high-pod-density sites. No further increase in predation took place beyond this level. Seed predation by C. serratus was found to be minimal (<5% of seeds infested) throughout the monitoring period. We suggest that seed predation by the bruchids, prior to pod drop and removal by cattle, is insufficient to cause major impacts on A. nilotica populations. Bruchid seed damage is likely to be significant only in situations with low cattle numbers, where cattle cannot remove the majority of fallen seed pods or where cattle are excluded. Exclusion of cattle from some areas within A. nilotica-invaded landscapes to potentially increase bruchid effectiveness is proposed. The demographic impact of optimal A. nilotica seed losses is discussed.  相似文献   

16.
Rotenone has been shown to induce many parkinsonian features and has been widely used in chemical models of Parkinson’s disease (PD). Its use is closely associated with α-synuclein (α-syn) phosphorylation both in vivo and in vitro. However, the mechanisms whereby rotenone regulates α-syn phosphorylation remain unknown. Protein phosphatase 2A (PP2A) has been shown to play an important role in α-syn dephosphorylation. We therefore investigated if rotenone caused α-syn phosphorylation by down-regulation of PP2A activity in mice. Rotenone increased the phosphorylation of α-syn at Ser129, consistent with the inhibition of PP2A activity by increased phosphorylation of tyrosine 307 at the catalytic subunit of PP2A (pTyr307 PP2Ac). We further explored the interactions among rotenone, PP2A, and α-syn in SK-N-SH cells and primary rat cortical neurons. Rotenone inhibited PP2A activity via phosphorylation of PP2Ac at Tyr307. The reduction in PP2A activity and rotenone cytotoxicity were reversed by treatment with the PP2A agonist, C2 ceramide, and the Src kinase inhibitor, SKI606. Immunoprecipitation experiments showed that rotenone induced an increase in calmodulin–Src complex in SK-N-SH cells, thus activating Src kinase, which in turn phosphorylated PP2A at Tyr307 and inhibited its activity. C2 ceramide and SKI606 significantly reversed the rotenone-induced phosphorylation and aggregation of α-syn by increasing PP2A activity. These results demonstrate that rotenone-reduced PP2A activity via Src kinase is involved in the phosphorylation of α-syn. These findings clarify the novel mechanisms whereby rotenone can induce PD.  相似文献   

17.
Metformin is widely used as a hypoglycemic agent for the treatment of type 2 diabetes. Both metformin and rotenone, an inhibitor of respiratory chain complex I, suppressed glucose-6-phosphatase (G6pc), a rate limiting enzyme of liver glucose production, mRNA expression in a rat hepatoma cell line accompanied by a reduction of intracellular ATP concentration and an activation of AMP-activated protein kinase (AMPK). When yeast NADH-quinone oxidoreductase 1 (NDI1) gene was introduced into the cells, neither inhibition of ATP synthesis nor activation of AMPK was induced by these agents. Interestingly, in contrast to rotenone treatment, G6pc mRNA down-regulation was observed in the NDI1 expressing cells after metformin treatment. Since NDI1 can functionally complement the complex I under the presence of metformin or rotenone, our results indicate that metformin induces down-regulation of G6pc expression through an inhibition of complex I and an activation of AMPK-independent mechanism.  相似文献   

18.
Environmental rotenone is associated with Parkinson's disease due to its inhibitory property to the complex I of mitochondrial respiration chain. Although environmental pollution has been postulated as a causal factor for the increasing prevalence of obesity, the role of rotenone in the pathogenesis of obesity has not been studied. We employed muscle-derived cell C2C12 as a model and shotgun lipidomics as a tool for lipid analysis and found that treatment with rotenone led to the profound deposition of intracellular triacylglycerol (TAG) in a time- and dose-dependent fashion. The TAG deposition resulted from complex I inhibition. Further studies revealed that rotenone induced mitochondrial stress as shown by decreased mitochondrial oxygen consumption rate, increased NADH/NAD+ ratio (i.e., reductive stress) and mitochondrial metabolites. We demonstrated that rotenone activated fatty acid de novo synthesis and TAG synthesis and ultimately resulted in intracellular TAG deposition. These studies suggested that increased mitochondrial stresses might be an underlying mechanism responsible for TAG accumulation manifest in obesity.  相似文献   

19.
Abstract The intensity of seed predation the invasive tropical legume Leucaena leucocephala by the bruchid Acanthoscelides macropthalmus was investigated in south‐eastern Queensland, Australia. The number of seeds damaged by A. macropthalmus as a proportion of total seeds available was found to increase the longer the pods remained on the tree. Seed predation ranged from a mean of 10.75% of seeds on pods that remained on the plant for 1 month and increased to 53.54% for pods that remained of the plant for 4 months. The low bruchid populations at high pod densities results in ‘predator satiation’. However, pods dehisce over time and the proportion of pods available over time to the bruchid correspondingly declines. By the time bruchid densities build up, most pods have dehisced and the seeds consequently escape predation. As a result the number of seeds lost to bruchid damage increases only marginally over time. Despite the levels of seed predation observed over the course of the study, the number of seeds in the soil seedbank almost doubled over time increasing from 8.5 seeds m?3 to 15.5 seeds m?3 over a 4‐month period. Levels of seed predation and addition of seeds to the soil seedbank were not correlated. The taxonomic (subspecies) status and apparency of host plants as measured by plant and patch traits (average plant height, density of podding plants and patch size) did not influence levels of seed predation. Pre‐dispersal seed predation studies need to take into account the pod/seed retention behaviour of the plant. The ability of the bruchid to regulate the invasiveness of Leucaena through influencing its demography is likely to be diminished if the insect populations cannot increase rapidly enough to use the seeds before pod dehiscence.  相似文献   

20.
为了解粉垄“145”模式在新植蔗上的应用效果,解析其生理生态基础,该研究以桂柳05136为材料,设置常规耕作(CK)和粉垄“145”模式(FL145)2个处理,通过大田试验研究粉垄“145”模式对土壤性质以及新植蔗农艺性状、光合特性和产量品质的影响,并分析其经济效益。结果表明:(1)与CK相比,FL145的0~20 cm、20~40 cm根区,土壤容重显著降低1.25%~5.98%,土壤孔隙度显著提高1.08%~4.77%,土壤含水量显著提高1.78%~8.23%。(2)FL145促进新植蔗根系生长,出苗效果和农艺性状表现良好,株高显著增加2.20%~7.86%。(3)FL145的新植蔗单株叶面积显著增大15.88%,叶绿素含量、净光合速率、气孔导度、胞间CO2浓度分别显著提高1.41%、6.84%、18.67%、10.06%,光合能力增强,单株干物质积累量显著增加9.26%。(4)收获时,FL145的新植蔗有效茎数、茎长、茎径显著增加,理论产量和实际产量分别显著提高5.07%和5.11%,蔗汁蔗糖分和锤度分别显著提高1.61%和1.50%,还原糖分显著下降12....  相似文献   

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