3-1047 January 1966 icrenccs in the Diapause Response Illoll Weevils irom the High Plains and Central Texas Significance oi this Phenomenon Revising Present Fall Insecticidal Control Programs W. L. Sterling, research associate, Perry L. Adkisson, professor, Department of Entomology TEXAS A&M UNIVERSITY _, Texas Agricultural Experiment Station E. Patterson, Director, College Station, Texas SUMMARY I Boll weevils on the High Plains of Texas enter diapause considerably earlier in the fall and in greater percentages than do those in Central Texas. This difference in response is due not only to differences in environment between the two areas, namely, earlier occurring more severe winters in the High Plains than in Central Texas, but also to a difference in the genetic makeup of the two weevil populations. The difference in genotype was found when the weevils were maintained in identical photoperiods. Short days induce diapause in both populations; however, significantly greater percentages of the High Plains weevils entered diapause under inductive photo- periods than did those from College Station or Pre~ sidio. It was concluded that the boll weevil is becoming adapted to the environment of the High Plains and a distinctive photoperiodic race of the species may have already evolved in this area. It is suggested that a new type of approach for reducing diapausing boll weevils might be particu- larly effective in the High and Rolling Plains. This approach would have two phases. Phase 1 would be directed towards killing the last reproductive genera- tion of adult females before they lay the eggs that give rise to the potentially overwintering adults. This would be accomplished with insecticidal applications made at 5-day intervals during September. Phase 2 would have as its objective the killing of any dia- pausing adults in October that may have survived earlier insecticidal treatments or that may have developed from eggs laid between applications in September. INTRODUCTION THE BOLL WEEVIL presently is expanding its geo- graphical distribution both southward and north- ward. It is suspected that the center of origin of the boll weevil was Mexico or Central America. The pest has recently entered Colombia and Venezuela to the south. Invasions have occurred to the north into the El Paso Valley and the High Plains of Texas and into the Sonora area of Northwestern Mexico and Arizona. These recent invasions should be sufficient evidence to point out the adaptability of the species to great differences in environment as found at these widely separated geographical loca- tions. The ability to diapause is evidently one of the mechanisms used by the weevil to expand its range of distribution. There may be considerable differences among widely separated insect populations in the nature of the diapause response to certain environmental stimuli such as photoperiod. Local variations in the characteristics of the diapause response may operate as important isolating mechanisms in the formation of geographical races and possibly in the development of new and distinct species. At the present time, insecticidal control programs are being directed at fall populations of boll weevils in the Presidio-El Paso Valley and the High and Rolling Plains area of Texas. These control pro- grams have as their objective the killing of the potentially overwintering populations before they leave the cotton fields for hibernating sites. The present experiments were designed to detect any basic differences in the diapause response of boll weevil populations from different geographical locations. This information is essential to the development of more efficient control programs. The knowledge of when diapause occurs in field populations and the environmental conditions which trigger the diapause is absolutely essential in determining when the insecti- cidal applications should be initiated to prevent the formation of potential overwintering populations. MATERIALS AND METHODS Source of Boll Weevils Frequent collections of adult boll weevils and infested squares were made during the fall of 1964 in cotton fields near College Station and near the junction of the High and Rolling Plains in the vicini~ ty of Afton, Texas (Dickens County). The seasonal incidence of diapause was determined in the weevils taken in these collections. In addition, for photo- periodic studies, adult weevils were collected from the two above locations and Presidio and cultured in the laboratory. The laboratory cultures were maintained on a cottonseed meal diet according to procedures described by Sterling et al. (1965). Henceforth in this paper the various populations will be designated 2 I Seasonal Incidence of as follows: College Station (CS), Afton (H A, Presidio (PD). x Handling of Weevils Since procedures for handling the weevils for each experiment, they will be presented A appropriate section of “Results.” a Determination of Diapause The diapause status of adult weevils termined according to the method outlined |§ zel and Newsom (1959). The adults were l under the microscope, the appearance of the t, was noted and size of the ovaries or testes s-f, Adults were classified as being reproductive, e mediate-diapause,” or “firm-diapause" acco ~1 the criteria furnished by Brazzel and Newsom j Photoperiod Studies All life history stages of the boll weevi; maintained in bioclimatic cabinets progra ~__ maintain selected photoperiods at 70° F. The I periods were programmed by time switches wi 3 daylight type 15 watt fluorescent lamps if cabinet. The timers were set to provide the -‘7 number of hours of light and dark per day. i RESULTS AND DISCUSSION Diapause in HP and CS Populations A. Occurrence 0f diapause in field-co adult boll weevils examined 0n date of coll Most authors who have studied diapause i’ populations of the boll weevil have report obtained by examination of adults shortly af_ insects had been collected (Brazzel and N.‘ 1959; Brazzel and Hightower, 1960; Lloy Merkyl, 1961; Beckham, 1963; Lloyd et al.,§ Mitchell and Mistric, 1965). A similar type study was made during the y“ investigation. Adult boll weevils were coll random in cotton fields near Afton (HP) and u‘ Station (CS). These insects were brought 3 laboratory, dissected, examined under a mi, L and their diapause status determined accor procedures described earlier in this report. ' Percentages of diapausing weevils were n5 the collections from both populations prior t0 September (Figure 1). Thereafter, the incid diapausing weevils gradually increased as the 1 advanced. However, the percentages of w" adults varied greatly between collection dates HP population having more variance than one. For example, in the HP collection of 15 there were only 12 percent diapausing adults.;__ days later, the HP collection contained 96 -_ i‘ diapause, an increase of 84 percent. Near of September, the percentage of diapausing g had decreased to approximately 20 percent. collection showed a similar type of variability it to such a great extreme. The extreme ' ity between successive collection dates is dif- to explain. Perhaps, during the fall, migra- of reproductive and diapausing weevils from a v field and from cotton fields to nearby over- g sites might be responsible for some of this A ce. Also, there may be inherent errors in lethod of sampling since it does not take into t the age structure of the population or dif- 1~~~ in behavior between reproductive and dia- ,;--_ weevils. he diapause curves plotted in Figure 1 clearly i‘ that the incidence of diapause after early Sep- 3.1 generally was considerably greater in the HP " in the CS population. As the season advanced t ber of diapausing weevils in both populations M d; however, the CS population lagged several (‘behind the HP weevils in this respect. It should lpected that the differences in the occurrence of use between the two populations will vary idling to seasonal differences in climate between , igh Plains and College Station. the northern hemisphere, as an insect species y» ds its area of distribution from south to north, orthern strains of the species should enter dia- _ " earlier in the season and in greater percentages those of the south. This is because the winters earlier, last longer and generally are colder an increase in latitude. i 3B. Incidence of diapause in field-collected fed bolls for 20 days prior t0 examination. use of the great differences in the percentages apausing weevils in collections made within a 1 of a few days, a second method of handling the ts was tested. By this method, adults collected _A ndom in fields near Afton (HP) and College ("on (CS) were brought to the College Station w atory and caged in an open insectary. These _ ts were fed small cotton bolls for 20 days before :3. g examined. The 20-day feeding period was ed since Brazzel et al. (1961) had suggested this jple time for an adult boll weevil to acquire all l“ characteristics of diapause, such as the accumula- fi of fat, atrophy of reproductive organs, and so V This technique should have allowed ample "e for all adults destined for diapause to attain state of “firm diapause" described by Brazzel and y some (1959) and ‘should minimize the numbers g weevils in “intermediate diapause” as defined by 1.. two authors. Thus, by this technique, differ- ; es between diapausing and reproducing individ- _ should be more clearly defined. The resulting ta then should furnish a more accurate estimate of "l9 , 0-0 HP 9 *-* CS g BU . Q 3f 5 an * a ' \ ' E ° I *o/ a w. / \.-. /\ m *_ “of ' * o * Z9 - ' /\ \ iv‘ c i: l‘ __/_l}-*-e r/ '/.\:/ i . l‘ . . . l5 l5 l5 l5 AUG lSEPT OCT . NOV Figure 1. Seasonal incidence of diapause in populations of boll weevils from the High Plains (HP) and College Station (CS). Diapause determinations were made on the date adults were collected in the field. The percentages of diapausing adults were greater on almost every date in the High Plains population than in the one from College Station. the percentages of potential overwintering weevils in the population than in the case of weevils examined on date of field collection. Diapause curves made from these data, presented in Figure 2, were considerably smoother than those reported for weevils examined on the date of collec- tion. These data indicate that approximately 20 percent of the adults in the field collections of HP weevils made in August were destined for diapause. Shortly after September 1, the percentages of diapaus- ing adults in the HP population increased steadily to a maximum in November when about 8O percent of the individuals were of the diapause type. The incidence of diapause in the CS population was small until late September. After this, the per- centages of diapausing weevils in this population increased rapidly achieving a maximum of 75 percent in mid-October. These data show even more clearly the difference in the seasonal incidence of diapause in the two popu- lations. Diapause occurred much earlier in the HP than in the CS population. Also, greater percentages of the HP adults entered diapause. A comparison of the response curves plotted in Figures l and 2 shows that greater percentages of diapausing weevils were detected in adults fed bolls for 20 days prior to examination than in those exam- ined immediately after collection. This method ap- parently furnished a better estimate of the potential diapausing population because all weevils in the collection that had been “triggered” for diapause had an ample opportunity to achieve a “firm” state of diapause before examinations were made. C. The incidence of diapause in square-reared and field collected adults. Data presented in Figures 3. 100 . 0-0 HP % w i *-* CS D é g 50 . 0- Z LLI U 4U , m o hi’ l \. * 2o. ° X’. n l n l n 1s 1s 1s 1s AUG SEPT OCT NOV Figure 2. Seasonal incidence of diapause in boll weevils from the High Plains (HP) and College Station (CS) which were fed bolls for 20 days prior to examination for diapause. Dates indicate date adults were collected from fields. The incidence of diapause was much greater in the High Plains population during August and September than in the one from College Station. l and 2 show that the percentages of diapausing weevils reported in a collection of field adults on any given date may be influenced to some degree by the method of handling insects after collection. The use of field-collected adults for studying the seasonal incidence of diapause in boll weevil populations may also have other disadvantages. First, the investigator using this method will have little or no information on the age structure of his collection. Thus, the adult collection, particularly those made early in the season, may contain a preponderance of old reproductives formed before the environment demanded any dia- pause in the population. This would bias the sample in favor of reproductive weevils and would make it difficult to detect the first formed diapausing weevils of the season since their numbers would be small. Secondly, field collections of adults do not take into account any differences in behavior between repro- ductive and diapausing weevils. Reproductive weevils seek fields having an abundance of uninfested squares or bolls for places to lay their eggs and for feeding. Diapausing weevils apparently do not seek squares or bolls for ovipositional purposes but only for food. Apparently, they do not discriminate as do reproduc- tive weevils between infested and non-infested squares and thus may not be influenced to migrate from heavily infested fields as readily as do the reproduc- tive forrns. Because of this difference in behavior, one might logically expect to collect a preponderance of reproductive weevils during late seasons in fields having an abundance of non-infested fruit. Alter- nately, heavily infested fields with few undamaged squares or bolls may be more heavily invested with weevils destined for diapause simply because the reproductive weevils have migrated to abundantly fruiting fields. Therefore, the incidence of diapaus- ing or reproductive weevils in a sample might be greatly influenced by the fruiting condition of the 4 field chosen for the sample. Results gained z such samples could lead an investigator to erron conclusions regarding the seasonal occurrence of f pause in the total boll weevil population. S samples might also be influenced by the past hist of insecticidal treatment of the field. Recently tre _ fields would have a preponderance of young add of the same generation while “iuntreated fields m contain adults of one, two or more previous gen tions. This could greatly affect the percentages; adults in “firm-diapause” in any given sample. l One method for overcoming any difficulty-f to sampling would be to collect squares infested the immature forms of the boll weevil, then hold g in an open insectary under conditions similar s’ those experienced in the field, and then collect i the adults as they emerge. By this method, one if have weevils of the same age group and would » able to eliminate any bias in the data from behavii differences between reproducing and diapa Q weevils. Also, this method would provide a m; representative of all the weevils being formed in population during a given span of time. This procedure was tested during the aw] study. The adult weevils were reared from inf, squares collected on various dates from mid-A to late-November in fields near Afton (HP) College Station (CS) The squares from both V“ tions were placed in the insectary at College s: and the adults were allowed to emerge. These dividuals were fed small bolls for 20 days before v sacrificed for diapause determinations. Y‘ Results are presented in Figures 3 and 4. _ these data are compared to results in Figure l; field-collected adults examined on date of collec, it is obvious that the incidence of diapause in sq 1 reared weevil samples was considerably greater f in samples of randomly collected field adults. l‘ samples show that the incidence of diapause in HP population in August and in the CS popular, in early September was much greater than one {f expect if only samples of field-collected adults ‘ examined. Data from square-reared weevils ap f to furnish a much better indication of the u.’ formation of the diapausing individuals than j" those collected by the other two methods. These show that in the High Plains a majority of the a emerging after September 1, 1964, were of the, pause type. Diapause occurred several days lat College Station, and it was not until late Sept == that more than 50 percent of the weevils emer‘ each date entered diapause. ; A comparison of square-reared and field-coll boll weevils showed that the incidence of dia was much greater in the square-reared weevils (F 3) . This probably was because the infested s a were collected when environmental conditions riate to “trigger” the diapause in the immature _ The field-collected boll weevils were taken i; early fall from fields which had not been Q- with insecticides and almost certainly were (‘ted with adults of one, two or three previous " tions formed at the time when environmental ions were inappropriate for diapause. 1 comparison of the curves plotted in Figure 4 a indicates the difference in reaction of the HP populations to the environmental stimuli g diapause. The onset of diapause occurred _' 50 days earlier in the season in the HP popula- fthan in the one from College Station. This ed even though the insects were held in the location (the College Station insectary) for part developmental period and all of their adult After October 1, the percentages of diapausing 51s in both populations ranged around 80 percent. data furnished clear evidence of a difference in diapause response of the two populations. The reaction of the boll weevil again shows that ‘(species spreads from south to north, the northern i lations will enter diapause earlier than those f} the south. This is as it must be since diapause is l adaptation by which insects survive the ‘er. If winter occurs earlier, the insects must r diapause earlier. Otherwise, they would not ble to survive from one growing season to the a f sona t of Daylength on Diapause Earle and Newsom (1964) reported diapause in boll weevil could be induced by daylengths, or - operiods, having daily light (L) - dark (D) peri- of LD 11:13. By the same token, diapause may juppressed by LD 13:11. This research led to the ent study to determine the photoperiodic re- _ es of boll weevil populations originally collected n near Afton (HP), Presidio (PD) and College ion (CS). Results, summarized in Figure 5, show that LD 6, LD 10:14 and LD 14:10 tended to induce dia- l e while LD 12:12 and LD 16:8 tended sup- , diapause. The incidence of diapause under g‘ t days was considerably greater in the HP than in _ CS and PD populations. The HP population wed a greater photoperiodic response than the er two populations. This difference in reaction _ests there is a difference in the genetic makeup t the three populations with regard to the photo- iodic response since all were held in identical ditions. v It logically might be suspected that boll weevils i the High Plains should enter diapause earlier in fall than those at College Station because the Sowing season in northwest Texas is considerably _.. 1> 3 I ‘i. SQQ O FIELD ‘Tt.’ \' " \ PERCENT DEAPAUSE '\ O Q3 Z O F19 6/ G3 \ \® A / ‘lifjg’ n a n c a I l | l5 NOV 15 - l5 l5 AUG SEPT OCT Figure 3. A comparison of the seasonal incidence of diapause in High Plains boll weevils reared from squares in the insectary or collected as adults in the field. Dates are for time of col- lection of infested squares or field adults. shorter. The average growing season at College Station is approximately 275 days while that at Cros- byton on the eastern edge of the High Plains averages about 206 days, or 69 days shorter than at College Station. Thus, it should be expected that the boll weevil on the High Plains would have to enter dia- pause about a month earlier than at College Station in order to avoid death because of an earlier freeze date. However, environmental differences between the two areas are only part of the story. When weevils from the two locations were reared in identical con- ditions and exposed to certain photoperiods, it was found that the HP population was significantly more responsive to short days than the CS population. In all diapause-inducing photoperiods tested, greater percentages of the HP population entered diapause than either the CS or PD populations. Thus, it 100 _ .._ HP 1-1 cs *.’"~."\. * $5 an . s \. / g I /* < /\ t “o” 60 _ 2 - ' a 4e _ ,/ LU Q- ‘k 2n, 15 1.5 {a i is ‘J AUG SEPT OCT NOV Figure 4. Seasonal incidence of diapause in adult weevils from the High Plains (HP) and College Station (CS) which were reared from squares held in the College Station insectary. Dates indicate the dates on which the squares were collected. The initial onset of diapause was approximately 30 days earlier in the High Plains population than in the one from College Station. 5 so P C o——-0HP sn *_"*Cs V‘) I 3 E 4o _ i Q +- % 30 - g X~\\\ til 20 _ \.\‘I\\\\\ 9 -‘\‘.:; l0 - l] a | 8 1U l2 l‘ 1'5 HOURS LIGHT PER 24 HouR cvcuz, 10°F Figure 5. The incidence of diapause in boll weevil populations from the High Plains (HP), College Station (CS), and Pre- sidio (PD) when maintained in photoperiods having from 8 to 16 hours of light per day. appears that the HP population does possess a differ- ent genotype than the other two groups of boll weevils. Apparently the HP population, through the process of natural selection, is becoming adapted to the environmental conditions found on the High and Rolling Plains of Texas. This adaptation has permitted the dispersal and survival of the boll weevil into cotton areas previously uninhabited by the pest. Importance of the Present Results in Improving the Efficiency of Diapause Boll Weevil Control Programs A review of the development of the diapause boll weevil control program presently being conducted in the High and Rolling Plains of Texas is a necessary prerequisite to a discussion of methods for improving the efficiency of said program. The first boll weevil invasion of any serious consequence in the High Plains occurred in 1962. First evidence of successful overwintering of the pest in this area was obtained during the winter of 1963-64. As a result of this threat, the producers of the area banded together, raised funds and requested that a diapause boll weevil control program be initiated. This program was to prevent the westward migration of the boll weevil into uninfested cotton areas. The control program was initiated in September 1964. This program was based on the premise by Brazzel (1961) that four applications of an effective insecticide made at l0 to 14-day intervals during the fall months would materially reduce or possibly eradi- cate overwintering boll weevil populations. This was provided that the last insecticidal application was timed to occur just prior to frost or to the chemical or mechanical destruction of food and breeding sites of the weevil. Accordingly, a four-application pro- gram was initiated on September 16, 1964. It soon became apparent that, because of cultural practices and environmental conditions in the control area, 6 seasonal onset of diapause in boll weevil populati .\ four applications would not extend control until. nonnal frost date. Therefore, two additional secticidal applications were made to the most hea infested part of the control zone. Lack of f _ prevented the additional treatment of the entire -' trol zone. ‘ Adkisson et al. (1965) evaluated the 1964 gram and found that althoughlisubstantial reduct‘ in overwintering populations had been obtain great many weevils survived the applications * successfully overwintered. These populations A of such size that serious crop damage in the c y‘ zone during 1965 was prevented only by commu‘ wide control efforts carried out from June to Sep ber (Rummel and Adkisson, 1965). These res showed the four- to six-application program H‘, soley at killing diapausing boll weevils before t leave cotton fields for overwintering sites was x adequate to obtain the desired population reduc ' A more efficient method of control was needed l, 1965. i Earlier, Knipling (1963) had advanced the - ment that a control program designed to kill the {j reproductive generation of boll weevils would considerably more effective in reducing overwin’ “ ing populations than a four-application sched aimed at killing diapausing adults before they ' cotton fields. However, in 1963, knowledge of _ was so limited that Knipling’s hypothesis could be adequately tested. Present results, particularly those presented; Figures 3 and 4 for square-reared weevils, furnish information needed to implement Knipling’s ‘o; These data show remarkably smooth curves for seasonal onset of diapause in the boll weevil pop- tions in the High Plains. Under conditions of l A the first diapausing boll weevils of the season d oped from eggs laid from mid-August to early Sept ber. The last reproductive generation, for the "5; part, occurred in September. The September fe f" produced the majority of the eggs which gave rise: the diapausing, or overwintering, adults. This formation provided the basis for revising the diapa f‘ control program used in 1965. f‘ The 1965 program was a two-phase pr g Phase 1 was aimed at controlling the last reprodu g generation of boll weevils by using three applicati of insecticide. These applications, made at g intervals beginning September 7, 1965, were desi to kill newly emerging adult females before _ t; could mate and lay eggs. The basic premise these applications was to have an “egg-free” pe during September when environmental conditi, are such as to cause the first boll weevils to diapa Phase 2 consisted of four applications of ins cide made at 10- to 14-day intervals beginning at _g letion of phase l. These applications were ‘ ed to (1) kill any adult weevils that may have {d from eggs laid before September 7 and (2) ny adults that may have survived the September 1' ‘cations. These four applications were scheduled “'1 any diapausing weevils that might have devel- l} in the control zone before they could leave the fields for overwintering sites. ' The combined effects of the two phased program, flculated by Knipling (1963), should be as fol- ‘i provided that the insecticide will kill at least ' rcent of the adults. V‘ 1. If 90 percent of the adults of the last repro- ductive generation are killed, there should be a - concomittant decrease in egg production of 90 ‘ percent. This means that for each 100 diapaus- .ing weevils that would have developed if there f had been no control, only l0 will be formed. This provides a 90 percent reduction in the potential overwintering population. 2. If the phase 2 treatments also kill 90 percent j of the adults (which by this time will be mostly of the diapausing type), for each 100 potentially f‘ overwintering weevils that would have formed if no control had been practiced, only one sur- vives to leave cotton for overwintering sites. The net effect of the two treatment schedules 'uld be to reduce the population by 99 percent. evils t0 survive the new program than the original This degree of reduction should be particularly i i, astating to boll weevil populations in the High F: Rolling Plains since the winters generally are so ere as to cause additional mortality in the popula- n of 90 percent or more. The combined effect of =~ diapause treatments and winter mortality should __ to reduce the boll weevil population to very small i bers. g _e importantly, there should be nine times fewer LITERATURE CITED Adkisson, P. L., J. W. Davis, W. L. Owen and D. R. Rummel. 1965. Evaluation of the 1964 diapause boll weevil control program on the High Plains of Texas. Tex. Agric. Expt. Sta. Dept. of Entomol. Tech Rpt. No. 1. Beckham, C. M. 1963. Seasonal occurrence of hibernation in the boll weevil. Ga. Agr. Exp. Sta. Mimeo. Ser. N. S. 164. Brazzel, J. R. 1961. Destruction of diapause boll weevils as a means of boll weevil control. Tex. Agric. Expt. Sta. Misc. Publ. 511. Brazzel, J. R. and L. D. Newsom. 1959. Diapause in Anthonomus grandis Boh. J. Econ.Entomol. 50: 603-611. Brazzel, J. R. and B. G. Hightower. 1960. A seasonal study of diapause, reproductive activity, and seasonal tolerance to insecticides in the boll weevil. J. Econ. Entomol. 53:41-46. Brazzel, J. R., T. B. Davich and L. D. Harris. 1961. A new approach to boll weevil control. J. Econ. Entomol. 54:723-730. Earle, N. W., and L. D. Newsom. 1964. Initiation of diapause in the boll weevil. J. Ins. Physiol. 10:131-139. Knipling, E. F. 1963. An appraisal of the relative merits of insecticidal control directed against reproducing versus diapausing boll weevils in efforts to develop eradication procedures. A letter dated January 28, 1963, addressed to members of the Cotton Insects Research Branch, Ent. Res. Div., Agr. Res. Ser., U. S. Dept. Agric. Lloyd, E. P., and M. E. Merkyl. 1961. Seasonal occurrence of diapause in the boll weevil in Mississippi. J. Econ. Entomol. 54:l214-1216. Lloyd, E. P., M. L. Laster and M. E. Merkyl. 1964. A field study of diapause, diapause control, and population dynamics of the boll weevil. J. Econ. Entomol. 57:433-436. Mitchell, E. R., and W. J. Mistric, Jr. 1965. Seasonal occurrence of diapause and hibernation of the boll weevil in North Carolina. J. Econ. Entomol. 58:309-312. Rummel, D. R. and P. L. Adkisson. 1965. Unpublished data. Texas AXcM Univ., College Station. Sterling, W. L., S. G. Wellso, P. L. Adkisson and H. W. Dorough. 1965. A cottonseed meal diet for rearing the boll weevil. J. Econ. Entomol. 58:867-869. ACKNOWLEDGMENTS This research was conducted in cooperation with the Entomology Research Division, USDA, under a memorandum of agreement 12-14-100-5607 (33). Texan 56M Univereity Texan’ Agricultural Eacperimeut Station College Station. 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