Speciation and adaptive radiation in the fig wasp , Parasitodiplagaster ( : ) in Panama George O. POINAR,Jr. and Edward A. HERRE Department of Entomological Sciences, University of California, Berkeley, CA 94720, USA and Snzithsonian Tropical Research Institute,Apartado 2072, Balboa, Republic of Panama,

SUMMARY

The following species of the hereby redefined Parasitodiplogaster are described from fruits of Ficus spp. in Panama : P. citrinema n. sp. from F. citrifolia P. Miller, P. duganema n. sp. from F. dugandii Standl., P. mminema n. sp. from F. maxima P. Miller, P. nymphanema n. sp. frorn E nymphaeifolia L., P. obtusinema n. sp. frorn E obtusifolia HBK, P. paranema n. sp. from F. paraensis (Miq.) Miq., P. pertanema n. sp. from F. pertusa L. f., P. popenema n. sp. from E popenoei Standl., P. trigonema n. sp. from E trigonata L. and P. yoponema n. sp. from E yoponensis Desv. Al1 of the above species, which are presumed to have evolved from a free-living diplogasterid stem species, represent alloparric species which are carriedby specific figwasp species to specific fig fruitspecies. The diverse morphological characters (especially those ofthe alimentary tract) exhibited among these ten new species of fig wasp represent examplesof selective adaptation to physical and nutritional conditions foundin the fruits (sycones) of the different fig species.

&SUME

Spéciation et divergence adaptative chez le nématode de la guêpe du figuierParasitodiplogaster (Rhabditida :Diplogasteridae) au Panama Le genre Parasitodiplogaster est redéfini et les espèces nouvelles suivantes, provenant de fruits de Ficus spp. au Panama, sont décrites :P. citrinema n. sp. surE citrifolia P. Miller, P. duganema n. sp. surE dugandii Standi., P. maxinema n. sp.sur F. maxima, P. Miller, P. nymphanenza n. sp. sur E nymphaeifolia L., P. obtusinema n. sp. sur F. obtusifolia HBK, P. paranema n. sp. sur F. paraensis (Miq.) Miq., P. pertanema n. sp. sur E pertusa L. f., P. popenenza n. sp. sur F. popenoei Standl., P. trigonema n. sp. sur F. trigonata L. et P. yoponema n. sp. surF. yoponensis Desv. Présumées avoir évoluéa partir d'unrameau de Diplogastérides libres, elles forment un groupe d'espèces alloparriques, transportées par des guêpes spécifiquessur des fruits de figuiers spécifiques. Les variations des caractères morphologiques présentes chez ces dix espèces (en particulier ceux liés au tractus digestif) représentent un exemple d'adaptation sélective aux conditions physiques et alimentaires des fruits (des sycones) des différentes espèces de figuier.

The genus Parasitodiplogaster Poinar, 1979, was paraensis (Miq.)Miq., E pertusa L. f., E popenoei described as a parasite of the fig wasp Elisabethiella Standl., E trigonata L. and E yoponensis Desv. by EAH stuckenbergi Grandi (Agaonidae : Hymenoptera) from in the vicinity of the Panama Canal, Republic of Pa- the fruits of Ficus burkei (Miq.) Miq. in Rhodesia. nama. The figs were placed in 70 % alcohol and sent to Several years ago, one of us (EAH) discovered ne- GOP, Jr. for determination. The preserved fruits were matodes in thefruits of different Ficus species in cut open and adult nematodes were removed from the Panama. These nematodes al1 belong to the genus Pa- cavity and processed to glycerin for taxonomic studies. rasitodiplogaster, which is the first record of this group In some cases, juvenile nematodes were also removed in the New World. The present study redefines the from adult fig wasps found inside the fruits. genus Parasitodiplogaster and describes ten new species The anterior pharynx was measured from the tip of from Panamanian fig fruits. the head to theline separatingthe anterior and posterior pharynx; the posterior pharynx was measured from the line separating the anterior and posterior pharynxto the base of the posterior pharynx. Specific names of the Materials and methods nematodes are based on thespecific names of the Ficus spp. in which the nematodes occurred. In illustrations Figfruits were collected from wild trees of Ficus of the pharyngeal regionof the variousParasitodiplogas- citrifolia P. Miller, E dugandii Standl., E maxima P. ter species, the pattern of vertical lines represents strong Miller, E nymnphaez$olia L., E obtusifolia HBK, E musculature,broken vertical linesrepresents weak

Revue Nématol. 14 (3) :361-374 (1991) 361 G. O. Poinar, Jr. & E. A. Herre musculature and fine wavy lines represents glandular Parasitodiplogaster citrineman. sp. tissue. (Figs lB, 3C) MEASUREMENTS Parasitodiplogaster Poinar, 1979 Adttlts :See Tables 1 and 2. DIAGNOSIS(emended) Diplogasteridae(Micoktzky, 1922) Steiner, 1929. DESCRIPTION Adults variable in size, ranging from 0.5 to 4.0 mm in Females :Medium to large nematodes with a well- length; cuticle smooth or with faint annulations, often developed stoma and very muscular anterior pharynx swollen aroundvulva; six lipspartly or completely wi& inconspicuous valve plates at base; base of anterior fused; amphids pore-like, on lateral lips; stomavariable, , pharynxdistinctly wider than proceedingposterior from reduced -to distinct;glottoid apparatus incon- pharynx; posterior pharynx short,also muscular; baseof spicuous or lacking; pharynx composed of an anterior posterior pharynx withvalve area; pharyngeal-intestinal portion containing faint to distinct crescentvalve plates valve well-developed; excretory pore opposite posterior near the base and a basal portion sometimes terminating pharynx;intestinal lumen distinct, wide, filled with in faintcrescent valve plates; ovaries paired, tips usually viscous material and containing both bacterial rodsand reflexed; four vaginal glands (variable in size) surround- cocci; vaginal glands large, cuticle expanded in region ing vagina; testis single, tip outstretched or reflexed; of vulva, especially on ventral side; tail bearing a fine spicules paired, separate, gubernaculum present, bursa mucron at tip. absent;pre-and post-cloaca1 genitalpapillae present; found in fig fruits (Ficus spp.) and fig wasps (Agaoni- Males : Generalfeatures same as describedunder dae : Hymenoptera). females:, testis outstretched; spicules short and wide;

Table 1 Morphometric data for femalesof Parasitodiplogaster species.

Species L Body Stoma Stama L. ant.pharynx Past. L. post. Ant. end Ant. end V L Width L Width width width L pharynxpharynx ht, pharynx to E. P. to N. R. tail tail egg egg (Pm)(Pm) (Pm) (Pm) Cm) (Pd (Pm) (Pm) (Pm) (Pm) (Pd (Pm) citrinelna n. sp. 1950 98 5 797 141 3'6 216 177 5032 9160 30 (N = 3) 1 800-2 040 45-120 5-8 75-81138-144 .52-.59 174-180 45-53 88-93 57-6330-36 dugonefnan. sp, 2 080 96 2 5 101 87 36 173 141 53 39 9758 36 (N = 4) 1 728-2 208 79-108 4-6 79-9298-105 31-.90 165-184 130-152 49-54 89-108 35-4157-5931-41 nwineam n. sp. 1 7892.5 47 695 159 .43 175 168 48 32 12654 28 (N = 3) 1 500-2 280 2-3 45-51 4-6 60-75150-168 .40-.47 165-180 159-180 46-51 102-150 24-3048-6024-39 rrvlllphanelm n. sp. 2 136 96 3 724 116 .61 197 137 50 132 35 (N = 10) 1 560-2 880 75-129 2-4 3-5 78-135 60-84 36-.69 135-273 96-159 44-63 75-183 24-48 obtirsinetm n. sp. 2 745 3.5 133 4.5 102 79 .78 256 150 51 81 168 124 48 (N = 2) 2 550-2 940 105-1623-4 4-5 99-105 78-81 ,7432 237-276 141-159 50-52 45-51120-129 72-90 paranerm? n. sp. 920 42 2 723 108 .66 75 129 43 69 20 (N= 1) pertaneau n. sp. 1747 109 , 6 668 127 .52 216 157 5436 8374 35 (N = 10) 1440-2 016 3-988-133 5-11 54-79120-136 .44-.63 159-279 133-174 38-73 63-98 32-3863-8625-48 popenenla n. sp. 2 001 87 4 825 164 .50 190 198 47 132 3237 66 (N = 7) 1740-2 190 3-5 75-93 4-7 144-183 69-102 .39-.60 150-240 159-240 37-52 105-162 33-4260-9024-36 tigonelna n. sp. 2 556 129 2 2 99 14 .73 210 128 5142 16274 52 (N = 10) 2 070-3 330 1-399-156 1-3 66-120 60-90 ,6588 171-279 90-159 48-55 129-180 42-6360-9039-45 yoponema n. sp. 1331 51 4 577 142 .39 122 117 5034 8764 24 (N = 5) 1088-1 600 4-5 47-54 6-8 41-73120-174 .30-.49 108-143 95-143 48-55 76-101 19-2954-7332-38

3 62 Revue Nimatal. 14 (3) :361-374 (1991) Parasitodiplogaster spp. in fig wasps

, Table 2 Morphometric data for males of Parasitodiplogaster species.

citrinenla n. sp. 1835 56 6 9 134 69 .52 23 1 176 92 46 29 8 18 4 (N = 5) 1 384-2240 45-76 4-8 6-13 126-143 60-82 .48-.57 171-260 153-193 77-10241-48 26-32 7-9 16-19 3-5 dugatmna n. sp. 1888 98 2 4 92 80 .87 187 129 83 42 45 9 20 4 (N = 3) 1760-1 98488-105 3-5 82-98 73-86 .86-.90 127-225 117-146 77-8840-43 39-48 8-10 17-21 3-5 tminema n. sp. 1098 30 3 5 144 65 .46 162 16025 50 36 6 22 4 (N = 5) 750-1350 21-42 2-3 4-6 117-156 57-75 .38-.59 144-198 138-180 19-2939-54 26-41 4-8 15-26 3-5 nylnphanenla n. sp. 761 150 3 4 120 68 36 158 13756 84 78 14 28 4 (N = 10) 960-1520 60-92 2-4 3-5 102-141 42-90 .41-.70 78-210 117-156 71-10551-64 66-84 12-15 22-33 3-6 obusinelna n. sp. 2670 109 3.3 3.6 106 85 .80 250 150 61 117 55 8 31 7 (N = 8) 1 920-3990 84-174 3-4 3-4 93-120 66-99 .65-.90 174-315 135-186 90-15353-75 48-68 6-9 24-36 4-8 paranetna n. sp. 43 633 3 3 117 66 .70 143 12629 47 25 7 11 3 (N = 3) 620-640 3948 3-4 2-4 63-69 .62-.75 135-150 115-132 39-5321-36 24-26 6-8 10-12 perlanenla n. sp. 1360 69 7 5 129 53 -43 208 15248 73 33 7 17 6 (N = 6) 1056-1696 63-76 6-8 4-6 115-133 47-70 .36-.50 153-225 135-171 59-8045-51 32-35 6-8 16-18 5-7 popene8la n. sp. 1984 78 3 5 181 98 54 195 185 56 110 52 IO 25 6 (N = 3) 1 600-2624 63-95 4-6 151-200 87-101 .51-.58 147-238 162-200 83-12551-61 47-53 8-13 24-27 5-7 trigonetna n. sp. 2528 99 2 2 108 75 .?O 218 14358 135 48 9 24 5 (N = 10) 2 020-3 O40 79-130 1-3 2-3 88-123 63-92 .57-.79 187-247 110-181 115-15248-64 36-56 8-13 21-32 3-8 yoponama n. sp. 43 789 2 8 132 46 .34 119 10728 48 41 5 25 4 (N = 6) 640-86435-48 1-3 3-11 120-142 38-60 29-.43 98-133 95-120 42-5326-30 38-45 4-6 24-26 3-5

gubernaculumstraight, narrow, genital papillae eight thickened inner gut Wall, well-developed vaginal glands, pairs, consisting of four pre-cloaca1 pairsand four short, broad spicules, narrow, long gubernaculum and post-cloaca1 pairs; two pairs at tip of tail;phasmids eight pairs of genital papillae. conspicuous (tubular)at tipof tail; tail tip bearinga fine The following keys separate this species from other mucron. Panamanian members of the genus.

TYPE HABITAT AND LOCALITY Foundin fruits of Ficuscitrifolia growing inthe Parasitodiplogaster duganerna n. sp. vicinity of Barro Colorado Island, Republicof Panama. (Figs 2C, 3F)

TYPEMATERIAL MEASUREMENTS Holotyfie (male) and allotype (female) depositedin the Adults :See Tables 1 and 2. nematology collection of the Department of Nemato- logy, University of California, Davis, California; para- DESCRIPTION types in the Laboratoire des Vers, Museum national d'Histoire naturelle, Paris. Females :Medium to large nematodes; stoma small; anterior and posterior pharynxes almost equalin length, bothwith distinct bulbs at base;anterior pharynx DIAGNOSISAND RELATIONSHIPS muscular, lumen expanded at base; distinct isthmus in P. citritzenza n. sp. is characterized by a well-developed glandular posterior pharynx; pharyngeal-intestinalvalve stoma, well-developed muscular anyrior and posterior inconspicuous; excretory pore opposite anterior part of pharynxes, well-developed pharyngeal-intestinal valve, intestine;intestine thin-walled, but walls expanded

Revue Névzatol. 14 (3) :361-374 (1991) 363 G. O. Poinar, 3. & E. A. Herre

- ~~ around pharyngeal bulb; vulva surrounded with a con- region of vulva; tail long, slender, witha small mucron tinuous or broken cuticular ring; vaginal glands dis- at tip. tinct, little cuticular modification at vulva; Young fully developed in eggs; tail tip pointed. Males :General features as described under females; testis outstretched, rarely reflexed; spicules long, nar- Males :General features as described under females; row, with a small manubrium (width less than that of testisoutstretched; spicules narrow, with a distinct corpus); gubernaculum long, slightly bent at proximal manubrium setoff from corpus; gubernaculumswollen end; genital papillaesix pairs, three pairspre-cloaca1 and in middle;genital papillae sevenpairs; three pairs three pairspost-cloacal; phasmids inconspicuous, tail tip preanal and four pairs postanal; two pairs at tip of tail;' pointed. phasmids conspicuous (tubular) at tip of tail; tail tip pointed. TYPE HABITAT AND LOCALITY Foundin fruits of Ficus maxima growing inthe 'TYI?E HABITAT AND LOCALITY vicinity of Barro Colorado Island, Republicof Panama. Found infruits of Ficus dugandii growing in the vicinity of Barro Colorado Island, Republicof Panama. TYPE MATERIAL. mEMATERIAL Holotype (male) and allotype (female) depositedin the nematology collection of the Department of Nemato- Holotype (male) and allotype (female) depositedin the logy, University of California, Davis, California; para- nematology collection of the Department of Nemato- types in the Laboratoire des Vers, Muséum national logy, University of California, Davis, California; para- d'Histoire naturelle, Paris. types in the Laboratoire des Vers, Muséum national d'Histoire naturelle, Paris. DIAGNOSISAND RELATIONSHIPS

DIAGNOSISAND RELATIONSHIPS P. maxinema n. sp. is characterized by a well-devel- opedstoma, slender, elongate anterior and posterior P. duganema n. sp. is characterized by a small stoma, pharynxes without noticeable bulbs, a thick-walled in- nearly equal length of the anterior and posterior phar- testine,reduced vaginal glands, no obvious cuticular ynxes, swoUen bases of both anterior pharynxes, reduced thickening at vulva, slender slightly-bowed spicules with pharyngeal-intestinal valve, intestinal cells surrounding a small rounded manubrium, slender, elongate guberna- pharyngeal base, large thin-walled intestinal cells, excret- culum and six pairs of genital papillae. orypore opposite intestine, slender, narrow spicules, The following keys separate this species from other wedge-shaped gubernaculum and seven pairs of genital Panamanian members of the genus. papillae. The following keys separate this species from other Panamanian species of the genus. Parasitodiplogaster nymphanema n. sp. (Figs 1C; 3A; 4C, DyE) Parasitodiplogaster maxinema n. sp. MEASUREMENTS (Figs 2E, 3H) Adults :See Tables 1 and 2. MEASUREMENTS DESCRIPTION Adults :See Tables 1 and 2. Females :Medium tolarge nematodes; stoma distinct, DESCRIETION longer than wide; both anterior and posterior pharynxes well-developed and muscular; both with short crescent Females :Medium tolarge, slender nematodes; soma valve plates at base, basal pharynx with distinct lumen longer than wide; anterior pharynx more than twice as in mid-region; excretory pore opposite posterior phar- long as posteriorpharynx; anterior pharynx faintly ynx; pharyngeal-intestinal valve distinct;intestine muscular, elongate, swollen only slightly at base which thin-walled, cells large,transparent; ovaries reflexed, contains slender crescentvalve plates; posterior pharynx sometimes double reflexed; vaginal glands well-devel- short, slender, glandular,slightly swollen basal bulb area oped, cuticle slightly thickened in region of vulva; tail with expanded lumen; pharyngeal-intestinal valve pres- bluntlyrounded, sometimes with a terminal minute ent; excretory pore usually opposite posterior pharynx; knob on the tip. intestine thick-walled, containing an amorphous massof material together with yeast-like cells; vaginal glands Males :General features as described under females; reduced; ovaries reflexed; body cuticle not thickened in testis reflexed or outstretched; spicules large, well-devel-

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oped, with a distinct manubrium,tips pointed and region; small pharyngeal-intestinalvalve; intestine with cuTed slightlyposteriorly; gubernaculum straight; large cells filled with corpuscles rangingfrom 3-6 Fm in genital papillae sevenpairs, four pre-cloaca1 in a row up diameter; ovaries reflexed; vaginalglands indistinct; the body and three post-cloacal; terminal tail papilla body cuticle not or little expandedat vulva; tailtip blunt, pair have two nerveendings each; tail tiprounded; with a small mucron. phasmids inconspicuous. Males :General features as described under females; Juveniles :Some juveniles were found in association testis outstretched; spicules large, slightly curved, with with adult wasps (Pegoscapus sp.) inside the fig fruits. manubrium slightly wider than the corpus, tips blunt; These juveniles rangedfrom 222-288 Pm in length, gubernaculumstraight, with a medialconstriction; 10-13 pm in width, the distance from the head to the genital papillae consist of three pre-cloacal pairs and base of the posterior pharynx was 80-92 pm and thetail four post-cloaca1 pairs (two on tailtip); phasmids length was 28-29 pm. It is assumed that these are dauer distinct, tubular, protruding; tail tip blunt, with a fine andpre-dauer that developed inthe fig from eggs mucron. deposited by the females. Similar dauer nematodeswere also removed from the genital system of female wasps. TYPE HABITAT AND LOCALITY

TYPE HABITAT AND LOCALITY Found in fruits of Ficus obtusifoolia growing in the vicinity of Barro Colorado Island, Republicof Panama. Found in the fruitsof Ficus nymphaeifolia growing in the vicinity of BarroColorado Island, Republic of TYPE MATERIAL Panama. Holotype (male) and allotype (female) depositedin the TYPE MATERIAL nematology collection of the Department of Nemato- logy, University of California, Davis, California; para- Holotype (male) and allotype (female) depositedin the types in theLaboratoire des Vers, Muséum national nematology collection of the Department of Nemato- d’Histoire naturelle, Paris. logy, University of California, Davis, California; paruty- pes inthe Laboratoiredes Vers, Muséum national DIAGNOSISAND RELATIONSHIPS d’Histoire naturelle, Paris. P. obtusinema n. sp. is characterized by a small but DIAGNOSISAND RELATIONSHIPS distinct stoma, well-developed, muscular anterior and posterior pharynxes with distinct swollen basal bulbs; P. nymphanema n.sp. is characterized by a well- exeretory pore opposite intestine, a faint pharyngeal- developed stoma, anterior and posterior muscular phar- intestinal valve, thin, but expandedintestinal cells ynxes with imperceptible bulb-areas, a distinct phar- containing small elliptical corpuscules, indistinct vaginal yngeal-intestinal valve, large, thin-walled intestinal glands,slender, slightly bentspicules, gubernaculum cells, well-developed vaginal glands, body cuticle swol- withmedial constriction and sevenpairs of genital len at.vulva,large, well-developed spicules with elongate papillae. manubria, spicule tips bent posteriorly and seven pairs The following keys separate this species from other of genital papillae. Panamanian members of the genus. The following keys separate this species from other #. Panamanian members of the genus. Parasitodiplogaster paranenta n. sp. Parasitodiplogaster obtusinema n. sp. (Figs lE, 3E) (Figs 2A, 3J) MEASUREMENTS MEASUREMENTS Adults :See Tables 1 and 2. Adults :See Tables 1 and 2. DESCRIPTION DESCNPTION Females :Small nematodes; stoma small; anterior and Femules : Largenematodes; stoma small, slightly posterior pharynxes slender, weakly muscled, base of longer than wide; anteriorpharynx well-developed, anterior pharynx swollen and containing crescent valve muscular, with crescent valve plates in the expanded plates; base of posterior pharynx slightly swollen, with basal portion, clearly set off from muscular posterior indistinct valve plates; excretory pore posterior opposite pharynx; base of posterior pharynx swollen, with faint pharynx; bulb of posterior pharynx inserted into anterior crescent valve plates; excretory porein anterior intestinal portion of intestine, pharyngeal-intestinalvalve present;

Revue Nématol. 14 (3) :361-374 (1991) 365 G. O. Poinar, Jr. & E. A. Herre intestine with distinct bush border (may be symbiotic expanded, tail bluntly pointed, sometimes terminating ); vaginal glandsgreatly reduced, cuticle not in a small button or mucron. thickened in region of vulva; tail tip pointed. Males : General features as described for females, Males :General features as described under females; exceptstoma is wider than long;testis outstretched, testis outstretched; spicules short andwide, with a large, spicules shortand broad, withwide manubrium; corpus distinct manubrium; corpus constricted in middle for- containing a proximal constriction resulting in a knob- ming two small ventral arches, tip bluntlypointed; bed tip; gubernaculum robust; eight pairs genital pa- gubernaculum short, wide at proximal end and pointed pillae, four pairs pre-cloaca1 and fourpairs post-cloacal; at tip;genital papillae sevenpairs, four pairs pre-cloaca1 tail tips pointed; phasmids inconspicuous. in a series and three pairs post-cloaca1 (one pair at tail tip); tail tip pointed; pliasmids inconspicuous. TYPE HABITAT AND LOCALITY Found in fruitsof Ficus pertusagrowing in thevicinity TYPEHABITAT AND LOCALITY of Barro Colorado Island, Republic of Panama. Found in fruits of Ficusparaensis growing in the vicinity of Barro Colorado Island, Republicof Panama. TYPE MATERIAL Holotype (male) and allotype (female) deposited in the TYPE MATERIAL nematology collection of the Deparmentof Nematology, Holotype (male) and allotype (female)deposited in University of California, Davis, California; paratypes in the nematology collectionof the Department of Nema- the Laboratoire des Vers, Muséum national d’Histoire tology, University of California, Davis, California; para- naturelle, Paris. types in the Laboratoire des Vers, Muséum national d’Histoire naturelle, Paris. DIAGNOSISAND RELATIONSHIPS Small to medium-sized nematodes characterizedby a DIAGNOSISAND RELATIONSHIPS stoma wider than long, well-developed anteriorand P. paranema n. sp. is characterized by a small stoma, posterior pharynxes with basalbulbs, an indistinct slender weakly muscled anteriorand posterior pharynxes pharyngeal-intestinal valve, small intestinal cells, form- with small basal bulbs; intestinalcells surrounding basal ing a tube-like intestinewith thickened walls;well bulb of posterior pharynx, intestine tubular, with thick developed vaginal glands; greatly expanded cuticle in inner walls, short, wide spicules with a distinctma- area of vulva; short and broad spicules, withmanubrium nubrium, a shortgubernaculum, with theproximal wider than corpus, gubernaculum with proximal portion portiongreatly swollen and seven pairs of genital bent posteriorly and eight pairs of genital papillae. papillae. The following keys separate this species from other The following keys separate this species from other Panamanian members of the genus. Panamanian members of the genus. Parasitodiplogaster popenenzan. sp. (Figs lA, 3G) Parasitodiplogaster pertaneman. sp. (Figs 1D; 3D; 4A, B) MEASUREMENTS

MEASUREMENTS Adults :See Tables 1 and 2. Adults :See Tables I and 2. DESCRIETION

DESCRIPTION Females : Medium-sizednematodes with stoma slightly longer than wide; anterior muscular pharynx Females :Small to medium-sized nematodes; stoma twice as long as posterior weakly rnuscledpharynx; longer than wide (often coated witha deposit); muscular anterior pharynx with faint crescent valves in unmodif- anterior and weakly muscled posterior pharynxes, both ied basal portion; posterior pharynx lackingvalve plates with a basal bulb (containing faint valve plates); excret- and only slightly swollen at base whichis surrounded by ory poreopposite posterior pharynx; intestine with intestinal cells; excretory pore in region of junction distinct Wall, containing yeast and bacterial cells in between anterior and posterior pharynxes; pharyngeal- lumen; vaginal glands large; cuticle greatly expanded intestinal valve inconspicuous; inner lining of intestine (from 32-70 Pm) in region of vulva, ovaries reflexed thick, outer walls thin;vaginal glands present, body (sometimes past ‘vulva), reflexed portion of ovary often cuticle slightly thickened in region of vulva; eggs deve-

366 Revue Nérnatol. 14 (3) :361-374 (1991) Parasitodiplogaster spp. in fig wasps

~~ ~~ ~~ lop to young juvenile stage in uterus; ovaries reflexed, ynx; intestinal walls thin, intestinal lumen wide; vaginal sometimes past vulva; tail tip pointed. glands well developed; cuticle thickened in region of vulva, ridged on the ventral side; ovaries reflexed for a Males :General features as described under females; short distance; tail rounded, sometimes with a small testis reflexed or outstretched;genital cone present; mucron on the tip. spicules curved, manubrium large; corpus bent at 22" angle, somesculpturing present on distalportion of Males :General features as described under females; corpus,tips acute; gubernaculum straight, proximal testisoutstretched; spicules straight, with a distinct portion swollen; genital papillae seven pairs, four pairs manubrium wider thancorpus; spiculetips bluntly pre-cloaca1 in a series, three pairs post-cloaca1 (pair of pointed; gubemaculum straight, wedge-shaped with double-nerved papillae on tail tip); phasmids conspi- proximal end swollen; genital papillae eight pairs, four cuous, tubular; tail tip with minute mucron. pairs pre-cloaca1 in a series up the body, four pairs post-cloacal (double pair of papillae on tail tip); phasm- TYPE HABITAT AND LOCALITY ids conspicuous; tail tip with or without mucron.

Found infruits of Ficuspopenoei growing in the TYPE HABITAT AND LOCALITY vicinity of Barro Colorado Island, Republicof Panama. Foundin fruits of Ficustrigonata growing in the TYPE MATERIAL vicinity of Barro Colorado Island, Republicof Panama. Holotype (male) and allotype (female) depositedin the TYPEMATERIAL nematology collection of the Department of Nemato- Holotype (male) and allotype (female) depositedin the logy, University of California, Davis, California; para- nematology collection of the Department of Nemato- types inthe Laboratoire des Vers, Muséum national logy, University of California, Davis, California; para- d'Histoire naturelle, Paris. types in the Laboratoire des Vers, Muséum national d'Histoire naturelle, Paris. DIAGNOSISAND RELATIONSHIPS DIAGNOSISAND RELATIONSHIPS Medium-sized nematodes withwell-developed stoma, Large nematodes with a small stoma, anterior and anterior pharynx approximately twice the length of the posterior pharynxes glandular, both with distinct basal posterior pharynx, excretory pore opposite the middle swellings, excretory pore normally opposite the intestine portion of thepharynx; base of posteriorpharynx (sometimes posterior pharynx), intestine composed of normally surroundedby anterior intestinal cells, intesti- large, thin-walled cells, vaginal glands well developed, nal cells thin on the outerside but thick-walled surroun- body cuticle expanded in region of vulva, straight spic- dingthe lumen; spiculeswith a largemanubrium, ules with a distinct manubrium, bluntly pointed tips, corpus witha distinct bend,the tips pointed; gubernacu- straight gubemaculum, swollen proximally, eight pairs lum straight, proximal portion swollen; seven pairs of of genital papillae, tail rounded, sometimes bearing a genital papillae and tail tip with a small mucron. small mucron. The following keys separate this species from ,other The following keys separate this species from other Panamanian members of the genus. Panamanian members of the genus.

Parasitodiplogaster trigonema n. sp. Parasitodiplogaster yoponeman. sp. (Figs 2B, 3B) (Figs 2D, 31)

MEASUREMENTS MEASUREMENTS Adults :See Tables 1 and 2. Adults :See Tables 1 and 2.

DESCRIPTION DESCRIPTION

Fetnales :Large nematodes with small stoma; both Females :Small nematodes with stoma longer than anterior and posteriorpharynxes glandular, muscle wide; anterior pharynx muscular, three times as long as fibersrestricted to swollen basalportion of anterior posterior glandular pharynx; basal bulb absent in anter- pharynx (with crescent valve plates); posterior pharynx ior pharynx but weakly formed in posterior pharynx; with basal bulb but valve plates absent; small phar- valve area in anterior pharynx located some distance yngeal-intestinal valve; excretorypore in anterior from the base; nerve ring encircles anterior pharynx; intestinal region or occasionally opposite posterior phar- excretory pore opposite base of anterior pharynx; base

Revue Nématol. 14 (3) :361-374 (1991) 367 G. O. Poinar, Jr. & E. A. Herre

E D A C l 54m I

Fig. 1. Anterior portion of male PanamanianParasitodiplogaster. A :P. popenema n. sp.; B : P. c trinema n. sp.; C : P. nymphanema n. sp.; D : P. pertanema n. sp.; E : P. paranerna n. sp. of posterior pharynx deeply embedded in anterior por- post-cloacal (two ventral pairson tail tip); tailtip bearing tion of intestine;pharyngeal-intestinal valve incon- a mucron; phasmids inconspicuous. spicuous, vaginal glands greatly reduced, vulva incons- picuous, body cuticle not swollen in region of vulva; tail TYPE HABITAT AND LOCALITY acutely pointed, with a small mucron on tip. From fruitsof Ficus yoponensis growing in thevicinity Mules :General features as described under females; of Barro Colorado Island, Republic of Panama. testisoutstretched or reflexed; spicules curved, ma- nubrium small, width less than that of corpus, tips blun- TYPE MATERIAL tly pointed, slightly expanded; gubernaculum relatively large, proximal portion curved anteriorly; distal portion Holotype (male) and ullotype (female) depositedin the encloses corpus of spicules, genital papillae eight pairs, nematode collection at the Departmentof Nematology, three pairs pre-cloacal, extending up body; five pairs University of California, Davis, California; purutypes in

368 Revue Nématol. 14 (3) :361-374 (1991) Parasitodiplogaster spp. in fig wasps

Fig. 2. Anterior portion of male Panamanian Parasitodiplogaster. A : P. obtusinema n. sp.; B : P. trigonema n. sp.; C : P. duganewza n. sp.; D : P. yoponenza n. sp.; E : P. nzaxinema n. sp. the Laboratoire des Vers, Muséum national d'Histoire cuticle could not be detected over the hypodermis in the naturelle, Paris. a withspecimens loose cuticle. These aresmall nematodes with a deeD stoma,a DIAGNOSISAND RELATIONSHIPS muscular anterior pharynx approximately -hee times the length of the posterior glandular pharynx, the base One interesting feature of both sexes is the tendency of the posterior pharynx Üsually sünounded by the for the cuticle to separate from the hypodermis. Initially,anterior intestinal cells, the excretory pore opposite the it was suspected that anextra molt occurred orthat the anterior pharynx, the intestinal cells large and thin, the final juvenile cuticle was retained. However, a second vaginal glands indistinct,swollen body cuticle absent in

Revue Nénzatol. 14 (3) :361-374 (1991) 369 G. O. Poinar, Jr. & E. A. Herre

Fig. 3. Lateral view of male tails of Panamanian Parasitodiplogaster. A : P. nynzphanema n. sp.; B : P. trigonema n. sp.; C : P. citrinema n. sp.; D : P. pertanema n. sp.; E : P. paranema n. sp.; F : P. dzcganema n. sp.; G : P. popenelna n. sp.; €3 : P. maxinema n. SP.; 1 : P. yoponema n. sp.; J : P. obtusinenza n. sp. For the purposeof space, the most anterior pre-cloaca1 papillae maynot be included in the illustrations.

the region of the vulva, narrow spicules, roundma- Key to thefemales of the Panamanian species nubrium, smaller thanthe corpus diameter,corpus the of Parasitodiplogaster bow-shaped, a narrow gubernaculum, curved up proxi- mally, eightpairs of genitalpapillae and the tail tip 1 - Ratio of posteriorpharynx to anteriorpharynx 0.68 or pointed. greater (0.65-0.90);pore excretory in region of intestine; The following keys separate this species from other soma reduced ...... 2 Panamanian members of the genus. - Ratio of posterior pharynx to anterior pharynx usually less

3 70 Revue Nématol. 14 (3) :361-374 (1991) Fig. 4. A :Lateral view of mid-region of Parasitodiplogaster pertanema n. sp. showing greatly swollen body cuticlein region of vulva (arrow); B : Bacterial rods in the intestinal lumen (L) of Parasitodiplogaster pertanema n. sp.; C : A pair of Parasitodiplogaster nynphanema n. sp. ‘‘ in copula ”. Two of the four vaginal glands (arrows) are visible; D : Dauer juvenile of Parasitodiplogaster nynzphanema n. sp. enclosed in previous juvenile cuticle (from fig fruit); E :Pre-dauer developing juvenile ofParasitodiplogaster nymphanema n. sp. (from fig fruit). (Bars equivalent :A = 100 pnJ B = 38 PmJ C = 78 pzJD = 45 ,unJ E = 52 p.) G. O. Poinar, Jr. & E. A. Herre

than 0.68 (0.30-0.69); excretory porein region of pharynx - Length of tail equal to or lessthan twice spicule length.. 6 or if in region of intestine and pharynx ratio is greaterthan 2 - Excretory pore opposite intestine; average ratio of pos- 0.68 (P. obtusinema n. sp.) then intestinal cells filled with terior pharynx to anterior pharynx between 0.70 and 0.80 numeroussmall globules ...... 3 (range from 0.57 to 0.90) ...... 3 2 - Base of posterior pharynx surroundedby anterior intesti- - Excretory pore opposite pharynx or intestine; average nal cells; tail length usually less than 100 pm ...... ratio of posterior pharynx to anterior pharynx between ...... P. duganema n. sp. 0.43 and 0.54 (range from 0.36-0.58) ...... 4 - Base of posterior pharynx not or only occasionally sur- 3 - Pairs of genital papillae = 8; corpus straight; gubernacu- rounded by anterior intestinal cells; tail length usually lum without medial constriction .... P. trigonema n. sp. greater than 100 pm ...... P. trigonema n. sp. - Pairs ofgenital papillae = 7;corpus slightly curved; 3 - Excretory pore in region of intestine; stoma reduced gubernaculum with medial constriction ...... P. obtusinema n. sp...... P. obtusinema n. sp. - Excretory pore in region of pharynx or at pharyngeal- 4 - Corpusof spicules bent at a 22' angle; borders of intestinal junction; stoma distinct ...... 4 manubriumangled; spicule tips pointed, forming an 4 - Anterior pharynx usually from two to three times the angle of 40" ...... P. popenema n. sp. length of the posterior pharynx (ratio of posterior phar- - Corpus slightly curved, not withan abrupt bend; borders ynx to anterior pharynx generally lessthan 0.50 [range = of manubrium rounded; spicule tips rounded or bluntly 0.30-0.601); vaginal glands reduced ...... 5 pointed, forming an angle of 60" or more ...... 5 - Anterior pharynx usually less than two times the length 5 - Stomawider than long;pharyngeal-intestinal valve of the posterior pharynx (ratio of posterior pharynx to reduced ...... P. pertanema n. sp. anterior pharynx generally greater than 0.50 [range = - Stomalonger than wide; pharyngeal-intestinal valve 0.41-0.661); vaginal glands well-developed (except in P. distinct ...... P. citrinema n. sp. pertanema n. sp. which has greatly expanded body cuticle 6 - Excretory pore opposite intestine; ratio of posterior to in the vulvar region) ...... 7 anterior pharynx = 0.87 (0.86-0.90) .. P. duganema n. sp. 5 - Excretory pore opposite posterior pharynx; base of pos- - Excretory pore opposite pharynx or rarely intestine; ratio terior pharynx rarely surrounded by anterior intestinal of posterior to anterior pharynx = 0.29-0.70 ...... 7 cells ...... P. maxinema n.sp. 7 - Excretory pore opposite anterior pharynx; average ratio - Excretory pore opposite anterior pharynx, posterior phar- of anterior pharynx to posterior pharynx = 0.34 (range ynx or pharyngeal-intestinal junction; base of posterior = 0.29-0.43); 8 pairs genital papillae .. P. yoponema n. sp. pharynx surrounded (sometimes only slightly) by anterior - Excretory pore opposite posterior pharynx or rarely an- intestinalcells ...... 6 teriorpharynx or intestine; average ratio of anterior 6 - Anterior pharynx approximately twice the length of the pharynx to posteriorpharynx = 0.57(range = posterior pharynx; body cuticle slightly swollen (and often 0.38-0.75);7 pairs genital papillae ...... 8 ridged) in region of vulva ...... P. popenema n. sp. 8 - Base of posterior pharynx surrounded by intestinal cells; - Anterior pharynx approximately three times the length of spicules short and wide, with rounded manubrium as the posterior pharynx; body cuticle not swollenin region wide as corpus width ...... P. paranema n. sp. of vulva ...... P. yoponema n. sp. - Base of posterior pharynx not surrounded by intestinal 7 - Vaginal glands inconspicuous; body cuticle not expanded cells; spicules long and relatively narrow; manubriumnot in region of vulva; base of posterior pharynx surrounded rounded, or if so then smaller than corpus width ...... 9 by anterior intestinal cells ...... P. paranema n. sp. 9 - Manubrium large, elliptical; width nearly equalto that of -Vaginal glands conspicuous; body cuticle expanded in corpus width; angle between corpus and manubrium = region of vulva; base of posterior pharynx free or only 45'; spicule tips pointed and curved downward ...... slightly surrounded by anterior intestinal cells ...... 8 ...... P. nymphanetna n.sp. 8 - Body cuticle greatly expanded(up to 1/2 body width)in - Manubrium small, rounded; width smaller than that of regionofvulva; phaqngeal-intestinal valve incon- corpus width; angle between corpus and manubrium = spicuous ...... P. pertanema n. sp. O; spicule tips blunt and straight .. P. maxinema n. sp. - Body cuticle slightly expanded in region of vulva (less than 1/4body width); pharyngeal-intestinalvalve distinct ...... 9 Discussion 9 - Stomashort (3-5 Pm); length anterior pharynx from 78-135pm ...... P. nymphanema n. sp. The ten species of Panamanian fig wasp nematodes - Stomalong (5-8 Pm); length anterior pharynx from described here present a remarkable picture of adapta- 138-144 p ...... P. citrinema n. sp. Ztive radiation from a single stem species (monophylo- geny). This stem species has not been found but m.ay Key to the males of the Panamanian species still exist and would be a microbotrophic, free-living of Parasitodiplogaster diplogasterid feeding on bacteria and other microorga- 1 - Length of tail twice or more than spicule length ...... 2 nisms in and around fallen fig fruits. The sequence of

3 72 Revue Nématol. 14 (3) :361-374 (1991) Parasitodiplogaster spp. in jïg wasps developmentthrough geologic historymight be the there are two distinct morphological typesof alimentary following. tracts. One of these would appear to be for the intake Through constant association with the fig fruit, the of solid particulate food (medium to large stoma, well nematodes encountered the fig wasps as they left some muscled pharynxes; gut cells small with distinct border of the fruit that had fallen prematurely. Or the nema- lining the lumen). The other type consists of a small todes couldhave been associatedwith the above-ground stoma, glandular pharynxes and large, thin-walled gut portions of thetree. This wouldbe possible under cells with indistinct borders. A range of intermediary conditions of high moisture asfound in thetropics. The types occur between these two extremes. Such modifkat- initial association with the fig wasp was probably phor- ionsundoubtedly reflect the type of nourishment etic, possibly external with the nematodes on or wrap- utilized in the fig fruit and the nature of the physical ped around thewasp’s body. Eventually the association environment (liquid, moist, or dry). became more complex, with the nematodes exhibiting In microbotrophic species of Diplogasteroidea, oneof internal phoresy in the ’s intestinal tract or reprod- which was undoubtedlyancestral to these fig wasp uctive system andthen invading the hemolymph where forms, the pharynx is usually modified into a robust development was initiated. anterior muscular portion and a smaller posterior gland- The dauer stage of the nematodes enter the female ular poftion. These portions are usually not delineated wasps when they emerge from the gall flowers of the as much as they are in Parasitodiplogaster (thus justif- fruit. They then matureinside the fig wasp and by the ying the terms anterior and posterior pharynx for the time the figwasp is prepared to oviposit in a fig flower, latter).Spirurid nematodes are also characterized by the nematodes have reached the adult orpre-adult stage. having the pharynx divided into anterior muscular and After the wasp enters the ostiole of the fig the nematodes posterior glandular sections (Poinar & Quentin, 1972). leave the wasp and enter the flower cavity where they The presence of glandular pharynxes may be a modifi- mate andegg development occurs.The adult nematodes cation for taking in liquid nourishment, since with Para- die andthe youngdevelop to the third stagedauer sitodiplogaster, those species with glandular pharynxes (resistant stage) which search for female wasps. The also tend to have an intestinecomposed of large, nematodes could reachthe female wasp by entering the thin-walled cells. In such species, the intestinal lumen gall through the hole the male wasp makes for mating was usually filled witha clear fluid probably originating or waiting until the wasps emerge. from the fig fruit. This product could itself serve as a Al1 ten species of Panamanian fig wasp nematodes source of nutrients orit could serve as a culture medium show differences in size, digestive system morphology formicroorganisms inside the nematode’s intestinal (stoma, pharynx and intestine) and primary and second- tract.Bacterial rods were obsenred in the intestinal ary reproductivestructures (vaginal glands, modified lumen of P. paranema n. sp. and P. pertanenaa n. sp. body cuticle at vulva, spicules,gubernaculum and (Fig. 4B), cocci in theintestine of P. yoponenza n. sp.and genital papillae). P. citrinema n. sp., undeterminedcorpuscles in P. This condition of adaptive radiation is analogous to obtusinelna n. sp. and yeast-like cells in P. maxinema n. the classical case of Darwin’s finches on the Galapagos SP. Islands. There is still discussion on whether the latteris Many of thePanamanian Parasitodiplogaster spp. a result of a competitive character displacement (Lack, possess modifiedcrescent-shaped valves inboth the 1947) or adaptation to an available food source (Bow- anterior and posterior pharynxes. At this time, the man, 1961). presence of a valve in the pharynx is considered to With the fig wasp nematodes, only one species of represent a recent modification related to the nema- nematode has been foundin each fig, so character tode’s behavior andnot an indicationthat thestem displacement cannot be used as an explanation for this species was a member of the Pseudodiplogasteroidea divergence. The fig trees and fig wasps represent sym- (Goodey, 1963) althoughthis possibility cannotbe patric species since their range overlaps. However each ruled out. figspecies is pollinated by a singlefig wasp species Further variable modification in female Parasitodi- (Wiebes, 1979), thus the nematodes in actuality rep- plogaster were the vaginal glands and body cuticlein the resentallopatric populations since physical overlap vulvar region. The vaginal glands varied from very large never occurs. in P. citrinema n. sp., P. pertanenaa n. sp., P. duganema What selective forcesresulted in thesedivergent n.sp. and P. nymphanema n.sp. (Fig. 4) to incon- morphological nematode species which obviously arose spicuous in P. yoponema n. sp., P. trigonema n.sp., from a common stem species? In the present situation, P. paranema n.sp. and P. maxinema n.sp. A thick it was probably conditions associated with reproduction yellowish deposit which pluggedthe vagina and adhered and feeding in the different fig fruits. The size of the to the outside of the vulva may have been produced by stoma,structure of the pharynx and intestineand these glands. A similar deposit frequently surrounded rnicroscopic contents of the gut are related to the type the male cloacal opening. of food utilized.In the Panamanianfïg wasp nematodes, The modification of the body cuticle in the region of

Revue Ne‘matol. 14 (3) :361-374 (1991) 313 G. O. Poinar, Jr. & E. A. Herre

~~ ~ ~____ the vulva varied from nonexistent or barely noticeable the genus Ficus (Müller, 1984). It is quite probable that in P. yoponema n. sp., P. paranema n. sp., P. obtusinema the relationship between figwasps and Ficus was estab- n. sp. and P. maxinema n. sp. to well developed in P. lishedby the mid-.Since South America pertanema n. sp. (Fig. 4A). The selective pressure for became separated from Africa in the late Cretaceous this excess cuticle expansion in P. pertanema n. sp. is (Cox & Moore, 1985), this would explain the presence unknown. of Parasitodiplogaster on both continents today. The Males varied considerably in the structure of the Diplogasteridae as a group are quite ancient and prob- spicules and gubernaculi.The spicules ranged in length ably arose in the early Paleozoic (Poinar, 1983). from 24to 84 Pm and varied in curvature, general shape, distal morphology and manubrium structure. During mating, the spicule corporewere inserted intothe vagina REFERENCES (Fig. 4C). The gubernaculi ranged from 10 to 36 pm among the species studied and varied in curvature and BOWMAN,R. 1. (196 1). Morphological variation and adaptation in the Galapagos fïnches.Univ. Cal8 Publ. Zool., 59 : 1-302. general shape. The number of genital papillae varied from six to eightpairs. Especially interesting was the pair BRUES,C. (1910). The parasitic Hymenoptera of the Tertiary on the ventral tip of the tail, which was single with a of Floressant, Colorado. Bull. Mus. comp. Zool., 54 : 1-125. single innervation in P. paranema n. sp., P. pertanema Cox, C. B. & MOORE,P. D. (1985). Biogeography. Boston, n. sp., P. obtusinema n. sp., P. duganema n. sp. and P. Blackwell Scientific Publications, 244 p. maxinema n. sp., single with two innervations in each GOODEY,J. B. (1963). Soi1 andfieshuater nematodes. London, papilla in P. nymphanema n. sp. and P. popenema n. sp. Methuen & Co., 544 p. and double, each with a single innervation in P. trigo- LACK,D. (1947). Darwin’sFinches. CambridgeUniversity nema n. sp., P. yoponema n. sp. and P. citrinema n. sp. Press, 380 p. Eight to nine pairs of caudal papillae are the normal number fordiplogasterids (Goodey, 1963). The range in MULLER,J. (1984). Signifîcance of fossil pollen for angiosperm history. Ann. Missouri Bot. Gard., 71 : 419-443. shape and size of the spicules and gubernaculi and the presence of vaginal glands and vulva cuticle expansion POINAR,Jr., G. O. (1979). Parasitodiplogaster sycophilon gen. must pertain to different mating behavioras reflected by n., sp. n. (Diplogasteridae :Nematoda), a parasite ofElisabe- different conditions in the fig fruits. thiella stzrckenbergi Grandi (Agaonidae : Hymenoptera) in Rhodesia. Proc. K. ned. Akad. Wetensch.,Ser. C, 82 : Representatives of the genus Parasitodiplogaster also 375-381. occur in association with African figs and fig wasps (Poinar, 1979) which brings up the question regarding POINAR,Jr., G. O. (1983). The Natural Histoy of Nematodes. Englewood Cliffs, New Jersey, Prentice-Hall, 323 p. the origin of this associationin time and place. Fossil fig wasps occur in the Oligocene Florissantshales of Colo- POINAR,Jr., G. O. & QENTIN, J.-C. (1972). The development rado (Brues, 1910) andin Dominican amber (A. P. of Abbreviata caucasica (Von Linstow) (Spirurida: Physal- Rasnitsyn, pers. comm.), whichranges from the opteridae) in an intermediate host. J. Parasitol., 58 : 23-28. Lower Miocene to Upper Eocene. This date roughly WIEBES,J. T. (1979). Co-evolution of figs and their insect coincides with the earliestfossil record (mid-Eocene) for pollinators. Am.Rev. Ecol. Syst., 10 : 1-12.

Accepté pour publication le 21 mai 1990.

3 74 Revue Nématol. 14 (3) :361-374 (1991)