SYSTEMATICS Leucochrysa (Leucochrysa) varia (: ): Larval Descriptions, Developmental Rates, and Adult Color Variation

EUZILENI MANTOANELLI,1 GILBERTO S. ALBUQUERQUE,1 CATHERINE A. TAUBER,2 2 AND MAURICE J. TAUBER

Ann. Entomol. Soc. Am. 99(1): 7Ð18 (2006) ABSTRACT The larval and biological characteristics of the large Neotropical genus Leucochrysa are poorly known. This report provides the Þrst taxonomic descriptions of the larvae (three instars) of a species from the subgenus Leucochrysa, Leucochrysa (L.) varia (Schneider). In doing so, it conÞrms and expands the generic diagnosis that was previously formulated on the basis of larvae in the other Leucochrysa subgenus, Nodita. Our report also provides biological data (developmental times and survival rates) from rearings under a range of temperatures (18Ð30ЊC). Finally, it describes the remarkable color variation of L. varia adults; this variation is neither maternally determined nor based on a simple (single-gene) mode of inheritance.

RESUMO As caracterõ´sticas biolo´gicas do grande geˆnero neotropical Leucochrysa, assim como suas larvas, sa˜o muito pouco conhecidas. Este trabalho fornece as primeiras descric¸o˜es taxonoˆmicas das larvas (treˆsõ´nstares) de uma espe´cie do subgeˆnero Leucochrysa, Leucochrysa (L.) varia (Schneider). A partir destas descric¸o˜es, conÞrma-se e expande-se a diagnose gene´rica formulada previamente com base no outro subgeˆnero de Leucochrysa, isto e´, Nodita. Este trabalho tambe´m fornece dados biolo´gicos (tempos de desenvolvimento e taxas de sobreviveˆncia) obtidos de criac¸o˜es sob uma se´rie de tem- peraturas (18 a 30ЊC). Finalmente, e´ descrita a nota´vel variac¸a˜o de cor dos adultos de L. varia; esta variac¸a˜ona˜oe´ determinada maternalmente e nem tampouco e´ baseada em um modo de heranc¸a simples (gene u´ nico).

KEY WORDS Leucochrysinae, larvae, phenotype, development, asymmetry

WITH APPROXIMATELY 190 DESCRIBED and numerous un- tant biological and larval morphological characters are described species, Leucochrysa constitutes the largest unstudied for the majority of species. and taxonomically most difÞcult genus in the green The lack of information on Leucochrysa larvae is lacewing family Chrysopidae. Its distribution is re- especially unfortunate because recent studies encom- stricted to the Western Hemisphere: seven species passing a wide range of chrysopid taxa reveal that occur in the United States (Tauber 2004), whereas larval morphology provides exceptionally distinctive, most are Neotropical (Brooks and Barnard 1990, stable characters for identifying chrysopids and for Freitas and Penny 2001). The genus is a prominent analyzing their phylogenetic relationships (Tauber member of the fauna in primary and secondary 1974, 2003, 2004; Dõ´az-Aranda and Monserrat 1995; Neotropical forests, where it is both abundant and Tsukaguchi 1995; Tauber et al. 2000, 2001). diverse. With the purpose of developing systematic and Leucochrysa presents multiple taxonomic chal- biological information on the genus Leucochrysa, we fo- lenges: 1) Most species are poorly described. 2) In- cused on Leucochrysa (Leucochrysa) varia (Schneider). traspeciÞc variation in adult morphology and color We did so for several reasons. First, this species is one commonly results in large differences among individ- of the most commonly collected green lacewings in uals; as a result, synonymies are numerous (Adams South American forests. Second, it has been reported 1977, 1987; Tauber 2004). 3) Adults have relatively from eucalyptus and rubber tree plantations in BrazilÕs simple genitalia that lack many of the structures that Atlantic region, where it may help suppress provide useful taxonomic characters in other genera pests (Freitas and Penny 2001). Third, adults exhibit (Adams and Penny 1986). And, 4) potentially impor- considerable variation in color, the extent and nature of which are largely unevaluated. This lack of infor- 1 Laborato´rio de Protec¸a˜o de Plantas, CCTA, Universidade Estadual mation seems to underlie numerous problems (e.g., do Norte Fluminense, Campos dos Goytacazes, Rio de Janeiro, Brazil synonymies) in the systematics literature. Last, the 28013-602 (e-mail, [email protected]). 2 Department of Entomology, Comstock Hall, Cornell University, larvae of only Þve species in the genus have been Ithaca, NY 14853Ð0901 (e-mail, [email protected]). described, and all Þve of these species belong to

0013-8746/06/0007Ð0018$04.00/0 ᭧ 2006 Entomological Society of America 8ANNALS OF THE ENTOMOLOGICAL SOCIETY OF AMERICA Vol. 99, no. 1

Nodita, one of LeucochrysaÕs two subgenera (Adams analysis software (Image-Pro Plus, version 3, Media 1987, Tauber 2004). Data from the second subgenus Cybernetics, Inc., Silver Spring, MD). (Leucochrysa), to which L. (L.) varia belongs, are Terminology for larval morphology follows stan- needed for comparison. dard use (head: Rousset 1966; thorax and abdomen: Our study here provides descriptions of L. (L.) Tauber and de Leo´n 2001, Tauber et al. 2001, Tauber variaÕs three instars and egg, as well as developmental 2004). For both the head and body, primary setae are data, and a characterization of the adult color variation designated with an “S” followed by a number; setae (its nature and possible modes of inheritance). We that also are associated with thoracic sclerites are used the larval descriptions, which are the Þrst for a designated with a setal number, followed by “Sc” and Leucochrysa Leucochrysa species in the subgenus ( ), the scleriteÕs number. Body segments are referred to to test and expand the current generic deÞnition of as T1 to T3 (thorax) and A1 to A10 (abdomen). Thus, Leucochrysa larvae (Tauber 2004). Taxonomic issues e.g., T1-S1Sc1 is the Þrst seta associated with sclerite related to the adult color variation will be dealt with elsewhere. 1 on the prothorax. The number of setae in rows on the thorax and abdomen is reported to the nearest round number; it should be noted that these Þgures are approximate because the left and right sides are often Materials and Methods asymmetrical. Descriptions. We obtained eggs and larvae from Leucochrysa setae are either smooth or thorny L. (L.) varia females collected at several forested sites (Tauber 2004). In L. (L.) varia, the setae are all smooth, inside or near the large continuous forest in the although occasionally some of the long setae on the Desengano State Park in northern Rio de Janeiro thoracic lateral tubercles of the third instar seem to State, Brazil (from September 1998 through February have very small, sparse thorns (barely visible under 2005; see Table 2 for speciÞc sites and coordinates). 200ϫ magniÞcation). We categorize these setae as Additional specimens were collected from a wooded smooth. area at the Sucupira farm (CENARGEN/EMBRAPA) We conÞrmed our identiÞcation of L. (L.) varia by near Nu´ cleo Bandeirante, Brasõ´lia, DF. comparing our specimens with SchneiderÕs type, Adults were transferred either to the Universidade which is deposited in the Museum of Comparative Zo- Estadual do Norte Fluminense (Campos dos Goyta- ology (Harvard University, Cambridge, MA). Voucher cazes, Brazil) or to the University of California (Davis, specimens from our study currently reside in the insect CA), where pairs were maintained in cages under collection at the Universidade Estadual do Norte Flu- controlled conditions (24 Ϯ 1ЊC, photoperiod of 16:8 minense and the research collection of M.J.T. and C.A.T. [L:D] h) and given a diet of yeast autolysate, fructose, and honey (1:1:1 volumetric mixture). We recorded Developmental Requirements. To examine the in- the color changes in 10 eggs from the Þrst laboratory ßuence of temperature on L. (L.) variaÕs developmen- generation (daily, oviposition to hatching). Another tal rates and survival, we conducted rearings at each Ϯ Њ 10 eggs were used for the measurements. of Þve constant temperatures (18Ð30 1 C; Table 1). Larvae were reared individually in 40-ml glass vials Twenty-Þve eggs in individual vials were placed in (sealed with plastic Þlm) on a diet of Anagasta kueh- each temperature regimen, and the immatures were niella (Zeller) eggs. Larval color was recorded during reared as described above. Development and deaths development. Specimens (10 per instar) were killed in were recorded daily. KAAD solution (Stehr 1987); after 30 min, they were Adult Color Variation. Most Þeld-collected adults transferred to 95% ethyl alcohol for preservation. Lar- could be assigned to one of four more-or-less discrete vae were killed at 48 h after hatching (Þrst instar) or color categories; specimens that typify the categories molting (second and third instars). Six specimens from were photographed and described. To examine the each instar were cleared in 10% KOH solution at 38ЊC nature of the variation, we made two sets of observa- for 1 d. Subsequently, they were stained with double tions: 1) We assessed whether the color variation is stain (pink lignin and fuchsin) for 1 to 4 d, depending age-related by observing the color patterns of adults on the instar; dehydrated in increasing concentra- from collection date to natural death in the laboratory tions of ethyl alcohol (70Ð100%); and preserved in (from 2 to 16 wk, average 7 wk). 2) We tested whether glycerin. Larvae were illustrated under a stereomi- laboratory-reared adults have phenotypes that are ma- croscope with a camera lucida (60ϫ magniÞcation ternally correlated (maternal effect) or mixed (pos- for second and third instars, 126ϫ magniÞcation for sible genetic involvement). F1 and a few F2 offspring Þrst instars). Measurements of head capsule width were made were reared from approximately three individual fer- across the widest part of the head and include the eyes; tile females from each category, that were collected head length was determined along the midline from in the forest (Tables 2 and 3). Rearing procedures clypeus to cervix. Body length was measured along the followed those above. We were able to rear only a curved midlateral line of distended specimens (tip of few second generation adults in the laboratory, and clypeus to tip of abdomen). Mandible length was mea- the low numbers precluded genetic analysis; how- sured along the center of the mandiblesÕ curved dorsal ever, the results provided evidence for several con- surface. All measurements were made with image clusions. January 2006 MANTOANELLI ET AL.: L. varia LARVAE AND COLOR VARIATION IN ADULTS 9

(Table 1. Duration of the immature stages and survival of L. (L.) varia reared under five constant temperature regimens (؎ 1°C (photoperiod of 16:8 ͓L:D͔ h)

Temp Stage 18ЊC21ЊC24ЊC27ЊC30ЊC Developmental time, x៮ Ϯ SD,d(n) Egg 10.5 Ϯ 0.76 (25) 8.9 Ϯ 0.64 (25) 6.2 Ϯ 0.37 (25) 5.5 Ϯ 0.51 (25) 5.0 Ϯ 0.00 (25) First instar 11.7 Ϯ 0.87 (24) 9.3 Ϯ 0.78 (22) 6.5 Ϯ 0.59 (24) 6.2 Ϯ 0.70 (21) 6.1 Ϯ 0.59 (15) Second instar 11.6 Ϯ 0.96 (19) 9.8 Ϯ 1.24 (13) 6.6 Ϯ 0.50 (16) 5.7 Ϯ 1.58 (18) 5.9 Ϯ 0.64 (8) Third instar 14.3 Ϯ 1.50 (9) 12.2 Ϯ 3.35 (5) 9.1 Ϯ 1.30 (11) 9.7 Ϯ 1.50 (9) 9.0 Ϯ 2.83 (2) Prepupa ϩ pupa 31.2 Ϯ 0.82 (5) 26.3 Ϯ 0.58 (3) 20.4 Ϯ 0.74 (8) (0) (0) Total development 79.8 Ϯ 1.09 (5) 67.0 Ϯ 1.00 (3) 48.3 Ϯ 0.89 (8) Survival, % (n) Egg to adult emergence 20.0 (25) 12.0 (25) 32.0 (25) 0 (25) 0 (25)

Generic Characteristics of Leucochrysa Larvae (ii) Thoracic tubercles with very long apical and lateral setae, apically arranged in a fan-shape (Figs. 2 The few Leucochrysa larvae that have been studied and 5). All three instars. to date (Smith 1926, Skorepa and Sharp 1971, Adams 1987, Tauber 2004), including L. (L.) varia, all carry (iii) Body: Anterior abdominal segments (A1ÐA5) large packets of trash on their dorsa. These larvae thicker (dorsum to venter) than thoracic segments express the morphological characteristics that are typ- but without an abrupt increase in height as is typical ical of trash-carrying chrysopids. That is, they have of Chrysopodes (metathorax and A1) and Ceraeochrysa gibbous, setose bodies; elongate thoracic tubercles; (A1 and A2) (Tauber et al. 2000, Tauber 2003). All well-developed abdominal tubercles; and elongate, three instars. serrated and/or hooked setae. (iv) Abdomen: Segments A2 and A3 with lateral Based on a comparative study of the larvae of four tubercles papiliform, bearing long setae on anterior North American species of Leucochrysa, subgenus surface (Fig. 3). In the four species described from Nodita, Tauber (2004) proposed a set of six features the United States, the trait is as stated, whereas in L. that distinguishes Leucochrysa larvae from other New (L.) varia the tubercles are papiliform (as described World chrysopid genera. L. (L.) varia larvae have the above), but the position of the long setae differs in entire set of six of characters, with one minor excep- that most are dorsal and apical. Second and third tion (as noted below in italics): instars. (i) Thorax with three pairs of very long lateral (v) Abdomen: Segments A4ÐA7 with lateral tuber- tubercles (i.e., the prothoracic lateral tubercles ex- cles slightly elongated, bearing long setae arising tend anteriorly at least to the midregion of the head; mainly from apex. All three instars. the meso and metathoracic tubercles are longer than (vi) Abdomen: Segments A7ÐA10 small, curved half the width of the respective segment) (Figs. 2 and ventrally and partially withdrawn into each other. All 5). All three instars. three instars.

Table 2. Color variation in field-collected L. (L.) varia

Collection site Phenotype (n, sex) (coordinates) Red, open Red, entire Black, open Black, entire Babiloˆnia farm 12 F, 25 M 7 F, 12 M 13 F, 26 M 18 F, 28 M (21Њ 51Ј S, 41Њ 48Ј W) Boa Vista farm 1 F, 1 M 1 F, 1 M 2 M 5 F, 2 M (21Њ 57Ј S, 41Њ 49Ј W) Pedrinhas farm 1M (21Њ 50Ј S, 41Њ 44Ј W) Santo Agustinho 1 F 2 M 1 F, 7 M 4 F, 4 M (22Њ 08Ј S, 41Њ 51Ј W) Santo Antoˆnio do Imbe´ 1F,6M 1M 1F,3M 2F,1M (22Њ 00Ј S, 41Њ 52Ј W) Sossego 1 F 1 M 1 M (21Њ 55Ј S, 41Њ 48Ј W) Terras Frias 2 F, 16 M 5 F, 7 M 5 F, 12 M 9 F, 5 M (21Њ 55Ј S, 41Њ 55Ј W) Sucupira farm 1 F, 1 M 1 M 1 F, 3 M (15Њ 54Ј S, 48Њ 01Ј W) Totala 65 (17 F, 48 M) 39 (15 F, 24 M) 72 (20 F, 52 M) 84 (39 F, 45 M)

All collection sites, except Sucupira farm (CENARGEN, near Nu´ cleo Bandeirante, Brasõ´lia, DF), are in or near Desengano State Park in northern Rio de Janeiro State, Brazil (Sept. 1998ÐFeb. 2005). a Sex ratio (F:M, total) ϭ 91:169. 10 ANNALS OF THE ENTOMOLOGICAL SOCIETY OF AMERICA Vol. 99, no. 1

Table 3. Color variation in L. (L.) varia rearings (24 ؎ 1°C, contiguous), conßuent with, but distinguishable from photoperiod of 16:8 [L:D] h) postfrontal markings; mesal section extending longi- tudinally from near mid-dorsal base of cranium almost Field-collected females to midregion of head; lateral section extending an- Maternal phenotype Offspring phenotype (n offspring) (n females) terolaterally from posterolateral margin of cranium to eye. Postfrontal marking prominent, dark brown me- Red, entire (2) Red, entire (2 females, 3 males) sally, brown distally, extending from anterior margin Red, open (3) Red, open (3 females, 1 male) Black, open (7 females, 2 males) of epicranial marking to antennal base. Anterior Black, entire (3) Black, entire (5 females, 4 males) (ϭfrontal ϩ intermandibular) marking brown, trian- Black, open (1 male) gular with sides extending from mid-region of head Black, open (3) Red, entire (2 females, 3 males) anteriorly almost to anterior margin of clypeus, with Black, entire (2 females, 2 males) Red, open (3 females, 5 males) pair of small dark brown markings on anterior tentorial Black, open (3 males) pits (mesal to base of antenna). Mandibles golden brown, with single lateral seta. Genae with small light Second generationa brown mark basally. Parental phenotype; Clypeus straight anteriorly, with right-angled cor- ϫ Offspring phenotype (n) female male (n pairs) ners; surface of corners rugose; dorsum with three Red, open ϫ red, entire (1) Black, open (1) pairs of apical setae (two lateral, one submesal). La- Red, entire ϫ red, open (1) Black, entire (1) brum heavily sclerotized and rounded anteriorly Red, entire ϫ black, Black, entire (2) entire (1) Black, open (1) (when everted, the labrum gives the anterior margin Black, entire ϫ black, Red, open (1) of the head a trigonate appearance). Tentorial pit entire (1) prominent. Setae S1ÐS12, Vx present, most brown; S11 long. Labial palpi amber; distal annulation of second a Twelve types of crosses were carried out, most with two pairs, but segment with one long, two short setae; remaining only four pairs produced offspring that reached the adult stage. annulations with approximately seven setae; basal seg- ment with one pale lateral, one brown mesal seta. Our current study leads us to propose Þve additional Palpiger with two brown ventral setae. Antenna with characters that further delineate Leucochrysa larvae scape brown; pedicel, ßagellum amber. Venter white, from other New World chrysopid larvae: unmarked, except ventral margin of head capsule (vii) Antenna: Terminal seta at least one-half length brown, junction of cardo and stipe, base of cardo with of ßagellum (Figs. 1 and 4). All three instars. small brown marks. Cervix cream colored, laterally (viii) Primary prothoracic setae S1, S3, S4, S5 with pair of very light brown marks, pair of brown present; S2 absent (Figs. 2 and 5). All three instars. setae. (ix) Abdomen: Segments A1ÐA5 each with pair of Thorax (Fig. 2). White to cream colored. Lateral laterodorsal tubercles each of which bears two long, tubercles (LTs) white to cream colored, sclerotized, smooth, hooked, laterodorsal setae (LDS) and a single arising from sclerotized bases, with spinules on sur- small, pointed microseta (Figs. 3 and 6). All three face, with long, pointed, smooth or very sparsely instars. thorned, golden-to-dark brown setae (LS). Dorsal se- (x) Abdomen: Segment A1 with single row of 8Ð16 tae smooth, pointed. Venter white to cream colored, (L3), 3Ð6 (L2), or four (L1) smooth, hooked, sub- unmarked; setae brown. Legs mostly white, except median setae (SMS) between the two laterodorsal coxopleural articulation with dark brown mark; base tubercles (Figs. 3 and 6). of coxa with very light brown shading; femur with light (xi) Abdomen: Segments A2ÐA5 each with three brown shading in dorsomedial band; tibiae, tarsi light (L3, L2) or two (L1) dorsal rows of smooth, hooked, amber. SMS, as follows. In each case, the posterior row of SMS Prothorax (T1): Anterior margin slightly convex. lies between the two laterodorsal tubercles. Pair of dorsal sclerites (Sc1) diamond shaped, light L3: Row 1 with 6Ð14 SMS; row 2 with 4Ð8 SMS; row brown laterally, slightly darker mesally; S1Sc1 long, 3 with 6Ð26 SMS (Fig. 3). thin, pointed; S2Sc1 short, anterior to S1Sc1. Sc2 trans- L2: Row 1 with 2Ð6 SMS; row 2 with four SMS; row parent (visible only with transmitted light), ovate, 3 with 2Ð10 SMS. approximately one-third length of Sc1, without asso- L1: Row 1 with four SMS; row 2 with two SMS (Fig. 6). ciated setae. LTs extending anteriorly to middle of eyes, each with 22Ð28 LS (10Ð14 apical, 12Ð16 lateral, dorsal). Primary seta S1, S3 intermediate size; S4, S5 Descriptions of Leucochrysa (L.) varia (Schneider) short; S2 absent. No secondary setae. Larvae Mesothorax (T2): Anterior subsegment with spi- Third Instar. Body. Cream colored; 6.9Ð8.1 mm in racles prominently raised, sclerotized, amber-colored length. Integument with numerous pointed spinules. dome-like cap apically. Spiracular chamber bulbous Setae golden to dark brown. distally, with basal stem; exterior surface of chamber Head (Fig. 1). Head 1.21Ð1.33 mm in width; 0.70Ð with horizontal rings; interior surface with scale-like 0.84 mm in length. Mandibles 1.32Ð1.44 mm in length. vertical striations. Sc1 small, transparent, on anterior Cream colored. Eyes black. Epicranial markings light margin, with three associated setae, one pair on the brown, entire (mesal and lateral sections on each side lateral side (S1Sc1, S2Sc1) and one on the mesal side January 2006 MANTOANELLI ET AL.: L. varia LARVAE AND COLOR VARIATION IN ADULTS 11

Fig. 1. L.(L.) varia head (third instar, dorsal). epi, epicranial marking; post, postfrontal marking; ant, anterior (ϭfrontal ϩ intermandibular) marking; S1Ð7, S11, S12, setae; Vx, three short posterior setae surrounding a pore.

(S3Sc1). Posterior subsegment with each LT bearing (A7ÐA10). LTs of segments 4Ð7 heavily sclerotized. Ϸ18Ð24 LS (10Ð12 apical; 8Ð12 lateral, dorsal); Sc2 A1ÐA7 each with pair of laterodorsal tubercles small transparent, on anterior margin, with two asso- (LDTs) bearing one pointed microseta, two long, usu- ciated setae (S1Sc2, S2Sc2); Sc3 large, mesal to LTs, ally hooked setae (LDS). transparent; S1Sc3 small; S2Sc3 absent; posterior re- A1: Spiracle prominent (raised, larger than those on gion with row (R1) of six, sometimes seven, long, thin, A2ÐA8), sclerotized, amber; SSp present. Spiracular hooked or pointed setae on chalazae. S1, S2 absent. chamber bulbous distally, stem-like basally; exterior Metathorax (T3): Anterior sclerites (Sc1) transpar- with horizontal rings throughout. LTs absent. Dorsum ent, small; S1Sc1 very small; no S2Sc1, but 1Ð2 small with single row of eight to 10 long, thin, hooked SMS secondary setae lateral to Sc1; mesolateral sclerites between LDTs. S1, S2 absent. (Sc2) small, transparent; S1Sc2 short; LTs each bear- A2ÐA4: LTs with seven to eight robust, long, hooked ing Ϸ20Ð23 LS (8Ð10 apical; 10Ð13 lateral); posterior LS apically and laterally, two to three short, pointed row (R1) of eight long, thin, hooked setae on chalazae. LS basally. Dorsum of each segment with three trans- S1 absent. verse rows of long, thin, hooked SMS, as follows: an- Abdomen (Fig. 3). Dorsum, venter white to cream terior row with six to eight (A2) or eight (A3, A4); colored, unmarked. Integument transparent. Dorsal mesal row with four; posterior row with 10Ð12 be- setae mostly long, thin, hooked (A1ÐA6) or pointed tween LDTs. SSp short, pointed. 12 ANNALS OF THE ENTOMOLOGICAL SOCIETY OF AMERICA Vol. 99, no. 1

Fig. 2. L.(L.) varia prothorax (A), mesothorax and metathorax (B) (third instar, dorsal). T1Ð3, thoracic segments; LT, lateral tubercle; LS, setae on lateral tubercle; S1, S3Ð5, setae; Sc1Ð3, sclerites; SxScx, setae associated with sclerites; Sp, spiracle; R1, row of setae.

A5: LTs each with Ϸ10 long LS laterally, three short anterior row with four straight SMS, posterior row LS at the base. Dorsum with three transverse rows of with two straight SMS between small LDTs. SSp short, SMS: anterior row with Þve to eight, mesal row with pointed. four, posterior row with four to eight setae between A8: LTs small, with three LS (two short, anterior; LDTs. SSp short, pointed. one slightly longer, posterior). Dorsum with two A6 and A7: LTs each with six long, robust LS lat- transverse rows of four and eight small SMS. SSp short, erally, four shorter LS basally. A6 with three trans- pointed. verse rows of dorsal setae: anterior row of two long, A9: Dorsum with three transverse rows of small hooked SMS, mesal row of four midsized, pointed setae (two anterior, four mesal, 6Ð8 posteromesal). SMS, posterior row of two to four SMS between small Venter with approximately six setae in two vertical LDTs. A7 with two transverse rows of dorsal setae: rows. January 2006 MANTOANELLI ET AL.: L. varia LARVAE AND COLOR VARIATION IN ADULTS 13

Fig. 3. L.(L.) varia abdominal segments 1Ð3 (third instar, dorsal). A1Ð3, abdominal segments; LT, lateral tubercle; LS, setae on lateral tubercle; LDT, laterodorsal tubercle; LDS, setae on laterodorsal tubercle; Sp, spiracle; SSp, seta associated with spiracle; SMS, submedian setae.

A10: Dorsum with two brown lateral sclerites; Ϸ10 (anterior row with two; mesal row with four; posterior short, straight, thin setae posteriorly, most on sides. row with two, between LDTs). A8: Dorsum with three Second Instar. Body. Body 4.3Ð6.1 mm in length. rows of small pointed setae (two in anterior row; four Similar to the third instar, with the following excep- in each of mesal, posterior rows). tions. First Instar. Body. Body 2.2Ð2.7 mm in length. Pre- Head. Head 0.76Ð0.80 mm in width; 0.47Ð0.51 mm dominantly white, with digestive tract visible through in length. Mandibles 0.81Ð0.94 mm in length. Dorsal transparent integument. Setae smooth; setae on head markings brown throughout, large, fused, covering and thorax pointed; abdominal setae pointed or entire surface except for an elongate, transparent hooked. Setae on thoracic and abdominal LTs dark (white) area basally. Genal markings extending from brown; remaining setae light brown to pale. cervical margin almost to eyes. Clypeus enlarged and Head (Fig. 4). Head 0.49Ð0.51 mm in width; 0.32Ð mildly trigonate in cleared, expanded specimens, but 0.35 mm in length; mandibles 0.50Ð0.55 mm in length. less so than in L3. Second segment of labial palpus with White, with brown markings; epicranial marking seven or eight setae: distal annulation with one long, paired, with darker slightly reddish brown pigmented two or three short, remaining annulations with four area beneath; frontal marking brown; intermandibular short. and postfrontal markings not distinguishable. Primary Thorax. LS smooth, dark brown, almost black. T1: cephalic setae present; S2, S5, S12 short; S1, S11 long. LTs with 20Ð22 LS (10Ð12 apical, Ϸ10 lateral, dorsal); Labial palpi cream colored basally, amber distally; a few basal LS on each LT hooked. Sc1 light brown distal annulation of second segment with one long, one throughout. T2: LTs with 15Ð17 LS (8Ð10 apical, short seta; remaining annulations without setae; basal approximately seven lateral, dorsal). T3: LTs with segment with two mid-length setae; distal segment 13Ð15 LS (approximately eight apical, six lateral, without setae. Mandibles golden, with single lateral dorsal). seta. Maxilla with very small setae apically. Antennae Abdomen. A1: transverse row with Þve to six SMS cream colored. Venter white. between LDTs. A2ÐA5: LTs each with approximately Thorax (Fig. 5). White; LTs elongate, white, scle- six LS. Dorsum of each segment with three transverse rotized, with sparse covering of small scale-like rows of long, hooked SMS as follows: anterior row with spinules; sclerites transparent, barely discernible on two; mesal row with four; posterior row with two to six cleared specimens. LS smooth, pointed. Prothorax between LDTs. A6, A7: LT with approximately seven (T1): LTs each with two long, dark brown to black LS, long LS; LDT sclerotized, light brown; dorsum of each one pointed microseta. Sc1 large; Sc2 not observed. segment with three transverse rows of pointed SMS S1Sc1 long, arising from chalaza. S1, S3, S4, S5 short, 14 ANNALS OF THE ENTOMOLOGICAL SOCIETY OF AMERICA Vol. 99, no. 1

Fig. 4. L.(L.) varia head (Þrst instar, dorsal). epi, epicranial marking; fr, frontal marking; S1Ð7, S11, S12, setae; Vx, three short posterior setae surrounding a pore. pointed; S4 very short; S2 absent. Mesothorax (T2): microsetae; LTs absent from A1. LDTs present on A1 Spiracles tuberculiform, prominent, with sclerotized, to A5, each with two long, hooked LDS, one short, dome-like apical cap; spiracular chamber bulbous. LTs pointed microseta. Spiracles present on A1ÐA8, each each bearing three long, dark brown to black LS. Sc1, with short, pointed SSp (except on A1). A1: Dorsum Sc2 small; associated setae (S1Sc1, S1Sc2, sometimes with single row of four long, hooked SMS arising from S2Sc1, S2Sc2) present, very small. Sc3 large, transpar- chalazae between LDTs. A2ÐA5: Dorsum of each seg- ent; S1Sc3 short to mid-sized. Posterior row with four ment with two transverse rows of long, curved, hooked setae, lateral-most large, arising from large chalazae. SMS: four in Þrst row; two in second row, arising from Metathorax (T3): LTs each with three long, dark chalazae between LDTs. SMS in anterior row of A5 brown to black LS. Sc1 very small, with associated seta small, LDS relatively long. A6: LTs relatively large. (S1Sc1) very small; Sc2 not discerned. Posterior row Two transverse rows of setae: anterior row with four (R1) with four long setae on chalazae. Legs setose; short, pointed SMS, posterior row with two long, white with dark mark at coxopleural articulation; coxa hooked SMS on chalazae, two very short, pointed with mottled light brown marking basally; tibia with SMS. A7: similar to A6, except dorsum without anterior two brown markings (basal and apical); claws brown; row of SMS; posterior row with SMS pointed. A8: LTs empodia cream. each with two LS: one very short, the other of inter- Abdomen (Fig. 6). LTs present on A2 to A7, each mediate size, both straight, pointed, dark brown to with two long, hooked, dark brown to black LS, no black. Dorsum with single row of four SMS. A9: six January 2006 MANTOANELLI ET AL.: L. varia LARVAE AND COLOR VARIATION IN ADULTS 15

Fig. 5. L.(L.) varia thorax (Þrst instar, dorsal). T1Ð3, thoracic segments; LT, lateral tubercle; LS, setae on lateral tubercle; S1, S3Ð5, setae; Sc1Ð3, sclerites; SxScx, setae associated with sclerites; Sp, spiracle; R1, row of setae.

Fig. 6. L.(L.) varia abdominal segments 1Ð5 (Þrst instar, dorsal). A1Ð5, abdominal segments; LT, lateral tubercle; LS, setae on lateral tubercle; LDT, laterodorsal tubercle; LDS, setae on laterodorsal tubercle; Sp, spiracle; SSp, seta associated with spiracle; SMS, submedian setae. 16 ANNALS OF THE ENTOMOLOGICAL SOCIETY OF AMERICA Vol. 99, no. 1 pairs of setae straight, pointed. A10: three pairs of L. (L.) varia required Ϸ1 wk longer to develop than straight, pointed setae, laterally. did Leucochrysa (N.) americana Banks, the only other Leucochrysa species for which developmental times have been reported. Development from oviposition to Species-Specific Characteristics of L.(L.) varia adult emergence for this species, reared at the same Larvae temperature, took 39Ð41 d (Tauber 2004), and as in L. Among the Þve species of Leucochrysa larvae that (L.) varia, development was not interrupted by dia- have been fully described (four in Tauber 2004 and pause. the one described here), L. (L.) varia larvae are the No L. (L.) varia individuals survived beyond the Њ least setose. For example, 1) the lateral tubercles on third instar at temperatures above 24 C, and few sur- Њ ϭ Њ the L. (L.) varia prothorax bear fewer than 30 setae, vived at temperatures below 24 C(n 5at18 and Њ compared with 38Ð44 on the other four species; 2) the three at 21 ) (Table 1). Given the high mortality rates posterior row of setae on the mesothorax has only six at all temperatures, we conclude that our rearing con- smooth, terminally hooked setae, compared with 12 or ditions, apart from temperature, were unsuitable for more on the other species; 3) secondary setae are larval growth and development. We fed our larvae the largely lacking, whereas most of the other species have eggs of A. kuehniella; we suspect that these relatively at least some, sometimes many secondary setae. L. (L.) small eggs did not offer sufÞcient nourishment (qual- varia larvae also are characterized by enlarged, tuber- ity and/or quantity) for L. (L.) variaÕs large second culiform spiracles on T2 and A1; these spiracles are and third instars. well sclerotized and have a bulbous, stemmed cham- ber. L. (L.) varia larvae also have distinctive head Adult Color Variation markings (Figs. 1 and 4), and setae that are golden to dark brown and always smooth. Descriptions of Phenotypes. L. (L.) varia adults Note on Asymmetry. We observed two asymmetri- have distinctive variation in the red and black mark- cal features of L. (L.) varia larvae. First, in 17 of 18 ings on the head (frons and vertex), meso- and met- larvae that we measured, the right mandible was anota, and Þfth and sixth abdominal tergites. The color slightly longer than the left (L3: left, 1.32Ð1.41 mm; of other parts of the body is relatively uniform among right, 1.37Ð1.44 mm; L2: left, 0.81Ð0.88 mm; right, 0.83Ð specimens. For example, the prothorax and abdominal 0.94 mm; L1: left, 0.50Ð0.54 mm; right, 0.53Ð0.55 mm). segments are generally yellowish to green, and with Second, there was often an uneven number of sub- the exception of abdominal segments 5 and 6, they do median setae on the dorsum of the abdomen; we did not have remarkable markings or color variation. Usu- not determine whether one side or the other consis- ally, younger (teneral) individuals are more yellowish, tently had the higher number. Our (C.A.T.) observa- whereas older individuals are darker green. The meso- tions of other taxa indicate that these types of asym- and metathoracic pleuron and venter have a light metry may not be uncommon among chrysopid larvae. cream color on all of our specimens. The color variation is particularly conspicuous on the meso- and metanota. All of the adults that we Egg collected or reared could be assigned to four catego- ries (Fig. 7) based on two character states for each of Ovoid, 0.87Ð0.93 mm in length; 0.50Ð0.56 mm in two characters related to the meso-metanotal mark- width; stalked; laid singly on the substrate, without any ings: 1) the predominant color of the markings (black speciÞc pattern; micropyle ßat, disc-shaped, white. or red) and 2) the pattern of the markings (entire or Stalk hyaline, 6.2Ð7.4 mm in length, without droplets. open). Oviruptor crescent shaped, without teeth. 1) Black, entire (Fig. 7A): All mesonotal and met- Eggs are light green when oviposited and they re- anotal sclerites black, except metascutellum light Ϸ main light green for 24 h. On the second day, three green or yellow, occasionally black. light brown bands develop; on the third day, the entire 2) Black, open (Fig. 7B): Same as black, entire center portion of the egg becomes brown, and the except that, in addition to the metascutellum, the poles assume a light yellow color. Subsequently, the mesoscutellum is also light green, yellow or white. developing larva is visible through the transparent 3) Red, entire (Fig. 7C): All meso- and metanotal chorion. sclerites rosy red, except metascutellum light green or yellow, occasionally rosy red. 4) Red, open (Fig. 7D): Same as red, entire except L.(L.) varia Biology both the mesoscutellum and metascutellum green, Development and Survival in Relation to Temper- yellow or white. Often, posterior part of mesopres- ature. At 24 Ϯ 1ЊC, eight of 25 individuals survived cutum and mesal section of mesoscutum also light from the egg to the adult stage; development (ovipo- green or yellow. sition to adult emergence) took 48.3 Ϯ 0.3 d (Table 1). Several characteristics of the color variation are At this temperature, the egg required Ϸ6 d, the larval noteworthy: 1) Although most specimens can be easily stage Ϸ22 d, and the cocoon Ϸ20 d. Most of the categorized as predominantly black or red, consider- mortality occurred after the Þrst instar. There was no able intergradation in color exists. In contrast, we evidence of diapause or arrested development. observed no intergradation in the coloration of the January 2006 MANTOANELLI ET AL.: L. varia LARVAE AND COLOR VARIATION IN ADULTS 17

Fig. 7. L.(L.) varia adult color variation. (A) Black, entire. (B) Black, open. (C) Red, entire. (D) Red, open. mesoscutellum; it was either completely colored red white frons, to a white frons surrounded by red, to a or black (entire) or unmarked green, yellow, or white completely red frons. The vertex varies from largely (open). 2) Both black and red individuals may have red to largely green. In general, individuals that fall either entire or open patterns. 3) In black individuals, into the black category have darker facial markings the open pattern mainly affects the mesoscutellum, than those in the red category, but the correlation is whereas in the red morphs the open pattern also may not without exception. extend anteriorly through the middle of the mesoscu- Field Collections. The black, entire phenotype was tum and posterior section of the mesoprescutum. 4) most common in our Þeld collections, followed by The markings on the Þfth and sixth abdominal tergites black, open and red, open (Table 2). All four catego- vary from entirely rosy red without any black, to dark ries included both males and females. It is noteworthy red with a black central spot. The variation in abdom- that for all sites except one, the sex ratio of our col- inal markings generally corresponds with the meso- lections was strongly biased toward males (combined and metanotal markings. That is, red, entire or open collections, F:M 1:2; n ϭ 260); however, the same bias individuals have red abdominal markings, whereas was not observed in our laboratory rearings (F:M black, entire or open individuals have the central spot 26:24; Table 3). We conclude that the male bias in our black. 5) The variation in facial color ranges from a Þeld collections resulted either from disproportionate 18 ANNALS OF THE ENTOMOLOGICAL SOCIETY OF AMERICA Vol. 99, no. 1 female mortality in the Þeld, male bias in the lower Zoology) for facilitating the loan of the L.(L.) varia type. The story of the forest where we sampled (within Ϸ10 feet following people helped collect specimens or facilitated our of the ground), or male bias in our collecting tech- Þeldwork and/or laboratory rearings: E. A. Silva, J. S. Multani, niques (netting specimens after we observed or dis- P. J. Tauber, A. J. Tauber, M. J. Tauber, E. M. G. Fontes, C. S. S. turbed them in the forest). Pires, E. R. Sujii, V. Becker, and P. S. Silva; it is a pleasure to acknowledge this help, and we express our gratitude to them all. Results from Rearing, Inheritance. Our rearings conÞrm that the color differences we observed in our Þeld-collected specimens are an expression of intraspe- ciÞc variation, rather than species differences. This con- References Cited clusion is consistent with previous Þndings of intraspe- Adams, P. A. 1977. of United States Leucochrysa ciÞc color variation in adults of other Leucochrysa species (Neuroptera: Chrysopidae). Psyche 84: 92Ð102. (Adams 1977, Tauber 2004), and it underscores the im- Adams, P. A. 1987. Studies in neotropical Chrysopidae portance of using characters in addition to adult color (Neuroptera) III. Notes on Nodita amazonica Nava´s and (e.g., male and female genitalia, larval characteristics) in N. oenops, n.sp. Neuroptera Int. 4: 287Ð294. identifying new species of Leucochrysa. Adams, P. A., and N. D. Penny. 1986. Faunal relations of Our rearings also provided some insight into the Amazonian Chrysopidae, pp. 119Ð124. In J. Gepp, H. nature of the variation. First, in all of our rearings, we Aspo¨ck, and H. Holzel [eds.], Recent research in Neu- never observed time- or age-related changes in the ropterology. Druckhaus Thalerhof, Graz, Austria. Brooks, S. J., and P. C. Barnard. 1990. The green lacewings adult thoracic markings; that is, time in the laboratory of the world: a generic review (Neuroptera: Chrysopi- (up to 16 wk) had no discernible effect on either the dae). Bull. Br. Mus. Nat. Hist. (Entomol.) 59: 117Ð286. color or the pattern of the thoracic markings. de Freitas, S., and N. D. Penny. 2001. The green lacewings Second, the color is not maternally correlated. That (Neuroptera: Chrysopidae) of Brazilian agro-ecosystems. is, individual black and red females produced both Proc. Calif. Acad. Sci. 52: 245Ð395. black and red offspring (Table 3). Similarly, individual Dı´az-Aranda, L. M., and V. J. Monserrat. 1995. Aphidophagous entire and open patterned females produced both predator diagnosis: key to genera of European chrysopid entire and open patterned offspring. larvae (Neur.: Chrysopidae). Entomophaga 40: 169Ð181. Third, the few offspring that we obtained from our Rousset, A. 1966. Morphologie ce´phalique des larves de Pla- nipennes (Insectes Ne´vropte´roõ¨des). Me´m. Mus. Nat. pairings of laboratory-reared adults indicate that the Hist. Ser. A 42: 1Ð199. color aspect of the variation is not a simple dominant- Skorepa, A. C., and A. J. Sharp. 1971. Lichens in “packets” of recessive trait. Our pairings of red females with red lacewing larvae (Chrysopidae). Bryologist 74: 363Ð364. males produced black offspring; similarly, the black Smith, R. C. 1926. The trash-carrying habit of certain lace- female and black male pairing produced a red off- wing larvae. Sci. Mon. 23: 265Ð267. spring. This Þnding was not unexpected, given the Stehr, F. W. 1987. Immature , vol. 1. Kendall/Hunt, continuous range of color variation (from red to inter- Dubuque, IA. mediate to black) that we observed in our specimens. Tauber, C. A. 1974. Systematics of North American chry- Fourth, the same pairings of laboratory-reared sopid larvae: Chrysopa carnea group (Neuroptera). Can. Entomol. 106: 1133Ð1153. adults show that the entire pattern is not a simple ϫ Tauber, C. A. 2003. Generic characteristics of Chrysopodes recessive trait (entire female entire male pairings (Neuroptera: Chrysopidae), with new larval descriptions produced both entire- and open-patterned offspring). and a review of species from the United States and Can- We did not have sufÞcient pairings to make a conclu- ada. Ann. Entomol. Soc. Am. 96: 472Ð490. sion regarding the open pattern. However, we plan to Tauber, C. A. 2004. The genus Leucochrysa (Neuroptera: pursue this and related questions about the adult color Chrysopidae) in the United States: a systematic review. variation in later studies. Ann. Entomol. Soc. Am. 97: 1129Ð1158. Tauber, C. A., and T. de Leo´n. 2001. Systematics of green lacewings (Neuroptera: Chrysopidae): larvae of Ceraeo- Acknowledgments chrysa from Mexico. Ann. Entomol. Soc. Am. 94: 197Ð209. Tauber, C. A., T. de Leo´n, N. D. Penny, and M. J. Tauber. 2000. Our work was supported by the National Science Foun- The genus Ceraeochrysa (Neuroptera: Chrysopidae) of dation (Grants INT 9817231, INT-9912449), the National America north of Mexico: larvae, adults, and comparative Geographic Society, the “Conselho Nacional de Desenvolvi- biology. Ann. Entomol. Soc. Am. 93: 1195Ð1221. mento Cientõ´Þco e Tecnolo´gico” (CNPq, Brazil, Grants Tauber, C. A., M. J. Tauber, and G. S. Albuquerque. 2001. 300504/96-9 and 466439/00-8), the National Research Initia- Plesiochrysa brasiliensis (Neuroptera: Chrysopidae): lar- tiveÐUSDA Cooperative State Research Education and Ex- val stages, biology, and taxonomic relationships. Ann. tension Service (Grants 9802447 and 2002-35316Ð12231), Re- Entomol. Soc. Amer. 94: 858Ð865. gional Project W-1185, Cornell University, and Universidade Tsukaguchi, S. 1995. Chrysopidae of Japan (Insecta, Neu- Estadual do Norte Fluminense. C.A.T. and M.J.T. thank L. E. roptera). S. Tsukaguchi, Aioi-cho 6-14-102. Nishinomiya- Ehler, R. E. Page, D. E. Ullman, S. Kelley, L. S. Kimsey, S. shi, Hyogo, 662 Japan. Soucy-Lubell, and the Department of Entomology, University of California, Davis, for help and cooperation in a variety of ways. We also thank P. D. Perkins (Museum of Comparative Received 21 April 2005; accepted 5 July 2005.