Systematic revision of Macrotingis and phylogenetic analysis of the genera Macrotingis and Ceratotingis (Heteroptera: Tingidae)

The Central American genus Macrotingis Champion, 1897 was revised and the morphological characters of the species of Macrotingis and those of its sister group Ceratotingis Montemayor, 2008 were subjected to a phylogenetic analysis. A species previ­ ously placed in the genus Tigava is now Ceratotingis spatula (Monte, 1945), new combination. Cladistic analysis indicates that both genera are monophyletic and defined the relationships between species. Keys to the species in these genera are provided together with illustrations of their main morphological characters and a map of their distribution in Central America.


INTRODUCTION
The lace bug genus Macrotingis Champion is restricted to Central America and currently includes M. biseriata Champion, 1897, M. novicis Drake, 1928, M. schaffneri Froeschner, 2003and M. uniseriata Champion, 1897. Recently Montemayor (2008) erected another Central American genus: Ceratotingis, which includes Ceratotingis zeteki (Drake, 1950), transferred from Macrotingis, and two new species, C. costarriquense Montemayor, 2008 andC. rafaeli Montemayor, 2008, andnoted the affinities between these two genera.The most comprehensive study of the genus is that of Froeschner (2003).In this study he describes a new species, M. schaffneri, elevates to species status M. novicis and provides a key to the species of Macrotingis.
In the present contribution Tigava spatula is transferred to the genus Ceratotingis, which constitutes the first record of the genus for South America.A comprehensive systematic revision of Macrotingis was carried out, the monophyly of Ceratotingis and Macrotingis confirmed and the relationships between species defined.Illustrations, photographs and a distributional map are provided.A new key to species of Macrotingis is presented, which includes new characters, and a key to species of Ceratotingis is adapted by incorporating C. spatula comb.n.

MATERIAL AND METHODS
The terminology for the morphology follows Drake & Davis (1960).Measurements (Table 1) were made using an ocular micrometer and are in millimetres, and "*" is used to indicate the measurement was repeated.When at least five specimens were measured minimum, maximum and mean values are provided; when four or fewer specimens were measured values for each individual are provided.Photographs were taken using a Sony DSC-W7 camera attached to a Wild M-5 stereomicroscope.The specimens studied belong to the Institut Royal des Sciences Naturelles de Belgique, Bruxelles, Belgique (IRSNB); The Museum of Natural History, Smithsonian Institution (NMNH); The Natural History Museum, London, England (BMNH) and The Museu Nacional de História Natural da Universidade Federal do Rio de Janeiro (MNRJ).The labels of examined specimens were transcribed in the section "Material examined", where "ND" indicates it was impossible to determinate the sex of the specimen.
Diagnosis.Antennae longer than body; segment I at least 4 times the length of the head; bases of segment I widely separated; segment IV spatulate; hood well developed, compressed; one long cephalic spine; pronotum with one or three low carinae and lacking areolae; lateral pronotal carinae when present only developed on posterior process; lateral margins of hemelytra serrate, with setae; discoidal area short, not reaching the middle of the hemelytra.
Redescription.Head short, broad.One median, unpaired, long cephalic spine.Bucculae in contact apically, moderately broad.Eyes oval.Rostrum reaching at least the posterior half of mesosternum.Antennae extremely long and slender, at least 1.1 times the length of the body; segment I very long, between 4.4 to 7.1 times the length of the head; segment II short; segment III very long; segment IV spatulate.
Pronotal disc convex, shiny.Collar raised in a high, compressed hood, projected forward.Paranota wide, margins with setae.One or three carinae very low, lateral carinae when present only developed on posterior process.Posterior process long, tapering gradually to a sharp point.Rostral laminae composed of one row of areolae, high, mesosternal laminae more widely separated anteriorly than posteriorly, metasternal laminae closed posteriorly.
Hemelytra narrow, widened distally, extending considerably beyond the apex of abdomen; margins serrated, with setae.Costal area wide, composed of one or two rows of areolae.Subcostal area subvertical, narrow, composed of a variable number of rows of areolae.Discoidal area short, not reaching the middle of the hemelytra.Sutural area narrow, with areolae becoming larger and more irregular posteriorly.Hypocostal area with one row of areolae.Legs yellowish, long, slender.
Discussion.Macrotingis as well as Ceratotingis are closely related to Tigava Stål, a genus distributed in Central and South America.The general appearance of Tigava is very similar to Macrotingis and Ceratotingis.In these three genera the body is long and slender, the legs and antennae are thin and long, and segment I is particularly long.This last character is very infrequent among Tingidae and is a synapomorphy shared by these genera.Macrotingis is the sister group of Ceratotingis and shares, among other characters, antennae longer than the body, segment IV spatulate and cephalic spines long.The main characteristics that distinguish these two genera are the number of cephalic spines and presence of pronotal carinae.Macrotingis has one cephalic spine, the pronotal carinae are low and the lateral carinae are absent or only present on the posterior process, whereas Ceratotingis has three cephalic spines, the pronotal carinae are high and the lateral carinae are present on the pronotum.The main characters that distinguish Tigava from Macrotingis and Ceratotingis are antennae shorter or a little longer than the body, segment IV cylindrical and short cephalic spines.(5.08) 5. 13-5.46 (5.34) 5.59-5.13 -5.53 5.19-5.33 -5.33-5.46 5.19-5.59 (5.41) 5.25-5.66 (5.39)Total length than those in inner row; thoracic sterna black; mesosternal rostral laminae converge gradually posteriorly; costal area with two rows of areolae; abdomen bicoloured, pre-genital segments black, genital segments red brown.

Key to species of Macrotingis
Redescription.Total body length (females N = 5 and males N = 5, respectively): 5. 25-5.66 (5.39) 5.19-5.59 (5.41).General colour yellow brown.Head red brown.Cephalic spine slender, erect, covered with disperse, long, erect setae (Figs 2A,3A).Clypeus reddish brown or dark brown.Bucculae yellow, with three rows of areolae, similar width over all its length.Rostrum yellow except for the brown apex, long, reaches the meso-metasternum.Antennae 1.2 to 1.3 times the length of the body; segment I between 5.0 and 6.1 times the length of the head, with abundant long, semi-erect setae; segment II subconic, glabrous; segment III with abundant long semidecumbent setae and some short semierect setae; segment IV with abundant long, semidecumbent setae and some scattered short, semierect setae.
Pronotal disc red brown, coarsely pitted, three carinae (Fig. 4A).Hood projects as far as anterior half of eyes, surface with long, semi-decumbent scattered setae (Fig. 3A).Callus dark brown, depressed, with abundant long, semidecumbent setae.Paranota sub-vertical, margins serrated with long disperse setae; with two rows of areolae, areolae in outer row bigger than those in inner row.Posterior process yellow.Thoracic sterna black.Rostral laminae: mesosternal laminae convex, maximum separation in anterior third, converge gradually posteriorly; metasternal laminae longer than space separating both laminae.
Hemelytra (Fig. 5A): costal area subhorizontal, with two rows of areolae, areolae big, especially posteriorly.Subcostal area with four rows of medium sized areolae.Discoidal area at widest part with four rows of small areolae.
Discussion.Macrotingis biseriata can be distinguished from the other members of the genus by its very long rostrum, which reaches the meso-metasternal suture, and it's entirely biseriated costal area.In the rest of the species the rostrum reaches the posterior half of the mesosternum and the costal area is uni-biseriated or entirely uniseriated.3B).Clypeus black.Bucculae yellow, with three rows of areolae, posteriorly wider.Rostrum yellow except the brown apex, moderately long, reaching up to posterior half of mesoesternum.Antennae 1.2 to 1.3 times the length of the body; segment I between 6.0 and 7.1 times the length of the head, with short, semierect setae; segment II subconic, with short, semierect setae; segment III with scattered long semidecumbent setae; segment IV with abundant long, semidecumbent setae and some scattered short, semierect setae.
Hemelytra (Fig. 5B): costal area subhorizontal, anterior and posterior thirds with one row of areolae, medial third with two, areolae larger posteriorly.Subcostal area with two rows of areolae, areolae of medium size.Discoidal area at its maximum width with five rows of small areolae.
Discussion.Macrotingis schaffneri has several unique features, such as one pronotal carina and the anterior paranotal margins forming an acute angle with the hood.The rest of the Macrotingis species have three pronotal carinae and the anterior paranotal margins form a right angle with the hood.
Redescription.Total body length 5.13.General colour yellow brown.Head reddish brown.Cephalic spine slender, erect, glabrus (Figs 2D, 3D).Clypeus dark brown.Bucculae yellow, with three rows of areolae, similar width over all its length.Rostrum yellow except the black apex, moderately long, reaching posterior half of mesoesternum.Antennae 1.2 times the length of the body; segment I 4.5 times the length of the head, with short, semierect setae; segment II subconical, glabrus; segment III with abundant long semierect setae; segment IV with abundant long, semidecumbent setae and some scattered short, semierect setae.
Pronotal disc dark brown, coarsely pitted, three carinae (Fig. 4D).Hood projecting as far as the middle of eyes, glabrus (Fig. 3D).Callus dark brown, depressed, with abundant long, semidecumbent setae.Paranota subvertical, margins serrated, glabrus; with two rows of areolae with those of the outer row bigger than those of the inner row.Posterior process yellowish brown.Pro and metasternum brown, mesosternum black.Rostral laminae: mesosternal laminae converge gradually posteriorly; metasternal laminae longer than space separating the laminae.
Hemelytra (Fig. 5D): costal area subhorizontal, with one row of areolae, towards the middle of this area there may be some extra areolae, areolae increase in size posteriorly.Subcostal area with three rows of areolae, areolae small.Discoidal area at its maximum width with three rows of small areolae.
Discussion.Macrotingis uniseriata, like M. novicis and M. biseriata, has a pronotum with three carinae and biseriate paranota, and has the same rostral length and unicoloured abdomen as M. schaffneri.It can be distinguished from the other members of the genus by its uniseriated costal area and red brown pro and metasternum and black mesosternum.

Genus Ceratotingis Montemayor, 2008
Ceratotingis Montemayor, 2008:  Diagnosis.Antennae longer than body; segment I at least 3 times the length of the head; bases of segment I widely separated, segment IV spatulate; hood well developed, compressed; three cephalic spines, occipital pair very long reaching the bases of segment I; pronotum with three high carinae and areolated; lateral pronotal carinae fully developed; lateral margins of hemelytra serrated and with setae., 31.i.1978, collected on Phaseolus vulgaris (NMNH).
Discussion.Ceratotingis costarriquense like C. rafaeli and C. spatula has a uniseriated costal area, but it can be distinguished from C. rafaeli by its wider paranota and shorter segment I and from C. spatula by its hump, higher hood and longer discoidal area.Diagnosis.General colour yellowish brown.Cuticle anterodorsal to eyes with same characteristics as the rest of the head.Occipital spines gradually diverging.Segment I approximately four times longer than the head.Hood moderately developed.Median pronotal carina same height along all its length except at the apex where it is lower.Paranota with one row of areolae.Mesosternal lamina subparallel.Costal area with one row of areolae.
Discussion.Ceratotingis spatula was previously placed in the genus Tigava.The members of the genus Tigava have the bases of segment I adjacent, a cylindrical segment IV, segment I and IV of the same length, median spine is short and directed forwards, occipital spines are subparallel and run close to the surface of the head, a generally short rostrum that does not extend beyond the prosternum, a low hood that scarcely projects over the head and low pronotal carinae.
In C. spatula, like in other species of Ceratotingis, the bases of segment I are widely separated, segment IV is spatuliform, segment I is much longer than segment IV, median spine is long and directed forward, occipital spines form an angle with the surface of the head, the rostrum is long and reaches at least the posterior half of the mesosternum, the hood is well developed, projects over the head and the pronotal carinae are high.Ceratotingis spatula is the sister group of C. costarriquense, C. rafaeli and C. zeteki; and like the first two species has a uniseriated costa on which, like the latter species, the occipital spines gradually diverge and a median pronotal carina of the same height along its entire length.
Ceratotingis spatula can be distinguished from the rest of the Ceratotingis by its rostrum, which reaches the metasternum, a lower hood, paranota with only one row of areolae, shortness and shape of the discoidal area and the portion of the inner discoidal vein that reaches the subcostal vein is curved whereas in all other Ceratotingis it is rect.
Up to now this is the only species of the genus in South America and there is a large gap in the distribution between the Central American species and C. spatula (Fig. 7).Probably this is a consequence of the lack of field work in this area as the range of the genus should be much wider than that recorded.(Drake, 1950) (Figs 2H, 3H, 4H, 5H and 6D) Macrotingis zeteki Drake, 1950: 299;Drake & Ruhoff, 1965: 294;Froeschner, 2003: 29.Ceratotingis zeteki : Montemayor, 2008: 448.Diagnosis.General colour yellowish brown.Cuticle anterodorsal to eyes with same characteristics as the rest of the head.Occipital spines gradually diverging.Segment I approximately five times longer than the head.Hood well developed.Median pronotal carina same height along its entire length.Paranota with two rows of areolae.Mesosternal lamina subparallel.Costal area with two rows of areolae.
Discussion.Ceratotingis zeteki can be distinguished from other members of the genus as it has a biseriate costal area.

CLADISTIC ANALYSIS
Phylogenetic analysis was performed on a matrix (Table 2) comprising 21 morphological characters and 10 taxa, five species of Macrotingis, five species of Ceratotingis, one species of Tigava and one of Vatiga.Morphological characters are those of the head, thorax and abdomen, and coloration patterns.Tree searches were performed in TNT (Goloboff et al., 2000).Exact searches were performed by implicit enumeration.All characters were considered to be of equal weight, with multistate characters being treated as unordered.Characters were polarised using the outgroup Tigava praecellans Stål and Vatiga vicosana Drake & Hambleton.

Cladistic analysis
From the parsimony analysis of the data matrix (Table 2) one most parsimonious tree (Fig. 8) was obtained with a length of 36, a consistency index of 80 and a retention index of 81.The analysis recovered Ceratotingis and Macrotingis as monophyletic taxa.Both genera are related and share six character states of the head, prothorax and hemelytra: antennae longer than the body (1:1); segment IV spatulate (2:1); bases of segment I widely separated (4:1); rostrum reaching posterior half of mesosternum (9:1); hood well developed (10:2); and medial areolae on costal area subrectangular (17:1).
Ceratotingis was recovered as a monophyletic group based on characters of the head and prothorax: median spine long and porrect (7:1); occipital spines divergent (8:1); pronotal carinae high (14:1).Within this clade C. spatula comb.n., which was previously placed in the genus Tigava, is a sister species of the remaining group, C. rafaeli and C. costarriquense constitute a clade whose sister species is C. zeteki.Ceratotingis costarriquense and C. rafaeli are related by one synapomorphy the presence of a hump on the median pronotal carina (15:1).This last clade is related to C. zeteki by one synapomorphy and one homoplasy, these are in order, the distal portion of the cubital vein that reaches the radius media vein is straight (18:1) and the discoidal area is 0.45 or more times the length of the hemelytra (19:0).
In most of the species analyzed (Fig. 5A-B, G, I, J) the distal portion of the Cu vein that reaches the Radius Media vein (RM) is curved.In the case of C. costarriquense (Fig. 5E), C. rafaeli (Fig. 5F) and C. zeteki (Fig. 5H) it is straight.20.Length of discoidal area: 0.45 or more times the length of the hemelytra (0); 0.40 or less times the length of the hemelytra (1).The discoidal area is long, 0.45 or more times the length of the hemelytra in V. vicosana, C. costarriquense, C. rafaeli and C. zeteki, and short, 0.40 or less times the length of the hemelytra in all the species of Macrotingis, T. praecellans and C. spatula.
Abdomen 21.Abdomen: unicoloured (0); bicoloured (1).Of the species studied only two have a bicoloured abdomen: M. biseriata and M. novicis, in both these species the pre-genital segments are black and the genital segments reddish brown.22. Length of pygophore relative to total abdominal length: 0.23 times or less the abdominal length (0); 0.26 times or more the abdominal length (1).The three species that have a pygopore that is 0.26 times or more the abdominal length are M. biseriata, M. novicis and M. schaffneri, whereas in all the other species it is 0.23 times or less the abdominal length.For T. praecellans and C. costarriquense the character was coded as "?" because we were only able to study female specimens.

TABLE 1 .
Measurements of species of Macrotingis and Ceratotingis spatula comb.n.

TABLE 2 .
). Black circles indicate unique changes and white circles homoplasies.Data matrix used in the phylogenetic analysis.