BLUMEA 51: 165–195 Published on 10 May 2006 http://dx.doi.org/10.3767/000651906X622427 ANALYSIS OF GENERIC RELATIONSHIPS IN ANACARDIACEAE B.S. WANNAN Environmental Protection Agency, P.O. Box 802, Atherton 4883, Queensland, Australia SUMMARY Cladistic analyses were undertaken of Anacardiaceae using non-sequence data (30 genera and 81 characters from morphology, anatomy, palynology and chemotaxonomy), sequence data (26 genera – rbcL) and a combined dataset of 16 genera. All analyses supported a group of genera which can be recognised at the subfamily level: Anacardioideae. Sequence data and combined datasets supported the recognition of a second subfamily: Spondiadioideae Kunth emend. Wannan. Both datasets also suggested that Buchanania lies outside both subfamily groups. Key words: Anacardiaceae, cladistics, phylogeny, rbcL sequence data. INTRODUCTION The Anacardiaceae is a well recognised world-wide family of mostly tropical trees which has historically been placed in the Sapindales or Rutales (Bentham & Hooker, 1862; Takhtajan, 1980; Dahlgren 1980, 1983, 1989; Cronquist, 1981; Angiosperm Phylo- geny Group, 1998; Judd et al., 1999). Cronquist (1981) placed it with the Julianiaceae and Burseraceae, the three being the only families in the Sapindales with biflavonyls and vertical intercellular secretory canals in the primary and secondary phloem. The close relationships of the Anacardiaceae and Burseraceae have been recently reiter- ated by analysis of rbcL and atpB sequence data (Gadek et al., 1996; Savolainen et al., 2000a). The Burseraceae are distinguished by having two epitropous ovules per locule, in contrast to the one apotropous ovule in the Anacardiaceae. The Burseraceae also fre- quently possess lobed cotyledons in contrast to entire cotyledons in the Anacardiaceae. Amphipterygium and Orthopterygium, once a separate family (Julianiaceae), are now considered part of the Anacardiaceae based on molecular and non-molecular data (Peterson & Fairbrothers, 1983; Wannan & Quinn, 1988, 1990, 1991; Angiosperm Phylogeny Group, 1998; Judd et al., 1999, Savolainen et al., 2000b). The Anacardiaceae is generally considered to constitute about 70 genera and 600 species which are concentrated in the tropics of Africa, Asia and America with a smaller number of species occurring in subtropical and temperate areas. A number of infrafamilial classifications have been proposed in the Anacardiaceae (Bentham & Hooker, 1862; Marchand, 1869, 1874; Engler, 1883, 1892), but the most widely used for the last 100 years has been the five tribes of Engler (1883, 1892, 1897) which are based on floral characters and leaf dissection. A recent division of the family into five subfamilies by Takhtajan (1987) appears not to have been well accepted. © 2006 Nationaal Herbarium Nederland, Leiden University branch 166 BLUMEA — Vol. 51, No. 1, 2006 A range of systematic studies have tested the applicability of Engler’s tribal classifi­ cation, the most comprehensive having used stem anatomy (Jadin, 1894) or wood anatomy (Heimsch, 1942; Dadswell & Ingle, 1948; Kryn, 1953). These found support, with some reservations, for Engler’s tribes. Young (1976) looked at the wood flavonoids of 16 genera of the Anacardiaceae (including Julianiaceae), in the tribes Anacardieae, Rhoeae and Spondiadeae, as well as representatives of the Burseraceae, Rutaceae, Simaroubaceae, Cneoraceae, Meliaceae, Sapindaceae, Aceraceae, Hippocastanaceae and Juglandaceae. He found that there was a range of 5-deoxyflavonoids which was restricted to the Anacardiaceae (including Julianiaceae), but there were no clear pat- terns at a tribal level. More recently, Wannan & Quinn (1990, 1991) described the pericarp and floral morphol- ogy in 30 genera sampling all tribes in the family. They found that the distribution of reproductive, vegetative and secondary product character states did not closely reflect the subfamily taxonomies of either Engler (1883, 1892) or Takhtajan (1987). Rather, they found support for two informal groups, but suggested that these required further study to confirm their status. The first (Group A) included Engler’s tribes Anacardieae (without Buchanania), Rhoeae (without Pentaspadon and Campnosperma), Dobineeae and Semecarpeae, and the second (Group B) included the Spondiadeae but with the addition of Buchanania, Pentaspadon and Campnosperma. Work by Von Teichman and associates has confirmed the importance of pericarp structure for illucidating generic affinities in the family (Von Teichman & Robbertse, 1986a, b; Von Teichman, 1987, 1990, 1991, 1992, 1993; Von Teichman & Van Wyck, 1988, 1994). Recent studies of seed anatomy (Pienaar & Von Teichman, 1998) and wood anatomy (Dong & Baas, 1993) have also provided support for Wannan & Quinn’s (1991) groups. Support for Wannan & Quinn’s (1991) two informal groups has also been provided by an unpublished analysis of anatomical, morphological and rbcL sequence data across 17 genera (Terrazas & Chase, 1996). Their conference abstract reported two clades, broadly corresponding to Wannan & Quinn’s groups. Some of their sequence data were included in a molecular analysis of the Sapindales (Gadek et al., 1996) which used 7 genera from the Anacardiaceae and three from the Burseraceae. This analysis showed the Anacardiaceae and Burseraceae as sister groups, and two main clades in the Anacar- diaceae corresponding to Group A and B but with Buchanania diverging prior to both. Other sequence data (rbcL: Chayamarit, 1997) from an analysis of 16 Thai genera has also provided some support for the informal groups of Wannan & Quinn (1991), but the absence of bootstrap or decay analysis made it impossible to assess the strength of support for their clades. Recent sequence data from the internal transcribed spacer region from the ribosomal DNA has provided an indication of relationships amongst genera referred to Engler’s Rhoeae or Wannan & Quinn’s (1991) subgroup A2 (ITS: Miller et al., 2001). American species of Rhus s.s. (subgenera: Lobadium and Rhus) were shown to be closely related and more distant from other genera of the Rhoeae including Actinocheita, Cotinus, Malosma, Schinus, Searsia and Toxicodendron. The relationships of some genera of the Rhoeae were also investigated by Aguilar- Ortigoza et al. (2004) using non-sequence data. Their main focus was on the 6 species of Pseudosmodingium from Mexico which were shown to be most closely related to Bonetiella, also from Mexico. However, the larger clade, corresponding to genera of B.S. Wannan: Analysis of generic relationships in Anacardiaceae 167 the Rhoeae, included Smodingium (Africa) and Mexican representatives of Cardena­ siodendron, Cotinus, Rhus and Toxicodendron. A subsequent paper by a similar team (Aguilar-Ortigoza & Sosa, 2004) combined sequence (rbcL) and non-sequence data for 22 genera of Anacardiaceae. Both separate datasets show good support for Wannan & Quinn’s Group A (18 genera, including Anacardium and Mangifera from Engler’s tribe Anacardieae) but less support for Group B (3 genera). In both analyses Buchanania is placed as a sister taxon to genera in Group A, but outside the clade with Group B genera. Interestingly, the paper compares the Anacardiaceae clade with a clade of hemipteran insects (Calophya spp.) which feed on the family; the later shows closely related spe- cies feeding on Spondias and Buchanania. A conference abstract (Pell & Urbatsch, 2001) describing analyses of sequence data from the chloroplast genome (matK, trnL and the intergenic spacer between the trnL exon and trnF) has strongly supported the two groups of Wannan & Quinn (1991). Pell & Urbatsch (2001) reported two major clades in the family: one with members of the tribes Rhoeae, Semecarpeae, Dobineeae and Anacardieae, and the other with members of the Spondiadeae and a few members of the Rhoeae. They also reported that the Anacardiaceae proved to be monophyletic. Thus, there are some data which support the proposed infrageneric classification of Wannan & Quinn (1991). As yet, however, there has not been any broad analysis of generic relationships using the range characters which are known for the family. This paper analyses the available morphological, anatomical, chemotaxonomical, cytologi- cal, palynological characters and the available sequence data (rbcL) and aims to test support for the two informal subfamily groups proposed by Wannan & Quinn (1991) and identify key data gaps in the family. METHODS Non-sequence data The terminal taxa used in this analysis are genera (Table 1) with characters scored from usually more than one species. The characters used are listed in Table 2 and Ta- ble 3a, b is the data matrix. A list of the autapomorphies is provided in Table 4. Many descriptions of character states were reviewed for each taxon. In a few cases where differing character states were argued in the literature, these were scored as multiple states with each source cited. In most cases, however, a single reliable authority has been cited in Table 3a, b following a critical analysis of the literature. Some unpublished data are included and are supported by vouchers in Appendices A and B. Sequence data Sequences for the chloroplast encoded rbcL gene were obtained for a subset of taxa either from GenBank or from the sources cited in Table 5. Sequences were aligned in PAUP* (Version 4.0b10; Swofford, 2002). Analyses Heuristic parsimony analyses were performed in PAUP* set for tree bisection reconnection branch swapping on the best trees. Multistate characters were treated as polymorphisms. Multiple replicates of random taxon addition were employed to search for multiple islands of trees, and the CONDENSE option was employed to delete 168 BLUMEA — Vol. 51, No. 1, 2006 duplicate trees. Support for clades was inferred using the bootstrap option in PAUP* (Felsenstein, 1985) with 500 replicates, and also by decay values (Bremer, 1988; Donoghue et al., 1992). Decay command files were created in MacClade version 4.05 (Maddison & Maddison, 2002) and executed in PAUP* using 10 replicates of random taxon addition on each constraint tree. Output trees from PAUP* were transferred to MacClade and manipulated to test other topologies and explore character state evolution. Table 1. Genera included in analysis. Taxon No. of Natural distribution Subfamily Group species Tribe1 Group2 ANACARDIACEAE Amphipterygium 4 Mexico Unplaced3 A Anacardium 11 Tropical America Anacardieae A Astronium 13 Tropical America Rhoeae A Blepharocarya 2 Australia Rhoeae A Buchanania 25 Asia­Pacific, Australia Anacardieae B Campnosperma 10 Tropical America, Madagascar, Rhoeae B Seychelles, S.E. Asia, Malesia Cotinus 4 Temperate northern hemisphere Rhoeae A Cyrtocarpa 4 Tropical America Spondiadeae B Dobinea 2 Himalaya, China Dobineeae A Dracontomelon 8 China, Malesia, Pacific Spondiadeae B Euroschinus 6 New Caledonia, Papua New Guinea, Rhoeae A Australia Harpephyllum 1 Southern Africa Spondiadeae B Lannea 40 Africa, Arabia, Tropical Asia Spondiadeae B Lithraea 4 South America Rhoeae A Loxopterygium 4 South America Rhoeae A Mangifera 35 Tropical Asia, Malesia Anacardieae A Pentaspadon 6 Tropical Asia, Malesia, Pacific Rhoeae B Pistacia 14 Eurasia, Malesia, Mexico, Africa Rhoeae A Pleiogynium 2 Malesia, Pacific, Australia Spondiadeae B Rhodosphaera 1 Australia Rhoeae A Schinopsis 8 South America Rhoeae A Schinus 27 South America Rhoeae A Semecarpus 60 Indo-Malesia, Australia Semecarpeae A Spondias4 8 Tropical America, Asia4 Spondiadeae B Swintonia 12 Burma, Malesia Anacardieae A Tapirira 15 Tropical America Spondiadeae B Toxicodendron 30 America, Indo-Malesia Rhoeae A BURSERACEAE Bursera c. 100 Tropical America Bursereae5 Canarium c. 100 Africa, Malesia, Pacific, Australia Canarieae5 Garuga 4 Asia, Malesia, Pacific, Australia Protieae5 1) Engler, 1892, 1897. 2) Wannan & Quinn, 1991. 3) Not placed in any family by Engler, 1883, 1897. 4) Does not include Solenocarpus with representatives in Tropical Asia. 5) Leenhouts, 1955; Forman et al., 1994. B.S. Wannan: Analysis of generic relationships in Anacardiaceae 169 Character polarities were determined by outgroup analysis (Maddison, Donoghue & Maddison, 1984). Branch lengths were calculated using the ACCTRAN optimisation. Only unambiguous character state changes were recorded on the branches in the final figures. Choice of taxa Ideally terminal taxa should be species, so that both generic concepts and intergeneric relationships could be tested in the cladistic analyses. Unfortunately, nonmolecular data are not available for most of the species in the family. In fact, the full range of characters has yet to be scored for a single species. The limited data that are available have been assembled piecemeal by many workers as suitable material serendipitously has come to hand. Hence, in order to obtain a preliminary estimate of the phylogenetic signal in the available data, genera are used as the terminal taxa in the nonmolecular analysis, with the data drawn from one or more species. Genera were chosen primarily on availability of data, preferably from a number of authors. Thirty genera of Ana- cardiaceae are included representing all five tribes (Engler, 1892) and both informal subfamily groups (Wannan & Quinn, 1991). The outgroup for the analysis comprised three genera from the Burseraceae (Bursera, Canarium and Garuga) representing the three tribes (Leenhouts, 1955; Forman et al., 1994). The details of the genera used are provided in Table 1. Sequence data were available for representative species of only 16 of the genera included in the nonmolecular analysis (Table 5). Exemplars of three outgroups, Burseraceae, Rutaceae and Sapindaceae, were included, with trees being rooted on the last two. Additional information is provided below for some characters (Table 2) (5) Leaves — The outgroup and most Anacardiaceae have imparipinnate leaves (b). Fewer genera have simple leaves (a), though in some genera there are species with both. Paripinnate leaves (c) are very uncommon, occurring in some species of genera that possess mostly imparipinnate leaves. Bipinnate leaves occur only as polymorphic character states in Spondias (S. bipinnata; Airy Shaw & Forman, 1967) and Bursera (B. bipinnata; Porter, 1970) and have not been scored. (9) Inflorescence structure — Few Anacardiaceae and Burseraceae have had their inflorescence structure analysed (sensu Briggs & Johnson, 1979; Barfod, 1988). Most descriptions do not accurately describe this character. (10) Flower sex — Most genera of the Anacardiaceae, and probably Burseraceae, have unisexual flowers (b), though frequently with the aborted remnants of the other sex present. Fewer have bisexual (a) or polygamous (coded a & b) flowers. Botanists have frequently confused the sex of flowers and it is likely that the flowers of some genera which have been recorded as polygamous, and which have not been closely studied, will be found to be unisexual. In many cases the male flowers are clearly unisexual (with an aborted smaller ovary) while the female flowers appear to be bisexual but the stamens have aborted anthers which are apparent only after sectioning (Wannan & Quinn, 1992). 170 BLUMEA — Vol. 51, No. 1, 2006 Table 2. Description of characters and their states. Ch a r ancutemrberCharacter Ch a r astctateres 1 Habit a = tree; b = shrub 2 Leaf duration a = evergreen; b = deciduous 3 Leaf phyllotaxis a = alternate; b = opposite 4 Rachis wings a = not winged; b = winged 5 Leaves a = simple; b = imparipinnate; c = paripinnate 6 Leaflet phyllotaxis – = simple; a = opposite or subopposite; b = alternate 7 Leaf margin dissection a = entire; b = dentate 8 Inflorescence position a = terminal; b = axillary 9 Inflorescence structure a = thyrsoid; b = panicle 10 Flower sex a = bisexual; b = unisexual 11 Calyx number a = 6; b = 5; c = 4; d = 3; – = absent 12 Corolla number a = 6; b = 5; c = 4; d = 3; – = absent 13 Calyx aestivation a = valvate; b = imbricate; – = absent 14 Corolla aestivation a = valvate; b = imbricate; – = absent 15 Stamen number a = 2 whorls; b = 2 whorls with antepetalous whorl reduced to staminodes; c = 1 antesepalous whorl 16 Anther orientation a = introrse; b = extrorse 17 Nectariferous disc a = intrastaminal; b = extrastaminal; c = absent 18 Floral axis a = hypogynous; b = part perigynous; – = no perianth 19 Androgynophore a = absent; b = present 20 Carpellode number a = 5; b = 4; c = 3; d = 2; e = 1; – = bisexual flower/absent 21 Carpellode position a = antepetalous; b = 3 arrangement; c = antesepalous; – = bisexual flower/absent 22 Carpel number (fertile or infertile) a = 5; b = 4; c = 3; d = 2; e = 1 23 Position of fertile carpel a = antepetalous; b = antesepalous 24 Number of locules at anthesis a = 5; b = 4; c = 3; d = 2; e = 1 (fertile or infertile) 25 Level to which carpels are a = base of ovary; b = top of ovary; c = mid style; connate d = stigma; – = n.a. ie 1 carpel 26 Carpel definition a = present; b = absent; – = 1 carpel (e.g. Dracontomelon) 27 Stylar insertion a = ventral; b = apical; c = dorsal 28 Stigma morphology a = Dracontomelon-type; b = capitate; c = spathulate; – = not as previous 29 Ovule orientation a = apotropous; b = epitropous 30 Number of ovules per locule a = 1; b = 2 31 Ovule insertion a = apical; b = apico-lateral; c = latero-basal; d = basal 32 Number of ovule integuments a = 2; b = 1 33 Microphyle orientation a = superior; b = inferior 34 Winged fruit a = absent; b = present 35 Postanthetic growth of hypocarp a = absent; b = present 36 Number of seeds in fruit a = 5; b = 4; c = 3; d = 2; e = 1 37 Operculum in fruit a = absent; b = present 38 Thickness of fruit exocarp a = 0–15; b = 16+ – no. of cells 39 Epidermis of fruit a = unlignified; b = lignified B.S. Wannan: Analysis of generic relationships in Anacardiaceae 171 40 Hypodermis of fruit a = absent; b = parenchymatous; c = lignified 41 Mesocarp – sclereid bands a = absent; b = present associated with resin canals 42 Mesocarp – inner parts lignified a = absent; b = present 43 Endocarp – discrete 4th layer a = absent; b = parenchyma; c = sclereids (outermost) 44 Endocarp – crystals in 4th layer a = absent; b = present (outermost) 45 Endocarp – discrete 3rd layer a = absent; b = palisade sclereids; c = sclereids; d = parenchyma 46 Endocarp – discrete 2nd layer a = absent; b = palisade sclereids; c = sclereids; d = parenchyma 47 Endocarp – discrete 1st layer a = parenchyma; b = sclereids; c = palisade sclereids (innermost) 48 Endocarp – 1st layer radially a = cells not radially elongated compared to adjacent elongated (innermost) layers; b = cells > 2 times as long 49 Embryo shape a = straight; b = curved 50 Testa consistency a = membranous; b = not membranous 51 Testa connection to endocarp a = free; b = adherent; c = labyrinthine 52 Cotyledons a = plano­convex or flat; b = lobed 53 Radicle a = superior; b = inferior 54 Germination a = epigeal; b = hypogeal 55 Seedling phyllotaxis a = opposite; b = alternate 56 Seedling leaf dissection a = imparipinnate; b = simple; c = trifoliolate 57 Seedling leaf margin a = entire; b = serrate 58 Resin canals in phloem a = present; b = absent 59 Resin canals in pith a = present; b = absent 60 Resin canals in cortex a = present; b = absent 61 Wood parenchyma-apotracheal a = present; b = absent 62 Wood parenchyma-paratracheal a = vasicentric and alliform/banded; b = vasicentric; c = absent 63 Wood ray width a = 1–6 cells; b = 1–10 cells 64 Wood ray type a = heterogeneous type IIB; b = heterogeneous type IIA; c = heterogeneous type III 65 Septate wood fibres a = present; b = absent 66 Resin canals in wood rays a = present; b = absent 67 Xylem vessels spirally thickened a = present; b = absent 68 Pollen a = Rhus type; b = Pistacia type; c = Dobinea type 69 Chromosome number (2n) a = 24; b = 26; c = 28; d = 30; e = 32; f = 36; g = 40; h = 42; i = 78; j = 104; k = 22; l = 14; m = 58; n = 60 70 Butein glucoside in heartwood a = present; b = absent 71 Sulphuretin in heartwood a = present; b = absent 72 Sulphur glucoside in heartwood a = present; b = absent 73 Fisetin in heartwood a = present; b = absent 74 Fis 7-0-B glucoside in heartwood a = present; b = absent 75 Fustin in heartwood a = present; b = absent 76 7,3,4’trihydroxyflavone in a = present; b = absent heartwood 77 Rengasin in heartwood a = present; b = absent 78 Agathisflavone in leaves a = present; b = absent 79 Amentoflavone in leaves a = present; b = absent 80 Cupressusflavone in leaves a = present; b = absent 81 Hinokiflavone in leaves a = present; b = absent 172 BLUMEA — Vol. 51, No. 1, 2006 (11) Calyx number — Most genera of the Anacardiaceae have a 5-partite calyx (b), with very few having a 6-partite (a), 4-partite (c) or 3-partite (d) calyx. Many genera are polymorphic. Only the flowers of Amphipterygium and female flowers of Dobinea and Campylopetalum (not in this analysis) have been interpreted as having no calyx (–). In genera with very strongly dimorphic unisexual flowers (i.e. parts absent in one sex) the number of parts has been scored from the sex in which they are present (e.g. Dobinea). In Amphipterygium the female flowers have no perianth. The male flowers are recorded as apetalous with a 5–8-partite calyx and a similar number of alternisepalous stamens (Hemsley, 1908; Hutchinson, 1959; Cronquist, 1981). Given the absence of alternisepalous stamens, and the frequent occurrence of antesepalous stamens elsewhere in the family, the perianth of the male flowers of Amphipterygium is here interpreted as representing a corolla. The Burseraceae are recorded with 3-, 4- or 5-partite calyces. (12) Corolla number — Most genera of the Anacardiaceae have a 5-partite corolla (b), but in a few it is 6-partite (a), 4-partite (c) or 3-partite (d). Many genera are polymor- phic. The Burseraceae are recorded as 3-, 4- or 5-partite. Pistacia has been recorded as having no corolla (–). In male flowers of Pistacia there is a single perianth whorl of mostly 4 (rarely 5) segments (pers. obs. on P. chinensis). The stamens are equal in number to, and stand opposite, the perianth segments suggesting that, as in other genera of the Anacardiaceae (e.g. Cotinus, Semecarpus), there is a single antesepalous whorl of stamens (and hence no corolla). This conclusion is supported by Bentham & Hooker (1862), Baillon (1878), Standley & Steyermark (1949), Rechinger (1969) and Siddiqi (1978). Payer (1857) and Eichler (1878) also considered that only the calyx is present in female flowers. The reporting of alternitepalous stamens in P. malayana (Hou, 1978) suggests that this character needs examination across all species attributed to this genus. In genera with strongly dimorphic unisexual flowers (i.e. parts absent in one sex) the number of parts has been scored from the flower in which parts are present (e.g. Dobinea). (17) Nectariferous disc — Most of the outgroup and Anacardiaceae have an intra- staminal disc (a). A small number of genera have an extrastaminal disc (b) or no sign of any disc at all (c). This character can be difficult to interpret in unisexual male flowers where there are no carpellodes. However, in most cases the disc is still apparent, or the character can be scored from assessment of the sterile stamens in the female flower. (18) Floral axis — The outgroup and most Anacardiaceae have an undeveloped floral axis with an hypogynous flower (a). In these, the nectariferous disc encircles the ovary with the stamens inserted on its outer edge (i.e. intrastaminal). Some genera in both families show a tendency towards perigyny (b), where the lower parts of the perianth are fused and the stamens are connate, or adnate to the fused perianth or to the disc, forming a circular column around the ovary (e.g. Canarium, Garuga, Melanochyla, Thyrsodium). In the Anacardiaceae, true epigyny (inferior ovary) is restricted to Drimy­ carpus and Holigarna, which are not included in this analysis. (19) Occurrence of androgynophore — These are absent (a) from most Anacardiaceae and Burseraceae. An androgynophore (b) occurs in some species of Mangifera, Swin­ B.S. Wannan: Analysis of generic relationships in Anacardiaceae 173 tonia and Dobinea (and also Gluta which is not included in this analysis). The occur- rence of a gynophore has not been scored as it occurs only in Garuga. (20) Carpellode number — Evidence of carpellodes varies considerably from well- developed carpellodes (with small locules) through to very small bulges of remnant carpellode tissue (Wannan & Quinn, 1991). Common states are those which reflect the number of carpels in the female flowers, i.e. 5 (a), 3 (c) or 1 carpellode (e). In some cases there appears to have been reduction of the vestigial carpels so much that none are evident (Amphipterygium, Astronium, Pistacia, Rhodosphaera, Semecarpus); this condition is scored as inapplicable (–). The character is also not applicable in bisexual flowers. This character has not been widely scored in the outgroup. (21) Carpellode position — The outgroup and most Anacardiaceae often have their carpellodes reflecting the position of carpels in the female flowers: either standing opposite the petals (a), three carpellodes in a flower with a 5-partite perianth (b), or standing opposite the sepals (c). This character is not applicable (–) in bisexual flowers and in those where none are evident (as above). (22) Carpel number — Ranges in the Anacardiaceae from 13 in Pleiogynium (an autapo- morphy not coded for in this analysis) to a single carpel (e). Abortion of carpels leading to pseudomonomery is widespread in the families. This character has been inferred from the number of styles or stigmas, often with anatomical support from vascular bundles in the ovary wall and/or abortive locules (Wannan & Quinn, 1991). (23) Position of fertile carpel — One of the features of the outgroup and many Anacar- diaceae is that, even in multicarpellary flowers, there is only one fertile seed produced. The remaining carpels abort following anthesis or were never properly formed (Wannan & Quinn, 1991). In the outgroup and many Anacardiaceae the fertile carpel is antepetal- ous (a). In other Anacardiaceae the fertile carpel is antesepalous (b). (24) Number of locules at anthesis — This character scores locules whether or not they are fertile and provides some indication of the degree of carpel abortion occurring dur- ing development. The greatest number of locules is in Pleiogynium, i.e. up to 12 (an autapomorphy not coded in this analysis); other genera have 1–5 locules. (25) Carpel connation — There is a varying degree of carpel connation across the Ana- cardiaceae from apocarpus gynoecia in Buchanania (a), to genera in which the ovaries and parts of the styles are fused (c). Most genera in the Anacardiaceae have gynoecia in which the ovaries are connate and the styles free (b). The Burseraceae have almost complete connation with even the stigmas partly fused (d). (26) Carpel definition — This character refers to the furrows between the individual carpels in the ovary (a) and is a feature of Buchanania, Dracontomelon and Spondias. Although not present (b) in most Anacardiaceae or Burseraceae it does occur widely in more distant outgroups such as the Rutaceae and Sapindaceae. (28) Stigma morphology — There is a wide variety of stigma morphologies in the family but capitate or clavate stigmas (b) are the most common condition in the Anacardiaceae. 174 BLUMEA — Vol. 51, No. 1, 2006 More restricted morphologies include: – an oval opening at the top of each carpel as in Dracontomelon (a), which is generally characteristic of those genera in which there is good carpel definition but where the style gradually merges with the ovary, and – spathulate (c), which appears to be a development of the capitate condition (e.g. Amphipterygium, Pistacia). There are a number of autapomorphs which have been coded as inapplicable (–, see Table 4). The stigma morphologies of the Burseraceae are not well known. Garuga has a Dracontomelon-type of stigma (a). There has been no anatomical investigation of the crown-like angular stigma in Canarium that appears to be derived from the Dracontomelon-type. It may occur more widely in the Burseraceae. (31) Ovule insertion — Robbertse et al. (1986) have suggested that the differing posi- tions of funicle insertion on the locule wall may be related to the abortion of carpels alongside the fertile carpel (they cite Mangifera). While this may be true in some genera, it appears not to be the case in Pistacia, where on occasions there are two fertile carpels produced both with basally attached ovules (fig. 6d in Grundwag, 1976), nor in Astro­ nium, where there is only a single fertile carpel (and two sterile carpels as evidenced from two other styles and stigmas) with an apically attached ovule. (32) Number of ovule integuments — This character has not been widely scored. The outgroup and most Anacardiaceae have two ovule integuments (a). Much less common is one (b), and very rarely two on the outside and one on the inside (Pistacia). There are suggestions that a single integument has been derived from the bitegmic condition, but there is little agreement on how this may have occurred. Robbertse et al. (1986), working on Mangifera, have suggested that the single integument condition may be a neotonic form, but Copeland (1962) indicated that the single integument in Anacardium exhibited features that indicated it was the fusion product of two integuments, and Von Teichman (1990) suggested that in Tapirira there has been reduction of the inner integument. Hence, the single integument state may not be homologous in all taxa. (33) Micropyle orientation — Micropyle orientation is upwards or superior (a) in the outgroup and most Anacardiaceae, but an inferior micropyle (b) occurs in some gen- era with a basally attached funicle. In Pistacia the micropyle is initially inferior but becomes superior during development (Marchand, 1869; Copeland, 1955). Both states were coded in this genus. (34) Winged fruit — In the analysis winged fruit (b) occur only in Loxopterygium and Schinopsis. The ciliate margins in the fruit of Blepharocarya and the membranous margin in Dobinea have been interpreted as unwinged. (35) Postanthetic growth of hypocarp — Occurs only in Anacardium and Semecarpus (b). The outgroup and most Anacardiaceae have no significant postanthetic growth of the perianth or floral axis (a). Postanthetic growth of the calyx occurs in Astronium (and also Parishia, not in this dataset). Postanthetic growth of the corolla occurs in Swintonia (and also Gluta, not in this dataset). These latter two conditions (calyx and corolla) were not coded as they are autapomorphs (see Table 4).