Monograph of Andira

Monograph of Andira (Leguminosae-Papilionoideae) Author(s): R. Toby Pennington Source: Systematic Botany Monographs, Vol. 64, Monograph of Andira (LeguminosaePapilionoideae) (Mar. 31, 2003), pp. 1-143 Published by: American Society of Plant Taxonomists Stable URL: http://www.jstor.org/stable/25027903 Accessed: 02/12/2009 12:12 Your use of the JSTOR archive indicates your acceptance of JSTOR's Terms and Conditions of Use, available at http://www.jstor.org/page/info/about/policies/terms.jsp. JSTOR's Terms and Conditions of Use provides, in part, that unless you have obtained prior permission, you may not download an entire issue of a journal or multiple copies of articles, and you may use content in the JSTOR archive only for your personal, non-commercial use. Please contact the publisher regarding any further use of this work. Publisher contact information may be obtained at http://www.jstor.org/action/showPublisher?publisherCode=aspt. Each copy of any part of a JSTOR transmission must contain the same copyright notice that appears on the screen or printed page of such transmission. JSTOR is a not-for-profit service that helps scholars, researchers, and students discover, use, and build upon a wide range of content in a trusted digital archive. We use information technology and tools to increase productivity and facilitate new forms of scholarship. For more information about JSTOR, please contact [email protected].

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MONOGRAPH OF ANDIRA (LEGUMINOSAE-PAPILIONOIDEAE) R. Toby Pennington Botanic Garden Edinburgh Royal 20a Inverleith Row Edinburgh EH3 5LR, United Kingdom The principally Neotropical 29 species; one genus Andira Lam. (Leguminosae) comprises two of which occur in Africa. Chloroplast DNA restriction site includes three subspecies, analysis allows phylogenetic placement of 21 species o? Andira, including five species that were not previously studied. These data are also used to investigate and to identify ac intraspecific chloroplast DNA polymorphism Abstract.

species,

cessions

A.

inermis,

of A. humilis with plastomes characters provides

12 morphological of cpDNA

restriction

A cladistic analysis of from probable introgressive hybridization. little phylogenetic resolution in comparison with a simultaneous analysis some of the clades discovered site data and morphology. Although by this analysis are derived

characters and have no morphological the "cryptic," i.e., they are supported only by molecular synapomorphies, results provide a reasonable estimate of the phylogeny of Andira and are used as the basis of discussions of char acter evolution, and biogeography. Previous of Andira classification, infrageneric infrageneric classifications were based on the nature of the indumentum of the gynoecium, but glabrous ovaries have evolved in parallel at least three times. Detailed

informative characters and analysis of fruit wall anatomy provides phylogenetically biology. There have been at least five instances of independent evolution of large, puta fruits and three of large, persistent stipules. The frequent changes in stipule morphology tively rodent-dispersed are perhaps due to mutation in a single gene. Evidence radi points to a relatively recent (possibly Pleistocene) insights

into dispersal

ation of species of Andira are illustrated. Five new macrocarpa,

A. praecox,

are fully described and their ranges are mapped, and 15 are proposed: A. chigorodensis, A. jaliscensis, A. subspecies and A. inermis subsp. glabricalyx.

in eastern Brazil. All

species

and one new

species A. taurotesticulata,

INTRODUCTION is a genus of 29 woody species (including three subspecies of A. inermis) dis tributed throughout tropical America; A. inermis also occurs in Africa. Andira is most Benth. (Bentham 1860; Polhill 1981), a genus of 10-12 closely related toHymenolobium species confined to South America. They are similar florally and vegetatively, and they Andira

share a root nodule morphology that is very rare in the Papilionoideae (de Faria et al. 1987; Sutherland et al. 1994). The principal difference between the two genera is in fruit has samaras and Andira drupes. The two genera also differ in their type; Hymenolobium mode of germination: Andira is cryptohypogeal, whereas Hymenolobium is phanerohy a Andira is considered with group pogeal. Thus, monophyletic synapomorphies of a dru paceous fruit and cryptohypogeal germination (Pennington 1995, 1996). The close relationship of Andira and Hymenolobium is confirmed by recent chloro trnL intron et nucleotide sequence data (Pennington al. 2001). The two genera form plast a well-supported monophyletic group, but their phylogenetic position is unresolved, and no clear sister group has been discerned; however, it is clear that they are distantly related to the other genera of tribe Dalbergieae to which they are currently assigned. Dalbergieae comprises 19 genera and is centered in tropical America; only Inocarpus J. R. Forst. & G. Forst, is endemic to the Old World (in tropical Asia; Polhill 1981). Other recent l

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systematic studies (Doyle et al. 1997; Lavin et al. 2001) indicate that Dal This confirms earlier suggestions, based upon consideration bergieae is not monophyletic. that Dalbergieae are not a natural group (de Lima of fruit, seed, and seedling morphology,

molecular

1991). This taxonomic revision

is based upon examination of ca. 3,000 herbarium specimens in five Neotropical countries from 1991 to 1997. Species de inAndira has been regarded as problematic, and the only recent revision (Mat

and field studies conducted

limitation tos 1979), covering the Brazilian species, is inadequate for species identification, particu coastal rain forest and coastal restinga forest of larly for taxa from the Atlantic southeastern Brazil (Lewis 1987), where Andira is both diverse (eight species) and abun dant. Three new species have been described recently from eastern Brazil (Pennington & de Lima 1995) and the Venezuelan Guayana (Pennington et al. 1997). Four of the five new species included in this monograph area of diversification for Andira.

grow around the fringes of Amazonia,

which

is an

frameworks for Andira based upon both chloroplast DNA (cpDNA) re Phylogenetic striction site data and morphology have been presented by Pennington (1995, 1996). The are drawn from these studies, but in this monograph phylogenetic analyses presented in coding of some characters are proposed based upon new observations of modifications pollination syndromes and fruit anatomy. Furthermore, new cpDNA restriction site data have provided phylogenetic placements for five more species and helped resolve some of the problems of intraspecific cpDNA polymorphism discussed by Pennington (1995, are used to re-evaluate infra The refined here 1996). phylogenetic hypotheses presented in Andira, to present the first biogeographic generic classification the evolution and diversification of the genus, and to investigate phological features, such as stipules, flowers, and fruits.

hypotheses to explain the evolution of mor

TAXONOMICHISTORY to the attention of European science through the use of extracts from the bark and seeds as anthelminthic drugs by indigenous peoples in the Neotropics. as an anthelminthic in Per Piso (1648) described the use of the seeds of A. fraxinifolia Andira

first came

nambuco, Brazil, and named the species "Andira Ibaiariba." The first species descriptions were included in accounts of the medicinal use of the bark and seed of A. inermis in Ja maica (Wright 1777) and A. surinamensis in Suriname (Bondt 1788). Both authors placed their species in Geoffroea Jacq., which shares Andira's unusual drupaceous fruit, but is distinct inmany floral and vegetative characters. the first valid publication of the name Andira has been problematic. It Determining had generally been assumed to be by Lamarck (1783), and as such was proposed (Harms 1904) and accepted (Briquet 1906) for conservation against Vouacapoua Aubl. Although these two genera are now considered taxonomically quite distinct, Lamarck had included Vouacapoua in his original concept of Andira, making the latter name illegitimate. Yet, as part of the Rickett and Stafleu revision of the lists of Nomina Conservanda, Cowan (1959) noted that Lamarck's

(1783) description of Andira racemosa provided no generic diagno sis, and since Lamarck cited Vouacapoua americana Aubl. (1775) in synonymy his genus and species could not be treated as new. Cowan decided that the long-standing provisions of the International Code of Botanical Nomenclature (ICBN) permitting a "descriptio to validate simultaneously the generic and specific names could not generico-specifica"

3

ANDIRA

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apply. He therefore concluded that neither Andira nor the species name, A. racemosa, were validly published by Lamarck, and proposed that the conservation of Andira be from its next usage, that by A. L. de Jussieu (1789), with A. racemosa Lam. ex J. St.-Hil., ap parently the first validation of that name, as type. R. K. Bmmmitt (pers. com.) noted that Lamarck did indeed validly publish Andira but as an illegitimate replacement (a nomen novum) of Vouacapoua Aubl., because he treated

Vouacapoua

as a synonym

of his

generic

name

Andira.

His

species

name

A.

race

is also illegitimate, as a nomen novum for V. americana. Under Art. 7.5 of the cur rent ICBN (Greuter et al. 2000), the type of Andira Lam. is that of Vouacapoua (i.e., the type of V. americana Aubl.), but because Vouacapoua is now recognized as a distinct cae salpiniaceous genus, this typification would be extremely disruptive.

mosa

The current conservation is of "Andira Juss.," not "Andira Lam." Jussieu (1789) cited no species in his account, and Cowan (1959) considered Andira Juss. to be typified by A. racemosa Lam. ex J. St. Hil.; however, Jaume St. Hilaire (1804) was merely using Lamarck's A. racemosa, so there is no case for treating his name as a later homonym under Art. 48.1 of the ICBN. If "Andira Juss." were to be maintained as the conserved name, the type requires correction to A. racemosa Lam., which is automatically typified by V. americana. Therefore, themaintenance of Andira Juss. would defeat the purpose of the conservation of the name Andira. For these reasons, a formal proposal to change the authorship of Andira to Andira Lam. nom. cons., and to provide it with a conserved type, A. inermis, was submitted and accepted (Pennington 2002). The name Andira inermis (Wright) DC. has been widely ap plied to the species that includes A. racemosa Lam., the type of the conserved Andira (i.e., excluding the Vouacapoua element). Thus, itwas appropriate to propose, under Art. 14.9 of the ICBN, that the type of A. inermis be the conserved type of Andira Lam. If the name A. racemosa Lam. were conserved with a new type, itwould displace A. inermis, the name traditionally used for the most

common, widespread,

and frequently

collected

species of

Andira.

Infrageneric Bentham

classification

1839, 1860, 1862) features prominently in the taxonomic history of studying the specimens of Martius, Pohl, and Schott from eastern Brazil, he the first major account of Andira (1837, reprinted in 1839). He placed the produced two in sections: sect. Lumbricidia, defined by a hairy ovary on a relatively short species (1837,

Andira. After

stipe, and sect. Euandira, defined by a glabrous ovary on a relatively long stipe. Walpers (1843) followed Bentham. In his later works (1860, 1862), Bentham abandoned both the indicate that the ratio of stipe length to ovary length stipe character (my measurements across all species of Andira varies very little) and the formal sections, but he did group species based upon the presence of hairy or glabrous ovaries. The next attempt at infrageneric classification was by Mattos (1979). She recog two sections, one with two subsections. Section Lumbricidia is defined by the presence of more than five leaflets (i.e., more than two pairs of leaflets). Subsection Lumbricidia includes species with a hairy ovary and subsect. Glabratae those with a nized

sections. Section Paucifoli glabrous ovary; these subsections correspond to Bentham's olatae is defined by presence of 1-3 leaflets, and therefore contains only A. unifoliolata and A. trifoliolata.

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MORPHOLOGY Habit. Species of Andira grow in a wide variety of habitats and have varied life forms (character 1 in cladistic analysis). Rain forest species, such as A. macrothyrsa, A. parvi flora, A. cori?cea, and A. anthelmia, are trees that can reach 40 m. Several species (in are small trees and shrubs abundant in cluding A. nitida, A. carvalhoi, and A. fraxinifolia) the sandy coastal restinga scrub and forest of eastern Brazil. Andira humilis is a geoxylic suffrutex, defined by White (1976) as plant with massive, woody, underground axes but only annual or short-lived shoots above ground. Its aerial shoots rarely exceed 50 cm. The species appears to be well adapted to survive the regular fires in the seasonally dry cer rado woodland of central Brazil by an ability to sprout from its underground rhizomes after burning. This growth form is unique to this species of Andira, but several other are able to species (A. verm?fuga, A. carvalhoi, A. legalis, A. nitida, and-A. fraxinifolia) to A. often This enable carvalhoi survive on may root-sprout (pers. obs.), extensively. steep and easily eroded white sand slopes in restinga scrub. Life form provides a useful field character for distinguishing A. ormosioides from A. fraxinifolia. The latter species has a broad, spreading crown and short bole when in open situations, whereas A. ormosioides has a long bole and small crown. Polhill (1969) indi cated that growth form may also be useful in distinguishing A. inermis subsp. rooseveltii. Field notes (from herbarium specimen labels) indicate it to have a narrow, conical crown, which contrasts with the broad, spreading crown of A. inermis subsp. inermis. Bark and slash. All species of Andira produce a little red ex?date when the bark is cut, a characteristic also of some other woody papilionoids (e.g., Dussia Krug & Urb. ex Taub., Geoffroea, Pterocarpus Jacq.). Bark color, bark texture, and slash color vary, al too few been made to assess their taxonomic significance. Pale observations have though grey-brown bark color characterizes A. fraxinifolia, whereas other eastern Brazilian tree species (A. ormosioides, A. legalis, A. anthelmia, and A. nitida) have darker bark (pers. slash of A. parviflora and A. cori?cea contrasts with the more obs.). The orange-brown reddish brown observed in other Brazilian species, such as A. cordata and A. surinamen sis. The thick, corky barks of A. cordata, A. cujabensis, and A. verm?fuga probably serve as protection against fire in the cerrado woodlands of central Brazil. Wood. The wood anatomy of eleven species of Andira (A. anthelmia, A. fraxinifolia, A. legalis, A. humilis, A. nitida, A. carvalhoi, A. verm?fuga, A. inermis, A. surinamensis, A. cori?cea, and A. cordata) was studied by Herridge (1992). All these species have very similar wood anatomical features, and any interspecific differences noted are quantitative and may be due to the nature of the sample (from a branch or the trunk), its age, or vari ation in the growth rate of different species. These difficulties in interpretation mean that these data are of little relevance to the species-level taxonomy of Andira. The wood is diffuse-porous and without growth rings. The storied vessels, which are of up to four, are large (98-234 mm), widely solitary or clustered in radial multiples spaced, and often contain deposits. The intervessel pitting is vestured, small, and alternate. The axial parenchyma is storied, aliform (winged to lozenge-shaped), and in tangential bands ranging from 3-20 cells wide. There are gelatinous prismatic crystals in chambered axial parenchyma cells. The rays range from 1-3 to 1-6 cells wide and 12-19 cells high, and are irregularly storied or non-storied. Multiseriate rays aremostly heterocelluar (A. in ermis is homocellular).

The fibers are non-septate

and usually very thick-walled.

ANDIRA

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5

Indumentum. When present, hairs are always simple and generally either flexuose and erect or short and appressed. The indumentum of different structures (twigs, stipules, rhachis, petiolules, leaflet undersurfaces, inflorescence, calyx, and gynoecium) varies in dependently, and the variation for each part is discussed separately below. This variation for species delimitation, (characters 5, 8, 10). is useful

and provides

three characters

for cladistic

analysis

Stipules. Four species (A. multistipula, A. grandistipula, A. anthelmia, and A. legalis) have large (to 7.5 cm long, 6 cm wide in A. grandistipula), persistent stipules, which are crowded at the shoot apices and around the base of the inflorescence (Fig. 1A; character 3 in cladistic analysis). These contrast with the more typical small stipules of all other Andira species (Fig. IB). This difference in stipule size distinguishes A. multistipula from the otherwise similar A. inermis, and A. anthelmia and A. legalis from A. ormosioides. The function of the large stipules is presumably to protect the shoot apices. There is no obvi ous correlation of the occurrence of large stipules with geography, habitat, or life form. Leaves. The leaves of all species of Andira are imparipinnate. The leaflets are gener ally opposite with their petiolules subtended by stipels. Leaf length varies from 2.5 to 60 cm and is an important character in species delimitation. The number of leaflets varies to up to 19 (nine pairs) in A. chigorodensis from one in A. unifoliolata (character 4 in cladistic analysis; Fig. 1C, D). Unifoliolate leaves are unique to A. unifoliolata (Fig. 1C). Andira

trifoliolata is the only species with uniformly trifoliolate leaves; A. tervequinata has both trifoliolate and 5-foliolate leaves. In the other species the number may be con stant (e.g., A. micrantha, 2 pairs; A. cori?cea, 3 pairs) or vary within certain limits (e.g., A. chigorodensis, 7-9 pairs). Leaflet shape provides characters that are diagnostic of certain species. For example, the obovate, blunt-ended leaflets of A. marauensis distinguish it from the otherwise simi larA. nitida. The most critical character of the leaflets for species delimitation is the na ture and density of the abaxial indumentum (character 5 in cladistic analysis). Some species, such as A. inermis (Fig. 2A), have glabrous leaflets. In the other species, three in dumentum types are present (Fig. 2B, C, D). Eleven species (including A. surinamensis, A. macrothyrsa, A. praecox) have an indumentum of short, tightly appressed hairs (Fig. 2B), which contrasts with the varying densities of vestures composed of longer, erect hairs these two (Fig. 2C) of six species (such as A. legalis and A.fraxinifolia). Distinguishing indumentum types (short appressed versus longer erect hairs) is useful for species identi fication, but variation between the two is continuous, and they are lumped into a single character state in the cladistic analysis. Yet, two species, A. cujabensis andA.jaliscensis, have leaflet undersurfaces with a distinct indumentum of dense, fine, pale, tangled, erect hairs (Fig. 2D), which is the character that separates them from otherwise similar species (A. cordata and A. inermis; Fig. 2A) that are glabrous. There is no association of indu mentum type with habitat. For example, A. micrantha (glabrous), A. macrothyrsa (short, tightly appressed hairs), and A. parviflora (long, erect hairs) are all rain forest species. Similarly A. cordata (glabrous), A. humilis (short, tightly appressed hairs), A. verm?fuga (long, erect hairs), and A. cujabensis (long, fine, pale, tangled hairs) are all species of the seasonally dry cerrados of central Brazil. The primary vein is prominently raised on the leaflet undersurface (Fig. 2C), and ei ther channelled (Fig. 2E) or plane (Fig. 2F) on the leaflet uppersurface (character 6 in cladistic

analysis). Venation

pattern varies continuously

from entirely eucamptodromous

6

SYSTEMATICBOTANYMONOGRAPHS

FIG. 1. Stipules and leaves = ca. 5 cm; (scale bar photo G. P. scale = inches; right-hand scale = leaves of A. legalis. (Based on: A,

in Andira (characters 3 and 4). A. Large, persistent stipules of A. anthelmia Lewis). B. Small stipules ("s" at arrows) of A. surinamensis (ruler left-hand leaves of A. unifoliolata. D. Large, imparipinnate cm). C. Small, unifoliolate R. T. Pennington 183; B, R. T. Penningfon 463; C, W. Rodrigues 11182; D, R.

T Penningfon

305.)

VOLUME 64

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y.y

7

s

FIG. 2. Indumentum and venation of leaflets of Andira abaxial leaflet (characters 5 and 6). A. Glabrous surface of A. inermis, showing plane secondary and tertiary veins. B. Abaxial leaflet surface of A. surinamensis with short, straight, tightly appressed hairs, prominently raised secondary vein, and slightly raised tertiary veins. leaflet surface of A. fraxinifolia erect hairs, prominently C. Abaxial with long, flexuose, raised primary and sec ondary veins, and slightly raised tertiary veins. D. Pale, long, fine, tangled hairs of the abaxial leaflet surface of A. cujabensis. E. Adaxial leaflet surface of A. surinamensis with channelled primary vein. F. Adaxial leaflet sur face of A. unifoliolata with plane primary vein. P = primary vein; s = secondary vein; t = tertiary vein. Scale bars = 1mm. 463; C, G. Hafschbach 13558; B, R. T. Penningfon (Based on: A, T. D. Pennington 55043; D, C. A. Cid Ferreira 6259; E, R. T. Penningfon 463; F, A. Ducke 35078; all from herbarium material except E, leaf from live plant growing

at Royal

Botanic

Gardens,

Kew.)

SYSTEMATICBOTANYMONOGRAPHS

8

VOLUME 64

to entirely brochidodromous tomixed eucamptodromous (terminology of Hickey, 1979). The number of secondary veins, their angle of divergence from the primary vein and then nature of curvature varies and can be useful for distinguishing species. For example, the contrast with those of A. inermis, uniformly curving secondary veins of A. macrothyrsa curve only as they approach the margin. The secondary and tertiary veins also vary from plane to sunken on the leaflet uppersurface, and plane (Fig. 2A) to raised (Fig. 2B, C) on the undersurface, but this variation is continuous and not useful for cladistic

which

analysis.

Inflorescences are terminal and axillary panicles, which vary in size, Inflorescence. shape, compactness, number of branches, angle at which the branches are held, and the and density of the flowers. Intraspecific variation is often extreme (e.g., in A. fraxinifolia A. inermis), and this limits the use of these inflorescence characters for taxonomic pur poses. Terminal inflorescences may not be obvious on herbarium sheets, but have been ob served in all species seen flowering in the field. Bracts

and

bracteoles.

Bracts

and

bracteoles

ducous. There is little variation in theirmorphology paired at the base of the calyx.

are

small,

narrow,

and

often

early

throughout Andira. The bracteoles

ca

are

length varies continuously from 5 to 23 mm (Fig. 3).Within species, the range of variation is relatively narrow, approximately 2 mm for small-flowered species and 6 mm for large-flowered species. Flower length is a useful character for separating similar pairs of species, such as A. inermis and A. cubensis, A. fraxinifolia and A. ormo Flowers.

sioides,

Flower

and A. macrothyrsa

and A.

chigorodensis.

The angle and depth of the calyx lobes varies widely in several species and is thus of limited utility in species delimitation; however, the calyx indumentum (character 8 in cladistic analysis) provides more useful characters. For example, the densely hairy calyx of A. cubensis distinguishes this species from A. inermis. In A. inermis, A. multistipula, and A. humilis, the calyx is either sparsely hairy (the hairs short and appressed) or and this variation shows of scription subspecific taxa inA. The corolla o? Andira consists but are firmly attached because of

geographical correlation, which is the basis for the de inermis (see discussion under the individual species). of five glabrous petals. The keel petals are not united two interlocking folds. The wing petals are free from the keel. Petal color (character 7 in cladistic analysis) is an important phylogenetic char acter. Species with flowers of less than 8 mm (with the exception of A. cubensis) have white to yellowish petals with the standard marked with red or purple. The larger-flow ered species have pink to purple petals with the standard bearing a central white mark. glabrous,

Lamellate

wing petal sculpturing (Fig. 4A; character 9 in cladistic analysis) is present in all species with flowers greater than 9 mm long, and also in A. cubensis where the flow ers are 7 to 8.5 mm long. Lamellate wing petal sculpturing is absent from the other small flowered species (Fig. 4B).

In all species of Andira, the stamens are diadelphous (9 + 1); the vexil Androecium. and A. macrothyrsa lary stamen is free (Fig. 3). Very rarely, individuals of A. fraxinifolia have one side of the vexillary filament united to the other nine. The filaments are unequal in length; those closest to the standard are shorter and united for one half to two-thirds of their length. The ratio of the length of the united and the length of the free filaments varies

ANDIRA

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FIG.

9

of Andira 3. Floral morphology (characters 8, 9, and 10). For each species, from left to right: full and gynoecium; showing calyx indumentum and wing petal sculpturing; dissection showing androecium C. A. taurotestic longitudinal section of gynoecium showing ovule number. A. A. cordata. B. A. chigorodensis. v = vexillary ulata. D. A. nitida. E. A. jaliscensis. F. A. carvalhoi. G. A. fraxinifolia. H. A. ormosioides. stamen, = 5 mm; all drawn to same scale. in in H. all but marked Scale bar present species, only flower

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SYSTEMATICBOTANYMONOGRAPHS

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B(i)

in Andira. Species with wing petal sculpturing: A, wing petal of A. ver FIG. 4. Wing petal sculpturing = lamellate S scale bar = 1mm; A(i) wing petal of A. fraxinifolia (16 mm long); A(ii) wing m?fuga, sculpturing, (15 mm long); A(iii) wing petal of A. nitida (11 mm long); A(iv) wing petal of A. jalis petal of A. carvalhoi censis (12 mm long). Species without wing petal sculpturing: B, wing petal of A. trifoliolata, scale bar = 1mm; B(i) wing petal of A. cordata (5.5 mm long); B(ii) wing petal of A. chigorodensis (5 mm long); B(iii) wing petal of A. taurotesticulata 228; A(ii) M. P. M. de (8 mm long). (Based on: A, J. Ratter 3595; A(i) R. T Pennington Lima et al. 20; A(iii) A. J. Ribeiro 62; A(iv) E. J. Lott 2544; B, J. J. Wurdack & L. S. Adderley 43368; B(i) G. . Hatschbach 39477; B(ii) /. Brand & A. Cogollo 114; B(iii) G. Lozano & G. Galeano 3963.)

little across all Andira species, which is further proof of the highly conservative floral or in Andira that appears to reflect a general bee pollination mechanism.

ganization

Pollen. The pollen morphology of two species, A. inermis and A. macrothyrsa, has been found to be uniform by B. Klitgaard (unpubl. data). This is unsurprising, given the morphological uniformity of pollen in the many tropical woody papilionoids (Ferguson & Skvarla 1981; Ferguson et al. 1994). It therefore appears unlikely that pollen characters

11

ANDIRA

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will provide taxonomic characters within Andira. The pollen grains of the two species ex amined have an even, minutely reticulate tectum, are 3-colporate, with the colpi extend ing only partly to the poles, and have granular colpal membranes. Gynoecium. The ovary of Andira is borne on a stipe and contains 1-8 ovules (Fig. 3). The fruit of Andira is usually single-seeded (see "Fruit" below), the result of ovule abor tion. The style is curved and is included within the keel petals. The stigma is ciliate (sensu Lavin and Delgado, 1990). The proportions of the stipe, style, and ovary are more or less invariant. The extent of the indumentum of the gynoecium is, however, very variable (Fig. 3; character 10 in cladistic analysis) and is a critical character for species delimitation. are single-seeded 2-3-seeded) (occasionally drupes with or and hard thin exocarps. They are diverse mesocarps, endocarps, fibrously fleshy

Fruit. woody

Fruit of Andira

in the color

of

exocarp

and mesocarp,

scent,

mesocarp

structure,

and

endocarp

structure.

There are two general size classes of fruit: 6 cm long or less, weighing ca. 10-20 g when to 40-300 g when dry. These dry, and much larger fruits up to 12 cm long and weighing in dispersal biology size differences reflect probable differences (see "Dispersal Syndromes"). Bentham

in (1860) and Lewis (1987) point to the probable utility of fruit morphology species delimitation and the lack of adequate fruit collection. Field observations and the inferences made from alcohol preserved fruit confirm this. I have made field ob servations of fruit of seven species, which suggest that coloration of themesocarp and ex ocarp are useful characters. For example, the green mesocarp of A. anthelmia and A. frax

Andira

inifolia contrasts with the pale green-white mesocarp of distinctness of A. ormosioides. Andira anthelmia and A. is green. ocarps, whereas that of A. fraxinifolia Detailed field notes of exocarp and mesocarp color fruits in alcohol should be a priority for future collectors

A. ormosioides, and confirms the ormosioides have dark brown ex

and structure, and preservation of of Andira. Better field data might some in Andira (such as that of of delimitation the clarify outstanding problems species of A. nitida; see notes under that species), would benefit studies of dispersal biology in Andira, and provide suitable material for studies of fruit anatomy. A preliminary study of Andira fruit anatomy (Pennington 1994) revealed two distinct types of endocarp in Andira; one composed of woody fibers, the other of stone cells. The endocarp of stone cells appeared to be a possible synapomorphy for a well-supported clade of Andira

species discovered by cpDNA data (Pennington 1995). Pennington and Gemeinholzer (2000) examined the pericarp anatomy of 25 species in search of further taxonomic characters, and a summary of this work is provided here. The mesocarp of all Andira species examined comprises parenchyma cells in which stone cells are embedded, either individually or in patches. The mesocarp of seven species is homogeneous, whereas in 20 species it is heterogeneous, having either patches of stone cells close to the exocarp, fibers near the endocarp, and/or ergastic materials distributed unevenly throughout the parenchyma. Despite this variation, itwas decided thatmesocarp structures could not be used cladistically, because intraspecific variation was too great. Starch grains are abundant, occurring either scattered throughout the tissue, as greyish cell content or as "tyloses-like" features (probably tightly packed starch grains in highly packed parenchyma cells with squashed cell walls). At maturity, the endocarp is divided into a thicker hard layer and a thin papery layer within it, which is the remnant of the "seed cushion." This latter structure, described by

12

SYSTEMATICBOTANYMONOGRAPHS

VOLUME 64

(1914), is characteristic of legume fruits and consists of thin-walled, juicy parenchyma cells inwhich the developing seeds are embedded. At fruit maturity it shrinks and dries out, forming flake-like shreds. The hard, thick part of the endocarp is variable and provides phylogenetically useful information. Three main structures are present: (i) dominated by parenchyma and/or col lapsed cell tissue; (ii) dominated by fibers; (iii) dominated by stone cells. This variation can be coded as amultistate cladistic character (Pennington & Gemein holzer 2000; character 10). This is a refinement of the coding of Pennington (1996), where

Fucsk?

the distinct was

not

state of the endocarp dominated

by parenchyma

and/or collapsed

cell tissue

recognized.

formed by parenchyma and/or col species have an endocarp predominantly most to cell often tannin. Twelve species have an en which is brown due tissue, lapsed dominated fibers. Small cells and sometimes squashed cell tissue by parenchyma docarp occur between the fibers. Air gaps were often observed, probably due to ripening or de in layers, but in A. ver generation processes. The fibers can be arranged predominantly Five

m?fuga and A. carvalhoi they form sub-rounded patches surrounded by parenchyma cells. Andira galeottiana, A. legalis, and A. anthelmia are characterized by a mixture of these arrangements. Nine species have an endocarp that predominantly comprises stone cells. This endocarp type is the hardest and most dense, and it is often very difficult to cut with a sliding microtome. The densely packed tissue, lacking intercellular space, consists ei ther of stone cells alone (three species) or of stone cells interspersed with small patches of parenchyma (six species). In a single species, A. jaliscensis, the hard layer of the en an two of thickness: inner of equal layer parenchyma and col docarp comprises layers an outer stone cell of is with an unique and cells. This coded tissue, lapsed layer species (autapomorphic) state for this character. The thicker, fibrous endocarps, and the endocarps of stone cells may offer greater me chanical protection against seed predators (Pennington & Gemeinholzer 2000). Janzen in A. inermis, and I have seen fruit of (1982) noted seed pr?dation by Cleogonus weevils A. macrothyrsa on a herbarium specimen (A. Gentry 37168) that have been attacked by weevils. Both of these species have weak endocarps composed Andira humilis has an endocarp of woody fibers, but this layer species examined, perhaps making it less of a barrier against Handro (1969) reported seed pr?dation by Cleogonus weevils

of parenchymatous tissue. is thinner than in any other burrowing seed predators;

in this species. Field obser of preserved fruit revealed no seed pr?dation inA. fraxinifolia, A. anthelmia, A. ormosioides, A. nitida, and A. carvalhoi, which have thicker, tougher, woody fibrous endocarps and in A. cujabensis, which has an endocarp of stone cells. vation and examination

These data suggest

that greater seed protection may be provided by these harder, thicker

endocarps.

Seeds of species of Andira can be considered "overgrown" (sensu Comer, because 1976), they have an undifferentiated testa and contain no endosperm. They fill the entire cavity of the fruit, and are more or less globose tomore elongated, 2-8 cm long and 1.5-6 cm wide. The testa is thickly chartaceous, generally dark reddish brown, and often remains attached to the walls of the seed cavity (overlying the remains of the seed cush Seeds.

ion) when the seed is removed. The cotyledons are virtually united, and a thin line can be seen in transverse section where they meet. The hypocotyl-radicle axis is small and ap parent as a fold at the apical end of the seed.

2003

ANDIRA

13

(de Lima 1991; pers. obs. for A. carvalhoi, Seedlings. Germination is cryptohypogeal A. cujabensis, A. fraxinifolia, A. nitida, A. surinamensis, A. cori?cea, A. anthelmia). Nu merous spirally arranged scale-like prophylls are produced before the first true leaves, which are trifoliolate or with two or more pairs of leaflets (even in the unifoliolate A. uni was not observed in pers. obs.). Leaflet nyctinasty (evening leaf movements) foliolata, seven of the of Andira this any seedling grown during species study. It appears possible that polyembryony?the presence of more than one embryo in an occur in Andira, because more than one shoot can be pro (Richards 1997)?may duced from a single seed o? A. fraxinifolia (P.Griffiths, pers. comm.). This raises the pos that the variation patterns of this species might be ex sibility complex morphological ovule

plained

in part by apomixis.

studies of this and other species are

Further developmental

necessary.

POLLINATIONBIOLOGY Frankie et al. (1976) recorded numerous species of bees visiting the flowers of A. in ermis in Costa Rica in large numbers. Other observations of floral visitors are anecdotal. I have made similar observations of a variety of bee species visiting A. fraxinifolia and A. carvalhoi (R. T. Pennington, unpubl.) in Bahia, Brazil. These are two species with pink purple flowers, less than 18mm long. Although the larger (19-24 mm long) flowers of A. and are not significantly larger in terms visited by large xylocopid bees, which suggests a different pollination biology. This also appears to be reflected in the phenology of A. anthelmia, where a high percentage of individuals were observed to be synchro nously flowering in one area (vicinity of Ilh?us, Bahia, Brazil, October 1994; R. T. Pen nington, unpubl.) in contrast to smaller-flowered species, such as A. fraxinifolia, A. car anthelmia of flower

have superficially similar morphology length, I have only seen these flowers

valhoi, and A. verm?fuga, where T. Pennington,

unpubl.;

J. A.

few individuals

Ratter,

pers.

comm.).

in a population Two

other

flower synchronously

species,

A.

legalis

and A.

(R. or

mosioides, have flowers of a correspondingly large size to those of A. anthelmia and may share the same pollination biology, but direct pollination observations are lacking. The completely different flower color of other species o? Andira (white to yellow, the standard petal with red or purple markings) suggests tion biology, although there are no field observations

that they, too, have distinct pollina to confirm this. James Ratter (herb,

label) has observed large numbers of bees visiting the small, white-flowered A. cujaben sis. This suggests that the pollinators of the small-flowered species may simply be smaller bees. The absence of wing petal sculpturing suggests a difference in pollinator of the small flowered species, because wing petal sculpture is thought to provide a foothold for floral visitors (Stirton 1981). Further evidence that these small flowers may attract different pol linators comes from the general correlation of the characters flower size, flower color, and presence of wing petal sculpturing. Bee pollination of all species o? Andira might provide an explanation for the covari ance in size of the floral parts throughout the genus. Yet, the probable pollination of A. an thelmia by different vectors (xylocopid bees), demonstrates that care must be taken in generalization. For example, all the small flowers may not be as similar as they appear; A. Ducke (herb, label) noted that the small flowers of A. macrothyrsa smell fetid, which con trasts with the sweet scent of the flowers of A. cujabensis noted by J. Ratter.

SYSTEMATICBOTANYMONOGRAPHS

14

FIG. 5. A. Small, fruits of A.

legalis

fruits of A. fraxinifolia (R. T. Pennington bat-dispersed (G. P. Lewis & H. C. de Lima 1196; photo G. P. Lewis.)

VOLUME 64

213). B. Large,

rodent-dispersed

These observations of pollination biology have influenced the coding of character 7, which now has three states, rather than the two described by Pennington (1996).

DISPERSAL SYNDROMES Janzen et al. (1976) observed dispersal of the fruits of A. inermis by bats of the fam in Costa Rica. It seems reasonable to assume thatmost Andira species ily Phyllostomidae also have bat-dispersed fruits, because their fruits are morphologically similar to those of A. inermis (Pennington & de Lima 1995; Fig. 5A). The fruits of these bat-dispersed species do not exceed 6 cm in length, weigh 10-20 g when dry, are green or occasionally yellow (A. humilis) when ripe, have a strong, sweet scent and a fibrous mesocarp. The bat dispersal of Andira fruits is well known by local people in South America; "Andira" means bat in the Tupi Amerindian language (Milliken et al. 1992). of Andira possess much larger, heavier fruits, up to 12 cm long and Eight species to 500 g when fresh (Fig. 5B; dry fruit of A. macrocarpa weigh weighing probably up 300 g). These fruit are too large and heavy to be carried by South American fruit bats, which can carry a maximum of 100 g (Fleming 1986). It is probable that they are adapted for dispersal by large rodents (van Roosmalen 1985; Pennington & de Lima 1995), such as paca (Agouti paca), agouti (Dasyprocta spp.), and acouchy (Myoprocta spp.), which are (Emmons 1990). Of these large important dispersers of large fruit in the Neotropics fruited species, I have only seen fresh fruit of A. carvalhoi, which are brown and odorless

ANDIRA

2003

15

when ripe, with a hard, non-fibrous, pale green to green-white mesocarp. When dry, the mesocarp becomes hard and finely granular. Dried fruit of the other large fruited species also have these characters, which suggests that they may be dispersed by similar means. No direct observations of rodent dispersal have been made, but if these large fruits are ro dent dispersed, then this represents a considerable novelty in legumes. The only reported are of Parkia mul cases of rodent dispersal in the estimated 650 genera of Leguminosae tijuga Benth. (Mimosoideae; Hopkins & Hopkins 1983) and Hymenaea courbaril L. (Cae 1986). Some species appear to be secondarily dispersed by salpinioideae; Hallwachs water. The Mexican endemic A. galeottiana has a large fruit of which the dry mesocarp is soft and spongy with air cavities, apparently adapting this species to dispersal by water; its fruits are regularly found on Mexican coastal beaches (Rovirosa 1890; Gunn & Den nis 1976). Andira anthelmia and A. surinamensis are small-fruited species that show river ine distributions and may also be secondarily dispersed by water (Pennington & Gemein holzer 2000). It also seems probable that small-fruited species may be secondarily dispersed by rodents, because their fruits fall to the ground (pers. obs.). Pennington and de Lima (1995) noted thatmode of dispersal appeared to have an ef fect on the distributions of Andira species, with the putatively bat-dispersed species gen erally more widespread. The best example of a wide distribution of a bat-dispersed species isA. inermis (see Figs. 14,15). Putatively rodent-dispersed species are often restricted en is confined to the vicinity of Manaus in central Amazonia, A. demics, e.g., A. micrantha to is the Pakaraima Mountains in Guyana, A. carvalhoi is endemic endemic grandistipula to southern Bahia in Brazil. These distributional differences may reflect the restricted home ranges of seed-dispersing rodents (Emmons 1990; Hallwachs 1986) and the long distances flown by Neotropical fruit bats from their sleeping roosts to feeding areas (up to 10 km; Williams & Williams 1967). Standard analyses of the composition of the dried mesocarps of five Andira species were made using the methods of AOAC (1990). These confirm that starch, sugars, pro tein, and fat are important nutritional components of these fruits (Table 1). It was hoped that the probable differences in dispersal of large and small Andira fruits might be re flected in the composition of their mesocarps. For example, Pannell and Koziol (1987) found wide differences in the nutritional composition of fruits of Aglaia (Meliaceae) dis the data in Table 1 are somewhat inconclu persed by birds and primates. Unfortunately, sive, and it should be noted that A. carvalhoi was the only large-fruited species for which material was available. First, the difference in fat content between the two accessions of A. fraxinifolia suggests that small interspecific differences in values must be treated with

Table 1.Mesocarp of five species of Andira. All figures are % by weight of the air-dried composition the maturity of the fruit was proven by the subsequent germination ex of seed from all accessions, mesocarp; All vouchers are collections of R. T. Pennington and deposited at CEPEC, FHO, and K. cept A. ormosioides. Fruit Speciespresumed A. surinamensis A. fraxinifolia A. fraxinifolia

RTP

364

RTP 202 RTP

213

size and disperser

Starch Sugars

6.6

3.4 32.6

7.3

7.23.3

9.5 5.3

11.6 2.8

5.3

2.0

6.8

A. anthelmia

RTP 227

A. carvalhoi

RTP 232

large, rodent

RTP 306

Fat

small, bat small, bat small, bat small, bat small, bat

A. ormosioides

Protein

8.9

8.6

1.62.2 5.6

2.3 26.2

0.6

13.7 28.7 8.8

SYSTEMATICBOTANYMONOGRAPHS

16

VOLUME 64

caution. Second, although A. carvalhoi has a particularly high starch content, which re the sim flects the large numbers of starch grains in the parenchyma cells of itsmesocarp, ilar value for A. anthelmia indicates that high starch content is not a unique feature of large fruit. Overall, there is little difference in composition between the two fruit types, and the inclusion of more rodent-dispersed species and more accessions of individual species are necessary to further investigate the small differences found. A notable aspect of these data is the high sugar content of the mesocarp of A. suri namensis. This sample was analyzed four weeks after collection in Guyana, whereas the other accessions were a year old; however, the sugars should remain more or less stable unless dried fruit suffer microbial contamination, of which there was no evidence. It ap pears that these fruit are much sweeter than the others. It is possible that these sweet fruits are attractive to different bat species from those that disperse other species of Andira. The fruit of A. humilis has a completely different scent from the fruit of the five other bat-dis persed species that I have collected, which may also be attractive to alternative bats. Andira fruit generally contain a single seed, so the large, putatively rodent-dispersed fruit contain a correspondingly large seed. It is possible that large seeds allow seedlings to tolerate shade for long periods in the rain forest understory (Leishman & Westoby 1994), which may explain why several rain forest species o? Andira (e.g., A. micrantha, A. cori?cea, A. taurotesticulata) have large fruit. In restinga vegetation, the large seed of A. carvalhoi may permit the immediate development of an extensive root system, which helps the seedlings to survive in the easily eroded, rapidly drained, white sandy soils. Seed of this species, grown at the Royal Botanic Gardens, Kew, produced extensive roots, and little above-ground growth, in contrast to themuch stronger shoot growth of the other four bat-dispersed

species germinated.

CHROMOSOMENUMBERS of chromosome numbers in Andira is restricted to those reported by Knowledge Goldblatt (1981). Both n = 10 and n = 11 have been reported in A. inermis. One of these counts (Fritsch 1970; n = 11) is from a tree from Cuba, where A. inermis is absent, and is therefore more likely to be of A. cubensis, the only Andira species in Cuba. Two other = 10. species (not named by Goldblatt) have n Further chromosome counts were attempted for A. anthelmia {R. T. Pennington 282) and A. fraxinifolia {R. T. Pennington 236), two species for which living plants were avail able at the Royal Botanic Gardens, Kew and Edinburgh. Root tips were collected and in distilled water before pre-treatment in a saturated solution of paradichloroben zene for four hours to overnight at room temperature. They were then fixed in ethanol:acetic acid (3:1 vol:vol). The root tips were hydrolyzed for 40 minutes in 5N hy

washed

acid at room temperature before staining with Feulgen (prepared after Fox, two for hours. They were then mounted in 45% acetic acid and observed under 1969)

drochloric

phase

contrast.

Perhaps because the root tips were not collected from entirely healthy, vigorously growing plants, the preparations showed few cells inmetaphase. Thus, the counts made should be regarded as preliminary. Counts for A. fraxinifolia show 2n = 22, and for A. an = 21 or 22. These results, together with those reviewed by Goldblatt (1981), thelmia, 2n indicate that there may be two basic chromosome numbers in Andira: n = 10 and n = 11.

ANDIRA

2003

17

AND INTRASPECIFTC cpDNA POLYMORPHISM POTENTIALHYBRIDIZATIONINANDIRA Introduction Intraspecific cpDNA variation is often low due to slow mutation rates, small effective population size, and the possibility of "selective sweeps" caused by the non-recombining nature of this genome (Ennos et al. 1999); however, Pennington (1995) reported intraspe in A. inermis, A. humilis, A. carvalhoi, and A. verm?fuga. cific cpDNA polymorphism inA. verm?fuga was dismissed, be Subsequently, the intraspecific cpDNA polymorphism cause the accession R. T. Pennington 250 does not belong to this species, but is an indi or perhaps a new species (Pennington 1996; see notes under A. vidual of A. fraxinifolia fraxinifolia). The intraspecific variation inA. inermis involves few restriction site changes and is likely to be due to processes of mutation within this species (Pennington 1995). In contrast, the haplotypes inA. humilis and A. carvalhoi are phylogenetically disparate. This can be explained by two processes: introgres type of intraspecific cpDNA polymorphism of an ancestral polymorphism. Unfortunately, sive hybridization and/or the maintenance the most basic information necessary to confirm introgression as a possibility inAndira? whether different species are interfertile?is missing. Yet, hybridization and introgression is perhaps a more

likely explanation than the maintenance of ancestral polymorphism be cause of the large number of restriction sites (7-10) that differentiate the haplotypes in A. carvalhoi and A. humilis, and the lack of intermediate haplotypes in these or other extant then this polymorphism has species. If ancestral polymorphism were the explanation, been successfully maintained over remarkably long periods, and perhaps we might of the intermediate haplotypes. expect to discover maintenance

also

Whatever the explanation for this infraspecific cpDNA polymorphism involving phy logenetically disparate haplotypes, it is vital to assess whether it is widespread, because processes of hybridization with subsequent introgression and lineage sorting (where an ancestral species is polymorphic for cpDNA, and these cpDNA types sort in subsequent species) can result inmisleading interpretations of phylogenetic relationships based upon cpDNA data (Doyle 1992). Fieldwork in Central Brazil, Costa Rica, and Ecuador pro vided a valuable opportunity to extend this survey to other species from different geo graphical areas. In particular, cpDNA variation was examined inmore populations of A. humilis in an attempt to discover how widespread is intraspecific cpDNA polymorphism in this species. Methods All methods

followed

(1995). Fourteen restriction site mutations Pennington (Ap for different clades of species (Figs. 7, 9) were selected for study, a pendix 1) diagnostic and Brunsfeld technique recommended by Doyle et al. (1990) and Rieseberg (1992). DNAs of accessions included in this previous study (A. humilis RTP 239, 269; A. ver m?fuga RTP 265; A. fraxinifolia RTP 213; A. inermis C. E. Hughes 1673) were included to facilitate identification of banding patterns. Forty-one accessions of eight species were screened (Appendix 2). These included 14 accessions from three populations of A. hu milis, 11 accessions from three populations of A. verm?fuga, and eight accessions from two populations of A. cujabensis.

18

SYSTEMATICBOTANYMONOGRAPHS

VOLUME 64

Results The results are summarized in Appendix 2, which also includes data reported by Pen nington (1995) in order to provide an overall summary of intraspecific cpDNA polymor involving phism in Andira. These data confirm that intraspecific cpDNA polymorphism phylogenetically disparate haplotypes is confined to A. humilis and A. carvalhoi. Wide geographic sampling of several species (e.g., eight populations o? A. fraxinifolia, five pop ulations of A. verm?fuga) has revealed no further instances of such polymorphism. This corroborates the conclusion (Pennington 1995, 1996) that intraspecific cpDNA polymor phism in Andira involving phylogenetically disparate haplotypes is restricted enough to treat the phylogenetic hypothesis derived from cpDNA restriction site data as an approx imation of the true species phylogeny. in seasonally dry cerrado vegetation in central Brazil, Andira humilis is widespread where it is sympatric with A. verm?fuga (Piastome Group I), A. cordata, and A. cujaben sis (both Clade II). Every accession sampled from the center of its distribution (Fig. 6) possessed the plastome type of Plastome Group I, indicating that the true phylogenetic po sition of A. humilis is in Plastome Group I; however, the accessions of A. humilis RTP 239, 246, and 247 have the plastome type of Clade III. These were sampled from populations in the state of Bahia at the edge of the species' range, where the cerrado vegetation meets vegetation of Atlantic coastal Brazil (Fig. 6); there are seven species of Andira, all with Clade III plastomes. It is only here, where these vegetation types meet, that A. humilis comes into close contact with several species of Clade III (e.g., A. fraxini the more mesic

folia, A. nitida, A. anthelmia, A. legalis), which are common elements of vegetation in coastal southeastern Brazil. It is thus tempting to hypothesize that hybridization and sub area. in This have occurred this sequent introgression hypothesis is supported by the ob servation that the individuals RTP 246 and 247 were

small trees, rather than geoxylic suf of A. the form humilis. frutices, typical growth Although A. surinamensis, a species of that border the northern Clade III, is distributed along the southern fringes of Amazonia cerrado (see Fig. 25), it does not occur at high density, and the records of A. humilis from the northern area of the cerrado are few (see Fig. 23). Introgression here seems less likely because of the low probability of contact between species, and itwould be interesting to screen populations of A. humilis from this area to test this. The case of A. carvalhoi ismore complex. Pennington (1995) screened two accessions informative restriction sites). One had the plastome of fully (i.e., for 37 phylogenetically Plastome Group I {RTP 229), and the other that of Clade III {RTP 233; see Fig. 6). Another accession screened for only two diagnostic mutations had one characteristic of Clade III and the other for Plastome Group I, indicating a third plastome type (Pennington 1995). Only one further accession of A. carvalhoi was available for the study reported here. This comes from the same locality as RTP 229, is identical to it in all mutations screened, and thus unequivocally has a plastome type of Plastome Group I. Therefore, the restricted data indicate A. carvalhoi has three different plastome types. Andira carvalhoi has a restricted coastal range in Bahia (Fig. 6) and is entirely sur rounded by species with the Clade III plastome type. If the true phylogenetic affinity of A. carvalhoi is with species of Plastome Group I and its "true" cpDNA type is that of Plas tome Group I, it is possible to explain the presence of the Clade III plastome type in ac cession RTP 233 by introgressive hybridization with a species of Clade III. If A. carval available

hoi's true phylogenetic position is in Clade III and its "true" cpDNA type is that of Clade in A. carvalhoi by recent III, it is difficult to explain the plastome type polymorphism

2003

andira

19

PLASTOMES: CLADEI CLADE II CLADE III(one species) PLASTOME GROUP I (one species)

-10?

PLASTOMESr CLADE II PLASTOME GROUP I

H 20? PLASTOME: CLADE IIIONLY

50?

Amazonian Rain Forest

Populations of A. humi/is with plastome type of Clade III

Seasonally Dry Vegetation

Range

FIG.

6. Location

polymorphism.

of populations The plastome

of Andira

humilis

types present

carvalhoi

A Populations of A. humilis with plastome type of Plastome Group

Atlantic Coastal Rain Forest

cpDNA

of A.

and A. carvalhoi

in each of the different

sampled vegetation

I

in a study of intraspecific types are indicated.

hybridization, because its coastal range is ca. 200 km from the closest localities of A. hu milis and A. verm?fuga, species with the cpDNA type of Plastome Group I. Itmight have been in closer proximity and hybridized with these cerrado species in times of drier cli mates during the Pleistocene, when species of both cerrado and restinga, which are to water have been more widespread. That the cpDNA type of acces stress, may adapted sion A. carvalhoi RTP 229 differs from those of A. humilis or A. verm?fuga by three re striction sites also indicates that recent introgression with these two species is unlikely to

VOLUME 64

SYSTEMATICBOTANYMONOGRAPHS

20

have been the source of its plastome. To unravel the reasons for the complex intraspecific inA. carvalhoi clearly requires sampling of more populations and cpDNA polymorphism full characterization of plastome types. It is tempting to ascribe some of the difficulties in species delimitation inAndira, such some specimens as A. fraxinifolia as the difficulties in distinguishing and A. ormosioides, of hybrids. Ifmorphological intermediacy may be taken as evidence for see are Bennett 1994, McDade 1990), then hy frequent exceptions; e.g., hybridity (there evidence bridization may be possible, but rare. I reported having seen no morphological of hybridity amongst sympatric species of Andira in southeastern Brazilian restinga forests (Pennington 1995). Subsequently, I have observed a single plant that appeared to to the occurrence

between A. anthelmia be intermediate in leaf and stipule morphology in the Poco das Antas Reserve in the State of Rio de Janeiro.

and A. fraxinifolia

CLADISTICANALYSES OF SPECIESRELATIONSHIPS Introduction Cladistic analysis of chloroplast DNA restriction site characters for species o? Andira are presented by Pennington and two outgroup species of Hymenolobium (1995). Pen site and that both restriction directly combining cpDNA morpho nington (1996) argued logical characters in a single cladistic matrix and analyzing both simultaneously provided the best phylogenetic hypothesis for Andira. Cladistic analysis of cpDNA restriction site characters alone failed to resolve relationships between closely related species. Cladistic characters produced a highly unresolved result because of the analysis of morphological lack of characters with discrete states that are suitable for cladistic analysis in Andira. In contrast, combining these data sets directly produced good phylogenetic resolution, be cause the different character sets appear to be providing resolution at different hierarchi cal levels: the cpDNA characters appear to be evolving slowly and delimit groups of characters provide resolution within these groups. The species, whilst the morphological same view is taken here: that a combined cladistic analysis of all available data represents the best phylogenetic sions of infrageneric

hypothesis for Andira, the hypothesis that will be used for discus classification, character evolution, and biogeography. Species

Concepts

were delimited largely using the framework of the phylo 1990; Luckow 1995) and die characters genetic species concept (e.g., Nixon & Wheeler and character states listed below. They are diagnosable entities, characterized by constant or consistent differences. Species

in this monograph

Methods Three cladistic analyses are presented that are based upon Pennington's studies (1995, These are: 1)morphological 1996), with small modifications. analysis; 2) combined mol ecular and morphological and combined molecular and morphological 3) analysis; analy a of subset restriction site data for species not included by Pen sis, including cpDNA nington (1995, 1996). It should be noted that "Andira sp. nov. 1" and "Andira sp. nov. 2" (Pennington 1995, 1996) have subsequently been named A. carvalhoi and A. cordata, re spectively (Pennington & de Lima 1995). "Andira sp. nov. 3" (Pennington 1995, 1996) is

2003

21

ANDIRA

Table

2. Data matrix

of morphological

characters

of Andira.

Character flavum nitidum

Hymenolobium

Hymenolobium A. inermis subsp.

inermis

0 0 0

subsp. rooseveltii

0

subsp. glabricalyx 0 A. jaliscensis A. multistipula 0

0

A.

inermis

A.

inermis

0

A. cubensis A. galeottiana A. macrothyrsa

0 0

A. verm?fuga 1 A. humilis 2 A. anthelmia

0

A. carvalhoi

1

A. fraxinifolia A. legalis 0

1

A. marauensis

0

A.

/u'f?fti 1

A. ormosioides

0

A. surinamensis A. cordata

0

0

A. cori?cea

0

0 A. cujabensis A. grandistipula A. micrantha

0

0

A. parviflora 0 A. praecox 0 A. trifoliolata A. unifoliolata

0 0

A. chigorodensis A. taurotesticulata A. tervequinata A. macrocarpa

0 0 0 0

here included inA. nitida, which is now polymorphic for several character states (see dis cussion under that species), and "Andira sp. nov. 4" (Pennington 1996) is included in A. fraxinifolia (see notes under that species). Data matrices were handled using Dada (Nixon 1993). Parsimony analyses were car ried out using Hennig86 (Farris 1988), using the mh*bb* command, and PAUP* 4.0, beta 2 (Swofford 1999). Character optimization and tree printing was achieved using Clados 1.2 (Nixon

1992).

Morphological analysis. Characters and character states. The data matrix is shown in Table 2. The criteria for selecting morphological characters used in the cladistic analysis follow those of Lavin (1993), and include the assumption that they are functionally and developmentally independent of one another, and that they are intrinsic attributes of the taxa. The characters should also show uniform and consistent occurrence or absence related. among the terminal taxa, which implies that they are not environmentally

22

SYSTEMATICBOTANYMONOGRAPHS

VOLUME 64

The view taken here is that characters with states that show overlapping variation are of doubtful cladistic significance or utility, because there is no objective means of delim 1987; Farris 1990; Stevens 1991; Gift & iting states within them (Pimentel & Riggins Stevens 1997; for alternative viewpoints see: Archie 1985; Chappill 1989; Thiele 1993). For this reason, characters such as leaf and leaflet size, which are useful for species de limitation, were excluded from the phylogenetic analyses. characters are included. These were

treated as unordered, thus min imizing the assumptions built into the analysis. In the list of characters and character states, notes have only been added where characters were not presented or where their coding differs from that described by Pen nington (1996). These differences are: (i) anatomical studies of fruit have refined the coding of the endocarp character; (ii) observation of the pollination of A. anthelmia has suggested a re-coding of the petal color character based upon presumed pollinator; and Several multistate

(iii) including all species (rather than just those for which cpDNA data are available) al lows a character state to be added to the leaflet indumentum character (5) and two new characters to be included: leaflet number (4) and nature of the primary vein (6). The fol lowing conventions are adopted: 1) The first number of each character refers to its po sition in the data matrix for the morphological cladistic analysis; the second number (in to refers its in the data matrix of the combined analysis. 2) The parentheses) position states of each character are described and assigned a code (the number in parentheses after

each

Vegetative

character

state).

characters.

1 (39). Tree (0); able to root-sprout and form amultistemmed shrub (1); geoxylic suf frutex (2). 2 (40). Germination phaneroepigeal (0); germination cryptohypogeal (1). 3 (41). Stipules small and caducous (0); stipules large and persistent (1). 4. Leaves with two or more pairs of leaflets (0); leaves trifoliolate only (1); leaves unifoliolate only (2).?This character was used by Mattos (1979) to define her sections Lumbricidia (defined by possession of five or more leaflets) and Paucifoliolata (defined by possession of trifoliolate or unifoliolate leaves). The character was run ordered and un ordered in separate analyses. The justification for treating it as ordered is ontogeny; seedlings of A. unifoliolata have multifoliolate eophylls (R. T. Pennington, unpubl.). Ap of Nelson's (1988) modification plying Weston's (1978) ontogenetic rule indicates that unifoliolate leaves are less general (i.e., derived) relative to multifoliolate leaves; how ever, until the ontogeny of A. trifoliolata is studied, the exact order of the character states cannot be entirely certain. 5 (42). Leaflet undersurface glabrous (0); leaflet undersurface with indumentum of short (0.2-1.0 mm), flexuose ? appressed to ? erect hairs (1); leaflet undersurface of pale, tangled, erect hairs (2). [Fig. erect hairs 2A, B, C, D]?The fine, pale, tangled, (state 2) of A. cujabensis and A. jaliscen sis are most distinct from the indumentum of all other species of Andira (Fig. 2D). 6. Primary vein channelled above (0); primary vein plane above (1). [Fig. 2E, F]? State 1 is found only in A. unifoliolata and A. trifoliolata. Floral

characters.

7 (43). Flowers yellow flowers (0); pink to purple,

to white,

the standard generally with red or purple markings the standard generally with a pale central marking, less than

ANDIRA

2003

23

18mm

long, pollinated by numerous species of small bees (1); flowers pink to purple, the standard with a pale central marking, 18-23 mm long, pollinated by large xylocopid bees (2).?The coding of this character reflects inferences of pollination biology described length inAndira varies continuously (Fig. 3), and alone it does not provide a good basis for the delimitation of character states. 8 (44). Calyx with indumentum (0); calyx glabrous (1). 9 (45). Wing petal sculpturing present (0); wing petal sculpturing absent (1). 10 (46). Gynoecium with indumentum (0); gynoecium totally glabrous or the ovary

above. Flower

with very few (1-3)

scattered hairs (1).

Fruit characters 11 (47). Fruit a samara (0); fruit a small, bat-dispersed drupe that dries smooth (1); fruit a small, bat-dispersed drupe that dries wrinkled (2); fruit a large, putatively rodent dispersed drupe, with non-fibrous mesocarp (large drupes with mesocarp drying finely granular) (3); fruit a large, putatively rodent-dispersed taining air spaces (A. galeottiana only) (4).

con

drupe, with fibrous mesocarp

12 (48). Endocarp absent (0); endocarp dominated by parenchyma and/or collapsed cell tissue (1); endocarp dominated by fibers (2); endocarp dominated by stone cells (3); character presents endocarp of fiber and stone cells (A. jaliscensis only) (4).?This an of because the lack outgroup species Hymenolobium problems, endocarp. Hawkins et al. (1997) argued that the most theoretically consistent way of coding such characters is by re-conceptualizing them as two characters (the first coding the presence and ab sence of the structure, the second coding the different types of structure, with the ter minal taxa that lack the structure scored as missing data). This is not a perfect solution, because it introduces problems of optimization?for the second character, species of for states of an endocarp, a structure that they do not is coded here as a single multistate character with ab coding as two characters, as recommended by Hawkins et

become optimized Hymenolobium have. For this reason the character sence as one state. Moreover, al. (1997), does not change lack the structure.

the outcome

of any analyses,

because

only the outgroup

taxa

and morphological analyses. Species for which cpDNA data are lacking (e.g., A. cubensis, A. chigorodensis) were not included in these analyses. The first analysis is identical to that presented by Pennington (1996) but includes the re-cod ing of the fruit characters and flower color character described above. The accession A. Combined

molecular

humilis RTP 239 has been identified as possessing a "foreign" plastome, probably derived from introgressive hybridization (see above), and is thus excluded from this and the fol lowing cladistic analysis. The data matrix is presented in Table 3. The second analysis includes a subset of cpDNA restriction site data for species not included by Pennington (1995, 1996). These are accessions o? A. jaliscensis, A. multistip ula,

A.

taurotesticulata,

A.

cujabensis,

and A. marauensis

that were

screened

for

the di

(Appendix 1) as part of the study of intraspecific and cpDNA polymorphism potential hybridization. Where cpDNA data were not avail were states character scored with question marks. The data matrix is presented in able, Table 3. agnostic mutations

described

above

SYSTEMATICBOTANYMONOGRAPHS

24

VOLUME 64

data for Andira. Characters 1-38 are re 3. Matrix of cpDNA restriction site and morphological a for accessions included in the molecular sites; characters 39-48 are morphological. species Multiple are abbreviated: AMC = A. M. de Carvalho; CEH = C. study are indicated by collector and number. Collectors de Lima; MC = M. Cheek; MS = M. Sugiyama; RTP = R. T. Pennington; TDP E. Hughes; HCL = H. Cavalcante = T. D. Pennington. Table

striction

Character

Hymenolobium

flavum nitidum Hymenolobium A. inermis CEH 1673 A.

inermis TDP

A.

inermis MC

A. macrothyrsa A. galeottiana

13558 3579 0 0

A. humilisRTP 269 A. fraxinifolia RTP 250 A. verm?fuga 0 A. anthelmia RTP 227,282 A. carvalhoi

RTP 229

A. carvalhoi

RTP 233

A. fraxinifolia A. fraxinifolia A. A.

RTP 213 MS

889

legalis RTP 307 legalis HCL s.n.

A. nitida RTP

300

A. nitida RTP 292 A. nifwfa RTP

301

A. ormosioides A. surinamensis A. cordata A. grandistipula A. parviflora A. unifoliolata inermis KY?

A.

512

A.

inermis RTP 580

A.

inermis RTP 589

A. multistipula A. jaliscensis A. taurotesticulata A. marauensis A. cujabensis A. carvalhoi AMC A

carvalhoi

RTP 217

12

3 4 5 6 7

1 1 1 1 1 1 ? 1 0 0 0 0 0 0

1 1 1 1 1 1 1 1 1

0

0

0

0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 ?

1 1 1 1 1 1 1 1 1 1 1 0 0 0 0 1

0 0 0 0 0 0 0 0 1 0 1 0 1 1 1 1 1 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 1 0 0 1

1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 ? 1 1 ? 1 0 0 0 ?

1 1 1 1 1 1 1 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 ? 1

1 1 0 0 0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 0

0 1 1 1 ? 1 0 ?

1 90123456789012345 1 1 1 1 1 1 1 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 ?

1 0 1 ? 1

1 1 1 1 1 1

0 0 0 ? ? 1

1 1 1 0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 0 0 0 1 1 1 1 ?

0 0 1 ? 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 ?

0 0 0 0 0 1 1 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 ?

? ? 1 1 1 0 0 0 1 0 1 0 1 1 1 1 1 1 1 1 1 1 1 0 1 0

0 0 0 ? 1 0 0 1 1 0 1 0 0 1 1 0 1 0 0 1 0 0 1 0 0 1 0 0 1 0 0 1 0 0 1 0 0 1 0 0 1 0 0 1 0 0 0 1 0 1 0 1 0 1 ? ? 1 0 1 0 ? ? 9 9 1 0 ? 0 ? ? 9 9 ? 0 1 0 1 ? 9 9? 1 ? 9 9 9 9 0 1 0 ? ? ? 1 ? ? ? ? ? ? 0 9 9 ? 0

2

? 0 0 0 0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 ? 1 1 0 1 1

1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 ? 0 0 0 ? 0

1 1 1 0 0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 ? 1 1 1 1 1

? ? ? ? ?

0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 ? 0 ? 0 1 1 1 1 0 0 0 ? ? 0 0 1 ?

0 1 0 1 1 1 1 1 1 1 0? 0 1 0 1 0 1 0 1 0 1 0 1 0 1 0 1 0 1 0 1 0 1 0 1 ? 1 0 1 0 1 0 1 0 0 0 0 0 0 0 0 1 ? 1 ? 1 ? 1 ? 1 ? 1 ? 0? 0 ? 0 ?

9

9

9

?

? 9

? 9

? ? 9 ? ? ? ? ? ? 9

9

1 1 1 0 0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 ? 1 1 1

0 0 1 1 1 0 0 0 0 0 0 0 0 0 0 ? 0 0 0 0 ? 0 1 1

? ? ?

9

? ? 9

9

? ? 9

9

? ? ? ? 9

9

Results Morphological analysis. Pennington (1996) showed that a cladistic analysis of mor characters of a subset o? Andira species provided little phylogenetic resolution. phological A cladistic analysis of all species, using the twelve morphological characters listed above, also produced little phylogenetic resolution. This lack of resolution reflects the low num ber of characters and the high levels of homoplasy. With all characters run unordered, the trees (the limit that the computer analysis resulted in 2966 equally most-parsimonious could store; CI = 0.57, RI = 0.79, length = 38). The strict consensus tree is entirely unre

ANDIRA

2003

Table

25

3 continued.

2

4

3

Character_67890123456789012345678

Hymenolobiumflavum Hymenolobiumnitidum A. inermisCEH 1673 A. inermisTDP 13558 A. inermisMC 3579 A. macrothyrsa A. galeottiana A. ?urni/frRTP 269 A. fraxinifolia RTP 250 A. verm?fuga A. anthelmia A. carvalhoi

RTP 227,282 RTP 229

A. carvalhoiRTP 233 A.yhmm/o/??RTP213 A.yhmm/0/uzMS889 A. legalisRTP 307 A.

legalis UCLs.n.

A. nitidaRTP 300 A. mttfo RTP 292 A. n?tidaRTP 301

0 1 ? 1 10?00010?0001100000 ?0?0001110000 01??????01 10010000100100011 010001 01000100010000100100011 10010000100100011 010001 1000101000011 10001 07001 1000101100142 10001 00001 10011 100210?1?0122 00001 1 101100012 ?00?1 10701 101 00001100111001101100722 00001101011010111200012 00001100011001100100032

00001101011011100100032 00001101011011101100012 70077777011011101100012 70071107011010111200032 00001101011010111200032

00001 10101 07701 10701 00001101011011107170772

101 101

A. ormosioides

7077777701

1010101200012

A. surinamensis

00001101011010101100022

A. cordata

10101100010100100011113

A. grandistipula

11101100010100111717033

A.parviflora

70177107017100101001013

A. unifoliolata

70101100010100101071013

A. inermisRTP 5\2 A. inermisRTP 5S0 A. intrm?RTP 589

77777777070700100100011 77777777070700100100011 77777777070700100100011

A. multistipula

77777777077700110100011

A. jaliscensis

????????0???00102100014

A.

77777777077700101001031

taurotesticulata

A. marauensis

1107170772 1 10?1?0??2

7777777707770010111017?

A.cujabensis

77777777070700102001013

A. carvalhoiAMC

77777777077701

A. ozraz//i0/RTP217

77777777777771100100032

100100032

solved, except for a clade comprising A. cordata, A. cori?cea, and A. micrantha. With the character of leaflet number ordered, the analysis resulted in the same number of equally trees (2966, CI = 0.57, RI = 0.80, length = 38), but the strict consen most-parsimonious sus tree resolved another clade comprising A. unifoliolata and A. trifoliolata. Combined molecular and morphological analyses. The first analysis resulted in five trees of length = 76; CI = 0.72; RI = 0.88. The strict consen equally most-parsimonious sus is shown in Fig. 7. Of the equally most-parsimonious trees, two can be rejected be cause they result from incorrect optimization of missing values in accessions of A. nitida for character 47. These accessions are assigned a character state for large, rodent-dis for the two persed fruit, when they do not have this state (this species is polymorphic states of bat-dispersed fruit). Rejection of these topologies does not change the topology

26

VOLUME 64

SYSTEMATICBOTANYMONOGRAPHS

-Hymenolobium

flavum

-Hymenolobium

n'rtidum

-A.

inermis

CEH

1673

-A.

inermis

TDP

13558

-A.

inermis MC

CLADE I

3579

-A. parviflora -A.

cordata

-A.

grandistipula

-A.

unifoliolata

-A.

carvalhoi

CLADE II

RTP 229

-A. macrothyrsa

PLASTOME GROUP I

-A.

galeottiana

-A.

humilis

-A.

verm?fuga

-A.

surinamensis

-A.

ormosioides

-A.

anthelmia

-A.

legalis

-A.

legalis HCL sn

-A.

fraxinifolia

-A.

carvalhoi

-A.

fraxinifolia

-A.

fraxinifolia

-A.

nitida

RTP 300

-A.

nitida

RTP 292

RTP 269

RTP 227,282

RTP 307

CLADE III RTP 250 RTP 233 RTP 213 MS

889

-A. nitida RTP 301 FIG. 7. Strict consensus cladogram of five equally most-parsimonious resulting from cladistic cladograms data. Collector and number are indicated analysis of combined Andira cpDNA restriction site and morphological when more than one accession for a single species was included in the molecular study.

27

ANDIRA

2003

of the strict consensus tree. When the same analysis was run on PAUP 4.0b2, where the fruit character (47) can be coded as polymorphic for A. nitida, 26 equally most-parsimo nious trees resulted, with an identical strict consensus. The greater numbers of equally trees reflect different arrangements of the accessions of A. fraxinifo most-parsimonious A. A. and carvalhoi {RTP 233). The semi-strict consensus tree ismore resolved, lia, nitida, with the accessions of A. nitida grouping with A. carvalhoi RTP 233. The only difference between this analysis and that presented by Pennington (1996) lies in the extra resolution in the strict consensus tree in Clade III (Fig. 7). Andira ormo sioides, A. anthelmia, and A. legalis form amonophyletic group, supported by the synapo morphy of larger flowers pollinated by xylocopid bees. Andira fraxinifolia, A. nitida, and A. carvalhoi are a monophyletic group diagnosed by the ability to root-sprout and form a multistemmed shrub. The combined analysis that included a subset of cpDNA restriction site data for species not included by Pennington (1995, 1996) resulted in ca. 2700 trees (the limit that the computer could store; length = 87; CI = 0.65; RI = 0.87). The strict consensus is shown in Fig. 8 and demonstrates that A. taurotesticulata, A. multistipula, and A. jaliscensis are members of Clade I, which previously only comprised accessions of A. inermis (Pen nington 1995, 1996). The affinities of A. multistipula and A. jaliscensis with A. inermis were suggested by their close overall similarity in morphology. Andira taurotesticulata, with its white-yellow flowers and large ridged fruit, is more divergent morphologically. Yet, it does have in common with A. inermis and A. multistipula an endocarp of parenchy matous

tissue, shown in the equally-most parsimonious tree (Fig. 9, character 48, state 1) to be the plesiomorphic state for Andira. Similarly, Pennington (1994) and Pennington and Gemeinholzer (2000) predicted that all species with an endocarp of stone cells would be members of Clade II. This is corroborated by the placement of A. cujabensis in this clade, and it is likely that A. cori?cea, A. trifoliolata, A. tervequinata, A. micrantha, and A. prae cox also belong here. Andira marauensis is a member of Clade III, placed as sister species to all other of this because it has the plesiomorphic state of restriction site characters clade, species 33 and 38. Clade III remains "cryptic" in the sense of Wojciechowski et al. (1993), be cause it is not marked by any morphological characters but well supported by cpDNA re striction Andira

site

characters. jaliscensis,

A.

multistipula,

A.

taurotesticulata,

A.

cujabensis,

and

A.

ma

rauensis have incomplete restriction site data, and therefore many missing values. Termi nal taxa with missing values are liable to occupy different topological positions in indi vidual equally most-parsimonious trees (Nixon & Davis 1991; Platnick et al. 1991), thus trees and decreasing the resolution increasing the numbers of equally most-parsimonious of consensus trees. Therefore, in an attempt to gain a more accurate idea of the relation ships of each of these species, separate analyses were run where all except one was ex cluded. In only one case, that of A. taurotesticulata, did this result in a different placement for the species in comparison with Fig. 8. In the analysis where A. taurotesticulata was in cluded alone, it is resolved as a sister species to the different accessions of A. inermis in Clade I in a strict consensus cladogram, because it possesses the plesiomorphic state for character 42 (leaflet undersurface indumentum). The results of this combined analysis that includes five extra species is used as the basis for the following discussion of character evolution and biogeography, despite the problem of the large number of equally-most parsimonious trees.

28

SYSTEMATICBOTANYMONOGRAPHS

VOLUME 64

-Hymenolobium

flavum

-Hymeno/obium

nrtidum

-A.

inermis

CEH

1673

-A.

inermis

TDP

13558 3579

-A.

inermis MC

-A.

inermis

RTP 512

-A.

inermis

RTP 580

-A.

inermis

RTP 589

CLADE I

-A. mu/tistipula -A.

jaliscensis

-A.

taurotesticulata

-A.

parvif/ora

-A.

cujabensis

-A.

cordata

-A.

grandistipu/a

-A.

unifoliolata

-A.

carvalhoi

RTP 229

-A.

carvalhoi

AMC

CLADE II

PLASTOME GROUP I

-A. macrothyrsa -A.

galeottiana

-A.

humilis

-A.

verm?fuga

-A. marauensis -A.

surinamensis

-A.

ormosioides

-A.

anthelmia

RTP 227,282

-A.

legalis

RTP 307

-A.

legalis

HCL sn

-A.

fraxinifolia

RTP 250

-A.

fraxinifolia

RTP 213

-A.

fraxinifolia

MS

CLADE III

889

-A.

n?tida RTP 300

-A.

n?tida RTP 292

-A.

n?tida RTP 301

-A.

carvalhoi

RTP 233

-A.

carvalhoi

RTP 217

FIG. 8. Strict consensus cladogram of 2996 equally most-parsimonious cladograms resulting from cladis tic analysis of combined Andira cpDNA restriction site and morphological of species data, including accessions not studied previously by Pennington Collector and number are indicated when more than one ac (1995,1996). cession for a single species was included in the molecular study.

ANDIRA

2003

29

CHARACTEREVOLUTION Stipules (Fig. 9, character 41). Large persistent stipules have pendently in Andira, and are autapomorphic for A. grandistipula synapomorphic for A. anthelmia and A. legalis. In Pisum sativum mutant, stipules-reduced {st) causes a marked reduction in stipule

arisen three times inde and A. multistipula and L. (pea), a single gene size from the large, fo

liaceous wild-type stipules (Marx 1987). It is tempting to ascribe the apparent lability of this character in Andira to a similarly simple genetic switch. Indeed, Andira could be a good model system to investigate the evolutionary significance of such a developmental mutation, especially in the case of A. multistipula, where it is the presence of these large, this species from A. persistent stipules that is the diagnostic character that distinguishes inermis. From the standpoint of the phylogenetic (Nixon & Wheeler concept species 1990), it was the fixation of the trait of large stipules in a population that represented the speciation

event.

Flowers (Fig. 9, character 43). White petals are shown to have arisen at least three times. They are a synapomorphy for Clade II, and an autapomorphy diagnosing both A. macrothyrsa and A. taurotesticulata. Other species not sampled in themolecular study (A. cori?cea, A. micrantha, A. praecox, A. tervequinata, A. trifoliolata) are known to have an endocarp composed of stone cells and to lack wing petal sculpturing, both synapomor phies for Clade II. Hence, all these species probably belong in this clade, which would then be defined by white flowers. Andira chigorodensis, the remaining species with white most to is A. similar and may be its sister species. petals, morphologically macrothyrsa Loss of wing petal sculpturing is a synapomorphy for Clade II, and an autapomorphy for A. taurotesticulata. Itmay have also been independently most is species closely allied with A. macrothyrsa.

lost by A. chigorodensis,

if this

Fruit type and dispersal (Fig. 9, character 47). The new restriction site data for A. tau rotesticulata demonstrate that it is a member of Clade I, and that there have been at least five independent origins of rodent dispersal from bat dispersal inAndira. This is one more independent origin than reported by Pennington (1996), confirming the prediction that four origins was an underestimate. Of the remaining species with large fruit, A. cori?cea and A. micrantha are likely to be members of Clade II based upon their floral characters of stone cells, but they share no great morphological similarity with A. the other species of that clade with large fruit. Andira macrocarpa has a woody endocarp, and its affinities are hard to guess, because it has never been collected in flower. Again, I suggest that further independent origins of rodent dispersal may have and endocarps

grandistipula,

occurred during the diversification

o? Andira.

Endocarp (Fig. 9, character 48). Examining the sequence of character state change on the cladogram (Fig. 9) shows a transition from weak endocarps of parenchyma to woody fibrous endocarps to endocarps composed of stone cells. This represents a sequence of in 2000; discussion above), and seed creasing seed protection (Pennington & Gemeinholzer have the been this pr?dation may pressure driving evolutionary sequence.

SYSTEMATICBOTANYMONOGRAPHS

30

r-fl-[]?Hymenolobium ?|?Hymenolobium

VOLUME 64

f/avum nitidum

1 7 13202530323642

fOttHf? 10 1 110

10 0

?A.

inermis

1673

CEH

1232 1824 rff--A.

4XH 0 ?

inermis

00

?-A.

CLADE I

13558

TDP 3579

inermis MC

parvif/ora 4246

I-A.

3 1012192122283637434548

ttOHUOHHH 000100101013

A.

cordata

CLADE M

16274147 0 113

A. grandistipu/a

A. unifoliolata

0 1111 394247 S-S? A.

RTP 229

carvalhoi

1? 3

4348

1423

0 1 4647 if 1 2

-A. macrothyrsa -??A. 3439

/H//W//S /?7P 269

-??A.

ft 1 1

HHKHKtH 0 0 0 10 2

PLASTOME GROUP I

galeottiana

?A.

verm?fuga

A. surinamensis A.

ormosioides A.

anthelmia

RTP 227r282 /?7P 307

4 6 8 3338

A.

/e#3//s

LKK}ft 10 0 11

A.

legalis HCL sn

A.

fraxinifolia

RTP 250

A.

fraxinifolia

RTP 213

-A. fraxinifolia 47 ?A,

MS

carvalhoi

CLADE III

889 RTP 233

A. n?tida RTP 300 A. n?tida RTP 292 A. n?tida RTP 301 FIG. 9. An equally most-parsimonious cladogram resulting from cladistic analysis of combined Andira data. The bars representing character changes are labelled above with cpDNA restriction site and morphological the character number and below with the character state. Solid black bars = unique changes, stippled black bars = and number are indicated when more than one accession for a single species changes. Collector homoplasious was included in the molecular study.

2003

ANDIRA

31

CRYPTICCLADES Pennington (1995) reported that none of themonophyletic groups, even the well-sup ported clades, discovered in themolecular analysis o? Andira had been recognized by pre vious workers. They lack major morphological innovations and hence are "cryptic" clades sensu Wojciechowski et al. (1993). The combined analysis demonstrates that new micro characters provide unambiguous support for some of these cryptic groups. morphological Clade II is supported by two unique micromorphological character states: lack of wing an and of stone cells petal sculpturing (character 45) (character 48). All species endocarp of Clade I have an endocarp of parenchyma, but this maps as a plesiomorphy for Andira rather than a synapomorphy. At a higher taxonomic level, micromorphological characters have also proved congruent with other clades that are well supported by molecular data in (Rudall 2000). In both cases, reciprocal illumination provided by new monocotyledons hypotheses of grouping from molecular data is important. For example, itwas only when the cpDNA restriction site data suggested the Clade II grouping of Andira species that I noticed

that endocarps of these species appeared somewhat paler and harder than in other species, which prompted the anatomical study reported above. Other clades, such as clade III, remain cryptic, supported by restriction site data but not by morphological characters. This raises the question of whether they should be rec ognized in formal classifications (Wojciechowski et al. 1993; Pennington & Gemeinholzer 2000), such as an infrageneric classification o? Andira.

INTRAGENERICCLASSIFICATIONSIN THE LIGHTOF THE CLADISTICANALYSES The cladistic analyses show that the infrageneric classifications proposed for Andira Bentham (1860, 1862) and Mattos (1979) are flawed. The only section that has been by that is sect. Paucifoliolata Mattos, comprising only A. may be monophyletic proposed A. amonophyletic and This is cladis unifoliolata trifoliolata. group in themorphological tic analysis, if the character of leaflet number is treated as ordered. The gynoecium indu mentum character used by Bentham to define his sections Lumbricidia and Euandira, and by Mattos to define corresponding subsections in her section Lumbricidia is homoplasious is paraphyletic and (Fig. 10); thus Bentham's section (Mattos's subsection) Lumbricidia section Euandira (Mattos's subsection Glabratae) polyphyletic. the cladistic framework presented here does not provide a clear route Unfortunately, to a new infrageneric classification o? Andira based upon monophyly. There are four well supported clades (Fig. 9): Clade I, Clade II, Clade III, and the group of Clade II, Clade III and Plastome Group I; however, giving these clades formal taxonomic recognition leaves the weakly supported "Plastome Group I," supported as monophyletic by a single, homo restriction site character. both Clade III and I remain cryptic, Clade Moreover, plasious restriction site but not data characters. Whilst there is no supported by by morphological reason to doubt that these cryptic clades reflect true phylogeny because they lack mor ultrastructural and characters may well phological support (indeed, micromorphological be found to support them, as was the case for Clade II), I question the practical utility of giving them formal taxonomic recognition. Because of the lack morphological characters, identification. Hence, I propose that sectional they cannot be keyed out for conventional classification within Andira should be abandoned. A further factor supporting this

SYSTEMATICBOTANYMONOGRAPHS

32

?Hymenolobium -Hymeno/obium

flavum nitidum

VOLUME 64

= 0 = 0 -A.

= 0

taurotesticulata ?A.

= 0

jalisensis

-A. 1-A -A.

inermis

TOP

-A.

inermis

-A.

inermisRTP

CEH

1673=

inermis

RTP 580=

-A.

inermis

RTP 589

?A.

= 0

0

512=

-A.

0

13558=

3579

inermis MC

0 0 = 0

= 0

multistipu/a -A.

= 0

cujabensis

= 0

-A. parviflora

cordata

=

-A.

grandistipula

A.

A.

c

A.

carvalhoi

RTP

A.

carvalhoi

AMC=

-j?A.

0

i-A.

=

humilis .

RTP =

verm?fuga

ormosioides= A.

A.

RTP

legalis

RTP 307

legalis

HCL

fraxinifolia

RTP

250

fraxinifolia

RTP

213=

fraxinifolia

MS

A.

n?tida RTP

A.

carvalhoi A.

0 0

0