Phylogenetic relationships of Caralluma R. Br. (Apocynaceae) Author(s): Peter V. Bruyns, Amina al Farsi and Terry Hedderson Source: Taxon, Vol. 59, No. 4 (August 2010), pp. 1031-1043 Published by: International Association for Plant Taxonomy (IAPT) Stable URL: http://www.jstor.org/stable/20773974 . Accessed: 09/02/2015 05:04 Your use of the JSTOR archive indicates your acceptance of the Terms & Conditions of Use, available at . http://www.jstor.org/page/info/about/policies/terms.jsp . 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 support@jstor.org. . International Association for Plant Taxonomy (IAPT) is collaborating with JSTOR to digitize, preserve and extend access to Taxon. http://www.jstor.org This content downloaded from 137.158.75.81 on Mon, 9 Feb 2015 05:04:47 AM All use subject to JSTOR Terms and Conditions TAXON 59 (4) August 2010: 1031-1043 Phylogenetic Bolus Herbarium, In this paper we trnH and trnT-F and Caralluma nopsis Hook. Pseudolithos some investigate invaded Keywords South Africa among two nuclear ITS regions one of these clades. lineages. of these clades One Caralluma Islands four times by species Apocynaceae; using Our contains species paleotropical while Caralluma', show Caralluma of Caralluma in the other clade of Caralluma. species its greatest DNA We INTRODUCTION Caralluma R. Br. belongs to a group of stem-succulents usually known as stapeliads. The approximately 330 species of stapeliads make up about half of the tribeCeropegieae of inApocynaceae (Endress & Bruyns, 2000). Asclepiadoideae are found They exclusively in theOld World, mainly inhabit areas inAfrica, Arabia and peninsular to arid semi-arid ing India. The major genera among the stapeliads areHuernia R. Br. (51 species, Bruyns, 2008), Orbea Haw. (56,Bruyns, 2005) and Caralluma (58).While all other largergenera of stapeliads are confined to theAfrican continent and southernArabia, Caralluma is unique as its distribution extends well beyond this region.Most of the species are found in the tropics of thenorthernhemisphere. They are known fromGran Canaria (Canary Islands) in thewest toMandalay in centralMyanmar (Asia) in the east and from as far north as 38?N in southern Spain inEurope to 6? S in central Tanzania inAfrica. Despite itswide distribution (much of itoutside the distribution of all other stapeliads), Caralluma has diversified most within the region where the other stapeliads occur and themajority (of over 30 species) is found inNorth-East Africa and southern Arabia. A unique feature of Caralluma is the furtherminor centre of diversity in the Indian region (includingMyanmar and Nepal), where 14 species are known. Gilbert (1990) defined Caralluma as those stapeliads with four (rarely six) laterally compressed angles along the stems without conical tubercles;with rudimentary, scale-like leaves; with inflorescence of very variable form at or near the apex of the stem;with corolla varying from deeply divided to tubu lar;with outer corona mostly conspicuously bi-lobed. On the basis of the colour of the stems, the development of the angles along the stems, the shape of the leaf-rudiments, the type and position of the inflorescence and the orientation of the fruit, Gilbert created four subgenera: C. subg.Boucerosia (Wight& consist is not monophyletic, with discuss and diversity data; two plastid markers: that the stapeliads and representatives subcontinent. sequence (Apocynaceae-Asclepiadoideae-Cerop the small genera in its distribution to the Indian R. Br. from four molecular analyses this lineage, inMacaronesia biogeography; of data Within among of Caralluma, analyses and ncpGS. Bally, while ismainly from the Canary for Caralluma relationships phylogenetic f. and Rhytidocaulon P.R.O. are embedded P.R.O. Bally of the clades. stapeliads, 7701 Rondebosch, the stapeliads, dominating two major among Town, of Cape Peter Bruyns, peter, bruyns@uct.ac.za the largest genus egieae), of Caralluma Phylogeny al Farsi & Terry Hedderson University for correspondence: Abstract al. R. Br. (Apocynaceae) of Caralluma relationships Peter V. Bruyns, Amina Author Bruyns & N.E. is the most widely We that the Indian is found inNorth-East distributed Africa Echid Dalz. and that characterize group subcontinent and with distributed Plowes, Br, Frerea features morphological show its species ofAnomalluma Edithcolea regions psbA of two lineages, among the has been southern Arabia. phylogeny Arn.) M. Gilbert (with 27 species), subg.Caralluma (21), subg. Desmidorchis (Ehrenb.) M. Gilbert (5) and subg. Urmalcala M. Gilbert (3). Of the 56 names listed by Gilbert, fourwere re duced subsequently to synonymyunder C. adscendens (Roxb.) R. Br. (Bruyns, 1992) and C. adenensis (Deflers) A. Berger (Bruyns& Jonkers, 1993).A furthersix new species (C. baradii Lavranos, C. flavovirens L.E. Newton, C. sudanica Bruyns and C. vaduliae Lavranos ofC. subg. Caralluma), C.faucicola Bruyns of C. subg. Boucerosia and C. lamellosa M. Gilbert & Thulin of uncertain affinity,were also described (Lavra nos, 1991, 1993; Newton, 1998; Bruyns, 2004, 2010; Gilbert & Thulin, 2005), bringing the total to 58. Plowes (1995) subdivided Caralluma into 17 genera, of which sixwere monotypic but this classification has not been accepted in any recent regional accounts of thegenus (Gilbert, 2003; Lavranos, 2006) and need not be considered furtherhere. In a preliminary phylogenetic investigation of the stape liads using the nuclear ITS region and the chloroplast trnT-F region (trnT-trnLintergenic spacer, trnL intronand trnL-trnF intergenicspacer),Meve & Liede (2002) considered thedelimi tationofCaralluma. They included 20 of the 56 known species of Caralluma (sensu Gilbert, 1990), as well as between one and three representatives of all other stapeliad genera except Baynesia Bruyns and Socotrella Bruyns. Their main findings were that: (1) the stapeliads aremonophyletic and nestedwithin Ceropegia L.; (2) representativesof generamainly fromAfrica south of the equator (including thewidely distributed genera Duvalia Haw., Huernia and Orbed) form a well-supported 'southernclade'; (3) this 'southernclade' is sister to awell-sup ported clade made up of the twomonotypic generaDuvaliandra M. Gilbert andWhitesloanea Chiov. fromSocotra and Somalia, respectively; (4)Duvaliandra + Whitesloanea + the 'southern clade' is one lineage among ten in an unresolved polytomy.The nine other lineages in thispolytomy consist of representatives of the remaining stapeliads, mainly fromAfrica north of the 1031 This content downloaded from 137.158.75.81 on Mon, 9 Feb 2015 05:04:47 AM All use subject to JSTOR Terms and Conditions Bruyns& al. Phylogeny of Caralluma equator toMyanmar, belonging to the genera Anomalluma Plowes, Caralluma (sensu Gilbert, 1990),Echidnopsis Hook, f, Edithcolea N.E. Br, Frerea Dalz, Pseudolithos P.R.O. Bally and Rhytidocaulon P.R.O. Bally; (5) Caralluma (sensu Gil bert, 1990) is notmonophyletic, since Caralluma peschii Nel is nested within the 'southern clade'. Apart fromCaralluma peschii, theother 19 species ofCar alluma sampled fell into seven lineages which, togetherwith twomore lineages (one containing species ofEchidnopsis and Rhytidocaulon and another containing Anomalluma maccoyi (Lavranos & Mies) Meve & Liede) made up the nine other lineages in the polytomy mentioned in (4) above. Using this polytomy of nine lineages,Meve & Liede (2002) distributed the species ofCaralluma among the six genera ApteranthesMikan, Boucerosia Wight & Arn, Caralluma, Caudanthera Plowes, Desmidorchis Ehrenb. andMonolluma Plowes. The monotypic genera Edithcolea andFrerea as well as Pseudolithos migiurti nus (Chiov.) P.R.O. Bally were nested in threeof these lineages, in each case as sister tovarious species ofCaralluma. Meve & Liede (2002) subsumedFrerea within Boucerosia but theother two small genera Edithcolea and Pseudolithos were not placed in synonymy.Thus, their conclusions were inconsistentwith theirresults.Gilbert (2009) has also questioned theirtreatment by suggesting thatthe level of samplingwas too low and the re sults too fragmented to justify any of theirtaxonomic changes. Using fourmolecular markers, we aim here to resolve this polytomy of ten lineages. In the process we aim to establish: (1) the relationships between the species of Caralluma (sensu Gilbert, 1990); (2) the relationships between the species of Caralluma (sensu Gilbert, 1990) and the lineage consisting of Duvaliandra + Whitesloanea + the 'southern clade'. MATERIALS AND METHODS ? For Preparation for scanning electron micrographs. SEM micrographs of leaf-rudimentsand associated structures, material was prepared as described inBruyns (1993). Taxon sampling. ? Taxa sampled are listed in theAppen dix. Since the stapeliads are nested within Ceropegia (Meve & Liede, 2002; Meve & Liede-Schumann, 2007), we have selected five species ofCeropegia as outgroup.Within the stapeliads, the lineage consisting ofDuvaliandra + Whitesloanea + the 'south ern clade' contains about 235 species. From this lineage, we choseDuvaliandra and Whitesloanea and, since itsmonophyly was well-supported inMeve & Liede (2002), we represented the 'southern clade' by Australluma peschii (Nel) Plowes, Ophi onella arcuata (N.E. Br.) Bruyns and Quaqua linearis (N.E. Br.) Bruyns. From the remaining stapeliads, we sampled two species each ofAnomalluma and ofEchidnopsis, threespecies of Rhytidocaulon and 41 out of a total of 56 species ofCaralluma (excluding C. peschii=Australluma peschii, referredtohereaf teras Caralluma s.l.),with all four subgenera ofGilbert (1990) well represented.Furthermore, threevarieties ofC. adscendens and duplicate accessions of each ofC. adscendens vdLX.fimbriata (one ofwhich represented the formerC. subulata fromArabia), C. arabica, C.flava and C. penicillata were included, bringing TAXON 59 (4) August 2010: 1031-1043 the totalnumber of accessions ofCaralluma s.l. sampled to 47. The twomonotypic genera, Edithcolea and Frerea as well as two representatives ofPseudolithos (P. caput-viperae, P. migi urtinus)were also included. DNA extraction, amplification and sequencing. ? For our investigations,we employed two chloroplast regions (psbA trnH, trnT-F) and two nuclear regions (ITS, ncpGS). The pro tocol ofDoyle & Doyle (1987) was followed for the extraction of DNA from livematerial or from pieces of stem dried in silica-gel. The psbA-trnH region was amplified with primers psbAF and trnHR (Sang & al, 1997). The trnT-Fregion (trnT trnL intergenic spacer, trnL intronand trnL-trnF intergenic spacer) was amplified using primers trnC and trnF (Taberlet& al, 1991). For the ITS region (ITS1 and ITS2 and the interven ing 5.8S region), primers ITS4 and ITS5 were used (Baldwin & al, 1995). For thencpGS regionwe used primers GScp687f and GScp994r (Emshwiller & Doyle, 1999). Initial PCR amplification of DNA was undertaken in a volume of 30 ul. Each reaction contained 3 ul of diluted (1:10) DNA template, 17.65 ul sterilisedPCR water, 3 ul 10xNH4 buf fer,3 ul 50 mM MgCl2, 1.2 ul dNTPs at 2.5 mM, 1 ul of each primer at 10mM strength,and 0.15 ul of Biotaq DNA Poly merase. Amplifications were performed on theGenAmp PCR system 2700, for 30 thermal cycles. Each cycle consisted of initialdenaturation at 94?C for2min, denaturation at 94?C for 1min, annealing of primers at 52?C for 1min, and extension of new strands at 72?C for2min. Each run also included a final extension of 7min. The PCR productswere checked on agarose gel for successful amplification. The amplified products were cleaned using a GFX PCR DNA purification kit (Amersham Biosciences Ltd). From the cleaned DNA, 2 ul was used for cycle sequencing with a reaction mix containing 3.68 ul of sterilisedwater, 2 ul ofABI BigDye Terminator Ready Reac tion (TRR), 1 jiiLof 5x sequencing buffer and 0.16 ul of each primer. This made a total volume of 10 ul of reagent,which was thencycle-sequenced in a PCR machine for25 cycles. The products were resolved on an ABI 3130XL Genetic Analyzer at theDNA sequencing unit. Sequences were aligned by eye inBioEdit (Hall, 1999). Indels were coded using the simple indel coding method of Simmons & Ochoterena (2000). A region of dubious alignment in thepsbA-trnHregion was excluded from the analyses. ? Parsimony analyses. Parsimony analyses were per formedwith PAUP V.4.0M0 (Swofford,2002). To detect poten tial conflicts between thedata, three sets of data were analyzed separately, namely combined cpDNA, combined nrDNA and totalDNA (theunion of the other two sets). For each analysis, all characterswere equally weighted. In each analysis a heuristic searchwas employedwith 1000 random sequence addition rep licates and TBR branch-swapping,with amaximum of 10 trees held at each replicate and 'Maxtrees' set to 10,000.Majority rule and strictconsensus treeswere calculated from the trees saved. For each set of data, branch support (Jackknifepercentage sup port, JK)was estimated by 1000 jackknife replicates (with the same settings as for the heuristic search),with 36.79% of the characters randomly deleted from the data in each replicate (Farris & al. 1996). For the analyses of the combined cpDNA 1032 This content downloaded from 137.158.75.81 on Mon, 9 Feb 2015 05:04:47 AM All use subject to JSTOR Terms and Conditions TAXON 59 (4) August 2010: 1031-1043 Bruyns and combined nrDNA, theconsensus treeswere inspectedvisu ally for stronglysupported conflict before the set of combined data was analysed. Taxa with missing data were included in the totalDNA analysis since, with the considerable number of parsimony-informative characters present in other regions, theywould not greatly affectresolution achieved (Wiens, 2003). ? Modeltest v.3.7 (Posada & Cran Bayesian analyses. dall, 1998) was used with theAkaike information criterion (Akaike, 1974) to determine the appropriate substitutionmod els for each of the four regions. The models foundwere used forBayesian Inference with MrBayes v.3.1.2 (Huelsenbeck & Ronquist, 2001) on the totalDNA dataset. The data were partitioned into the four regions, with parameters (statfreq, revmat, shape, pinvar) all unlinked between the partitions. was implemented for each partition. The model GTR+G+I Several analyses were conducted with fourMarkov chains in each of two independent runswith the following settings: 106 generations, trees sampled every 100thgeneration, four chains, branch lengths saved. After 106generations itwas found that the standard deviation of split frequencies was below 0.01 and the analysis was discontinued. In each analysis, by examining the decrease in the standard deviation of split frequencies, it was found that chains had achieved stabilitywithin the first quarter of the samples so burn-in was set to 2500. From the remaining 7501 trees a 50% majority-rule consensus tree and posterior probabilities (PP) were calculated. RESULTS Statistics, including thenumbers of variable positions, in formative positions and indels for each of the fourmolecular markers, are given inTable 1. The strictconsensus trees (not shown) obtained fromanal ysis of combined cpDNA and combined nrDNA were poorly resolved. Nevertheless, no hard incongruence between these two sets of data was found (asmeasured by Jackknifepercent ages) and so the two datasets were analysed together. Table 1. Statistics for themarkers used and for parsimony of taxa Aligned length Included characters Variable characters Parsimony informative characters Score of best treewithout Number gaps of informative gaps of trees retained PhylogenyofCaralluma The strictconsensus tree fromparsimony analysis of total DNA (not shown) was highly resolved. It differs but slightly from themajority-rule consensus tree fromBayesian inference (Fig. 1),which is also highly resolved, with most of the nodes supported. DISCUSSION Analysis of our data leads to resolution of thepolytomy of ten lineages ofMeve & Liede (2002), as well as clarifying the relationshipsbetween the species ofCaralluma. Furthermore,re lationshipsbetween Caralluma and theremaining stapeliads are also resolved.According to our results, the stapeliads consist of twomajor lineages.One of these lineages is thebranch consisting ofDuvaliandra + Whitesloanea + the 'southernclade',which has radiated especially in southernAfrica andMadagascar, where there are 182 species (Bruyns, 2005). Within this lineage the generaDwva/za,Huernia and Orbea (Meve & Liede, 2002) have diversified in southernAfrica and also have, in total, some 53 species inAfrica north of the equator and in southernArabia. The other lineage, inwhich themajor elements are Caral luma s.l, Echidnopsis and Rhytidocaulon, contains a total of about 95 species, mainly inAfrica and Arabia but only north of latitude 6?S. Within this lineage, the species of Caralluma s.l. group into twomajor clades (A and B in Fig. 1).Clade A is part of a lineage along with Anomalluma and Echidnopsis + Rhytidocaulon and consequently Caralluma s.l. is notmono phyletic. Clade B is sister to the lineage that includes clade A and includes species of Caralluma s.l. as well as Edithcolea, Frerea andPseudolithos, which are nested among the species of Caralluma. Our sampling is sufficientlydense to show clearly thatnone of the subgenera Boucerosia, Caralluma, Desmidor chis and Urmalcala of Gilbert (1990) ismonophyletic (Table 2) and we have established thatall except C. subg. Caralluma are contained within clade B. Species ofCaralluma s.l. are verywidely distributed in an East-West direction, fromMyanmar inAsia to theCanaries in conducted. ITS nrDNA 54 68 68 68 732 1227 2758 691 1178 2537 230 81 311 510 cpDNA 67 68 68 632 899 1531 495 492 867 1359 487 120 79 199 69 (14%) 29 (3%) 98 (7%) 36(7%) 143(21%) 179(15%) 298(12%) 172 103 290 112 447 570 5 7 12 2 7 9 10000 10000 10000 10000 ncpGS 318 Lengthofbest treewith gaps Number al. trnT-F psbA-trnH Number analyses & 10000 cpDNA+nrDNA 21 581 918 10000 480 Consistencyindex(CI) Retentionindex(RI) 0.7442 0.8058 0.7013 0.8036 0.6331 0.6575 0.6591 0.7991 0.9071 0.8051 0.8911 0.8041 0.8202 0.8057 Rescaled consistencyindex(RC) 0.5947 0.7309 0.5691 0.7161 0.5091 0.5393 0.5311 1033 This content downloaded from 137.158.75.81 on Mon, 9 Feb 2015 05:04:47 AM All use subject to JSTOR Terms and Conditions Bruyns& al. Phytogeny of TAXON Caralluma Macaronesia (Figs. 2-3), thoughwith very low levels of local endemism. Members of Caralluma s.l. are the only stapeliads found in the Indian subcontinent, but they are complemented thereby considerable diversity (including succulent species) in the closely related genus Ceropegia. Similarly, theyare theonly stapeliads inMacaronesia and here, too, theyare complemented by two succulent species of Ceropegia (Bruyns, 1986b). Our results show thatCaralluma s.l. has colonised the Indian sub continent four times, once within clade A and thrice in clade B and that these invasions exhibit two distinct patterns. One pattern is thatexhibited by clade A, where the species are found only in the Indian subcontinent and inNorth-East Africa. This suggests that the 'Indian subclade' of clade A has arisen after an event of long-distance dispersal, since the stapeliads (with a maximum age of approximately eightmillion years; Rapini & Fig. 1.Majority-rule tree from Bayesian exhibited bers above consensus analysis, as a phylogram. Num or below a branch are posterior probabilities/Jack for knife support percentages the branch. Only cases where at least one of these is 'strong' are shown. Thickened lines also indicate other cases of support wherePP > 0.97.On theright hand side theclades discussed in the text are indicated. C. = = Cer. Caralluma, Ceropegia, = ads. ascendens. 1.00/100 Cer. bulbosa August 2010: 1031-1043 (4) al, 2007) are too young to have been presentwhen peninsular India and theAfrican continentwere near one another. The same pattern seems to be true forFrerea and the seven species of the 'IndianBoucerosia group' (ifone assumed thattheircom mon ancestor with C. edulis + C.sinaica existed in theNorth East Africa-Arabian region) and it isknown in other succulent groups. Examples of this are several sections of Adenia (De Wilde, 1971: fig. 5) of thePassifloraceae, Euphorbia sect.Eu phorbia (Bruyns& al, 2006) ofEuphorbiaceae as well as in the closely related genus Ceropegia (as seen in the relationships of C.juncea Roxb. to species fromNorth-East Africa andArabia; Meve & Liede-Schumann, 2007). A second pattern is found in subclade III of clade B, where C. tuberculata has reachedNW India via an almost continuous distribution from theArabian Peninsula and thisphenomenon is repeated inCaralluma edulis. Cer. juncea 1.00/100 59 1.00/100 -Cer. occidentalis Cer. cimiciodora] Cer. radicans Outgroup -Ophionella arcuata -Australluma peschii 1.00/100 1.00/731 linearis -Quaqua -Duvaliandra dioscoridis crassa -Whitesloanea 1.00/87L -C. adenensis lavranii 1.00/ rC. C. flava Acc. 1 C. awdeliana C. acutangula 0.99/ C. flava Acc. 2 C. arabica Acc. 2 1.00/75 C. arabica Acc. 1 C. penicillata Acc. 1 C. penicillata Acc. 2 C. edithae 1.00/100 C. speciosa 1.00/100 i-Pseudolithos caput-viperae Pseudolithos migiurtinus C. hexagona C. faucicola 1.60/100|_|? 1.00/891?C. solenophora C. munbyana 1.00f91 1.00j [9V C. europaea 1.00/95 r-C. burchardii . H.00/9i ^C.joannis staintonii i-C. I tuberculata 1.00/98'-C. Frerea indica C. procumbens <-C. crenulata i89 LOO] '91 C. truncato-coronata 1.00/100|_rC. indica II Subclade 1.00/ C. pauciflora h .00/98 _r? C. diffusa LOO/TOO^C. umbellata _I? C. edulis 1.00/100 L- C. sinaica -Edithcolea grandis C. socotrana 1.00/100 1.00/100 I Subclade C. cicatricosa L C. 1.00/100 quadrangula rC. ads. var. attenuata [C. sarkariae 0.99/60 1.00/100 |? C. - stalagmifera C. aofs. var. adscendens Indian C1 bhupinderiana ads. var. fimbriataAcc. 1 1.00/1001 C. aafs. var. fimbriataAcc. 2 ^Southern Clade Subclade IV Clade B Subclade III subclade -C. 0.99)67 furta j C. priogonium arachnoidea 1.00/93|_ I-C.r H C. 0.99/511 ? C.dicapuae \ J pec/c/7 1.00/100 ? C. flavovirens NE African subclade C. turned -Anomalluma dodsoniana -Anomalluma maccoyi leachii 1.00/67r -Echidnopsis Echidnopsis scutellata 1.00/65 Rhytidocaulon fuller! H r Rhytidocaulon ciliatum 1.00/100 hytidocaulon macrolobum ^-Rhytid 1.00/67 1.00/99 r 1034 This content downloaded from 137.158.75.81 on Mon, 9 Feb 2015 05:04:47 AM All use subject to JSTOR Terms and Conditions Clade A TAXON 59 (4) August 2010: 1031-1043 2. Clades Table were obtained recovered are indicated Bruyns & in our analyses and the generic = in bold. 'C Caralluma. treatment of their species of Classification Clades in recent synopses Gilbert (1990) and species of Caralluma. al. Species of Classification Phytogeny of Caralluma forwhich sequences Classification of Meve & Liede (2002) Plowes (1995) CladeA Indian subclade C. adscendens C (Roxb.) R. Br. Caralluma subg. Caralluma Caralluma Caralluma Sarkaria Caralluma subg. Caralluma Caralluma Caralluma bhupinderiana Caralluma subg. Caralluma Caralluma Caralluma Caralluma subg. Caralluma Caralluma Caralluma Caralluma subg. Caralluma Spathulopetalum Caralluma subg. Caralluma Somalluma Caralluma subg. Caralluma Spathulopetalum C dicapuae (Chiov.)Chiov. Caralluma subg. Caralluma Spathulopetalum Caralluma C. edwardsiae(M.Gilbert)M. Gilbert Caralluma subg. Caralluma Spathulopetalum Caralluma Caralluma subg. Caralluma Saurolluma Caralluma subg. Caralluma Spathulopetalum Caralluma Caralluma subg. Caralluma Spathulopetalum Caralluma Caralluma subg. Caralluma Spathulopetalum Caralluma Caralluma subg. Caralluma Spathulopetalum Caralluma Caralluma subg. Caralluma Spathulopetalum Caralluma Caralluma subg. Caralluma Spathulopetalum Caralluma Caralluma subg. Caralluma Spathulopetalum Caralluma Caralluma subg. Desmidorchis Monolluma Caralluma subg. Desmidorchis Monolluma Caralluma subg. Desmidorchis Lavranos C. sarkariae C & Frandsen C.E.C. stalagmifera Fischer NE African subclade (P.R.O. Bally) M. C. arachnoidea Gilbert C. baradii Lavranos P.R.O. C. congestiflora P.R.O. Bally K. C. gracilipes Schum. C longiflora M. Gilbert Chiov. C mogadoxensis C moniliformis P.R.O. C. peckii C. sudanica Bally Bally K. C. priogoniunt C P.R.O. Caralluma Caralluma Caralluma Caralluma L.E. Newton C. flavovirens Cfurta Bally Chiov. Plowes Schum. Plowes Caralluma Bruyns turneri E.A. Bruce Clade B Subclade I (Defl.) N.E. C. cicatricosa C Edithcolea Br. (Forssk.) N.E. quadrangula C. socotrana Br. (Balf. f.) N.E. grandis N.E. Br. Edithcolea Br. N.E. Br. Sanguilluma Plowes Monolluma Monolluma Plowes Monolluma Edithcolea Edithcolea Subclade II Slender-stemmed C widespread e</?/fs (Edgew.) group Benth. & Hook. f. Caralluma subg. Caralluma Cryptolluma Spiralluma Lavranos C. mireilliae C. sinaica (Decne.) Benth. & Hook. f. Caralluma subg. Caralluma Caudanthera Caralluma subg. Caralluma Caudanthera Plowes, Plowes Caudanthera Plowes Caudanthera Caudanthera Indian group Frerea C Frerea indica Dalz. crenulata Wall. N.E. Br. C. rfijjTiisa (Wight) C i>i</ica (Wight & Arn.) N.E. Br. N.E. Br. G pauciflora (Wight) C procumbens C Gravely truncato-coronata C. umbellata Haw. & Mayuran. (Sedgew.) Gravely & Mayuran. Boucerosia Frerea Dalz. Wight & Arn. Boucerosia Caralluma subg. Boucerosia Boucerosia Caralluma subg. Boucerosia Boucerosia Boucerosia Caralluma subg. Boucerosia Boucerosia Boucerosia Caralluma subg. Boucerosia Boucerosia Boucerosia Caralluma subg. Boucerosia Boucerosia Boucerosia Caralluma subg. Boucerosia Boucerosia Boucerosia Caralluma subg. Boucerosia Boucerosia Boucerosia 1035 This content downloaded from 137.158.75.81 on Mon, 9 Feb 2015 05:04:47 AM All use subject to JSTOR Terms and Conditions Bruyns & Table al. TAXON 59 (4) August 2010: 1031-1043 of Caralluma Phylogeny 2. Continued. Classification Clades of of Classification Gilbert (1990) and species Classification of Plowes(1995) Meve & Liede (2002) Subclade III C. burchardii N.E. G (Guss.) N.E. europaea C.joannis Br. Br. Maire C munbyana (Decne.) Br. N.E. C. staintonii Hara C N.E. tuberculata Br. Caralluma subg. Boucerosia Apteranthes Mikan Apteranthes Caralluma subg. Boucerosia Apteranthes Apteranthes Caralluma subg. Boucerosia Apteranthes Caralluma subg. Urmalcala Borealluma Caralluma subg. Urmalcala Borealluma Apteranthes Caralluma subg. Urmalcala Borealluma Apteranthes Caralluma subg. Boucerosia Sulcolluma Caralluma subg. Boucerosia Apteranthes Plowes Apteranthes Subclade IV Bruyns C.faucicola C. hexagona C Lavranos Lavranos solenophora Pseudolithos caput-viperae Lavranos (P.R.O. Bally) P.R.O. R cubiformis P horwoodii R migiurtinus C acutangula C adenensis C arabica P.R.O. (Decne.) N.E. Bally Pseudolithos Pseudolithos Pseudolithos Pseudolithos Pseudolithos Pseudolithos Pseudolithos Pseudolithos Pseudolithos (Deflers) A. Berger Caralluma subg. Boucerosia Crenulluma Caralluma subg. Boucerosia Crenulluma Desmidorchis Caralluma subg. Boucerosia Crenulluma Desmidorchis Caralluma subg. Boucerosia Desmidorchis Desmidorchis Caralluma subg. Boucerosia Crenulluma Desmidorchis Caralluma subg. Boucerosia Desmidorchis Desmidorchis Caralluma subg. Desmidorchis Crenulluma Desmidorchis Caralluma subg. Boucerosia Desmidorchis Desmidorchis Caralluma subg. Boucerosia Crenulluma Desmidorchis Caralluma subg. Boucerosia Desmidorchis Desmidorchis Caralluma subg. Boucerosia Desmidorchis Desmidorchis Br. Br. Br. Bruce (Deflers) N.E. Br. Lavranos C. somalica Pseudolithos Pseudolithos Desmidorchis E.A. C. speciosa Monolluma subg. Boucerosia C. lavranii Rauh & Wertel C. petraea Bally Monolluma Caralluma edithaeNE. C. penicillata P.R.O. Cylindrilluma Plowes Br. (Deflers) A. Berger C. flava N.E. C. foetida (Chiov.) N.E. C. awdeliana C Bally Lavrabos Pseudolithos Plowes N.E. Br. (N.E. Br.) N.E. Br. Ehrenb. Plowes Desmidorchis Desmidorchis of uncertain position Species C lamellosa M. Gilbert & Thulin C. vaduliae Lavranos Evolutionary relationships inCaralluma support for clades morphological s.l. and Clade A.? Apart fromC. edulis and C. sinaica, clade A corresponds to Caralluma subg. Caralluma ofGilbert (1990) and itwas recognised as thegenus Caralluma byMeve & Liede (2002)(Table2). Clade A consists of a poorly resolved 'Indian subclade' and awell resolved 'North-EastAfrican subclade'.Morphologically the species of the 'Indian subclade' are very similar and can only be distinguished on subtle differences in the orientation of the flowers and thepresence or absence ofmarginal cilia on the lobes. This lack ofmorphological differentiationis reflected in the phylogram (Fig. 1),which shows very fewmolecular changes across the subclade and poor resolution. This subclade contains species from peninsular India, northern Sri Lanka and the dry parts of centralMyanmar (Fig. 2). Here we paid particular attention toC. adscendens var.fimbriata, which has been recorded fromMyanmar and peninsular India via Arabia (where itwas formerlyknown as C. subulatd) toWest Africa (where itwas previously called C. dalzielii) (Bruyns, 1992). Among our samples, we included two accessions of C. ad scendens var.fimbriata, one from India and one fromArabia. Our analyses show them to be closely related sisters (PP = 0.97). This suggests that, since this plant iswidely cultivated formedicinal purposes (Bruyns, 2005), itspresent-day distri bution has been greatly extended by human activity and so this subclade is native to the Indian region. 1036 This content downloaded from 137.158.75.81 on Mon, 9 Feb 2015 05:04:47 AM All use subject to JSTOR Terms and Conditions TAXON 59 (4) August 2010: 1031-1043 As the phylogram shows, the species of the 'North-East African subclade' are farmore diverse genetically. This is accompanied by substantial morphological diversity such as considerable range in the size of the plant, the thickness of the stems and the size of the flowers. This subclade ismainly concentrated around theHorn ofAfrica but is represented nei ther inArabia nor inWest Africa (Fig. 2). The members of clade A are perhaps the easiest group within Caralluma s.l. to characterise morphologically. In all species the stems are longitudinallymottled with purple or darker green on a much paler background. The leaf-rudiment is lanceolate and bears (except inC. furta) a cluster of small hairs in a stipular posi tion on either side at its base (Fig. 4A-B). When flowering, the4-angled stems elongate to a slender, less clearly angled to cylindrical, false raceme. In thisdistinct flower-bearing region, many few-flowered inflorescences develop, each alongside one of the leaf-rudiments(Wertel, 1976),with bracts thatare similar in shape to the leaf-rudiments. Clade B.? Clade B contains C. subg. Boucerosia, subg. Desmidorchis and subg. Urmalcala of Gilbert (1990) and, as shown inTable 2, clades within itwere recognised as a total of eight genera byMeve & Liede (2002). For claritywe divide the species of clade B intofour subclades, defined as inFig. 1. With thebroadest distribution in an East-West direction of any lineage within the stapeliads, clade B (Fig. 3) has diversi fied across Africa northof centralTanzania and has reached the westernmost as well as the easternmost limits of the distribu tion of the stapeliads in theCanary Islands and centralMyan mar respectively.Furthermore, ithas penetrated the temperate parts of the northern hemisphere inWestern Europe and the Nepalese Himalaya and inboth it is the only representative of the stapeliads. ? At a first Subclade I. glance, subclade I appears tobe an taxa of and, historically, therehas been no unlikelymiscellany that the suggestion monotypic Edithcolea (e.g.,Gilbert, 1990) could be so deeply nested within Caralluma s.l.Vegetatively Edithcolea, with itsmat-forming habit and spike-tipped tu bercles on the stems usually with a single hair on either side roughly in a stipularposition (Fig. 5F-G) looks very different from the others,which are erect shrubs rooting by a central stem only, with rounded tubercles and minute, soft leaf-ru diments. There is also wide floral diversity,with very large flowers inEdithcolea (60-125 mm diam.) and much smaller flowers in C. cicatricosa, C. quadrangula and C. Bruyns & al. PhytogenyofCaralluma (perhaps only introduced), fromwhich C. socotrana is ab sent.The stronglysupported sistersCaralluma cicatricosa and C. quadrangula are themost local members of the subclade, endemic to themountainous parts of tropicalArabia. They are also impossible to distinguish unless in flower. In C. cicatri cosa the flowers are dark purple tomaroon, rugose and evil smelling while in C. quadrangula they are yellow towhite, smooth and sweetly scented (Noltee, 1988). This would suggest differentpollination syndromes in these otherwise structurally very similar and apparently very closely related species. In the firstdiverging branch of subclade Subclade II.? C. edulis is II, especially widespread and is known fromRa in western India and Pakistan toWest Africa (Bruyns, jistan & Ricanek Hanacek, 2001). That it is reputedlycultivated 1989; as a vegetable was mooted as a possible reason for itswide distribution (Gilbert, 1977). The other species in this branch, C. sinaica, ismore local (from Israel toYemen, mainly along the eastern flank of theRed Sea; Bruyns, 1987b). In both, the leaf-rudiments (Fig. 4C-E), inflorescences and flowering branches are extremely similar to those in clade A andmust be a parallel development of these features. Caralluma sinaica is unique inCaralluma s.l. for thegreatly elongated, sterileanther appendages and the strangely-shapedpollinia (Bruyns, 1987b) but our results show that it isvery closely allied toC. edulis, in which these unusual features are absent. The remaining eight species are found inpeninsular India, northern Sri Lanka and centralMyanmar. The firstdiverging branch among these is themonotypic Frerea. This is the only stapeliad thatbears true leaves, each on a clearly defined peti ole, with minute and distinctively shaped stipules (Fig. 4F-H) and the tubercles are not organized intoangles along the stems. These features, sharedwith less succulent relatives such asmost members ofCeropegia, led toFrerea being considered the 'liv ing ancestor' of the stapeliads (White & Sloane, 1937; Good, 1953), fromwhich themore highly succulent, essentially leaf less species were assumed tobe derived. Frerea is of restricted occurrence along the dry, eastern flank of theWestern Ghats socotrana (12-22 mm diam.). The considerable morphological differ ences between Edithcolea and the others is reflected in the comparatively longbranch thattheothers are placed on relative toEdithcolea. Nevertheless, all species share the following features: (1) tubercles thatare fused intoobtuse, often unclear angles; (2) single-flowered inflorescences; (3) each pedicel is subtended by a single, small, slender bract not shaped like the leaf-rudimentsand (4) follicleswith a distinct lateral ridge along each side. The first two diverging species (C. socotrana, Edithco lea grandis) are themost widely distributedmembers of this subclade, found inEast Africa and theHorn ofAfrica. Both are plentiful on Socotra butEdithcolea is very rare inArabia of clade A. The numbers Fig. 2. Distribution species found in the respective areas. shown indicate endemic 1037 This content downloaded from 137.158.75.81 on Mon, 9 Feb 2015 05:04:47 AM All use subject to JSTOR Terms and Conditions of Bruyns& al. Phylogeny Caralluma between 17?and 20?N in India,where itgrows in shallow pock ets of soil on and below rock faces, receiving often enormous amounts of rainfall during the summer (Tetali& al, 1997). Frerea is sister to seven species (the 'Indian Boucerosia group' ofGilbert, 1990) thatare vegetatively diverse, from the large clumps of robust stems often severalmetres indiameter of C. diffusa,C. procumbens and C. umbellata to the small, highly rhizomatous, slender-stemmed and extremely inconspicuous plants of the remaining four species. Caralluma diffusa,C. pro cumbens and C. umbellata are found in the tropical, southern portion of peninsular India and northern Sri Lanka and all of TAXON 59 August 2010: 1031-1043 (4) them occur on exposed slabs of granite in shallow pockets of soil. The remaining fourare endemic to tropical, southern India and the low-lying,dry parts of centralMyanmar. They occupy an entirelydifferenthabitat-niche, growing in flat, oftenheav ilygrazed areas under spiny bushes, where theirrhizomatous habit provides an efficientmethod of expansion and of surviv ingboth predation and prolonged dry periods. The seven species of the 'Indian Boucerosia group' have fairly conspicuous leaf-rudimentsoftenwith relatively thick marginal hairs (Fig. 5B-E) and sporadicallywith small stipules (Fig. 5C, E). They exhibit great diversity in thegeometry of the 30? 40? 50c 0? of clade B. Fig. 3. Distribution in the respective areas. I, subclade I; II, subclade II; III, subclade III; IV, subclade 20? IV. The numbers I_1_1_I_I_I_l_l_l_L_I 20? 60? 40? shown 1038 This content downloaded from 137.158.75.81 on Mon, 9 Feb 2015 05:04:47 AM All use subject to JSTOR Terms and Conditions indicate endemic species found TAXON 59 (4) August 2010: 1031-1043 corolla and the shape of thepollinia. It appears that thisdiver sity inpollinia has arisen inparallel with thevariable geometry of the corolla tube since broad, round pollinia correspond to a broad, flat corolla with fully exposed corona and narrow pol linia correspond to a more tubular corolla inwhich the corona is contained within a narrow tube. The same phenomenon has been observed inStapelia L. and TromotricheHaw. inSouthern Africa (Bruyns, 2005). Gilbert (1990) placed C. edulis and C. sinaica inC. subg. Caralluma, maintained Frerea as a distinct genus and placed the remainder of subclade II in the C. umbellata-C. indica Bruyns & al. PhytogenyofCaralluma group of C. subg. Boucerosia. Meve & Liede (2002) recog nised two genera for subclade II, namely Caudanthera and Boucerosia, but did not provide diagnoses for either of them. According to their key, Caudanthera was said to have leaf rudimentsaccompanied by stipularglands, which isonly some times true ineitherof the species (Bruyns, 1987b, 1989),while Boucerosia was characterized by 'leaf rudiments (broadly) lanceolate, without stipular formings [sic]', a diagnosis that could apply equally well tomany other species inCaralluma s.l. andwhich does not apply to eitherBoucerosia frerei (Frerea indicd), inwhich the leaves are not lanceolate and stipular Fig. 4. Leaf-rudiments stipules in Caralluma. and A, C. adscendens, Bruyns B, C. adscendens, Bruyns 5937 (BOL), leaf-rudiment; 5937 (BOL), stipularstructures around base of leaf-rudiment; C, C. edulis, Ricdnek & Handcek 305 (BRNM), leaf rudiment; D, C. edulis, Ricdnek & Handcek 305 (BRNM), small stipule at base of leaf-rudiment; Bruyns 2484 (K), leaf-rudiments showing small stipules; F,Frerea indica, E, C. sinaica, Bruyns5925 (BOL, E), small stipule at base TV? of leaf; G, Frerea indica,Bruyns5925 (BOL, E), small stipuleat base of leaf; H#Frerea indica, Bruyns 5925 (BOL, E), hair alongmargin of leaf. This content downloaded from 137.158.75.81 on Mon, 9 Feb 2015 05:04:47 AM All use subject to JSTOR Terms and Conditions Bruyns& al. Phylogeny of Caralluma rudiments are present or toB. procumbens and B. pauciflora, inwhich stipules are sometimes present. Subclade III.? Subclade III contains all of C. subg. Ur malcala as well as C. burchardii, C. europaea and C.joannis from the informalC. europaea-C. hexagona group ofC. subg. Boucerosia (Gilbert, 1990).Meve & Liede (2002) included all of these species inApteranthes, even though theyfell into two separate lineages in an unresolved polytomy. They character ized Apteranthes by 'stems not pungent, podaria not grooved, leaf-rudiments subsessile to petiolate and sometimes swollen TAXON 59 (4) (A. tuberculatd),with stipular glands or stipular formings ab sent'.No evidence of petiolate or 'swollen' leaf-rudiments is known (Bruyns, 1986a, forC. tuberculatd). Stipular glands and small tubercle-like stipules are known inC. staintonii (Bruyns, 1989) and inC. tuberculata (Bruyns, 1986a) but are absent in the three species associated with southern Europe (Bruyns, 1987a). Thus Apteranthes isvery difficult to characterize. Nev ertheless, we have now established for the first time that the genus, as itwas proposed byMeve & Liede (2002), is a strongly supported,monophyletic entity.On theother hand, itshould be and stip Fig. 5. Leaf-rudiments ules inCaralluma. A, C. umbel lata, Bruyns 5939 (E), leaf-rudi B, C. umbellata, Bruyns ment; 5939 (E), hairalongmargin of leaf-rudiment; C, C. procum bens,Bruyns5887 (BOL, E, K, MO, etc.), leaf-rudiment; D, C. indica,Bruyns5876 (K,MO), leaf-rudiment; E, C. pauciflora, Bruyns5884 (BOL, E, MO); F, Edithcolea grandis, Bruyns 8681 (K), leaf-rudiment; G, Edithcolea grandis, Bruyns 8681 (K), hair instipularposi tion; H, C. arabica, (SQUH), August 2010: 1031-1043 Al Far si 235 leaf-rudiment. 1040 This content downloaded from 137.158.75.81 on Mon, 9 Feb 2015 05:04:47 AM All use subject to JSTOR Terms and Conditions TAXON 59 (4) August 2010: 1031-1043 Bruyns noted thatGilbert's C. subg. Urmalcala is notmonophyletic, since two of its species are nested among other species of this subclade,while theother (C. munbyana) is sister to all theother members of the subclade. The members of this subclade have thewidest distribu tion of all within Caralluma s.l. and are found from theCa nary Islands along the southern shores of theMediterranean to Iran, Pakistan, North-West India andWest Nepal (Fig. 3). Inmany of their localities, theyare the only representatives of the succulent Apocynaceae and they reach the furthest from the equator of any stapeliads (Bruyns, 1987a). The twomost widely distributed are C. europaea, found around the perim eter of theMediterranean fromMorocco to Jordan (Bruyns, 1987a,b;Meve & Heneidak, 2005) and C. tuberculata, whose distribution is almost continuous from Jordan toNorth-West India (Bruyns, 1986a; Ricanek & Hanacek, 2001). ? Members of subclade IV have been most Subclade IV. confusingly treated in the literature (Table 2). Gilbert (1990) kept Edithcolea and Pseudolithos separate from Caralluma and placed the species of Caralluma in C. subg. Boucerosia and a few inC subg.Desmidorchis. According to our results, subclade IV includes allmembers ofC. subg.Desmidorchis as well as most of the species of C. subg.Boucerosia thatGilbert (1990) did not place in any of his informalgroupings.Meve & Liede (2002) also kept Edithcolea and Pseudolithos separate fromCaralluma (despite the fact that theywere nested among the species ofCaralluma) and placed the species ofCaralluma in two genera,Desmidorchis andMonolluma. Thus theyrecog nised four genera for subclade IV. Their analyses showed that C. hexagona was nested between Edithcolea and C. cicatricosa + C. quadrangula. Consequently they included itand C. sole nophora, which they did not sample, inMonolluma, together with C. cicatricosa, C. quadrangula and C. socotrana. We have not been able to corroborate thisposition forC. hexagona, which may have been a consequence ofmis-identified mate rial. According to our results, their concept ofMonolluma is polyphyletic. This group is themost species-rich inCaralluma s.l. Car alluma acutangula iswidely distributed inAfrica across the arid sahel fromSenegal toEritrea and toOman in theArabian Peninsula. The others are confined to theHorn ofAfrica and tropicalArabia, where up to four of its species may be found growing in close proximity to one another. The large, shrubby species in this subclade are not specific in theirrequirementsof habitat and are found on gravelly plains and rocky slopes, often in extremely exposed situationswithout any significant cover fromother shrubs. It isnoteworthy thathere, as in subclade III but in contrast to subclades I and II, it is not themost widely distributed species thatdiverged earliest. Characters uniting themembers of this subclade are the extremely reduced leaf-rudiments,which usually form a min ute ridge at the apex of the tubercle, usually with a cluster of small hairs around itsbase and sometimes along itsmargins (Fig. 5H). The bracts in the inflorescence are slender and of very different shape to the leaf-rudiments. In all species ex cept C. lavranii (where each inflorescence consists of a single flowerwith 3-4 bracts around the base of the pedicel), dense & al. PhylogenyofCaralluma umbels of more or less simultaneously opening flowers are produced at the apices of the stems. The first diverging branch within subclade IV consists of C. faucicola, C. hexagona and C. solenophora, endemic to themountainous parts of tropicalArabia. They share a mat forminghabit, inwhich the soft and fleshy stems root readily where thebranches touch the ground and no rigid and slightly woody, central stem develops. The young tubercles are longi tudinally grooved on the surface between the leaf-rudiment and the base of the next tubercle. Florally C. faucicola and C. hexagona are similar,with small, almost rotate flowers and fairlyprominent outer and innercorona lobes.Nevertheless, the molecular data suggest thatC. faucicola and C. solenophora are more closely related. This is somewhat surprising, since C. solenophora has remarkable, cylindrical flowers (the longest relative to theirbreadth in any species of Caralluma s.l.), no innercorona lobes, very prominent, spreading guide-rails (the largest inCaralluma s.l.) and large,unusually-shaped pollinaria (again the largest inCaralluma s.l.). The next diverging branch within subclade IV consists of Pseudolithos caput-viperae and P. migiurtinus. These are very distinctive vegetatively from any species of Caralluma s.l. (Bruyns, 1990), a fact that is corroborated by the long branch subtending them inFig. 1.They occur only in eastern Ethiopia and Somalia (Gilbert, 2003), in areas of exceptional aridity, growing almost entirely in the open among sparse, low shrubs and stones in shallow ground overlying sheets of limestone (Bally & al., 1975).Here theirgreatly reduced, globu lar growth-formprotects themwell. Similar globular growth forms have developed independently inLarryleachia Plowes in the 'southern clade' mainly along the edges of theNamib Desert in response to similar stony habitats, where shelter is minimal (Bruyns, 2005). The remaining nine species of subclade IV are grouped into a poorly resolved branch inwhich there is relatively little variation in the gene-regions investigated. They are all found in tropical southernArabia and North-East Africa, with sev eral restricted to themountains of southernArabia. All are shrub-formingplants with more or less erect branches rooting only by a basally eventually somewhat woody central stem. There is considerable variation in the geometry of the corolla (fromrotate inC. arabica and C. lavranii to deeply campanu late inC adenensis and C. speciosa). The notable differences between the inflorescence and flower in C. lavranii and re lated species is not matched by significant genetic variation (as found forC. solenophora too). CONCLUSION In this studywe have shown that the stapeliads consist of twomajor lineages. One of these contains Caralluma s.l.,with Anomalluma, Echidnopsis, Edithcolea, Frerea, Pseudolithos and Rhytidocaulon nested within it.The other lineage ismade up ofDuvaliandra + Whitesloanea + the 'southern clade'. We have shown thatmost of themonotypic genera that have been recognised among the stapeliads outside southern 1041 This content downloaded from 137.158.75.81 on Mon, 9 Feb 2015 05:04:47 AM All use subject to JSTOR Terms and Conditions Bruyns& al. Phytogeny of TAXON Caralluma Africa (especially the numerous genera created by Plowes, 1995) are not isolated entities and we have identified the broader monophyletic groups towhich they belong. We have also shown that, although the genera recognised byMeve & Liede (2002) are much closer to the ideal of recognising only monophyletic groups, they too have recognised toomany gen era andwere unable to characterise the entities theyrecognised bymorphological features. Our results allow us to conclude that species ofCaralluma have invaded the Indian subcontinent four times and throughat least two distinct routes.We have found thatone monophyletic group of species has the largest distribution in an East-West direction of any such group among the stapeliads. Nevertheless the greatest diversity of Caralluma follows the pattern of all other stapeliad genera found in the northern hemisphere and occurs inNorth-East Africa and the adjacent southernArabia. While our analyses have resolved the relationships be tween the species of Caralluma s.l. and other stapeliads and suggested thatwithin Caralluma s.l. just two entities, clade A and clade B, should be recognised taxonomically, we refrain at this stage frommaking taxonomic changes based on these newly established relationships. In the firstplace, while clade A is easily characterisedmorphologically, clade B ismore diffi cult to characterise, though one possibility is thevery reduced, usually broadly ovate to absent leaf-rudiments thatare differ ently shaped to the slender bracts in the inflorescences, with a reversal in C. sinaica. and C. edulis more However, impor tantly,themonophyletic stapeliads are embedded within the approximately 160 species of a paraphyletic Ceropegia (Meve & Liede-Schumann, 2007) and the taxonomic consequences of this paraphyly must be resolved before furthernames are changed among the stapeliads. 59 August 2010: 1031-1043 (4) of the genera Pseudolithos 30: 31-36, 88-93. and Whitesloanea. Natl. Cact. Succ. J. notes on Ceropegieae P.V. 1986a. Miscellaneous (Asclepiada tuber culata. Bradley a 4:33-37. ceae). 3. The identity of Caralluma on the Canary Islands (Ascle Bruyns, P.V. 1986b. The genus Ceropegia Beitr. Biol. Pflanzen 60: 427-458. piadaceae?Ceropegieae). Bruyns, notes on Stapelieae P.V. 1987a. Miscellaneous (Asclepiada 5: 77-87. ceae). 5. The stapeliads of Europe. Bradleya R. Br. (Asclepiadaceae) Bruyns, P.V. 1987b. The genus Caralluma Bot. 36: 73-86. Israel. Israeli Bruyns, Bruyns, P.V. notes on Stapelieae 1989. Miscellaneous in (Asclepiadaceae) 7: 63-66. 7, 8. Bradleya Bruyns, P.V. 1990. Miscellaneous notes on Stapelieae (Asclepiadaceae) 8: 33-37. Bradleya notes on Stapelieae (Asclepiadaceae) Bruyns, P.V. 1992. Miscellaneous adscendens and its synonyms C. subulata 11.A note on Caralluma 9. A review of Pseudolithos. and C. dalzielii. Bruyns, Bradleya 1993. A revision P.V. P.V. 2008. Notes species ofHuernia 38: 83-85. Bothalia Bruyns, P.V. of southern 2010. A new P.V. & Bradleya from tropical on African Africa and Madagascar. A new plants. Apocynaceae. from Angola. (Asclepiadoideae-Ceropegieae) species of Caralluma from the Yemen. clepiadoideae-Ceropegieae) 76: 249-251. Bruyns, (Asclepiada 41:76-79. Aloe P.V. 2005. Stapeliads Bruyns, Hatfield: Umdaus Press. Bruyns, and Lavrania Bot. Jahrb. Syst. 115: 145-270. Two new species of Apocynaceae ceae?Stapelieae). P.V. 2004. Bruyns, Africa. 10: 95-96. of Hoodia (Apocynaceae-As S. African J. Bot. 1993. The genus Caralluma Jonkers, H.A. 11: 51-69. in Oman. T. 2006. A new subgeneric R. & Hedderson, Bruyns, P.V, Mapaya, for Euphorbia based on molecular classification (Euphorbiaceae) data. Taxon 55: 397-420. De Wilde, W.J.J.O. (Passifloraceae). 71(18): 1-281. of the genus Adenia Forsk. 1971. A monograph Meded. Wageningen Landbouwhogeschool J.J. & Doyle, J.L. 1987. A rapid isolation procedure for small of fresh leaf tissue. Phytochem. Bull. 19: 11-15. E. & Doyle, J.J. 1999. Chloroplast-expressed Emshwiller, glutamine Doyle, quantities synthetase(ncpGS):Potentialutilityforphylogeneticstudieswith ACKNOWLEDGEMENTS to thank the Director, The first author wishes India, Ram Gandhi, by assistance possible Department from the Royal of Environment reidi of Taiz, Yemen. We formaterial Klak & Vinod during travel in India. Gathering assistance Yemen and Meena R. Haresh of several for contributing zalo Aguilar species for assistance forDNA Research Edinburgh, and Dennis extraction. We Research Committee, the de Kock thank Gon research was Foundation, University partially Town. LITERATURECITED information. IEEE J.M., Wojciechowski, B.G., Sanderson, M.J., Porter, M.J. 1995. The ITS region C.S. & Donoghue, M.F., Campbell, source of evidence on an of nuclear ribosomal DNA: A valuable Baldwin, giosperm P.R.O., Bot. Gard. 82: 247-277. phylogeny. Ann. Missouri J. J. 1975. A monograph F.K. & Lavranos, Horwood, Molec. Phylog. Evol. 12: M. & Cladistics 12: 99-124. 1977. Caralluma Gilbert, M.G. Succ. J. 32: 26-31. M.G. 1990. A Bradleya Gilbert, M.G. 8: 1-32. Gilbert, sect. Caralluma review of Caralluma inEthiopia. Natl. Cact. R. Br. and its segregates. P. 183 in: Hedberg, S. I, Edwards, and Eritrea, vol. 4(1). S. (eds.), Flora of Ethiopia The National Herbarium. 2003. Pseudolithos. & Nemomissa, Addis Ababa: 2009. [Book Review]. Edinburgh J. Bot. 66: 201-203. Gilbert, M.G. lamellosa (Apocynaceae), Gilbert, M.G. & Thulin, M. 2005. Caralluma a remarkable new species from Somalia. Nord. J. Bot. 23: 523-525. R. 1953. The geography of theflowering plants, ed. 2. London: Green & Co. Longmans, sequence alignment Hall, T.A. 1999. Bioedit: A user-friendly biological 95/98/NT. Nucleic Acids editor and analysis program forWindows Good, Akaike, H. 1974. A new look at the statistical model 19: 716-723. Trans. Automatic Control Bally, (Oxalidaceae). Apocynaceae South Africa of Cape from Oxalis P.V. 2000. A revised classification of the Bruyns, 66: 1-56. s.l. Bot. Rev. (Lancaster) D. & Kluge, Farris, J.S, Albert, V.A, Kallersjo, M., Lipscomb, A.G. 1996. Parsimony jack-knifing outperforms neighbor-joining. Endress, al Khu are grateful to C. to our data and we with the figures. This Survey and Abdul Wali Bosma of Singh for generous inYemen was made Gardens & Tourism sequences funded by grants from theNational and from theUniversity of data Botanic also thankWiebe eleven Botanical an example 310-319. Symp. Ser. 41: 95-98. F. 2001. MrBayes: J. & Ronquist, Huelsenbeck, Bayesian 17: 754-755. trees. Bioinformatics of phylogenetic inference J.J. 1991. Succulent plant records from Somalia: Description Lavranos, 63: 167-172. of new taxa. Cact. Succ. J. (Los Angeles) 1042 This content downloaded from 137.158.75.81 on Mon, 9 Feb 2015 05:04:47 AM All use subject to JSTOR Terms and Conditions TAXON 59 (4) August 2010: J.J. Lavranos, Stapelieae) 247. 1993. A new from Somalia. 1031-1043 species Cact. of Caralluma Succ. Ricanek, (Asclepiadaceae 65: 246 J. (Los Angeles) J. Linn. Soc. Bot. Meve, 149: 419-432. S. 2002. A molecular U. & Liede, phylogeny and generic rear Ascle rangement of the stapelioid Ceropegieae (Apocynaceae PI. Evol. 234: 171-209. piadoideae). Syst. U. & Liede-Schumann, S. 2007. Ceropegia (Apocynaceae, but still taxonomically Ceropegieae, Stapeliinae): Paraphyletic Bot. Gard. 94: 392-406. sound. Ann. Missouri Noltee, 1998. Two new stapeliads 1988. Caralluma cicatricosa L.E. Newton, F. (Amsterdam) 67: 51-55. 1995. A reclassification Plowes, D.C.H. the model stapeliad of species from Iran. setts: Sinauer. Taberlet, P., Gielly, L., Pautou, G. & Bouvet, J. 1991. Universal prim ers for amplification of three non-coding regions of chloroplast DNA. PI. Molec. 17: 1105-1109. Biol. D.K. & Kumbhojkar, M.S. 1997. P., Tetali, S., Kulkarni, Studies on the status and conservation ofFrerea indica. J. Bombay Nat. Hist. S. 2007. Diversifi Rapini, A., Van den Berg, C. & Liede-Schumann, in theNew World. Ann. cation of Asclepiadoideae (Apocynaceae) Missouri Bot. Gard. 94: 407-422 Appendix. P. 2001. Two Soc. 94: 115-121. 1976. Vergleichend-morphologische Wertel, H.P. schichtliche Untersuchungen tiber Bau R. Br. (Stapelieae: 3: 49-70. Haseltonia Asclepiadaceae). D. & Crandall, K.A. 1998. Modeltest: Posada, Testing DNA substitution. Bioinformatics 14: 817-818. PhytogenyofCaralluma H. 2000. Gaps as characters in sequence Simmons, M. & Ochoterena, based phylogenetic analyses. Syst. Biol. 49: 369-381. 2002. PAUP*: Phylogenetic Swofford, D.L. analysis using parsimony (*and other methods), version 4.0 beta 10. Sunderland, Massachu Tetali, 74:23-25. Asklepios (Defl.) N.E. Br. Succulenta & Hanacek, 82: 21-26. al. D. J. & Stuessy, T.F. 1997. Chloroplast DNA phy Sang, T., Crawford, of Paeonia togeny, reticulate evolution and biogeography (Paeo niaceae). Amer. J. Bot. 84: 1120-1136. inKenya. of Caralluma M. Asklepios J.J. 2006. Caralluma. Lavranos, Pp. 174-178 in: Thulin, M. (ed.), Flora Kew: Botanic Gardens. Somalia. Royal of and U. & Heneidak, S. 2005. A morphological, Meve, karyological chemical study of theApteranthes europaea (Caralluma) complex. Meve, & Bruyns und entwicklungsge und Stellung der Inflo reszenzen bei einigen Stapelieen-Gattungen (Fam: Asclepiada 17: 1-70. ceae). Trop. Subtrop. Pflanzenwelt 1937. The Stapelieae, A.C. & Sloane, B.L. ed. 2. Pasadena, White, California: Abbey San Encino Press. Wiens, J.J. 2003. Missing data, incomplete Syst. Biol. 52: 528-538. taxa, and phylogenetic accuracy. Taxa and vouchers for species sampled. Species, country of origin, collector and number of collection (herbarium), ITS GenBank - = cession no., trnT-FGenBank accession no.; * = downloaded fromGenBank; missing accession no., GS GenBank accession no., psbA-trnH GQnBmk ac sequence). *, -, HM475404, *; Caralluma acutangula, Ethiopia, Bruyns 8518 (BOL), *, HM475351, peschii, Namibia, ex hort. Bosma (MSUN, UBT), 300 (BOL), * HM475352, HM475406, *; C. adenensis, Yemen, Ricdnek & Handcek *; C. adscendens var. adscendens, India, Bruyns 5870a C. adscendens var. attenuata (Wight) Gravely & Mayur., India, Bruyns 5881 (E), HM475309, (K, M, MO), HM475308, HM475353, HM475407, HM475470; var.fimbriata C. adscendens HM475354, HM475408, HM475471; (Wall.) Gravely & Mayur. Acc. 1, India, Bruyns 5920 (BOL), HM475310, HM475355, HM475409, HM475472; C. adscendens var.fimbriata Acc. 2, Yemen, Bruyns 10906 (BOL), HM475311, -, HM475410, HM475473; C. arabica Acc. 1,Oman, Al-Farsi 235 (SQUH), HM475312, HM475356, HM475411, HM475474; C. arabica Acc. 2, Oman, Al-Farsi 235a (SQUH), HM475313, HM475357, HM475412, var. arachnoidea, Tanzania, Bruyns 8708 (E, MO), *, HM475358, HM475413, C. arachnoidea *; C. awdeliana, Yemen, Bruyns 10286 (E, M), HM475475; Australluma HM475405, C. bhupinderiana, India, Bruyns 5885 (K), HM475315, HM475360, HM475415, HM475477; C. burchardii, HM475314, HM475359, HM475414, HM475476; Morocco, Ricdnek & Handcek 210 (BOL), HM475316, HM475361, HM475416, HM475478; C. cicatricosa, Yemen, Bruyns 10896 (E), HM475317, HM475362, C. crenulata, Myanmar, Bruyns 8808 (BOL), *, HM475363, HM475418, HM475417, HM475479; *; C. dicapuae, Ethiopia, Bruyns 8452 (E), HM475318, C. diffusa, India, Bruyns 5868 (BOL, E, K, M), HM475319, HM475365, HM475420, HM475481; C. edithae, Somalia, HM475364, HM475419, HM475480; Thulin & al. 9418 (UPS), HM475320, HM475366, HM475421, HM475482; C. edulis, Iran, Ricdnek & Handcek 305 (BRNM), *, HM475367, HM475422, *; C. europaea, Tunisia, Ricdnek & Handcek sub De Kock 1996, *,HM475368, HM475423, *; C.faucicola, Yemen, Bruyns 10263 (BOL, E, M), HM475321, -, HM475424, HM475483; C.flava Acc. 1,Oman, Al-Farsi 185a (SQUH), HM475322, HM475369, HM475425, HM475484; C.fiava Acc. 2, Oman, ex hort. SQU Botanic Garden, HM475323, HM475370, HM475426, HM475485; C. flavovirens, Kenya, Dodds s.n. (BOL), HM475324, HM475371, HM475427, HM475486; C. hexagona, Yemen, Lavranos 30726 (BOL), *,HM475373, C.furta, Ethiopia, Bruyns 8489 (BOL, E, K), HM475325, HM475372, HM475428, HM475487; HM475429, *; C. indica, India, Bruyns 5876 (K, MO), HM475326, HM475374, HM475430, HM475488; C.joannis, Morocco, Ricdnek & Handcek s.n. (BOL), HM475327, HM475375, HM475431, HM475489; C. lavranii, Yemen, Frank 119 (BOL), *,HM475376, HM475432, *; C. munbyana, Spain, Bruyns 2404 (E), *, HM475377, HM475433, *; C. pauciflora, India, Bruyns 5884 (BOL, E, MO), HM475328, HM475378, HM475434, HM475490; C.peckii, Ethiopia, Bruyns 8453 (E), HM475329, HM475379, HM475435, HM475491; C. pen kill at a Acc. 1,Oman, Al-Farsi 223 (SQUH), HM475330, HM475380, HM475436, HM475492; C. sub Al-Farsi 280 (SQUH), HM475331, HM475381, HM475437, HM475493; C. priogonium, Tanzania, Bruyns 8677 (S), *, penicillata Acc. 2, Oman, McLeish HM475382, HM475438, India, Bruyns 5887 (BOL, E, K, RSA), HM475332, HM475383, HM475439, HM475494; C. quadrangula, Oman, Al *; C. procumbens, Farsi 251a (SQUH), * HM475384, HM475440, *; C. sarkariae, India, Bruyns 5880 (E, K), HM475333, HM475385, HM475441, HM475495; C. sinaica, Yemen, * Noltee 1158 (E), HM475386, HM475442, *; C. socotrana, Ethiopia, El-Azzouni s.n., *,HM475387, HM475443, *; C. solenophora, Saudi Arabia, Collenette C. speciosa, Ethiopia, Bruyns 8441 (BOL), HM475335, HM475389, HM475445, HM475497; 8168 (BOL, E), HM475334, HM475388, HM475444, HM475496; C. staintonii, Nepal, Bruyns 2515 (K), HM475336, HM475390, HM475446, HM475498; C. stalagmifera, India, Bruyns 5880a (BOL), HM475337, HM475391, C. truncato-coronata, C. tuberculata, India, Bruyns HM475447, HM475499; India, Bruyns 5935 (K, MO), HM475338, HM475392, HM475448, HM475500; 5841 (BOL), *,HM475393, HM475449, *; C. turneri var. turneri, Ethiopia, Bruyns 8466 (E), HM475339, HM475394, HM475450, HM475501; C. umbellata, India, Bruyns 5874 (MO), *,HM475395, HM475451, *; Ceropegia bulbosa Roxb., Oman, Al-Farsi 203 (SQUH), HM475340, HM475396, HM475452, HM475502; C. cimiciodora Oberm., South Africa, Bruyns 2089 (NBG), HM475341, -, HM475453, HM475503; C.juncea Roxb., India, Bruyns 5889 (BOL), HM475342, -, HM475504; C. occidentalis R.A. Dyer, South Africa, Bruyns 7592 (BOL, E, K), HM475343, -, HM475455, HM475505; C. radicans subsp. smiV/iii dioscoridis (Lavranos) M. Gil (M.R. Henderson) R.A. Dyer, South Africa, Bruyns 2162 (BOL, K, MO), HM475344, -, HM475456, HM475506; Duvaliandra leachii Lavranos, Tanzania, Bruyns 8652 (BOL, E), HM475345, *; Echidnopsis bert, Yemen, Socotra, Smith & Lavranos 438 (K, PRE, Z), *, -, HM475457, HM475397, HM475458, HM475507; E. scutellata subsp. dhofarensis Bruyns, Oman, Al-Farsi 186 (SQUH), HM475346, -, HM475459, HM475508; Edithcolea * *; Frerea indica, India, Bruyns 5925 (BOL, E, M), HM475399, HM475461, *; grandis, Tanzania, Bruyns 8673 (BOL, E, NBG), *,HM475398, HM475460, ex South 7050 Pseudolithos hort. Africa, Bruyns *; (BOL, MO), *,-, HM475462, Ophionella arcuata, (BOL), HM475347, HM475400, caput-viperae, Somalia, dodsoniana HM475463, HM475509; Anomalluma (Lavranos) Plowes, Somalia, Lavranos 7326 (FT), HM475348, HM475401, HM475464, HM475510; A. mac HM475454, coyi (Lavranos & Mies) Meve & Liede, Oman, Miller 9376 (E), HM475349, -, HM475465, HM475511; P. migiurtinus, Somalia, ex hort. W. Bosma (UBT), *, -, HM475466, *; Quaqua linearis, South Africa, Bruyns 4552 (BOL, K, MO), HM475350, HM475402, HM475467, HM475512; Rhytidocaulon ciliatum Hanacek & Ricanek, Yemen, Ricdnek & Handcek 265 (BRNM), *,HM475403, HM475468, *;R. fulleri Lavranos & Mortimer, Oman, Collenette 8439 (UBT), *,-, -, *;R. crassa (N.E. Br.) Chiov., Somalia, Lavranos s.n. (BOL)*, -, HM475469, macrolobum Lavranos, Yemen, Noltee 1667 (MSUN, UBT), *,-, -, *;Whitesloanea *. 1043 This content downloaded from 137.158.75.81 on Mon, 9 Feb 2015 05:04:47 AM All use subject to JSTOR Terms and Conditions