516 S.-Afr.Tydskr. Plantk. , 1989, 55(5): 516-519 A new species of Lycium (Solanaceae) from

Andor M. Joubert* and H.J.T. Venter

Department of Botany, University of the Orange Free State, Bloemfontein, 9300 Republic of South

Accepted 22 May 1989

Lycium grandicalyx Joubert & Venter, a new species from Namibia is described. This species differs from other Lycium species by the enlarged and broadly urceolate calyx, which encloses the berry completely in the fruiting stage and by the presence of bicornate, stellate hairs. Florally L. grandicalyx most resembles L. campanu!atum E. Mey., L. ferocissimum Miers, L. pilifolium C.H. Wr. , L. prunus-spinosa Dun. and L. schizo­ calyx C.H. Wr.

Lycium grandica/yx Joubert & Venter, 'n nuwe spesie van Namibie word beskryf. Hierdie spesie verskil van ander Lycium-spesies in die vergrote en wyd-kruikvormige kelk, wat die bessie heeltemal omsluit tydens die vrugstadium, en in die teenwoordigheid van tweehoringrige, stervormige hare. L. grandicalyx stem ten opsigte van blombou meeste met L. campanulatum E. Mey., L. ferocissimum Miers, L. pilifolium C.H. Wr., L. prunus-spinosa Dun. en L. schizocalyx C.H. Wr. ooreen.

Keywords: Lycium, new species, Solanaceae, taxonomy

*To whom correspondence should be addressed

Introduction 20-40 X 8-15 mm, succulent, young leaves sparsely While revising Lycium in Africa the authors found three hirsute with gland-tipped hairs and bicornate stellate specimens, in the collection of the Windhoek Herbarium hairs, older leaves glabrescent; base cuneate to obtuse; (WIND) that differ conspicuously from the known apex obtuse to acute. Flowers solitary, pedicels 2-10 species. Podlech & Roessler (1969) noted these unfamil­ mm long. Calyx enlarged and broadly urceolate, iar specimens, but they only knew the Namibian species indumentum as in leaves; tube 6-8 X 7-10 mm; lobes 5, and were therefore hesitant to describe them as a new unequal in size , deltate, 3-4 X 4 mm, apices acute. species. The present authors are revising the genus on a Corolla trumpet-shaped; tube creamy white, 10--13 mm much larger scale with the rest of Africa included, but no long, exterior glabrous, interior pilose between stamen type specimens or descriptions (in the absence of type . bases; lobes 5, reflexed , broadly ovate with apices specimens), agree with this taxon , obtuse, 3-5 x 3-5 mm, violet, margins ciliated, apices Description obtuse. Siamens 5, epipetalous, inserted 4-5 mm above base of tube, clearly protruding from corolla mouth; Lycium grandicalyx Joubert & Venter sp. nov. filaments unequal in length, 10--13 mm long; pilose at Frutex erectus, densus, spinosus. Folia fasciculata ; lamina the base; anthers elliptic; laterotrorse, 2 x 1.5 mm; spathulata, obovata, vel ovata vel elliptica, 20-40 x 8- 15 pollen grains 3-colporate, striate. Ovary very broadly mm , succulenta, sparsim hirsuto-glandulosa, pilis bicornibus ovoid, 2 x 2 mm ; style 17-18 mm long, protruding from stellaribus. Flores solitarii. Calyx amplificatus et late urceo­ corolla mouth; stigma obtuse. Fruit a berry; ovoid to latus, indumentum ut in foliis , tubus 6-8 X 7- 10 mm; lobi 5, deltati, 3-4 x 4 mm. Corolla buccinatus; tubus 10-13 mm orbicular, bilocular, placentation axile, ovules numer­ ous. Seed obliquely discoid; embryo curved; testa longus; lobi 5, recurvati, late ovati, 3-5 X 3- 5 mm , violacei. Stamina 5, epipetalosa, exserta ex ostio corollae. Ovarium late ruminate. (Figures 1, 2 & 3) . ovoideum; stylus exsertus ex ostio corollae. Fructus bacca. The known distribution stretches from the Witpiitz Semina oblique discoidea; testa ruminata. area to the Canyon in southern Namibia (Figure 4) . L. grandicalyx is a component of an open TYPUS.- Namibia: Witpiitz, Schwarzkalkrand, 9.6 km north xerophytic scrub community in the desert and succulent of Witpiitz Police Station, 02/09/1963, Merxmuller & Giess 3447 (WIND, holotypus; BLFU, isotypus). steppe (Giess 1970). Very high summer temperatures prevail and the little rain received falls in winter. It An erect, dense, much-branched thorny shrub of up to seems to be restricted to black lime terraces [a dark grey 1 m high. Stems with woody thorns of 15-30 mm long; to blueish grey dolomitic limestone (SACS 1980)] , that lateral branches spiny-tipped; older bark greyish and are common in the southern Namib. The species is striated; younger bark whitish. Leaves clustered on usually found in rock crevices of the terrace slopes, but stems and thorns, often solitary and alternate on young may also occur in dry stream beds that drain the terraces. stems, fasciculate on older stems, subsessile or shortly The known flowering period extends from May to petiolated; lamina spathulate, obovate, ovate or elliptic, August. S.Afr.J. Bot. , 1989, 55(5) 517

A

~ IOmm

5mm

Figure I Lycium grandicalyx. A & B, flowering twigs; C, flower; D , flower with calyx opened; E, corolla opened. [A from Meyer 2 (WIND), B-E from Merxmii.ller & Giess 3447 (WIND)]. 518 S.-Afr.Tydskr. Plantk. , 1989, 55(5)

Figure 2 Bicornate stellate hairs of Lycium grandicalyx. A , one complete bicornate hair amongst pinhead glandular hairs; B, one bicornate hair enlarged showing its basal part. [A & B, scanning electron micrographs of leaf material from Merxmiiller & Giess 3447 (WIND)]. Scale bars equal 100 and 10 f.Lm respectively in A and B.

Discussion hairs, but only L. grandicalyx and L. hirsutum have L. grandicalyx is a rather unusual Lycium with its ordinary, non glandular hairs as well. However, this is enlarged, urceolate calyx and bicornate, stellate hairs. In no positive indication that the two species are related. L. campanulatum E. Mey. ex C.H. Wr. a widened and The flowers of L. grandicalyx mostly resemble those rather large calyx occurs, but it is campanulate in shape found in L. campanulatum E . Mey. , L. ferocissimum and always clasps tightly around the corolla tube. In L. Miers, L. pilifolium C.H. Wr. , L. prunus-spinosa Dun. grandicalyx, however, the calyx is much wider than the and L. schizocalyx c.H. Wr. However, it is difficult to corolla tube and clearly separated from it. decide, on morphological characteristics, to which In Africa only L. grandicalyx and L. hirsutum Dun. species L. grandicalyx is most related. have compound hairs, but there are marked differences Podlech & Roessler (1969) regarded L. grandicalyx as in the hair structure of the two species. The hairs of L. closely related to L. afrum L. and L. rigidum Dun. on hirsutum are variously branched, but are never bicornate the basis of the inflated calyx and large corolla. and stellate as in L. grandicalyx (Figures 2 & 5) . Some However, L. afrum belongs to the group of Lycium species outside of Africa also have compound hairs but species in which the anthers are more or less included not the bicornate type (Hitchcock 1932; Bernardello within the relatively long corolla tube. L. grandicalyx, in 1986) . L. grandicalyx seems to be unique in the genus, contrast, belongs to the other group in which the corolla with regard to calyx structure and the bicornate, stellate tube is relatively short and with the anthers all fully hairs. exserted from the corolla mouth. Therefore no close All the African species of Lycium have gland-tipped

26° 28· lO· ~_L V ,-' r . , ~I \ '1 \ I ! 1 l ' i\f I 1"- 0' , , '1, ~ , l\ I I ( I I I f---... y 1 '\) , ! I 2 \ \ I r r--\, " 'i .l , , , I v I I \ I 1, " \ I I .1 I VI 7- \ I \ tI-r---. ) .. I I /1 j: 26 .: -- '"'-- \\ . ~ .\ t-/ : I 1',T'- J,,- H" '! " I \ \ i I: v If . ' ~,- ~ \ ' \\ , I 1 F'T* J I ' V>- Al/ I I 1 1 y 32 \ I T -j f\1 \ ~I T 1 I lliNti 1 "

Figure 3 Pollen micrograph of Lycium grandicalyx. [Scan­ ning electron micrograph from Merxmiiller & Giess 3447 Figure 4 Known geographical distribution of Lycium grandi­ (WIND)]. Scale bar equals 1 f.Lm . calyx. S.Afr.J. Bot., 1989, 55(5) 519

(- DA), Merxmiiller & Giess 3447 (BLFU, WIND). -2717 (Chamaites): 3 km south of border of Farm Uitsig, LU 82 (-CA), Giess & Muller 14268 (WIND). -2718 (Grunau): Fish River Canyon Nature Reserve (- DA), Meyer 2 (WIND).

Acknowledgements We acknowledge financial support from the Council for Scientific and Industrial Research and the University of the Orange Free State which has made this publication possible. We also thank the director and staff of the Herbarium of Windhoek (WIND) for the loan of speci­ mens and Dr R.L. Verhoeven, Department of Botany, University of the Orange Free State for the pollen and vestiture micrographs.

References Figure 5 Compound hairs amongst pinhead glandular hairs BERNARDELLO, L.M. 1986. Revision taxonomica de las on Lycium hirsutum. Scale bar equals 10 fLm. especies sudamericanas de Lycium (Solanaceae). Boln. R. Acad. Cordoba 57: 174-356. GIESS, W. 1970. Vegetation map of South West Africa. John relationship exists between these two species. L. rigidum Meinert (Pty) Ltd, Windhoek. Dun. is a later homonym of L. rigidum Thunb. and is HITCHCOCK, C.L. 1932. A monographic study of the genus therefore an illegitimate name. However, L. rigidum Lycium of the western hemisphere. Ann. Mo. bot. Gdn. 19: sensu Dunal is a synonym of L. ferocissimum which was 179- 374. already shown to have flowers resembling those of L. PODLECH, D. & ROESSLER, H. 1969. Solanaceae: grandicalyx. Lycium. In: Prodromus einer Flora von Stidwestafrika, Vol. 124, pp. 2-8, Verlag von J. Cramer, Lehre. SOUTH AFRICAN COMMITTEE FOR STRATIGRAPHY Specimens examined (SACS) 1980. Stratigraphy of , Part 1, Comp. -2716 (Witptitz): 9.6 km north of Witptitz Police Station Kent, L.E., Handb. geol. Surv. S. Afr. 8. 520 S.-Afr.Tydskr. Plantk. , 1989,55(5): 520-522 Alkaloids of the genera AspaJathus, Rafnia and Wiborgia (Fabaceae - Crotalarieae) I B-E. van Wyk* and G.H. Verdoorn Department of Botany, Rand University, P.O. Box 524, Johannesburg and Department of Chemistry and Biochemistry, Rand Afrikaans University, P.O. Box 524, Johannesburg, 2000 Republic of South Africa

Accepted 7 June 1989

The presence of alkaloids in three southern African genera of the tribe Crotalarieae is reported for the first time. Sparteine, lupanine and nuttalline were detected in several species of Aspa/athus L., Rafnia Thunb. and Wiborgia Thunb., but rarely in more than trace quantities. Except for very large quantitative differences, the alkaloids are almost identical to those found in the genus Lebeckia Thunb. Available evidence suggests that the three genera are more closely related to Lebeckia than to any other genus of the tribe. The divergence of these genera seems to be linked to an almost total loss of the ability to produce alkaloids.

Die teenwoordigheid van alkal6iede in drie Suider-Afrikaanse genera van die tribus Crotalarieae word vir die eerste keer gerapporteer. Sparte'ien, lupanien en nuttallien is waargeneem in verskeie spesies van Aspa/athus L., Rafnia Thunb. en Wiborgia Thunb. maar slegs by uitsondering in meer as spoorhoeveelhede. Afgesien van baie groot kwantitatiewe verskille, is die alkal6iede feitlik identies aan die wat in die genus Lebeckia Thunb. aangetref word. Beskikbare getuienis dui daarop dat die drie genera nader verwant is aan Lebeckia as aan enige ander genus van die tribus. Die divergensie van hierdie genera hou skynbaar verband met 'n bykans totale verlies aan die vermoe om alkal6iede te produseer.

Keywords: Aspa/athus, generic relationships, quinolizidine alkaloids, Rafnia, Wiborgia

*To whom correspondence should be addressed

Introduction All reference samples used in analytical TLC and GC The high incidence of convergence and intricate relation­ were fully authenticated by IH_ and 13C NMR ships amongst the Cape Crotalarieae have been spectroscopy and mass spectrometry. Identifications by discussed by Dahlgren (1963) and Polhill (1976) . analytical TLC and GC were confirmed by GC-MS Lebeckia Thunb. is considered to be the least specialized studies of two extracts (Aspalathus longifolia and Rafnia genus of the tribe and morphological similarities with racemosa). other genera of the tribe are taken as evidence of a close relationship. The distinct pattern of alkaloids observed Results and Discussion in the genus Lebeckia (van Wyk & Verdoorn 1989a) The distribution of alkaloids in 20 species of Aspalathus, indicated to us that alkaloids may provide valuable infor­ Rafnia and Wiborgia is shown in Table 1. Small amounts mation about the generic affinities of three other genera of sparteine, lupanine and nuttalline were present in of the tribe, namely Aspalathus L., Rafnia Thunb. and several of the samples (a-isolupanine in a few of them), Wiborgia Thunb. These three genera are considered to but rarely in more than trace quantities. We have also be particularly closely related to Lebeckia, as is found some indications of the presence of evidenced by their sequence in the latest generic and ammodendrine (a piperidyl alkaloid) but this could not tribal revision (Polhill 1976, 1981). be confirmed. No evidence of (a-pyridone alkaloids or An alkaloid tentatively identified as sparteine has pyrrolizidine alkaloids were found in any of the extracts. been reported previously from one species of Wiborgia Yield figures were extremely low, so that relative (van Wyk et al. 1988a) but, except for Lebeckia, no other yields are not given except for one species of Aspalathus information is available for any of the genera. Our aim (A . nivea) . The latter is a large shrub with flowers not with the present study was, therefore, to provide unlike those of some species of Lebeckia. The almost alkaloid data for a comparison of above-mentioned total absence of alkaloids in Aspalathus (if taken to be a genera. derived character state) indicates a more basal position for A. nivea than the sequence of species in Dahlgren's Materials and Methods (1988) revision would suggest. However, Dahlgren A total of 22 samples, representing 11 species of Aspala­ explicitly stated that the affinities of this species are thus, 6 species of Rafnia and 3 species of Wiborgia were uncertain and it is interesting to note that it was placed in studied in this survey. The species, authorities for names a monotypic infrageneric group. and voucher specimens are listed in the appendix. Except for the large quantitative difference observed, Methods of extraction and identification were as the alkaloids of all three genera are virtually the same as previously described (van Wyk et al. 1988a, b; van Wyk those found in the genus Lebeckia, and the diversity is & Verdoorn 1988). Gas chromatography conditions much less than expected. A comparison between various were the same as used by van Wyk & Verdoorn (1989a). genera of the Crotalarieae is shown in Table 2. Rothia S.Afr.J.Bot., 1989,55(5) 521

Table 1 Approximate yields and distribution of alkaloids in 20 species of Aspalathus, Rafnia and Wiborgia. (Authorities for names and voucher specimen details are given in appendix 1, tr indicates trace amounts)

Distribution of major alkaloids (% of total yield) Approximate yield------(fl.g/g dry wt) spartein e lupanine isolupanine nuttalline

Aspalathus A. capitata 4 tr tr A. carnosa 2 A. chortophila 2 tr tr A. cordata 20 tr tr A. hirta 14 tr tr A. juniperina 17 tr tr A. linearis 3 tr A. longifo!ia 4 tr tr A. nivea 2587 47 45 2 A. perfoliata 2 tr tr tr A. spinosa < 1 tr? Rafnia R. angulata 3 tr tr tr R. capensis 10 tr? R. elliptica 14 tr tr tr R. opposita 12 tr tr R. perfoliata 9 tr tr R. racemosa 4 tr tr Wiborgia W. fusca 21 tr tr tr W. obcordata ( I) IS tr tr tr W. obcordata (2) 31 tr tr tr W. obcordata (3) 26 tr tr tr W. sericea 11 tr tr tr? tr

Table 2 Distribution of alkaloids reported from the genera Lebeckia, Wiborgia, Aspalathus, Rafnia, Pearsonia, Rothia, Lotononis and Grata/aria. [Data for Lebeckia from van Wyk & Verdoorn (1 989a), Pearsonia from van Wyk & Verdoorn (1989b), Rothia from Hussain et at. (1 988) and Lotononis from van Wyk & Verdoorn (1 989c)]

Lebeckia Wiborgia Aspalathus Rafnia Pearsonia Rothia Lotononis Crotalaria

Tetracyclic quinolizidine alkaloids: nuttalline +++ tr tr tr tr + + sparteine +++ tr + tr + + lupanine +++ tr + tr ++ ++ tr isolupanine + tr? tr + tr tr Piperidyl alkaloids: ammodendrine tr? tr? tr? tr? + ++ tr Esters of quinolizidine alkaloids: ++ ++ Macrocyclic pyrrolizidine alkaloids: ++ ++

Occurs as a major component in: +++all species/samples, ++ most species/samples, + at least some species/ samples (tr = trace amount) 522 S.-Afr.Tydskr. Plantk., 1989,55(5)

Pers., Pearsonia Dummer, Lotononis (DC.) Eckl. & chemotaxonomic survey of major alkaloids in Lotononis Zeyh. and Crotalaria L. have also been included but and Buchenroedera. Biochem. Syst. Ecol. 17 (in press). Argyrolobiurn Eckl. & Zeyh., Dichilus DC., Melolobiurn VAN WYK, B-E., VERDOORN, G.H. & SCHUTTE, A.L. Eckl. & Zeyh. and Polhillia Stirton, all of which contain 1988a. Observations on the occurrence and distribution of (\'-pyridone alkaloids such as cytisine, N-methykytisine, alkaloids in some genera and species of the tribe anagyrine and thermopsine have been excluded from the Crotalarieae (Fabaceae). S. Afr. 1. Bot. 54: 75- 79. comparison. The summary of available data in Table 2 VAN WYK, B-E., VERDOORN, G.H. & SCHUTTE, A.L. leaves little doubt that Aspalathus, Rafnia and Wiborgia 1988b. Chemotaxonomic value of alkaloids in the genus are more closely related to Lebeckia than to any of the Dichilus. Biochem Syst Ecol. 16: 471-474. other genera. The alkaloid pattern shows a striking degree of confor­ mity with presumed evolutionary trends based on Appendix 1 Plant material of Aspalathus, Rafnia and morphological evidence. A relatively recent common Wiborgia used for alkaloid extraction. Voucher ancestry with Lebeckia is suggested, and the subsequent specimens are all in the Rand Afrikaans University divergence of Aspalathus, Rafnia and Wiborgia seems to Herbarium (JRAU) have been accompanied by the almost complete absence of alkaloids. Aspalathus capitata L.: Top of Constantiaberg, Cape Peninsula, Van Wyk 2754; A. carnosa Berg.: Top of Acknowledgements Constantiaberg, Cape Peninsula, Van Wyk 2152; A. chortophila Eckl. & Zeyh.: Zuurberg National Park, E. We wish to thank Dr L. Fourie (Department of Chemis­ Cape, B. & M. Van Wyk 1436; A. cordata (L.) Dahlg.: try, Potchefstroom University) for GC-MS analyses Between Constantia berg and Vlakkenberg, Cape and Miss A.L. Schutte (Department of Botany, Rand Peninsula, Van Wyk 2759; A. hirta E. Mey. subsp. hirta: Afrikaans University) for assistance with extraction N. side of Garcia's Pass, S. Cape, Van Wyk 2802; A. work. Continued financial support from the Rand juniperina Thunb. subsp. juniperina: Near top of Afrikaans University is acknowledged. Constantiaberg, Cape Peninsula, Van Wyk 2756; A. linearis (Burm. f.) Dahlg.: Eselbank, Cedarberg, Van References Wyk 2829; A. longifolia Benth.: Garcia's Pass, S. Cape, DAHLGREN, R. 1963. Studies on Aspalathus and some Van Wyk 2799; A. nivea Thunb.: Kabeljous River near related genera in South Africa. Bot. Notiser 9: 254-268. Humansdorp, E. Cape, Van Wyk 2813; A. perfoliata DAHLGREN, R. 1988. Aspalathus. In: Flora of Southern (Lam.) Dahlg. subsp. perfoliata: Jonkershoek near Africa, ed. Leistner, O.A., Vol. 16 (3,6), Botanical L. Research Institute, Pretoria. Stellenbosch, Van Wyk 2786; A. spinosa subsp. HUSSAIN, R.A., KINGHORN, A.D. & MOLYNEUX, R.l. spinosa: Top of Rooiberg Pass, Oudtshoorn district, Van 1988. Alkaloids of Rothia trifoliata and Rothia hirsuta. 1. Wyk 2800. Nat. Prod. 51(4): 809-811. Rafnia angulata Thunb.: Somerset West, Van Wyk 2788; POLHILL, R.M. 1976. Genisteae (Adans.) Benth. and R. capensis (L.) Druce: Near top of Constantiaberg, related tribes (Leguminosae). Bot. Syst. 1: 143-368. Cape Peninsula, Van Wyk 2757; R. elliptica Thunb.: POLHILL, R.M. 1981. Tribe 29. Crotalarieae (Benth.) Zuurberg National Park, E. Cape, B. & M. Van Wyk Hutch. In: Advances in legume systematics, eds Polhill, 615; R. opposita Thunb.: Garcia's Pass, S. Cape, Van R.M. & Raven, P.H., Vol. 1, pp. 399-402, Royal Botanic Wyk 2798; R. perfoliata E. Mey.: Jonaskop near Gardens, Kew. Villiersdorp, Van Wyk 2067; R. racernosa Eckl. & VAN WYK, B-E. & VERDOORN, G.H. 1988. The chemotaxonomic significance of integerrimine in Zeyh.: Between Seweweekspoort and Laingsburg, Buchenroedera and Lotononis section Krebsia. Biochem. Cape, Van Wyk 2171. Syst. Ecol. 16: 287-289. Wiborgia fusca Thunb. subsp. fusca: 56 km from Cape VAN WYK, B-E. & VERDOORN, G .H. 1989a. Town on West Coast Road, Van Wyk 2686; W. Chemotaxonomic significance of alkaloids in the genus obcordata Thunb.: Near Rondeberg farm on West Coast Lebeckia. Biochem. Syst. Ecol. 17 (in press). Road, Van Wyk 2691 (sample 1), between Paarl and VAN WYK, B-E. & VERDOORN, G.H. 1989b. Wemmershoek, Van Wyk 2678 (sample 2), at Eilandia, Chemotaxonomic significance of alkaloids in the genus between Worcester and Robertson (sample 3); W. Pearsonia. Biochem. Syst. Ecol. 17 (in press). sericea Thunb.: Laingsburg, SW Cape, Van Wyk 2193. VAN WYK, B-E. & VERDOORN, G.H. 1989c. A