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Taxonomy of Cyanastroideae (): A Multidisciplinary Approach

Article in Kew Bulletin · January 1998 DOI: 10.2307/4118869

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R. K. BRU~IMITT~,HANNAH BANKS', MARGARET A.T. JOHN SON^, KATHARINEA. DOCHERTY"~ITHJONES~, MARK W. CHASE^ & PAULA J. RUDALL~

Summary. Cyanastrum has previously been considered to include four species, all from the African continent. One of these, C. hostifolium from eastern Tanzania and northern Mozambique, is now transferred to a new , Kabuyea (K. hostifolia). This differs from Cyanastrum in several respects: vegetative morphology, perianth colour, anther dehiscence, pollen grain morphology, seed structure, chromosome number and karyotype. Karyotype analysis indicates that the individuals examined of the new genus are triploids. Analysis of rbrL DNA sequences for Cyanastraceae and Trcophilaeaceae places K. hostifolia as sister to C. cordifolium, but the level of sequence divergence between these two species is similar to that between genera in Trcophilaeaceae. The presence of a 'chalazosperm' in seeds of Cyanastrum was the primary reason for former recognition of the monogeneric family Cyanastraceae, but this structure is absent from seeds of Kabujea. Relationships of Cjanastrum and Kabuyea with other Tecophilaeaceaeare discussed.

General introduction ...... 769 Formal taxonomy (R. K. Brummitt) ...... 770 Leaf anatomy (PaulaJ. Rudall) ...... 781 Pollen morphology (Hannah Banks) ...... 783 Cytology (Margaret A. T. Johnson, Katharine A. Docherty & Keith Jones) . - 785 Seed structure (Paula J. Rudall) ...... 791 Molecular data (Mark W. Chase) ...... 795 General discussion ...... 798 Acknowledgements ...... 799 References ...... 800

Cyanastrum Oliv. has been considered to consist of four species from tropical Africa since the revision by Carter (1962). Her species concept differed from that of Clausen (1940) who had earlier recognised seven species. Although Cornu (1896), apparently unaware of Oliver's description of Cyanastrum six years earlier, published the name Schoenlandia for the same genus and species, nobody has hitherto suggested that the species currently referred to Cyanastrum should be placed in more than one genus.

Accepted for publication May- 1998. Herbarium, Royal Botanic Gardens, Kew, Richmond, Surrey, W93hB, U.K. jodrell Laboratory, Royal Botanic Gardens, Kew, Richmond, Surrey, TW9 3AB, U.K.

769 770 KEW BULLETIK VOL. 53(4) One of us (R. K. B.) selected Cjanastraceae as a small (monogeneric) family to write up as a sample treatment for the Species Plantarum Project, aiming to bring together all the taxonomic information down to species level and below on a world basis. This coincided with a current research program at Kew on Lilianae (e.g. Chase et al. 1995, Rudall & Cutler 1995, Rudall & Chase 1996, Chase et al. 1996,Johnson et al. 1996, Rudall et al. 1997, 1998). In writing the account, R. K. B. found that one species (C. hostifolium) was so different from the other three that it appeared that it could not be retained in the same genus. Further investigations soon confirmed that this species differs from Cyanastrum not only in gross morphology but in other respects as well. This paper provides a description of the new genus Kabuyea, together with a review of the characters supporting its separation. As previous papers have indicated (e.g. Chase et al. 1995, Rudall et al. 1997, Simpson & Rudall 1998), particularly on the basis of molecular data from rbcL, Cyanastrum is embedded within Tecophilaeaceae,with which it shares many characters, such as apical anther dehiscence (see General Discussion). Cyanastrum and the new genus Kabuyea are sister taxa within Tecophilaeaceae. The main aim of this paper is to assess species and genus-level systematics. A further publication is planned on the systematics of Tecophilaeaceaeusing a combined morphological and molecular analysis.

FORMALTAXONOMY

The species concepts of Carter (1962) are retained here, and material in major herbaria is unlikely to be misidentified at species level. In the specimen citations below, and in preparation of the distribution maps, data have been included from the Brussels (BR), Nairobi (EA) and Lisbon (LISC) herbaria according to information kindly supplied (see Acknowledgements below) without the material having been seen by me.

Subfam. Cyanastroideae Engl. in A. Engler & K. Prantl, Nat. Pflanzenfam., Nachtr. 1 zu I - IV: 70 (1897), sub Pontederiaceae. Type: Cyanastrum Oliv. Perennial herbs with up to 5 superposed discoid to shortly cylindric corms up to 2.5 cm diameter; up to 50 cm high; glabrous. Corms without a membranous or fibrous tunic, producing on their upper surface sheathing scarious cataphylls (referred to subsequently as just 'cataphylls') which envelope the emerging aerial parts; cataphyll-like bracts also sometimes present on the scape and intergrading upwards into inflorescence bracts. Leaves basal, longitudinally rolled in bud, entire, linear-lanceolate to heart-shaped, more or less petiolate. Scape either in the middle of the leaf rosette or borne separately from the leaves; inflorescence a simple raceme or the flowers rarely paired at the nodes, bracteate or not; flowers pedicellate; bracteole sometimes present. Receptacle funnel-shaped, persistent after flowering. Flowers hermaphrodite, actinomorphic, trimerous. Tepals 3+3, petaloid, shortly connate at the base into a short tube constricted above, white or blue. Stamens 3+3; filaments linear, inserted at the top of the perianth tube; anthers basifixed, dithecal, thecae oblong to linear, yellow, dehiscing by a terminal pore or a subapical clavate introrse slit. Pollen spheroidal, monosulcate, inaperturate, with perforate or rugulose to granular sculpturing. Ovary semi-inferior in the funnel of the receptacle, the 3 carpels evident as shallow lobes on the surface when young, these swelling above the receptacle after flowering so that the ovary later becomes deeply lobed; ovules 2 in each loculus; style inserted between the lobes of the ovary, filiform, exceeding the stamens; stigma not clearly demarcated. Fruit a capsule, often with only 1- 3 seeds; seeds ovoid to spherical, with or without endosperm, with or without a pitted chalazosperm which may comprise up to half the seed. 2 genera, 4 species, in tropical Africa.

KEYTO GENERA

1. Foliage leaves c. 4 per corm, forming a rosette closely enfolding the base of the scape, attenuate to a relatively indistinctly defined petiole; perianth white; anthers dehiscing by a short clavate introrse slit near their apex . .I. Kabuyea 1. Foliage leaves usually solitary on each corm, not enfolding the base of the scape, cordate or at least abruptly constricted at the base and distinctly petiolate; perianth blue; anthers dehiscing by an apical pore . - . 2. Cyanastrurn

The difference between the two genera in the arrangement of their leaves and scapes as they arise from the upper surface of the corm may be clearly appreciated by comparison of the excellent illustrations provided by Engler (1900) which are reproduced here as our Figs 1 & 2, his Taf. IX, figs. A & H showing the scape emerging from the middle of the leaf rosette in Kabuyea (our Fig. 1) in contrast with that of Cyanastrum in Taf. X, fig. A (our Fig. 2) in which the bare scape emerges alongside the solitary foliage leaf in C. goetzeanum.

1. Kabuyea Brummitt gen. nov. A Cyanastro foliis scapum amplectentibus vix abrupte petiolatis, perianthio albo, thecis antherarum apicem versus rima brevi introrsum dehiscentibus, et seminibus chalazospermum deficientibus differt. Typus: K. hostifolia (Engl.) Brummitt (Cyanastrum hostifolium Engl.) . The genus is named after Christine H. S. Kabuye, who retired as Botanist-in- Charge of the East African Herbarium, Nairobi, in 1995, in appreciation of her services to African botany over many years. Corms usually shortly cylindric or bucket-shaped and as broad as deep, or sometimes discoid. Foliage leaves and inflorescence emerging from the same cataphyll on the corm; foliage leaves c. 4 per corm, closely enfolding the base of the scape, the inner cataphyll up to 9 cm, sometimes also with shorter outer ones; scape not bearing cataphyll-like sheathing bracts. Foliage leaves attenuate at the base into an indistinctly defined petiole usually shorter than the lamina. Scape occasionally with a solitary sterile bract. Inflorescence bracts linear-triangular, scarious, caducous; pedicel with a small scarious bracteole near its base. Tepals white. Anther thecae oblong, dehiscing by a short clavate introrse slit near their apex. Pollen grains with perforate tectum. Seeds spherical, with pitted testa with microprickles in the pits, with endosperm, lacking chalazosperm. KE\V BULLETIN \'OL. 53(4)

FIG.1. Kobzcppn hostifolia. Reproduction of Engler's (1900) illustratio~lof Cyunn~tr-urnho~trfub~~?n. FIL.2. Cynnastr-urn.Reproduction of Engler's (1900) illustration of C. gu~tzranurnand C rurdzfohum. 774 KEW BUI.LETIK VOL. 53(4) One species in eastern tropical Africa (eastern Tanzania and northern Mozambique) (Map 1).

MAP 1.Distribution map of Kahujea hostifolia

1. Kabuyea hostifolia (Engl.) Brummitt comb. nov. Cyanastrum hostifolium Engl., Bot. Jahrb. Syst. 28: 358 (1900). Types: Tanzania, Uluguru Mts, Stuhlmann 1894 (syntype B, not seen); Tanzania, zwischen Khutu und Uhehe, Dec. 1898, Goetze 394 (syntype B, not seen). Cjanastrum bussei Engl., Bot. Jahrb. Syst. 38: 88 (1905). Type: Tanzania, Lindi Distr., Makondeland, Seliman - Mamba, May 1903, Busse 2667 (holotype B, not seen).

Leaves (including petiole) 10 - 45 x 3 - 9 cm, lamina elliptic to linear-oblanceolate, attenuate at the base to an indistinctly defined petiole about one third as long as the lamina, acute at the apex. Scape 15 - 35 cm including inflorescence, sometimes with one foliaceous bract below the lo~r~ermostflower. Inflorescence 2 - 10 cm .with 10 - TAXONOMY OF CYNASTROIDEAE (TECOPHILAEACEAE) 775

25 flowers; bracts up to 3 mm; pedicels 2 - 8 mm. Tepals 7 - 12 x 2 - 5 mm, white. Anthers c. 3 mm. ILLUSTRATIONS.Engler (1900, Taf. IX), reproduced here as Fig. 1; Carter (1966: her Fig.1). DISTRIBUTION.Eastern Tanzania, northern Mozambique (Map 1); in shady damp places in riverine forest or woodland; alt. 0 - 700 m. All specimens seen are cited below. TANZANIA.T6: Morogoro Distr., 1100 ft., 17 Dec. 1932, Wallace 579 (K); Kiberege, 5 March 1959, Haerdi 223/0 (EA, K, WAG 2 sheets); Ifakara Funge, 9 Dec. 1959, Haerdi 223/0 (BR, K); Morogoro Distr., Turiani, 22 Nov. 1955, Milne-Redhead & Taylor 7356 (BR, EA, K, LISC). T7: Karenga, near Kidatu, 28 March 1970, Harm's €9 POCS4263 (EA); Iringa Distr., Kidatu, 9 Jan. 1971, Mhoro 118 (EA, K). T8: Kilwa Distr., Dec. 1900, Busse 499 (EA); Tendaguru, 7 Dec. 1925, Migeod 22 (BM) and 29 April 1930, Migeod 687 (BM) and 18 Dec. 1930, Migeod 1056 (BM); 100 km W of Lindi, 27 Oct. 1934, Schlieben 5137 (BM); Rondo (Mvera) Plateau, Mikima, 24 Oct. 1934, Schlieben 5537 (BR, LISC); Rovuma Valley, 30 miles [48 km] E of Newala, 15 Jan. 1959, Hay 27 (K); Kilosa Distr., 53 km S of Kilwa Kivinje turn-off on Dar - Lindi road, 25 Nov. 1966, Gillett 18023 (BR, EA, LISC); Lumumwe, 7 Dec. 1969, Paget- Wilkes 687 (EA); Selous Game Reserve, 7 km NW of Kingupira, 27 Nov. 1975, Vollesen MRC3049 (C, EA, K); Mtwara Distr., 43 km on Mnazimoja to Mtwara road, 5 March 1991, Bidgood, Abdallah & Vollesen 1784 (EA, K) . MOZAMBIQUE.Cabo Delgado Prov.: Mouth of Msalu [Messalo] R., Nov. 1911, C. E. R Allen 91 (K); Macondes Distr., 12 km Nantulo to Mueda, 30 Dec. 1963, Torre & Paiva 9787 (LISC); Metoro Distr., Namacuto, 30 Jan. 1984, Croenendijk, Maite & Dungo 868 (K, WAG); Ancuabe, Metoro, Nematuca, Monte Namatiri, 1 Feb. 1984, Maite, de Koning & Dungo 193 (K, WAG). Niassa Prov.: Marrupa, 20 km from Matiquite [Matikite] towards Mecula, 10 Feb. 1981, Nuvunga 490 (BR, K); Matikite, 20 km from Marrupa, 18 Feb. 1982, Jansen L3 Boane 7844 (K). Nampula Prov.: near Malema, 25 Nov. 1931, Gomes e Sousa 879 (K); Nampula, rio Monapo, 23 Nov. 1935 (LISC); Nampula, 15 Dec. 1936, Torre 1282 (LISC); Monapo Distr., 6 km from Namialo to Metocheria, 24 Nov. 1963, Torre & Paiva 9258 (LISC); Monapo, property of Sr. Wolf, 11 Feb. 1984, de Koning, Croenendijk & Dungo 9571 (K,WAG). Zambezia Prov.: Campo, 19 Dec. 1904, le Testu 565 (BM); Namagoa Distr., Dec. 1943 -Jan. 1944, Faulkner 330 (K); Massinjere, Lugela, Feb. 1946, Pimenta 9 (LISC); Namacurra Distr., road from R. Licungo to Macuzi, 27 Jan. 1966, Torre & Correia 14200 (LISC); Namacurra, 4 km on road from Nicuadala, 31 Jan. 1966, Torre & Correia 14276 (LISC); Altomolocue, km 57 on road to Gile, 1 Dec. 1967, Torre & Correia 16308 (LISC); Campo, 18 km on road to Raposo, 28 Dec. 1967, Torre & Correia 16751 (LISC). Unlocalised: 1911, Stocks s.n. (K, 3 sheets). USES.An annotation on Pimenta 9, above, notes that in Mozambique the tubers are eaten in times of famine.

2. Cyanastrum Oliv., Icon. P1. 20: t. 1965 (1891). Type: C. cordijolium Oliv. Schoenlandia Cornu, Bull. Soc. Bot. France 43: 24 (1896). Type: S. gabonensis Cornu (= Cyanastrum cordijolium). 776 KEW BCLLETIN VOL. 53(4) Corms usually discoid and about twice as broad as deep. Foliage leaves usually solitary and emerging on the corm from a separate cataphyll from the inflorescence, thus not enfolding the base of the scape, the surrounding cataphyll up to 13cm, sometimes also with shorter outer cataphylls; inflorescence surrounded by a cataphyll up to 7 cm, the scape devoid of foliage leaves but often bearing 1- 3 cataphyll-like sheathing bracts. Leaves with cordate base and usually conspicuous basal lobes, or at least abruptly constricted at the base, the petiole distinct and equalling or exceeding the midrib in length. Inflorescence bracts absent or subulate to foliaceous; pedicel with or without a bracteole. Tepals blue, sometimes with a lilac tinge. Anther thecae oblong to linear, dehiscing by a terminal pore. Pollen grains with imperforate rugulose to granular tectum. Seeds ovoid, with a pitted chalaza1 end and a smooth micropylar end, without microprickles in pits, without endosperm, with a large chalazosperm occupying about half the seed. 3 species in tropical Africa from Nigeria to Tanzania and Mozambique (Map 2).

MAP2. Distribution map of Cjanastrum species: C. johnstoniz x ; C. goetzeanum 0;and C. cordlfolzume. The name Cjanastrum Oliv. was proposed for conservation by Buchheim (1961) on account of the existence of the earlier CyanastrumCass. 1829 in Compositae. The latter, however, was proposed by Cassini only in anticipation of possible need for the name in the future and was not validly published, and conservation is therefore not necessary (Rickett 1963).

KEYTO SPECIES OF CYA~\~~~STRI;IM

1. Leaf lobes up to one third as long as the midrib, leaf apex rounded to occasionally bluntly acuminate; pedicels not subtended by bracts; inflorescence 3 - 13 cm from lowest flower to apex; anthers 5 - 7 mm ...... 1. C. johnstonii 1. Leaf lobes half as long as to about equalling the midrib, leaf apex acute to bluntly acuminate; pedicels subtended by small to large bracts; inflorescence 1 - 6 cm from lowest flower to apex; anthers 3 - 5 mm ...... 2 2. Pedicels subtended by inconspicuous triangular to linear bracts 0.1 - 0.8 x 0.1~~...... 2. C. goetzeanum 2. Pedicels subtended by conspicuous ovate to lanceolate bracts up to 2 x 1 cm ...... 3. C. cordifoliurn

1. Cyanastrum johnstonii Baker, F1. Trop. Mr. 7: 336 (1898). Type: Zambia (?), between Lakes Tanganyika and Nyassa, Nov. - Dec. 1889, H. H. Johnston s.n. (holotype K). C. verdickii De Wild., Ann. Mus. Congo, Skr. 4, Bot. 1: 5 (1902). Type: Zaire, Lukafu, Nov. 1899, Verdick 275 (holotype BR, not seen). C. hockii De Wild., Repert. Spec. Nov. Regni Veg. 11: 517 (1913). Type: Zaire, Elizabethville, 1911, Hock s.n. (holotype BR, not seen). C. johnstonii var. cuneifolium S. Carter, Kew Bull. 16: 194 (1962). Type: Tanzania, Mpanda Distr., Kapapa Camp, 29 Oct. 1959, Richards 11638 (holotype K).

Leaf lamina (including lobes) 9 - 24 x 6 - 17 cm, broadly elliptic to occasionally suborbicular or heart-shaped, cordate at the base with a somewhat parallel-sided sinus, the basal lobes up to a quarter as long as the mibrib and more or less posteriorly directed, or rarely shortly cuneate without a sinus or basal lobes, the apex rounded to bluntly acuminate; petiole 6 - 22 cm. Scape 10 - 35 cm including inflorescence; cataphyll-like bracts on scape 1 - 3, the lower 1 - 2 sheathing and up to 6 cm long, the upper one usually not sheathing and linear- oblong up to 5 mm long. Inflorescence 3 - 13 mm from lowest flower to apex, with 3 - 17 flowers, ebracteate; pedicels 2 - 17 mm. Tepals 12 - 21 x 2 - 6 mm. Anthers 5 - 7 mm. ILLUSTRATIONS.Carter (1962, fig.1, 10 - 13);Carter (1967, t. 3641). DISTRIBUTION.South-eastern Zaire to Tanzania, Zambia and northern Mozambique; shady places in woodland or on river banks; 780 - 1530 m alt. In specimen citations below, only a selection of those seen from Zai're is included. 778 KEN' BULLETIN VOL. 53 (4)

ZAIRE. Shaba: Lubumbashi - Kiniama km 30, riv. Kafubu, Bodenghien 311 (BR); 28 km NE Lubumbashi, R. Luiswishi, Bulaimu 717 (BR); Kilata, Mokembo - Teha km 15, d'Hose 33 (BR) and 40 (BR); Parc Nat. Upemba, rive gauche Lufira, Oct. 1947, de Witte 3026 (BR, K); R. Kahela, Desenfans 4684 (BR); Lubumbashi, Hock s.n. (BR); Kiniama, 11 Dec. 1970, Lisowski 66 (BR, K); near Tshinshenda, Lisowski 67 (BR); Lubumbashi, Kasapa, Lisowski 87685 (BR); Luanza R., Kyanga Fall, 4 km WSW of Kabiashia, Malaisse 6072 (BR, K); 5 km E of Mangombo, Malaisse 8819 (BR); Kasenga, Mortelmans 148A (BR); Kapanga, Overlaet 783 (BR); Katuba, environs d'Elizabethville, Nov. 1934, Quarre 4332 (BR, K); 80 km SSE of Lubumbashi on Kasumbalesa road, Schaijes 1194 (BR); Keyberg, Schmitz 1057 (BR); 18 km SW of Lubumbashi, Schmitz 4798 (BR); Pweto, Schmitz 5042 (BR); Kabelenge, Sandoa - Dilolo km 12, Schmitz 8062 (BR). TANZANIA.T4: Near Kigoma, Gomba Mkenke Watu path, Clutton-Brock 346 (EA); Kigoma Distr., Mtunda Hill Forest Reserve, Nov. 1954, Richards s.n. (K); Mpanda Distr., Kapapa Camp, 29 Oct. 1959, Richards 11638 (K); reserve N of Ihigoma, 4 Nov. 1960, Morris Goodall 107 (EA); Buha Distr., Kasakela Reserve, 17 Nov. 1962, Verdcourt 3344 (BR, EA, K); Kasakati basin, 50 miles [80 km] S of Kigoma, 17 Feb. 1964, Ztani I6 (EA); Gombe Stream Reserve, Mkenke stream, 15 March 1964, Pyrozynski 562 (EA, K); Kasoge Mahali, 26 Jan. 1974, Uehara 162 (EA). T6: Kilossa [Kilosa], 13Jan. 1922, Swynnerton 549 (BM). T7: Iringa Distr., Kidatu, 9 Jan. 1971, Mhoro 119 (EA). ZAMBIA.Northern Prov.: between Lakes Tanganyika and Nyassa, Nov. - Dec. 1889, H. H. Johnston s.n. (K); Abercorn [Mbala] Distr., Jan. 1935, A. H. Gumwell225 (BM); Abercorn [Mbala] Distr., Ulungu, near Kankonde village, Glover in Bredo 6154 (BR) and 6155 (BR); escarpment above Mweru Wantipa, 22 Oct. 1949, Bullock 1353 (K); Kalambo Falls, 2 Nov. 1952, R. G. Robertson 201 (K); Chilongowelo, side of Katula Gorge, 12 Nov. 1954, Richards 2209 (K); Mweru, 18 Nov. 1955, M. Jones 28 (BM); Mbala Distr., Balymain - Luanza Valley, 19 Nov. 1956, Richards 7018 (BR, EA, K); Kawamba, 9 Nov. 1957, Fanshawe 3900 (BR, K); Chinakila, near Lufubu, 26 Oct. 1959, Lawton 654 (K); Mbala Distr., Lufubu R., Iyendwe Valley, 10 Dec. 1959, Richards 1195 1 (K); Mporokoso Distr., 13 Nov. 1960, Richardson, Livingstone CY Silas 47 (EA, K); Kalambo Falls, 15 Nov. 1960, Richards 13567 (K); Kalambo Falls, 9 Nov. 1963, Carmichael1005 (EA). MOZAMBIQCE.Cabo Delgado: Ancuabe, Metoro, Namatuca, 29 Jan. 1984, Maite €9 de Koning 176 (K); ibid. 29 Feb. 1984, Maite, de Koning CY Dungo 176 (K, WAG) [These two sheets appear to represent the same collection, apparently with an error of date on one.] Nampula Prov.: 8 km from Imala towards Mocuburi, 16 Jan. 1964, Torre & Paiva 9993 (LISC). The type of C. johnstonii was collected by Harry H.Johnston, British administrator in what is now Malawi between 1889 and 1896. It might therefore be expected that the type was collected in Malawi, where Johnston did make occasional collections himself as well as encouraging others such as Alexander Whyte to do so. The specimen bears a label in Johnston's own hand which includes only descriptive notes, and another label reading "Between Lakes Tanganyika & Nyasa, c. alt. 5000 ft, Nov. - Dec./89" in the hand of Daniel Oliver of Kew. The date shows that it was collected only a few weeks after Johnston arrived in this part of Africa. His own account (Johnston 1897: 94 - 95) shows that after his first encounter with the Arab slave trader Mlozi, whom he later defeated and hanged, he left Karonga on 10 November 1889 and travelled north into Tanzania, west to the south end of Lake Tanganyika, and back to Lake Nyasa (Lake Malawi) via the Mission Station at Fwambo, now in Zambia. The collection was listed as 'Cyanastrum sp.' by Burkill (1897), together with a collection by Nutt (not now found at Kew) who is known to have collected in this part of Zambia near the Tanzanian border. It seems very probable that Johnston's specimen was also collected in the area near Fwambo, now in Zambia. Th s ecies has never been recorded from Malawi (nor, surprisingly, has 9p any other species of Cjlanastrum or Kabuyea) . This species usually has short rounded lobes to the leaves, and var. cuneifolia was described by Carter (1962) for two specimens with no lobes at all and the lamina merely cuneate at the base. The type of the varietal name is actually leafless, but corms collected with the specimen were grown at Kew and produced leaves lacking basal lobes. It might be thought that this is an abnormal condition induced by cultivation. However, the other specimen cited in the protologue was collected in the wild in the same general area and has similar leaves with no lobes. At the time of description these were the only collections of the species known from Tanzania, and the difference might well have been significant geographically. More recent collections from the same region of Tanzania, however, have the shortly lobed leaves found elsewhere, and the variety is not maintained here. Further comments on this species are given by Carter (1967).

2. C. goetzeanum Engl., Bot. Jahrb. Syst. 28: 359 (1900). Type: Tanzania, Iringa Distr., Uhehe, Lofia R., Jan. 1899, Goetze 438 (holotype B, not seen). Leaf lamina (including lobes) up to 14 x 14 cm, broadly heart-shaped to sagittate, deeply cordate at the base with the sinus with widely divergent sides, the basal lobes slightly more than half as long as the midrib and diverging from each other, the apex acute; petiole up to 22 cm. Scape 12 - 26 cm; cataphyll-like bracts on the scape 1- 2, the lower one sheathing, mostly 2 - 5 cm. Inflorescence 2 - 10 cm from lowest flower to apex, with 3 - 12 flowers, some usually paired at nodes; bracts 1 - 10 mm, shortly triangular to linear, scarious; pedicels 3 - 10 mm. Tepals 10 - 14 x 2 - 4 mm. Anthers 4 - 5 mm. ILLUSTRATION.Engler (1900, Taf. X, figs. A - C), reproduced here as Fig. 2. DISTRIBUTION. South-central to southeast Tanzania, apparently'uncommon; shady places; 125 - 600 m alt. All specimens seen are cited below. TANZANIA.T6: Ifakara, Machipi, Nakatimbo, 11 Dec. 1959, Haerdi 391/0 (BR, EA, K, WAG 2 sheets). T7: see type above. T8: Selous Game Reserve, Kingupira, 20 Dec. 1976, Vollesen 4243 (EA, K, LISC, WAG) and 15 Jan. 1977, Vollesen 4326 (EA, K) . Specimens numbered Haerdi 391/0 do not appear uniform, and one of those at WAG bears a date 1964. They may have been collected separately and all numbered as one collection. 780 KEW BULLETIN VOL.53 (4) 3. C. cordifolium Oliv., Icon. P1. 20: t. 1965 (1891).Types: Cameroon, Ambas Bay [Limbe], 1861, Mann 769 (K, chosen here as lectotype); Nigeria, expedition of the interior of Yoruba, 11 May 1890, Millson 89 (lectoparatype K); Cameroon, near shore, comm. Veitch May 1878, Kalbreyer 89 (lectoparatype K); Gabon, Sierra del Crystal, May 1862, Mann 1644 (lectoparatype K) . Schoenlandia gabonensis Cornu, Bull. Soc. Bot. France 43: 21 (1896).Type: cult. hort. Cornu ex Gabon, Libreville, 1887, Pierre s.n. (holotype BR, not seen). C. cordifolium var. breuipedunculatum R. T. Clausen, Gentes Herb. 4: 297 (1940).Type: Cameroon, Bipinde, 26 Oct. 1908, Zenker 3669 (holotype US, not seen; isotypes BM, K). C. cordifolium var. compactuln R. T. Clausen, Gentes Herb. 4: 298 (1940).Type: cult. Cornell Univ., R. 7: Clausen H42 (BH, not seen, photo in protologue).

Leaf lamina (including lobes) 7 - 20 x 6 - 15 cm, sagittate to heart-shaped, deeply cordate at the base, the sinus with parallel to slightly divergent sides, basal lobes half as long as to about equalling the mibrib, the apex acute to bluntly acuminate; petiole 7 - 32 cm. Scape 6 - 17 cm including inflorescence; cataphyll-like bracts 1 - 3, mostly 2 - 4 cm long. Inflorescence 1 - 4 cm from lowest flower to apex, with 1 - 7 flowers; bracts up to 2 x 1 cm, ovate to lanceolate, scarious to foliaceous, caducous or persistent; pedicels 1 - 4 mm. Tepals 10 - 14 x 2 - 6 mm. Anthers 3 - 4 mm. ILLUSTRATIONS.Oliver (1891, t. 1965);Hepper in F1. W. Trop. Mr., ed. 2, 3: 109 (1968);Cronquist (1981,p. 1207);Cheek (1996,colour photos). DISTRIBUTION.Southern Nigeria to Gabon; on floor of evergreen forest; alt. 0 -1000 m. A selection of specimens seen is cited below. NIGERIA.Interior of Yoruba, Sept. 1890, Millson 89 (K, also BM ?); Adeyanba, 15 April 1909, Dennett 224 (K); Oban, 1912, Talbot 91 1 (BM, K) ; Gambari Forest, [I928- 301, MacGregor 598 (K); Ehor, March 1934, Fairbairn 47 (BM); Ijeba Prov., Shasha F.R., 6 Feb. 1935, I? W Richards 3083 (BM); Sapoba, 1935, Kennedj 2713 (K); Benin Prov., Okomu Forest Res., 10 Dec. 1947, Brenan 8462 (BM, BR, K); Ibadan south, 24 July 1948, Ahmed & Chizea FHI 20009 (K); Ijebu-Ode Prov., SNR No.1 Etemi Omo F.R., 6 May 1961, Onjeachusim & Binuyo FHI 43375 (K); Knob Hill, School of Agriculture, Akure, 3 April 1962, Swarbrick TS 2634 (K);15 April 1969, Lowe 1785 (K, WAG); Ogun State, 1 km from Omi elerangun to Erunwon, 29 May 1993, Daramola 168 (K). C,LMEROON.Arnbas Bay [Limbe], 1861, Mann 769 (K, lectotype); near shore, 13 March 1877, Kalbrqer-89 comm. Veitch (BM, K); Bipinde, 1908, Zenker3669 (BM, K); 15 - 35 km NE of Victoria, Oct. 1928, Mildbraed 10583 (K); Kumba Distr., S. Bakundu, near Barombi, 10 Feb. 1956, Binujo & Daramola FHI 35520 (K); Mamfe Distr., opposite Eyo sawmill site, 12 Feb. 1958, Tiku FHI 29416 (K);Res. forest. des Eaux et Forets, 8 km SW of Makak, 27 March 1974, WJ.J. 0. de Wilde & de Wilde- Duyfies 2239 (WAG); km 81 route Kribi - Ebolowa, Nkolessan, near Mbanga, 26 April 1968, Letouzq 9440 (K, WAG); 12 km from Kribi, between Ebolowa road and Kienke road, 30 April 1969, Bos 4444 (WAG); Chutes Ekom, Nkam R., 19 April 1972, Leeuwenberg €3 Berg 9663 (BR, WAG); SE of Lac Tissongo 35 km SE of Eira, 9 Jan. 1974 Letouzey 12650 (BR, K); env. Munyengue 20 km NW of Muyuka, May 1976, Satabie 284 (BR, K); Dept. Fako, Bakingili, 12 June 1984, Thompson & Rawlins 1376 (K); 5 km NW of Kumba, Lake Barombi Mbo, 9 Nov. 1986, Manning 793 (K). GABOS. Sierra del Crystal, July 1862, Mann 1644 (K); Mfoa, 85 miles [I40 km] E of Gaboon, Oct. 1896, Bates 540 (BM, K); Ahmey R., Corisco Bay, Schlechter 12830 (BR); Rabi, Shell Oil concession, 300 m SW of Rabi 46,J J. de Wilde, Arends, Louis €3 Wieringa 9655 (WAG); Cristal Mts, Tchimbele, 13 Dec. 1989, J. J. de Wilde, Arends, Louis €3 Wieringa 10035 (WAG). ZAIRE.Mbanza Ngungu, Bequaert 7763 (BR). USES.Cheek (1996) has recorded that in Cameroon the tubers may be pulverized and mixed with oil as a treatment for sore throats, and also that they are used by women to decorate their bodies with black lines.

Material examined Cjanastrum cordzfolium (HK 1990-1936), C. cordifolium (HK 1976-1515), Kabuyea hostifolia (HK 1957-41605; K). HK indicates material cultivated at Kew.

Description Both species are similar except where otherwise indicated.

LEAFSURFACE (Fig. 3C, D). Epidermal cells polygonal, with fine surface striations, especially in K. hostifolia. Stomata present on abaxial surface only, slightly protruding; paracytic (with a more or less distinct subsidiary cell on either side of guard cells). Hairs and papillae absent. LEAFTS LAMINA (Fig. 3A, B). Leaf thickness c. 260 pm in C. cordifolium, slightly thinner in K. hostifolia (c. 230 pm). Epidermis: epidermal cells with domed inner periclinal wall; adaxial epidermis c. 50 pm thick; abaxial epidermis slightly thinner. Stomata: slightly raised above surface, with outer cuticular ridge. Mesophyll: consisting of 4 - 6 layers of abaxial spongy mesophyll cells and 1 - 2 layers of short adaxial palisade mesophyll cells; interspersed with occasional secretory ducts, each surrounded by 1 - 2 rings or epithelial cells. Occasional indigo-blue anthocyanin cells present ("qanocysten": Solereder (1917), or "anthoqanoplasts": Peckett & Small (1980)). Vascular bundles: in one row in centre of leaf, collateral, consisting of adaxial xylem and abaxial phloem; lacking sclerenchyma; with bundle sheath 1 - 2 layers of parenchymatous cells. Crjstals absent. LEAFTS PETIOLE.Outline: in K. hostifolia a shallow U-shaped arc; in C. cordifolium ranging from a deeper U-shape to a cylinder. Vascular bundles in 1 - 3 arcs, closing almost to a cylinder in C. cordifolium, lacking sclerenchyma. Ground tissue parenchymatous, interspersed with secretory ducts as in lamina.

Discussion Evidence from leaf anatomy supports a close relationship between Cyanastrum and Kabuyea since absence of crystals and presence of secretory canals are synapomorphies linking Cyanastrum and Kabuyea. Leaf anatomy in Cyanastrum was examined previously by Solereder (1917), Russell (1938) and Arroyo (1986), although their accounts differ in some respects. 782 KEW BULLETIN VOL. 53(4)

.>; -- Ex-.:

FIG. 3. Leaf anatomy (A, D Kabuyea hostifolia. B, C Cyanastrum cordifolium).A, B Cross sections of leaves, with secretory ducts arrowed. C, D Stomata on abaxial leaf surfaces. Scalebars = 100 pm.

The most unusual feature of both C. cordifolium and K. hostifolia leaves is the presence of secretory glands with a ring of epithelial cells (Fig. 3A, B) in lamina and petiole. These are visible with a hand lens as frequent pale dots in the leaf and were recorded by Solereder (1917) as "interzellularen schizogenen Sekretbehalter" in C. cordifolium, although Arroyo (1986) did not record them in either C. cordifolium or K. hostifolia (as C. hostifolium). They are absent from other Tecophilaeaceae (Arroyo 1986) and not recorded in leaves of other asparagoid families, although Solereder (1917) observed that similar glands are present in Dilatris, a genus of Haemodoraceae, with which Cyanastrz~mwas formerly associated. The lack of any type of calcium oxalate crystal (e.g. raphides or styloids) in Cyanastrum and Kabujea is also an unusual feature, also recorded by Solereder (1917) and Goldblatt et al. (1984) for Cyanastrum but not by Arroyo (1986), who reported crystals present. The presence of raphides is a plesiomorphic character for (Rudall & Cutler 1995). Arroyo (1986) reported raphides present in most species of Tecophilaeaceae. Arroyo (1986) recorded anomocytic stomata (without subsidiary cells) in Cyanastrum, although Solereder (1917) reported the paracytic type, as also found here in both genera (Fig 3 C, D). This probably represents a difference in interpretation rather than variation in the material; in some cases the subsidiary cells are indistinct (resembling other epidermal cells). Anomocytic stomata are the plesiomorphic type for Asparagales, although paracytic and tetracytic types occur in some families.

iVaterials and methods Buds from living specimens were plunged into 2% glutaraldehyde in cacodylate buffer with a pH of 7.2, then dissected in further buffer solution. Pollen was post- fixed in a 2% solution of osmium for two hours, rinsed in cacodylic buffer, and secured inside dialysis tubing, then taken through an ascending alcohol series from a 10% solution to absolute alcohol. After two rapid changes in absolute alcohol, the pollen was given two changes in acetone (transfer medium), then critical point dried in a Balzers CPD 030 using C02 as the transitional fluid. For scanning electron microscopy (SEM), pollen was dusted onto a stub and sputter-coated for two minutes with platinum in a Balzers Union SPD 030 sputter coater, then examined using a Hitachi S - 2400 SEM. For light microscopy, pollen was mounted in glycerol jelly and examined using a Nikon Optiphot microscope with an oil immersion x 100 magnification objective. Where fresh material was unavailable, buds were removed from herbarium sheets, fixed in osmium and critical-point dried. For surface-only detail it was found that simply putting the pollen through an ethanol series was sufficient, although this treatment did cause the delicate aperture membrane to rupture. Pollen was taken from herbarium material (K) or living material (HK) at the Royal Botanic Gardens (RBG) Kew, and all pollen preparations are held in the Palynology Unit there.

Descn$tions Pollen grains of all species are similar in being more or less spheroidal in shape, circular in outline, monosulcate with a granular membrane (Fig. 4 A, B). Tectal sculpturing and grain sizes differ between species as detailed individually below.

Kabuyea hostifolia (K: Haerdi 233/ 0, Tanzania) Surface ornamentation psilate-perforate (Fig. 4 E). Measurements; P: (35) - 39.3 - (44) pm, E: (37) - 39.3- (42) pm, P/E = 1.00. 784 KEW BULLETIN VOL. 53(4) Cyanastrum johnstonii (K: Bullock 1353, Zambia) Surface ornamentation rugulose to granular (Fig. 4 F). Measurements; P: (30) - 35.5- (39) pm, E: (30) - 33.5 - (35) pm, P/E = 1.07.

Cyanastrum goetzeanum (K. Vollesen 4243, Tanzania) Surface ornamentation rugulose to granular (Fig. 4 D). Measurements; P: (30) - 36.8 - (40) pm, E: (30) - 37 - (39) pm, P/E = 0.995.

Cyanastrum cordifolium (HK & K: Swarbnck TS2634, Nigeria) Surface ornamentation rugulose (Fig. 4 A - C). Measurements; P: (34) - 37.1 - (45) pm, E: (35) - 37.5 - (40) pm, P/E = 0.99.

Discussion Pollen grains of K. hostifolia are on average slightly larger than those of the Cyanastrum species, but the most significant variation in pollen morphology is in surface ornamentation. Whereas the three Cyanastrum species all have granular to rugulose tectal sculpturing, the surface of K. hostifolia pollen is psilate-perforate. The slight variation in the surface ornamentation of the Cyanastrum species (Fig. 4 C, D, F) is due to the length of the individual surface elements, which are more rounded in C. johnstonii, more elongate in C. cordifolium and intermediate in C. hostifolia. Amongst earlier works on Cyanastrum pollen, Schulze (1983) gave light microscope (LM) descriptions of pollen of C. cordifolium and K. hostifolia (as C. hostifolium), and Simpson (1985) considered only Cyanastrum cordifolium (SEM). Schulze's (1983) interpretation of surface patterning in K. hostifolia is slightly different from ours, but comparison between LM and SEM observations is difficult. He recorded frequent trichotomocolpate (= trichotomosulcate) grains amongst colpate (= sulcate) grains in K. hostifolia, and this is not inconsistent with records in other 'lower' asparagoids, where trichotonlosulcate pollen is associated with simultaneous microsporogenesis (Rudall et al. 1997). Both Cyanastrum and Kabuyea lack an operculum on the sulcus, which is present in other Tecophilaeaceae (Simpson 1985).

This account combines new data obtained by M. A. T. J. and K. A. D. with earlier unpublished work by K. J. The original study revealed striking cytological differences between Cyanastrum hostifolium (now Kabuyea hostifolia) and other species of Cyanastrum, which were confirmed by the more recent investigations. Some other species of Tecophilaeaceae available in the Living Collections Division at Kew have been included in the study to put the observations on Kabuyea and Cjanastrum into a broader context.

FIG.4. Pollen morphology (SEM). A, B, C Cyanastrum cord~olium(HK); D C. goetzeanum (Vbllesen 4243); E Kabuyea hostzfolia (Haerdi 233/0);F C. johnstonii (Bullock 1353). A whole grain. B whole grain showing aperture. C - F exine surface. Scale hars: in A, B = 10 pm, in C - F = 2.3 pm.

TAXONOMY OF CYNASTROIDEAE (TECOPHILAEACEAE) 787 Materials and methods Localities of the living material studied are shown in Table 1. Suitable rapidly growing root-tips were removed from potted maintained in the Living Collection Division at the Royal Botanic Gardens, Kew. Roots were pre-treated using a 0.002M solution of Ebhydroxyquinoline (OQ) for 3 hours at lg°C, prior to the preparation of slides using a standard Feulgen squash method (Johnson 1982). For meiotic preparations, buds were fixed in 1:3 acetic-alcohol and stained in 1% propionic carmine.

FIG. 5. Somatic cells of A Cyanastrum curdifolium (Cyt. ref. 62-1682), 2n = 22; B C. cordifolium (Cyt. ref. 94181), 2n = 22; C C. johnstonii var. johnstonii (Cyt. ref. 62-1779), 2n = 22; D Kabuyea hostifolia (Cyt. ref. 94180), 2n = 36; E TecqPhilaea cyanocrocus (Cyt. ref. 95-66) 2n = 24; F Walla'a gran'lis (Cyt. ref. 61-164), 2n = 24. Scale bar = 10 pm. 788 KEW BULLETIN VOL. 53(4) Permanent slides are retained in the Cytogenetics Section of the Jodrell Laboratory, and voucher specimens deposited in the Kew Herbarium (K). Chromosome photographs were taken on Zeiss photomicroscopes I or 111, the latter using PAN F film, and the karyotype drawings prepared from enlarged photocopies of photographs which were then cut out, arranged in size order and finally retraced.

Results Chromosome counts are given in Table 1,with new records asterisked. Fig. 5 shows somatic cells and Fig. 6 karyotypes of material investigated. Cyanastrum. All the accessions of Cyanastrum species were diploid with 2n = 22, x = 11 with relatively small meta- or submetacentric chromosomes of similar size (Figs 5 A, B, 6 A, B). Kabuyea. The chromosomes of the two collections of K. hostifolia are more than twice the size of the Cyanastrum (Figs 5 A, B, C), (Figs 5 E, 6 D) or Wallena chromosomes (Figs 5 F, 6 E). Both collections of this species revealed very similar results, with a chromosome number of 2n = 36. Karyotype analysis (Fig. 6 C) shows the 36 chromosomes of similar size, arranged in sets of threes. The karyotype comprises 33 metacentric or submetacentric chromosomes plus three acrocentric chromosomes (positioned last in Fig. 6 C). The first six chromosomes in Fig. 6 C and the last three all have prominent secondary constrictions and can be divided morphologically into three easily identifiable groups of three differently sized

(I 8 c~ iiil ir ~vt lib(a 111 r tn Ar xtr 4'1'r

FIG.6. Kary0t)~edrawings of A Cjanastrum cordfolium (Cyt. ref. 94181), 2n = 22; B C. johnstonii var. johnstonzi (Cyt. ref. 62-1779), 2n = 22; C Kabuyea hostfolia (Cyt ref. 94180), 2n = 36; D (Cyt. ref. 95- 66), 2n = 24; E gracilis (Cyt ref. 61-164), 2n = 24. Scale bar = 10 pm. chromosomes. The first two of these chromosome sets have median to submetacentric centromeres, but the sets differ in the position of the secondary constrictions which in the first set lie nearer the centre of the short arms. In the second set, the secondary constrictions are closer to the end of the shorter chromosome arms. The third set, positioned last in the drawing, shows that each of the three chromosomes has a short arm roughly one third the length of the long arms, with the secondary constrictions in the short arms delimiting distal segments about one half of its length. These three groups of chromosomes are easily distinguished in Fig 6 C, serving as useful markers. Tecophilaea. Both collections of 7: qanocrocus studied have 2n = 24, n = 12 (Figs 5 E, 6 D), with relatively small meta- to submetacentric chromosomes though they are slightly larger in overall size than those of Walleria grucilis (Figs. 5 F, 6 E). No secondary constrictions were seen in this diploid species, which is based on x = 11. Walleria. Material of W gracilis has 2n = 24 (n = 12) chromosomes (Fig. 5 F), which are slightly smaller than those of Tecophilaea qanocrocus. The karyotype of this diploid species, also based on x = 12, consists of twelve submeta- or metacentric chromosomes, with one pair of secondary constrictions in the long arms of the last chromosome pair (Fig. 6 E).

Meiotic analysis in Kabuyea hostijolia In K. hostijolia only four cells could be analysed for pairing behaviour at meiotic metaphase which gave the following complete cell pairing patterns: (1) 12111, (2) 1VI t 9111 t 111 t 11, (3) 2VI t 7111 t 111 t 11, (4) 2VI t 5111 t 311 t 31 (Fig. 7).

FIG.7. Metaphase I drawing of ~neiosisi11 KaDujea hostfolia (Cyt ref. 63-1493) 2n = 36, (L-R) 2V1, 5111, 311, 31. Total of 34 chiasmata.

Meiotic anaZysis in C. johnstonii ('uax cuneijolium') Meiosis was examined in a few flower buds of C. johnstonii ('var. cuneijolium'), in which pairing was irregular with no cells showing complete pairing. The number of bivalents ranged from two to only eight per cell with most bivalents associated by a single chiasma, giving an overall mean of 6.88 chiasmata per cell. During the meiotic study of this , a persistent nucleolus-like body was observed. This appeared at zygotene and leptotene stages as a large spherical structure positioned near the nuclear membrane and without any visible connection with the chromosomes which were clustered to one side of the nucleus. At pachytene and diakinesis there was some contact between the structure and several chromosome associations. In cells approaching metaphase it contracted in size and became very densely stained, and at this stage vacuolation (which had first 790 KEMTBULLETIN VOL. 53 (4) been observed during prophase) became more pronounced. The structure persisted at metaphase, becoming more highly vacuolated and lying outside the spindle. From metaphase to anaphase, a variable number from 14 to 20 or more minute bodies appeared scattered across the spindle which were thought to be micronucleoli or nucleoloides. From metaphase onwards the staining intensity diminished until at first telophase in every cell, the outlines of vacuolated spheres could barely be detected. The partial asynapsis gave rise to lagging of univalents at anaphase I and at the onset of telophase before segregation, which in some cases resulted in partial restitution, with the formation of dyads as well as tetrads at the end of meiosis. At that time telophase nuclei could be seen to possess one or more nucleoli of normal appearance. Pollen fertility in this plant was low.

Discussion All the Cyanastrum material studied here has 2n = 22, which agrees with the count recorded by Sat6 (1942), who studied material identified as C. cordifolium. However, Nietsch (1941) recorded 2n = 24, apparently for the same species. Ornduff (1979) suggested that Tecophilaeaceae have a basic number of x = 12. La Cour (1955) reported a count of 2n = 24 for Tecophilaea cyanocrocus, but there is no published karyotype for this species. The count of 2n = 24 obtained here for Wallem'a gracilis agrees with previous counts made for W mackenzii J. Kirk and W nutans J. Kirk by Goldblatt & Manning (1989). Cyanastrum cordifolium and C. johnstonii (typical and 'var. cuneifolium') have similar karyotypes comprising relatively small chromosomes with the same diploid basic number of x = 12. The irregular pairing in C. johnstonii 'var. cuneifolium' and its persistent nucleolus are interesting phenomena. No second division cells were available to examine the ultimate fate of the nucleolus-like body, and there was no evidence of its persistence in root-tip mitoses. The resulting high level of pollen abortion found in this plant is probably untypical of the species as a whole. Chromosome analysis of Kabuyea hostifolia reveals that it is triploid with 2n = 36 (x = 12) relatively large chromosomes. All six accessions studied of Cyanastrum species have much smaller chromosomes and are diploid (2n = 22) with a basic number of x = 11. Although Kabuyea shares the basic chromosome number of x = 12 with Tecophilaea and Walleria, its large chromosomes bear no karyotype resemblance to either genus, nor to Cyanastrum (based on x = 11) in which both karyotype and chromosome size are more similar to Tecophilaea and Wallem'a. Both collections of K. hostifolia were found to have remarkably similar karyotypes. Although the meiotic sample of K. hostzfolia was small, the high level of multivalent pairing with a high degree of structural homology between its constituent sets suggests that the plant is a polyploid. Trivalents were the most common configuration and in some cells all the chromosomes are associated in this way. This behaviour is typical of an autotriploid. On the other hand, the presence of hexavalents raises the possibility that the plant could be hexaploid, with trivalent formation as a consequence of an inadequate number of chiasmata for high levels of hexavalent pairing. It is also possible that hexavalents could be the result of heterozygosity for some interchanges in a triploid plant. Karyotype analysis has been helpful in deciding between these two possibilities. There are three distinctive types of cl-lromosomes with secondary constrictions each present three times, which suggests three homologous chromosome sets and supports the concept that the plants are triploids with a basic number of x = 12, rather than hexaploids based on x = 6. The small amount of heteromorphy seen in some of the chromosome triplets in the karyotype drawing, Fig. 6 C, indicates that this individual is more likely to be of allotriploid origin. The discovery that this new genus has triploid individuals is surprising, since triploid plants are rare and sterile unless seeds are produced asexually by apomixis or reproduction is by vegetative means. Examples of successful triploids have been reported in Lilium ticgrinurn by Takenaka & Nagamatsu (1930), in naturally occurring Lycoris hybrids by Kurita (1986, 1987) and two other cases have been discovered in ililuscari by Kar1i.n (1984) and Johnson et al. (1996). These papers reported only triploid individuals, (although with an apparent absence of plants at other ploidy levels in the populations examined), so it is impossible to be certain if every plant is always triploid throughout every population, or if the phenomenon is limited only to the individuals studied. The comprehensive study of 113 plants from a wild population of Ranunculusficarin (n = 8) by Marchant & Brighton (1974) found 29% of the plants investigated to be presumed triploids, 31% diploids and 40% tetraploids. Further studies are required to determine the frequency of triploidy in Knbuyea as it could be that our limited study has by chance only examined isolated triploid individuals occurring within diploid populations. Holvever, if K. hostifolio is a triploid species it ~vouldbe able to survive successf~lllyin its native habitat by non- sexual reproduction of its tubers, or pseudo-sexually by apomictic seed production.

SF.EDSTKL'CTURE &Iat~nnl~xamzned Material was obtained from Kew herbarium material (K): Cynnastru7n cor(i2folzum (K: Sabatz~284, 28 May 1976), C, gohnstonii (K: Lzsozuskz 66, 11 Dec. 1970), Kabuyea hostqolza (K: Bzdgood, Ahdnlluh & Ibllrsen 1784, 5 March 1991).

L)escrifition Seeds of Cjnnastrum and Kabuyea are large and yellow (lacking in phytomelan). Seeds of C. cordifolium (Fig. 8 A - C) and C. johnstonii (Fig. 8 D - F) are more or less ovoid (c. 9 mm x 5 mm across) ~vithhvo distinct parts: a pitted chalazal end (more deeply pitted in C. john.rtonii than in C. cor(liJb1iu~r~)and a smooth micropylar end. In C. cordifolium (Fig. 8 A) the smooth ~nicropylarend, containing the embryo, is slvollen. Tillich (1995) also illustrated a seed of C. corclifoliurn. The pitted chalazal region comprises about half the seed in C. cordijolium (Fig. 8 A) and about a third of the seed in C. john~lonii(Fig. 8 D). In sectioned material, the pitted chalazal region ('chalazosperm') is shown to contain loosely packed strands of parenchymatous cells with air spaces behveen (Fig. 10 E, F), as described by Fries (1919). Endosperm is entirely lacking in these two species, also as described by Fries (1919). In C. johnstonii testa1 epidermal cells at the chalazal end are thick walled, with spiral thickenings (Fig. 10 D), except in the deep surface pits. 792 KEW BULLETIN VOL.. 53(4)

FIG. 8. Seeds with pitted chalazal end on left hand side. A, B, C Cyanastrum cord~ulium,testa at chalazal end shallowly pitted; D, E, F Cyanmtrum johnstonii, testa at chalazal end deeply pitted. B, C, E, F Scanning electron micrographs. Scale bars: in A, B, D = 1 rnrn; in C, E = 500 pm; in F = 200 pm.

Seeds of K. hostijolia (Fig. 9 A) are roughly spherical, c. 4 mm diameter, without two distinct regions. The testa is thick and deeply pitted over the entire surface. The pits contain rows of tiny microprickles (Fig. 9 B - D, 10 A), which are absent from Cyanastrum seeds (Fig. 10). The microprickles are short pointed unicellular papillae (Fig. 10 B). There is no 'chalazosperm' region of loosely packed strands of parenchymatous cells, as in Cyanastrum. Furthermore, Kabuyea seeds contain both endosperm (Fig. 10 C) and embryo, in contrast to Cyanastrum, which lacks endosperm in the mature seed. TAXONOMY OF CYNASTROIDEAE (TECOPHILAEACEAE) 793

FIG. 9. Kabuyea hostijolia, A entire seed with deeply pitted surface with microprickles in pits. B, C, D Scanning electron micrographs. Scale bars: in A, B = 1 mm; in C = 500 pm; in D = 200 pm.

Discussion The swollen chalazal region of the seed which Fries (1919) called a 'chalazosperm' is present in both species examined of Cyanastrum but absent from Kabuyea; it is therefore an autapomorphy for Cyanastrum. (Fries described only C. cordifolium and C. johnstonii.) The function of this region is by no means certain. It has similarities with the elaiosomes found in seeds of other asparagoid taxa, and it could well have a role in seed dispersal, as Fries (1919) conceded. However, Fries considered it most likely to be a storage tissue because mature seeds of Cyanastrum lack endosperm. It is unusual for a seed storage tissue to occur outside the vascular bundle, i.e. not adjacent to the embryo sac. However, the thick-walled chalazal epidermis, especially in C. johnstonii, argues against the elaiosome theory, as it would make the food reserves less accessible to dispersal agents. Indeed, the presence of endosperm and concurrent lack of chalazosperm in Kabuyea seeds seem to support the storage tissue theory of chalazosperm in Cyanastrum. 794 KEW BULLETIN VOL. 53 (4) TAXONOMY OF CYNASTROIDEAE (TECOPHILAEACEAE) 795 The presence of conspicuous large yellow seeds with deeply pitted seed coats in both Cyanastrum and Kabuyea is evidence of a close relationship between them, although detailed morphology varies. Tecophilaeaceae have relatively small seeds, which are sometimes black (phytomelaniferous, e.g. in ) and always lacking the characteristic deep testa1 pitting of Cyanastrum and Kabuyea (Huber 1969; Rudall, unpublished). Seeds of Wallem'a are relatively large (although not as large as those of Cyanastrum) and dark brown with a 'warty' testa described by Huber (1969). Testa1 pitting is most pronounced in K. hostqolia, in which it occurs over the entire surface rather than just the chalazal end, and only K. hostifolia has microprickles within the pits (Fig. 9 B - D). The adaptive significance of the deep pits and microprickles, both highly unusual structures, is extremely obscure, although we can speculate that they may trap air, possibly as a flotation aid for seed dispersal.

MOLECULARDATA

Procedures for DNA extraction and PCR have been covered in previous papers (Chase et al. 1995; Rudall et al. 1998). Taxa for which rbcL sequences were analysed here as well as vouchers and electronic database accession numbers are listed in Table 2. Selection of taxa was guided by previously published rbcL analyses, particularly those cited above. Campynema was specified as the outgroup because previous studies have shown it to be a member of , whereas the other families included here are members of the 'lower' Asparagales. The rbcL gene was amplified using the 1F and 136813 primers (IF: 5'- ATGTCACCACAAACAGAAAC-3'; 1368R: 5'-CTTTCCAAATTTCACAAGCAGC A - 3'), and these plus two internal primers were then used for sequencing (636F: 5'- GCGTTGGAGAGATCGTTTCT-3'; 724R: 5'-TCGCATGTACCYGCAGTTGC-3'). Out of the standard length of 1428 basepairs (bp), we thus used 1347 bp of sequence for most taxa. Of this, 344 positions were variable but only 182 were potentially informative. Using PAUP 3.1.1 (Swofford 1993), we performed 1000 replicates of random taxon-entry, using tree bisection-reconnection (TBR) swapping and keeping multiple most-parsimonious trees (MULPARS on). We also used successive approximations weighting (SW; Farris 1969) to eliminate the effects of positions that change too frequently; based on the shortest trees obtained, eight positions change seven or more times. To evaluate internal support, we performed 1000 replicates of the bootstrap (Felsenstein 1985), both with and without the weights derived from SW. The single shortest tree from SW is shown with its equally weighted lengths (ACCTRAN optimization) above the branches and the two bootstrap percentages below (equal weight/SW; Fig. 11). Branches not present in the strict consensus tree of the equally weighted analysis are indicated by arrows.

FIG.10. Light micrographs of seed sections. A, B, C Kabuyea hostifolia; A deeply pitted testa with microprickles in pit; B microprickles; C endosperm; D Cyanastrum johnstonii, thick-walled chalazal epidermal cell, with spiral thickenings; E, F Cyanastrum cordifolium, loosely packed files of chalazosperm cells, containing starch. Scale bars: 50 pm. 796 KEW BULLETIN VOL. 53(4) TABLE2. Taxa, authors, vouchers (or previous published use of the sequence), plus database accession number. Taxon Voucher/Previous publication Database accession number ASTEI.L~~AE Milligania stylosa Chase et al. 1995 BLANDFORDIACEAE Blandfordia punicea Chase et al. 1995 BoRY.+c:E.@ Alania endlieha' Chase et al. 1993 Borya septentrionalis Rudall et al. 1997 CA~V~PY~YE~LXCE~ZE Campynema linearis Chase et al. 1995 DORYANTHACEAE Doryanthes excelsa Chase et al. 1995 H~~OXIDACEE Hypoxis l@tocarpa Chase et al. 1993 Rhodohypoxis millioides Chase et al. 1995 IRIDACE~ Chase et al. 1993 Anomatheca laxa Chase et al. 1995 Aristea glauca Rudall et al. 1997 Eleuthm'ne latifolia Rudall et al. 1997 Freesia alba Rudall et al. 1997 Gladiolus guienzii Duvall et al. 1993 Iris Xgermanica Chase et al. 1995 Zsophysis tasmanica Rudall et al. 1997 Zxia latqolia Chase et al. 1995 'Viuenia cory mbosa Chase et al. 1995 Orthrosanthus polystachyus Duvall et al. 1993 Sisyrinchium micranthum Rudall et al. 1997 1x101IRIACEAE Ixiolirion tartaricum Chase rt al. 1995 ~NARL~cEAE Lanaria lanata Chase et al. 1995 ORCHIDACWE Chase et al. 1995 Apostasia stylidioides Chase et al. 1993 &pripedium irapeanum Chase et al. 1995 Epipactis helleborine Chase et al. 1995 Spiranthes cmnua Cameron et al. in press TECOPHILAE~CEAE campanulata Chase et al. 1995 Conanthera trimaculata IMW Chase 1788,K; this paper Cjanastrum cordiJolium Chase et al. 1993 Rudall et al. 1997 Kabuyea hostifolia MW Chase 1378, K; this paper hartwegii Chase et al. 1995 Tecophilaea cyanocrocus Chase et al. 1995 Tecophilaea violijlora MW Chase 1498, K; this paper Walleria mackenzii unvouchered*; this paper elegans MW Chase 1575, K; this paper

* DNA extracted from wild-collected corm originating in Malawi; all plants in this collection died before flowering. Results and Discussion The 1000 rounds of random taxon-entries found 24 shortest trees of 812 steps, consistency index (CI) = 0.58, retention index (RI) = 0.63. SW found a single shortest tree, 332,94219 SW steps, CI = 0.87 and RI = 0.87 (equally weighted length 812 steps; the single SW tree is one of the 24 shortest trees found with equal weights). Most changes were at third positions (523 steps, 65%), least at second positions (96 steps, 12%),but the CI and RI were not proportionally different for each codon position(C1 = 0.55, 0.67, 0.58 and RI = 0.63, 0.59, 0.65, respectively).

Anomatheca lxia Freesia Gladiolus Nivenia Aristea lridaceae Otthrosanthus Sisyrinchium Ueutherine Iris lsophysis Walleria mackenzii Cyanella hyacinthoides Odontostomum harhvegii Tecophilaea violiflora Conanthera trimaculata Tecophilaeaceae Conanthera campanulata Zephyra elegans Ka buyea cordifolia Cyanastrum hostifolium

lxiolirion bdoliriaceae Lanaria Lanariaceae HYPOidaceae Boryaceae

Asteliaceae Blandfordia Blandfordiaceae Campynema Campynemaceae

FIG. 11. The single most parsimonious tree found from parsimony analysis of rbcL sequences. The numbers above the branches are optimised substitutions (ACCTRAN optimisation); numbers below the branches are the bootstrap percentages (equal weights/SU7). This tree is one of the 24 trees found with equal weights; its length is 812 steps, CI = 0.58, RI = 63 (SW 332,94219 steps, CI = 0.87, RI = 0.87). Branches not present in all 24 equally weighted trees are marked with an arrow. 798 KEW BULLETIN VOL. 53(4) There appears to be little doubt that Cyanastrum and Kabuyea are members of Tecophilaeaceaebased on analysis of rbcL sequences. Within the context of the lower asparagoids, Tecophilaeaceae appear to be mostly closely related to Dorjanthaceae, Iridaceae and Ixioliriaceae (Chase et al. 1995, Rudall et al. 1997, 1998). In the analysis here, Tecophilaeaceaeare strongly supported (97/99) as monophyletic, with the pair, Cyanastrum and Kabuyea, supported (82/87) as sister taxa and embedded within the family, perhaps as sister to all other species except Tecophilaea qanocrocus. This relationship is present in all equally weighted trees but not supported by the bootstrap. Tecophilaea has moderate to strong support as polyphyletic because T. violifZora clusters with other South America taxa, Conanthera and Zephyra, rather than with 7: qanocrocus. The African members of the family, Wallem'a and Cyanella, form a supported (69/97) pair that falls sister to the Californian endemic, Odontostomum, in all trees (but with only moderate support from SW, 73). Lanaria, sometimes included in Haemodoraceae or Tecophilaeaceae (Dahlgren et al. 1985) is supported (67/89) as a member of a clade composed of Asteliaceae, Blandfordiaceae, Boryaceae, Hypoxidaceae and Orchidaceae (see Rudall et al. 1998, for an analysis of the relationships of these families); it should be considered the sole genus of Lanariaceae.

Segregation of Kaabuyea from Cyanastrum As demonstrated here, there is much evidence to support separate generic status for Kabuyea, although it is clear that Cyanastrum and Kabuyea are closely related. The two genera differ in several respects. Cyanastrum has distinctly petiolate leaves which are usually solitary on each corm and arising separately from the scape (Fig. 2), whereas Kabuyea has indistinctly petiolate leaves, about four per corm, surrounding the scape (Fig. 1). Cyanastrum has a blue perianth and apically dehiscent anthers, whereas Kabuyea has a white perianth and anthers dehiscing by a short clavate introrse slit. The pollen surface sculpturing of the two genera differs (Fig. 4), but they both lack an operculum in the sulcus (unusually for Tecophilaeaceae: Simpson, 1985). They have different base chromosome numbers (Table l), with larger chromosomes in Kabuyea. Seed structure (Figs. 8 - 10) also differs in that Kabuyea lacks the 'chalazosperm' structure of Cyanastrum and has microprickles (Fig. 9). Analysis of molecular sequence data from rbcL (Fig. 11) indicates that the two genera are a sister pair embedded within Tecophilaeaceae. Several shared characters support a close relationship between them, since within Tecophilaeaceae and other Asparagales, secretory canals (Fig. 3) are otherwise uncommon, absence of raphide crystals is relatively unusual and deeply pitted seed coats apparently unique. Tillich (1995) noted an unusual seedling structure in C. cordifolium, but Kabuyea is unknown in this respect.

Relationships of Cyanastrum and Kabuyea Oliver (1891) originally referred Cyanastrum to Haemodoraceae and was followed by Baker (1898) and Brown (1901). Cornu (1896) placed it (as Schoenlandia) in Pontederiaceae, and Engler (1897) proposed a new subfamily, Pontederiaceae subfam. Cyanastroideae, and was followed by Dalla Torre & Harms (1900). Later, Engler (1900) raised the subfamily to family status as Cyanastraceae. De Wildeman (1902) initially followed this but later (1913) reverted to placing Cyanastrum in Pontederiaceae. The family Cyanastraceaewas accepted again by Lotsy (1911) and Fries (1919). Hutchinson (1934) noted a close affinity of Cyanastrum with Cyanella L. from southern Africa and placed it in Tecophilaeaceae. Carter (1962, 1966), Simpson (1985) and Arroyo (1986) favoured its inclusion in Tecophilaeaceae, but Clausen (1940), Nietsch (1941), Melchior (1964), Cronquist (1981), Schulze (1983), Dahlgren et al. (1985), Takhtajan (1987), Dahlgren & van Wyk (1988)and Tillich (1995) all favoured a separate family. Takhtajan (1997) considered Cyanastraceae an isolated family perhaps deserving recognition as a separate order. On the other hand, Thorne (1983) referred it to a broad concept of Liliaceae in his subfam. Tecophilaeoideae, but later (1992) raised this to family level again, but taking Cyanastrum out as a separate family Cyanastraceae well removed from Tecophilaeaceae and in a different suborder. Nearly all observations have been based on one or two species, particularly the well known C. cord~oliumfrom west tropical Africa which is also widely cultivated, and only Clausen (1940) and Carter (1962) seem to have looked closely at all four species together. Separate family status was initially accorded to Cjanastrum by Engler (1900) largely on the basis of a perisperm. However, Fries (1919) correctly pointed out that the swollen chalazal region of starch-rich tissue is not a perisperm, as it is not derived from the nucellus, which rapidly degenerates, but from the chalazal region outside the raphal bundle. He coined the new term 'chalazosperm', which has been frequently cited, because he considered this starch-rich, loosely packed tissue to have a nutritive role for the developing embryo, mainly due the lack of endosperm in the mature seed (although it may be an elaiosome: see above). Nietsch (1941) came to similar conclusions. A perisperm, although unusual, is present in some other Asparagales, but a chalazosperm is apparently a unique structure for Cyanastrum, and therefore an autapomorphy, although a similar, possibly homologous structure occurs in Odontostomum, a Californian genus of Tecophilaeaceae (Cave 1952). Whether or not Cyanastroideae should be maintained is outside the scope of this paper, but there is little evidence that this is appropriate from the DNA sequence analysis presented here (Fig. ll),which would require establishment of another two or three subfamilies. This would seem to be an excessive amount of taxonomic hierarchy for such a small group of genera. The differences in morphology, anatomy and cytology of Cyanastrum and Kabuyea in addition to their level of DNA sequence divergence make it appropriate to separate Kabuyea from Cyanastrum, but whether these two should be recognised as a distinct subfamily requires further research.

R. K. B. is indebted to Paul Bamps for kindly listing all material held in the Brussels Herbarium (BR), Geoffrey Mungai for doing the same for the East African Herbarium, Nairobi (EA), and Maria Adelia Diniz for doing the same for the Lisbon Herbarium (LISC), and to the Director of the Wageningen Herbarium (WAG) for sending their holdings on loan. H. B. gratefully acknowledges the assistance and 800 KEM' BULI.ETIX VOL. 53(4) advice of Madeline Harley and Marie Kurmann, especially in the working out of a successful pollen preparation technique. In the cytological study, we would like to thank Jim B. Smith and Peter Brandham. Thanks are also due to Susan Holmes (nee Carter) for helpful comments based on her work of 30 years earlier.

Arroyo, S. (1986). Leaf anatomy in the Tecophilaeaceae. Bot. J. Linn. Soc. 93: 323 - 328. Baker, J. G. (1898). Haemodoracea~.In: W. T. T. Dyer (ed.), Flora of Tropical Africa 7: 331 - 336. Reeve & Co., London. Brown, N. E. (1901). Pontedem'aceae. In: W. T. T. Dyer (ed.), Flora of Tropical Africa 8: 1- 6. Reeve & Co., London. Buchheim, G. (1961). Proposal for the conservation of 926 Cyanastrum Oliver. Taxon 10: 243 - 244. Burkill, I. H. (1897). List of the known plants occurring in British Central Africa, Nyasaland and the British territory north of the Zambezi. In: H. H. Johnston, British Central Africa: 233 - 284. Methuen, London. Cameron, K. M., Chase, M. W., Whitten, W. M., Kores, P. J., Jarrell, D. C., Albert, V. A., Yukawa, T., Hills, H. G. & Goldman, D. H. (1999). A phylogenetic analysis of the Orchidaceae: evidence from rbcL nucleotide sequences. Amer. J. Bot. 86: in press. Carter, S. (1962). Revision of Wallem'a and Cjanastrum ( Tecophilaeaceae). Kew Bull. 16: 185- 195. -(1966). Flora of Tropical East Africa: Tecophilaeaceae. 7 pp. Crown Agents, London. -(1967). Cyanastrum johnstonii Baker. Hooker's Icon. Pl., V, 7: t. 3641, pp. 1 - 2. Cave, M. S. (1952). Sporogenesis and gametogenesis in Odontostomum hartwegzi Torr. Phytomorphology 2: 210 - 214. Chase, M. W. and 42 others (1993). Phylogenetics of seed plants: an analysis of nucleotide sequences from the plastid gene rbcL. Ann. Missouri Bot. Gard. 80: 528 - 580. -, Duvall, M. R., Hills, H. G., Conran, J. G., Cox A. V., Eguiarte, L. E., Hartwell, J., Fay, M. F., Caddick, L. R., Cameron, K. M., & Hoot, S. (1995). Molecular systematics of Lilianae. In: P. J. Rudall, P. J. Cribb, D. F. Cutler & C.J. Humphries (eds.), : Systematics and Evolution, 109 - 137. Royal Botanic Gardens, Kew. -, Rudall, P. J. & Conran,J. G. (1996). New circumscriptions and a new family of asparagoid lilies: genera formerly included in Anthericaceae. Kew Bull. 51: 667 - 680. Cheek, M. (1996). C~lanastraceae.The World of Plants 113: 150- 151. Asahi Shimbun, Japan. Clausen, R. T. (1940). A review of the Cyanastraceae. Gentes Herb. 4: 293 - 304. Cornu, M. (1896). Note sur un genre nouveau de Pontederiacees d'Afrique: Schoenlandia (Sch. gabonensis Cornu). Bull. Soc. Bot. France 43: 21 - 24. Cronquist A. (1981). Cjanastraceae. An Integrated System of Classification of Flowering Plants: 1206 - 1208. Dahlgren, R. M. T., Clifford, H. T. & Yeo, P. F. (1985). Cyanastraceae. The Families of the Monocotyledons, Structure, Evolution and Taxonomy: 167 - 168. Springer Verlag, Heidelberg etc. -& van Wyk, A. E. (1988). Structures and relationships of families endemic to or centred in southern Africa. In: P. Goldblatt & P. P. Lowry, Modern Systematic Studies in African Botany [Proceedings of the Eleventh Plenary Meeting of AETFAT]. Monogr. Syst. Bot. Missouri Bot. Gard. 25: 1- 94. Dalla Torre, C. G. & Harms, H. (1900). Genera Siphonogamarum 1: 59. Engelmann, Leipzig. De Wildeman, E. (1902). Cyanastraceae. In: ~tudessur la Flore du Katanga. Ann. Mus. Congo, Ser. 4, Bot. 1: 5 - 6. -(1913). Decades novarum specierum florae katangensis, 1 - 7. Repert. Spec. Nov. Regni Veg. 11: 501 - 524. Duvall, M. R., Clegg, M. T., Chase, M. W., Clark, W. D., Kress, W. J., Hills, H. G., Eguiarte, L. E., Smith, J. F., Gaut, B. S., Zimmer, E. A. & Learn, G. H. (1993). Phylogenetic hypotheses for the monocotyledons constructed from rbcL sequence data. Ann. Missouri Bot. Gard. 80: 607 - 619. Engler, A. (1897). Pontederiaceae. In: A. Engler & K. Prantl, Die natiirlichen Pflanzenfamilien, Nachtr. 1 zu 2 - 4: 70. W. Engelmann, Leipzig. -(1900). Cyanastraceae. In: A. Engler (ed.), Die von W. Goetze und Dr Stuhlmann im Uluguru-gebirge, sowie die von W. Goetze in der Kisaki- und Khutu-Steppe und in Uhehe gesammelten Pflanzen. Bot. Jahrb. Syst. 28: 357 - 359, Taf. IX - X. Farris, J. S. (1969). A successive approximations approach to character weighting. Syst. Zool. 18: 374 - 385. Felsenstein, J. (1985). Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39: 783 - 791. Fries, T. C. E. (1919). Der Samenbau bei Cyanastrum Oliv. Svensk Bot. Tidskr. 13: 295 - 304. Goldblatt, P., Henrich, J. E. & Rudall, P. (1984). Occurrence of crystals in Iridaceae and allied families and their phylogenetic significance. Ann. Missouri Bot. Gard. 71: 1013- 1020. -& Manning, J. (1989) Chromosome number in Walleria (Tecophilaeaceae).Ann. Missouri Bot. Gard. 76: 925 - 926. Huber, H. (1969). Die Samenmerkmale und Verwandtschaftsverhaltnisse der Liliifloren. Mitt. Bot. Staatssamml. Miinchen 8: 219 - 538. Hutchinson, J. (1934). Tecophilaeaceae. The Families of Flowering Plants, 2, Monocotyledons: 102 - 104. MacMillan, London. Johnson, M. A. T. (1982). Karyotypes of some Greek species of Allium. Ann. Mus. Goulandris 5: 107 - 119. -, ozhatay, N. & Garbari, F. (1996). The genus Muscari in Turkey: taxonomy, distribution and chromosome analysis. In: M. oztiirk, 0. Se~men,& G. Gijrk (eds.). Plant Life in Southwest and Central Asia, 34 - 53. Ege University Press, Izmir, Turkey. Johnston, H. H. (1897). British Central Africa. Methuen, London. Karlkn, T. (1984). Karyotypes and chromosome numbers of five species of Muscari (Liliaceae).Willdenowia 14: 313 - 320. Kurita, S. (1986). Variation and evolution in the karyotype of Lycoris, , 1. General karyological characteristics of the genus. Cytologia 51: 803 - 815. 802 KEM7 BULLETIN VOL. 53(4) -(1987). Variation and evolution in the karyotype of Lycovis, Amaryllidaceae, 2. Karyotype analysis of ten taxa among which seven are native in China. Cytologia 52: 19- 40. La Cour, L. F. (1955). In: C. D. Darlington & A. P. Wylie, Chromosome Atlas of Flowering Plants. Macmillan, New York. Lotsy, J. P. (1911). Cyanastraceae. Vortrage iiber Botanische Stammesgeschichte, 3: 793. Fischer, Jena. Marchant, C. J. & Brighton, C. A. (1974). Cytological diversity and triploid frequency in a complex population of Ranunculus Jicaria L. Ann. Bot. (London) 38: 7 - 15. Melchior, H. (1964). A. Engler's Syllabus der Pflanzenfamilien, 2, Angiospermen. Borntraeger, Berlin. Nietsch, H. (1941). Zur systematischen Stellung von Cyanastrum. Oesterr. Bot. Z. 90: 31 - 52. Oliver, D. (1891). Cyanastrum cordifolium Oliv. Hooker's Icon. P1. 20: t. 1965. Ornduff, R. (1979). Chromosome numbers in Cyanella (Tecophilaeaceae ). Ann. Missouri Bot. Gard. 66: 581 - 583. Peckett, R. C. & Small, C. J. (1980). Occurrence, location and development of anthocyanoplasts. Phytochemistry 19: 2571 - 2576. Rickett, H. W. (1963). Report of the Committee for Spermatophyta: conservation of generic names, 5. Taxon 12: 235 - 238. Rudall, P. J. & Cutler, D. F. (1995). Asparagales: a reappraisal. In: P. J. Rudall, P. J. Cribb, D. F. Cutler, & C. J. Humphries (eds.), Monocotyledons: Systematics and Evolution, 1: 157 - 168. Royal Botanic Gardens, Kew -& Chase, M. W. (1996). Systematics of Xanthorrhoeaceae sensu lato: evidence for polyphyly. Telopea 6: 629 - 647. -, Furness, C. A,, Chase, M. W. & Fay, M. F. (1997). Microsporogenesis and pollen sulcus type in Asparagales (Lilianae). Canad. J. Bot. 75: 408 - 430. -, Chase, M. W., Cutler, D. F., Rusby, J. & de Bruijn, A. (1998). Anatomical and molecular systematics of Asteliaceae and Hypoxidaceae. Bot. J. Linn. Soc. 127: 1 - 42. Russell, W. (1938). Recherches sur la structure de Cyanastrum cordifolium. Bull. Mus. Natl. Hist. Nat., skr. 2, 10: 438 - 439. Sat6, D. (1942). Karyotype alteration and phylogeny in Liliaceae and allied families. Jap. J. Bot. 12: 57 - 161. Schulze, W. (1983). Beitrage zur Taxonomie der Liliifloren. XI. Tecophilaeaceae und Cyanastraceae. Wiss. Z. Friedrich-Schiller-Univ., Jena, Math.-Naturwiss. Reihe 32: 957 - 964. Simpson, M. G. (1985). Pollen ultrastructure of the Tecophilaeaceae. Grana 24: 77 - 92. -& Rudall, P. J. (1998). Tecophilaeaceae. In: K. Kubitzki. The families and genera of vascular plants. 111. Flowering plants. Monocotyledons. Springer-Verlag. Solereder, H. (1917). ~berdie Cyanocysten von Cyanastrum cordifolium Oliv. Beih. Bot. Centralbl. 33: 289 - 302. Sterling, C. (1974). Comparative morphology of the carpel in the Liliaceae: Baeometra, Burchardia and Walleria. Bot. J. Linn. Soc. 68: 115 - 125. Swofford, D. L. (1993). PAUP: phylogenetic analysis using parsimony, version 3.1.1. Smithsonian Institution. Takenaka, Y. & Nagamatsu, T. (1930). On the chromosomes of Lilium tip'num Ker- Gawl. Rot. Mag. (Tokyo) 44 : 386 - 391. Takhtajan, A. (1987). Cyanastraceae. Systema Magnoliophytorum: 291. Nauka, Leningrad. -(1997). Cyanastraceae. Diversity and Classification of Flowering Plants : 494. Columbia University Press, New York. Thorne, R. F. (1983). Proposed new realignments in the angiosperms. Nordic J. Bot. 3: 85 - 117. -(1992). Classification and geography of the flowering plants. Bot. Rev. (Lancaster) 58: 225 - 348. Tillich, H.-J. (1995). Friichte, Samen und Keimpflanzen bei den Cyanastraceae Engler 1900 und einigen vermuten Verwandten. Feddes Repert. 106: 483 - 493. http://www.jstor.org

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(81) Proposal for the Conservation of 926. Cyanastrum Oliver in Hook. Icon. Pl. 20: sub t. 1965. 1891 (nom. cons. prop.), T.: C. cordifolium Oliver (Cyanastraceae vel Tecophilaeaceae) versus Cyanastrum Cassini, Dict. Sci. Nat. 58: 458. 1829 (nom. rej. prop.) (Compositae) G. Buchheim Taxon, Vol. 10, No. 8. (Oct. - Nov., 1961), pp. 243-244. Stable URL: http://links.jstor.org/sici?sici=0040-0262%28196110%2F11%2910%3A8%3C243%3A%28PFTCO%3E2.0.CO%3B2-9

A Phylogenetic Analysis of the Orchidaceae: Evidence from rbcL Nucleotide Sequences Kenneth M. Cameron; Mark W. Chase; W. Mark Whitten; Paul J. Kores; David C. Jarrell; Victor A. Albert; Tomohisa Yukawa; Harold G. Hills; Douglas H. Goldman American Journal of Botany, Vol. 86, No. 2. (Feb., 1999), pp. 208-224. Stable URL: http://links.jstor.org/sici?sici=0002-9122%28199902%2986%3A2%3C208%3AAPAOTO%3E2.0.CO%3B2-E http://www.jstor.org

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Phylogenetics of Seed Plants: An Analysis of Nucleotide Sequences from the Plastid Gene rbcL Mark W. Chase; Douglas E. Soltis; Richard G. Olmstead; David Morgan; Donald H. Les; Brent D. Mishler; Melvin R. Duvall; Robert A. Price; Harold G. Hills; Yin-Long Qiu; Kathleen A. Kron; Jeffrey H. Rettig; Elena Conti; Jeffrey D. Palmer; James R. Manhart; Kenneth J. Sytsma; Helen J. Michaels; W. John Kress; Kenneth G. Karol; W. Dennis Clark; Mikael Hedren; Brandon S. Gaut; Robert K. Jansen; Ki-Joong Kim; Charles F. Wimpee; James F. Smith; Glenn R. Furnier; Steven H. Strauss; Qui-Yun Xiang; Gregory M. Plunkett; Pamela S. Soltis; Susan M. Swensen; Stephen E. Williams; Paul A. Gadek; Christopher J. Quinn; Luis E. Eguiarte; Edward Golenberg; Gerald H. Learn, Jr.; Sean W. Graham; Spencer C. H. Barrett; Selvadurai Dayanandan; Victor A. Albert Annals of the Missouri Botanical Garden, Vol. 80, No. 3. (1993), pp. 528-548+550-580. Stable URL: http://links.jstor.org/sici?sici=0026-6493%281993%2980%3A3%3C528%3APOSPAA%3E2.0.CO%3B2-W

Phylogenetic Hypotheses for the Monocotyledons Constructed from rbcL Sequence Data Melvin R. Duvall; Michael T. Clegg; Mark W. Chase; W. Dennis Clark; W. John Kress; Harold G. Hills; Luis E. Eguiarte; James F. Smith; Brandon S. Gaut; Elizabeth A. Zimmer; Gerald H. Learn, Jr. Annals of the Missouri Botanical Garden, Vol. 80, No. 3. (1993), pp. 607-619. Stable URL: http://links.jstor.org/sici?sici=0026-6493%281993%2980%3A3%3C607%3APHFTMC%3E2.0.CO%3B2-5

A Successive Approximations Approach to Character Weighting James S. Farris Systematic Zoology, Vol. 18, No. 4. (Dec., 1969), pp. 374-385. Stable URL: http://links.jstor.org/sici?sici=0039-7989%28196912%2918%3A4%3C374%3AASAATC%3E2.0.CO%3B2-X

Confidence Limits on Phylogenies: An Approach Using the Bootstrap Joseph Felsenstein Evolution, Vol. 39, No. 4. (Jul., 1985), pp. 783-791. Stable URL: http://links.jstor.org/sici?sici=0014-3820%28198507%2939%3A4%3C783%3ACLOPAA%3E2.0.CO%3B2-L

Occurrence of Crystals in Iridaceae and Allied Families and their Phylogenetic Significance Peter Goldblatt; James E. Henrich; Paula Rudall Annals of the Missouri Botanical Garden, Vol. 71, No. 4. (1984), pp. 1013-1020. Stable URL: http://links.jstor.org/sici?sici=0026-6493%281984%2971%3A4%3C1013%3AOOCIIA%3E2.0.CO%3B2-V http://www.jstor.org

LINKED CITATIONS - Page 3 of 3 -

Chromosome Number in Walleria (Tecophilaeaceae) Peter Goldblatt; John C. Manning Annals of the Missouri Botanical Garden, Vol. 76, No. 3. (1989), pp. 925-926. Stable URL: http://links.jstor.org/sici?sici=0026-6493%281989%2976%3A3%3C925%3ACNIW%28%3E2.0.CO%3B2-8

Chromosome Numbers in Cyanella (Tecophilaeaceae) Robert Ornduff Annals of the Missouri Botanical Garden, Vol. 66, No. 3. (1979), pp. 581-583. Stable URL: http://links.jstor.org/sici?sici=0026-6493%281979%2966%3A3%3C581%3ACNIC%28%3E2.0.CO%3B2-D

Report of the Committee for Spermatophyta: Conservation of Generic Names V H. W. Rickett Taxon, Vol. 12, No. 6. (Jul., 1963), pp. 235-238. Stable URL: http://links.jstor.org/sici?sici=0040-0262%28196307%2912%3A6%3C235%3AROTCFS%3E2.0.CO%3B2-J

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