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INTRASPECIFIC POLYPLOIDY IN CONIOGRAMME FRAXINEA (PTERIDACEAE) OF JAVA

Titien Ngatinem Praptosuwiryo Center for Plant Conservation – Bogor Botanical Gardens, Indonesian Institute of Sciences. Jl. Ir. H. Juanda No. 13 Bogor, West Java, Indonesia. E-mail : [email protected]

Titien Ngatinem Praptosuwiryo. 2011. Poliploidi Intraspesifik Coniogramme fraxinea (Pteridaceae) di Jawa. Floribunda 4(3): 70–74. — Penelitian sitologi Coniogramme fraxinea (D. Don) Diels telah dilakukan dengan mengamati 15 nomor koleksi yang berasal dari enam lokasi di Jawa. Pengamatan kromosom somatik memperlihatkan bahwa C. fraxinea mempunyai dua sitotipe berdasarkan jumlah kromosom dasar x = 30, yaitu 2n = 60 (diploid) dan 2n = 120 (tetraploid). Ini merupakan catatan sitologi pertama di Jawa. Dua varian yang dilaporkan di sini dikenali terutama dengan warna daun, yaitu ental yang memiliki lembaran daun bergaris putih (frond with white striped lamina, WSL) dan yang memiliki lembaran daun hijau penuh (frond with full green lamina, FGL). Tipe diploid lebih umum dijumpai, tersebar luas dan terdiri dari dua varian (WSL dan FGL). Tipe tetraploid (tiga individu) merupakan varian WSL. Kata kunci: Coniogramme fraxinea, kromosom, poliploidi intraspesifik, morfologi.

Titien Ngatinem Praptosuwiryo. 2011. Intraspecific Polyploidy in Coniogramme fraxinea (Pteridaceae) of Java. Floribunda 4(3): 70–74 — Cytological study on Coniogramme fraxinea (D. Don) Diels has been car- ried out by observing 15 collection numbers of Javanese specimens collected from 6 localities. Somatic chromosome observation revealed that C. fraxinea of Java comprises of two cytotypes based on the basic chromosome number x = 30, viz.: 2n = 60 (diploid) and 2n = 120 (tetraploid). It is first cytological record of C. fraxinea in Java. Two variants reported here are recognized mainly by the coloration of frond, viz. frond with white striped lamina (WSL) and those with full green lamina (FGL). The diploid type is more common, widely distributed and consists of two variations (WSL and FGL). The tetraploid (three individuals) are WSL. Keywords: Coniogramme fraxinea, chromosome, intraspecific polyploidy, morphology.

Coniogramme Fée (Pteridaceae) consists of nae extending almost from the base of the veins to about 20 species or more, and distributed from within 2–5 mm of their apices. tropical Africa east to the Himalaya, China, Japan, Polyploids, which contain more than two and Japan, and in the Pacific to the Hawaiian Is- sets of chromosomes number per cell, have lands and Samoa (Tryon in Kramer & Green important in plant evolution (Stebbins 1971; Lewis 1990). In Java, Backer & Posthumus (1939) re- 1980). Polyploidization and mode reproduction cognized four species of Coniogramme, namely C. types has played an important role in the geogra- intermedia Hieron., C. macrophylla Hieron., C. phical distribution and evolution of ferns. Up to fraxinea (D. Don) Diels and C. serrulata Fée 95% of pteridophytes may be polyploid (Grant which occurs from 900 m to 3200 m, from West to 1981) and recent polyploids may comprise 45% of East Java. It is terrestrial fern that is common extant homosporous fern (Vida 1976; Haufler & growing on moist soil of shady place in the forest, Soltis 1986). Polyploidy may lead to significant mainly near the streams and often amongst rocks. reproductive isolation (Ramsay & Schemske As described by Holttum (1966), Conio- 1998), and change in gene expression (Adams et gramme fraxinea is recognized by these characters: al. 2003), and ecological interactions (Segraves et fronds simple pinnate with 3–6 pairs of pinnae, the al. 1999). lowest pair of pinnae again pinnate with 1–3 pairs Cytological observations on C. fraxinea of pinnules below a terminal terminal lamina; tex- have been reported by Verma in Mehra (1961) ture thin; surface glabrous; veins slender, distinct from Himalaya, Kato et al. (1992) from Yunnan, on both surface, at an angle of more than 45º to the and Praptosuwiryo (2003) from Lombok Island. midrib, ending well within the margin in conspicu- However, there is no cytological information of ous elongated hydathodes. Sori in fully fertile pin- Coniogramme from Java. Cytological observa-

71 Floribunda 4(3) 2011 tions were carried out to: 1) to recognize the varia- Herbarium = BOHB). Cytological investigations tion of the chromosome number of Coniogramme were carried out by observing the somatic fraxinea of Java by counting its somatic chromo- chromosomes at the root tips following a proce- some; 2) to determine a correlation between va- dure developed by Manton (1950) and modified by riation on chromosome number and morphological Darnaedi (1991). Root tips were pre-treatment in 2 variation within the species. mM 8-hydroxyquinoline solution for 5–20 hours at 3–4º C. They were fixed in 45% acetic acid for 10 MATERIALS AND METHOD minutes and macerated in a mixture of CH3COOH 45% : I N HCl = 1 : 3 at 60º C for 3–4 minutes. Plants were collected from six localities in The fixed roots were stain-ed and squashed in Java (Table 1, Figure 1). Living material were aceto-orcein solution. Chromosomes observation planted in Bogor Botanical Gardens. Voucher was carried out under the microscope by using specimens are housed at the Herbarium of Bogor 1000x magnification. Botanical Gardens (Bogor Hortus Botanicus

Table 1. Frond variation, somatic chromosome numbers, vouchers, and localities of Coniogramme fraxinea from Java

Frond Chromosome Voucher Locality Variation number (2n)/ Specimens Level Ploidy WSL 60/Diploid TNgP 1062 Mt. Gede, Gede Pangrango National Park, West Java. Ca. 1200 m. WSL 60/Diploid TNgP 1132 Mt. Slamet, Central Java. 860 m. WSL 60/Diploid TNgP 1264 Mt. Pangrango, Gede Pangrango National Park, West Java. 1060 m. FGL 60/Diploid TNgP 1145 Track to Cibeureum Water Fall, Gede WSL 60/Diploid TNgP 1144 Pangrango National Park, West Java. Ca. 1400 –1450 m. WSL 60/Diploid TNgP 1570 Mt. Gede, Gede Pangrango National Park, West Java. 1200 m. WSL 120/ TNgP 1686 Mt. Wilis, East Java. 1230 m. Tetraploid WSL 120/ TNgP s.n. Mt. Gede, Gede Pangrango National Park, West Tetraploid Java WSL 60/Diploid TNgP 1730 Mt. Halimun, Halimun-Salak National Park, West Java. Ca. 1000 m. WSL 60/Diploid TNgP 2524 Cisekek forest, Cibeureum, Wangunharja 60/Diploid TNgP 2625 village, Lembang District, Bandung Regency, West Java. Ca. 1300 m FGL 60/Diploid TNgP 2627 WSL 60/Diploid TNgP 3097a Mt. Gede, Gede Pangrango National Park, West TNgP 3097e Java. 1487 m. WSL 120/ TNgP 3124 Cibogo forest, Mt. Salak, Halimun-Salak Tetraploid National Park, West Java . 1325 m.

Note: WSL = white striped lamina; FGL = full green lamina; TNgP = Titien Ngatinem Praptosuwiryo

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1 2 3 4 5 6

diploid tetraploid

Figure 1. Geographic distribution of ploidy level types of Coniogramme fraxinea in Java. 1. Mt. Halimun. 2. Mt. Salak. 3. Mt. Gede Pangrango. 4. Cisekek forest, Bandung. 5. Mt.Slamet. 6. Mt. Wilis.

RESULTS AND DISCUSSION tetraploid, respectively. Coniogramme fraxinea of Java reveals two ploidy level, namely diploid and The chromosome counts obtained are sum- tetraploid (Figure 2 a-c). marized in Table 1. Fifteen collections number of Previous studies and recently data on Coniogramme fraxinea have been examined their geographical distribution of ploidy level types of somatic chromosome number. Twelve individuals C. fraxinea in the world showed that diploid and showed chromosome number 2n = 60 and three tetraploid type are new records for Java. In plants have 2n = 120. Himalaya this species was found to have diploid At present the basic chromosome number of and tetraploid levels (Verma in Mehra 1961). Coniogramme is still in a question. Löve et al Praptosuwiryo (2003) reported the diploid type (1977) presumed that this genus may be have basic from Lombok Island. Kato et al (1992) found the chromosome namber x = 15 or x = 30. However diploid type in Mengla, Yunnan. there are not any record on chromosome number of Javanese C. fraxinea shows two morpho- Coniogramme with 2n = 30. Previous cytological logical variations of lamina recognized mainly by studies on seven species of Coniogramme showed the coloration of lamina, namely: the white stripe the lowest number with 2n = 60, viz. C. affinis lamina (WSL, Figure 3) and the full green lamina Hieron. (Mehra & Verma 1960), C. fraxinea (FGL, Figure 2 d). The WSL variant consists of (Verma in Mehra 1961, Praptosuwiryo 2003), C. two ploidy level, viz. diploid and tetraploid type, gracilis Ogata (Kurita 1972), C. indica (Verma & whereas those FGL has diploid only. However the Goloknath in Love 1967), C. intermedia Hieron. other morphological features distinguishing the var. glabra Ching (Verma & Khullar 1965 ), C. diploid and tetraploid type are still unclear. There- intermedia Hieron. var. villosa Ching (Verma in fore further examination on chromosome number Mehra 1961). Thus, it is presumed that the basic and its mode reproduction type from all the distri- chromosome number for Coniogramme as x = 30. bution range to clarify the correlation between Therefore plants with 2n = 60, 2n = 90 and 2n = variation on chromosome number and morphologi- 120 are determined as diploid, triploid and cal variation within the species is neccessary.

Figure 2. a-c. Somatic chromosome of Coniogramme fraxinea . a. 2n = 60 (TNgP 1132); b. 2n = 60 (TNgP 1062); c. 2n = 120 (TNgPs.n.). Bar = 3 μm for a-c. d. Fronds of C. fraxinea with FGL, diploid type (TNgP 2627). Bar = 60 mm.

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Figure 3. Coniogramme fraxinea with white striped lamina (WSL). A. Tetraploid, 2n = 120 (TNgP 3124). B. Diploid, 2n = 60 (TNgP 3097a). A1. and B1. Lamina. A2 and B2. Part of pinna. Bar = 50 mm for A1 and B1, 10 mm for A2 and B2

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