植物研究雑誌 J. J. Jpn. Bo t. 79:241-246(2004) 79:241-246(2004)

Chromosome Numbers for Eight in Five Five Genera of

Heng ・Chang W ANG ， Ai-Ping MENG ，Ji an-Qiang LI* and Zi-Can HE

Wuhan Wuhan Botanical Garden ， Wuhan Institute of Botany ，the Chinese Academy of Sciences ，Wuhan ， 430074 CHINA *E-mail *E-mail totheauthorforcorrespondence:jianqiangli@hotmai l. com (Received (Received on September 13 ，2003)

Chromosome numbers of 8 species in 5 genera of Menispermaceae from China 紅 e reported: reported: Tinospora sagittata (Oliv.) Gagnep. 2n = 4x = 52 ， and T. hain αnensis Luo & Z. X. X. Li 2n=26; D 伊loclisia glaucescens (B 1.) Diels 2n = 26; Stephania yunnanensis Luo 2n 2n = 26 ，組 d S. dielsiana Wu 2n = 26; orbiculatus (L.) DC. 2n = 4x = 52 ，組d C. C. laurifolius DC. 2n = 26; dahuricum DC. 2n = 4x = 52. The chromo- some numbers of the first five species 紅 e here reported for the first time. All eight spe-

cies cies studied have a basic chromosome number x= 13 ， and chromosome numbers 紅 e generally generally consistent with previous reports the for same species and genera.

Key words: Chromosome number ，Cocculus ，Diploclisia ，Menispermaceae ，Tinospora.

Introduction Introduction Although cytological studies in Menis- Menispermaceae ，consisting of five tribes ， permaceae have been published by various 71 71 genera and about 450 species ， is a authors (Darlington and Wylie 1955 ， pantropical pantropical family usually confined to the Fedorov 1974 ， Goldblatt 1984 ，1985 ，Hong lowlands. lowlands. All Menispermaceae are typically 1990 ，etc.) ，chromosome counts have been unisexual unisexual and dioecious. Most 紅 e lianas and made in only a few species in each of seven many have pharmaceutical properties and are of the 25 Malesian genera ， viz. Tiliacora ， poisonous poisonous (Kessler 1993). Tinospora ， ，Cocculus ，Cissampelos ， Nineteen Nineteen genera and 78 species of Cyclea and Stephani α(Forman 1986). Menispermaceae 訂 e recognized in China. The Chinese species of Menispermaceae ， They 訂 'e mainly distributed in the southem however ，訂e poorly treated in most standard regions regions of the Yangtze River ，and especially references. Chromosome counts for only two in in South China and Southwest China (Luo species from China ，Cocculus trilobus (= C. 1996). 1996). orbiculatus) reported by N ak 吋ima (1937) as Chromosome numbers can sometimes pro- 2n = 50 and Menispermum dauricum from vide vide information on the relationship of gen- North China observed by Langlet (1928) as era era or even of families (Forman 1986). 2n = 52-54 ，have been reported. In this Raven (1975) used chromosome information paper ， eight species in five genera of

to to postulate a relationship between Menis 四 Menispermaceae from China were studied to permaceae and Lardizabalaceae. Forman determine chromosome number and other (1 986) proposed Sabia Colebr. (Sabiaceae) cytological characteristics. to to be a near alI y of Menispermaceae ， based on chromosome data.

-241- 242 植物研究雑誌第79 巻第4号 平成16 年8月

Material Material and Methods Results and discussion The investigated were collected Chromosome numbers of the species stud- from from the original locality ， but now they have ied are given in Table 1. been been cultivated in South China Botanical 1. Tinospora Miers - According to Garden ， the Chinese Academy of Sciences Kessler (1 993) ，there 紅 'e about 32 species in (CAS) and Wuhan Botanical Garden ，CAS Tinospora. Seven are in 佐opical ，two (Table (Table 1). Voucher specimens were depos- in Madagascar ， and twenty-three 紅 'e from ited ited in the herbarium of the Wuhan Institute Asia to Australia and the Pacific. Six species of of Botany ， Chinese Academy of Sciences and two varieties have been recorded from (HIB). (HIB). China (Luo 1996). For For observations of somatic chromo- Two species had been examined previ- somes ， growing shoot apices were pre 佐eated ously. Tinospora tomentosa Miers was re- in in a saturated solution of p- Dichlorobenzene ported as 2n = 26 (Chattopadhyay and at at about 18-20 oC for 3.5 -4 hours ， fixed in Sharma 1989). Joshi (1 934) and Nanda Carnoy Carnoy I (glacial acetic acid: absolute etha- (1962) examined T. cordifolia Miers. from nol nol = 1: 3) at the same temperature for 24 and reported it to be 2n = 24 ， while hours ， and then transferred to 70 % alcoho l. Abraham (1 942) ， Bir et al. (1 987) ，Mathew The shoot apices were hydrolyzed in a1 :1 (1 958) ，Sharma and Bhattacharyya (1 955) ， mixture mixture of 1N hydrochloric acid (HCL) and Sharma and Sharma (1957) ， and Sarkar et al. 45% acetic acid at 60 oC for 6- 8 minutes ， (1 980) reported it to be 2n = 26. and and then stained in carbol fuchsin for 30 Two species ， T. sagittata (Oliv.) Gagnep. minutes minutes and squashed in 45 % acetic acid. and T. hainanensis Lo & Z. X. Li are re- More than 10 metaphase cells were exam- ported here for the first time. All five popu- ined ined to determine the chromosome number lations of T. sagittata were found to be of of each species. tetraploids with 2n = 4x = 52 (Fig. 1，1) ，

Table 1. Chromosome counts and source of materials studied in the family Menispermaceae

Chromosome Species Species Locality Sex Voucher* Previous (2n) reports number (2n)

Tinospora Tinospora sagittata Guangdong and Hainan (SCBG) “ male 03-008 52 none T. T. hainanensis Li ngao ，Wenchang County ， Hainan male 03-009 26 none (SCBG) Cocculus M t. Dinghu ，Guangdong (SCBG) male 03 ・004 26 26 (Bowden 1945 ， Gill et et a1. 1981 ， Sandhu and Mann 1988) C. C. orbiculatus Li ngao ， Wenchang and Wanlin male 03 ・001 52 52 (Bowden 1945) (= (= C. tribolus) County ， Hainan (SCBG) 50 (Nakajima 1937 ， from from Japan and China)

Diploclisia Diploclisia glauces- Hainan (SCBG) female 03 ・006 26 none cens cens

Menispermum dahu- Shennongjia ， Hubei (WHBG) 村* male 03 ・010 52 52-54 (Langlet 1928 ， ncum from N. China) &句 phania dielsiana Hebao Island ，Guangdong (SCBG) female 03-002 26 none S. S. yunnanensis Yunnan and Guangxi (SCBG) male H. C. Wang 26 none 03-007 03-007

*Collection *Collection number of H. C. Wang. * * SCBG = South China Botanical Garden ， the Chinese Academy of Sciences (CAS).

キ**WHBG = Wuhan Botanical Garden ，CAS. August August 2004 Journal of Japanese Botany Vo l. 79 No. 4 243

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Fig. Fig. 1. Chromosomes of eight species of Menispermaceae. 1. Tinospora sagittata (Oliv.) Gagnep. 2. T. hainanensis hainanensis Lo & Z. X. L i. 3. (L.) D C. 4. C. laurifolius DC. 5. Menispermum dα huricum D C. 6. Diploclisia glaucescens (Blume) Diels. 7. Stephania yunnanensis Luo (Arr ow indi- cates cates B-chromosome). 8. S. dielsiana Wu (Arrow indicates B-chromosome). 244 植物研究雑誌第79 巻第4号 平成16 年8月 while while six populations of T. hainanensis were with 2n = 4x = 52 (Fig. 1，5) ， which is con- 2n = 26 (Fig. 1 ，2). sistent with Kessler's report (1 993).

2. 2. Cocculus DC. - Eight species of 4. Diploclisia Miers - Two species D. Cocculus Cocculus are found in N orth and Central affinis (Oliv.) Diels and D. glaucescens America ，Africa ，Madagascar ， and eastward (Blume) Diels occur in tropical Asia (Kessler from from the Indian subcontinent to the 1993) ，and both are in China (Luo 1996). We Philippines Philippines (Kessler 1993). Two species and found D. glaucescens to be 2n = 26 (first re- one one variety have been recorded from China port) (Fig. 1 ，6). (Luo (Luo 1996). Chromosomes of C. pendulus ， C. villosus ， 5. Stephania Lour. - Stephania is some- C. C. carolinus and C. hirsutus were examined what confused taxonomically and the num- by Baquar et al. (1 965) ， Bir and Thakur ber of species is uncertain. According to (1984) ，Bowden (1 945) ， Gill et al. (1 981) ， Kessler (1 993) ，there 紅 e 30 species ，includ- Hagerup Hagerup (1932) ， Joshi (1934) ， Lewis et al. ing five in tropical Africa and 25 from south- (1 962) ， Sarkar et al. (1 973) ，Sharma and em India to central and southem China and Bhattach 紅 yya (1 955) ，Sharma and Sharma New Guinea. According to Luo (1996) ，there (1 957) and Sidhu et al. (1 983). 訂 e about 60 species with 39 species and 1 Cocculus Cocculus laurifolius was reported by variety in China. Bowden (1945 ，materials from The chromosome numbers of Stephania and and E. Asia) and Sandhu and Mann (1 988) are various ， with reports of 2n = 22 ，24 ， and as as 2n = 26 ， and by Gill et al. (1 981) as n = 26 (D 紅 lington and Wylie 1955 ， Fedorov 13 13 + 0-3B. They are generally consistent 1974 ，Goldblatt 1984 ，1985 ，Hong 1990). with with our report here 2n = 26 (Fig. 1， 4). Some Chinese species including S. elegans While While C. trilobus (Thunb.) DC. (= C. Hoo k. f. & Thomson (2n = 22) ，S. gracilenta orbiculatus orbiculatus (L.) DC.) was reported by Miers. (2n = 26) ， S. hernandifolia (Willd.) Bowden (1945 ， materials from the Walp. (2n = 22) and S. japonic α(Thunb.) Philippines) Philippines) as 2n = 52 ，by Nakajima (1 937 ， Miers. (2n = 22) ， were examined (Bir and materials materials from Japan and China) as 2n = 50 ， Thakur 1981 ， 1984 ， Chattopadhyay and we counted 2n = 4x = 52. We believe the Sharma 1989 ，Guha 1979 ， Sarkar et al. 1980 ， basic basic chromosome number of the tetraploid Sharma and Bhattacharyya 1955 ，Sharma to to be x = 13 (Fig. 1 ，3). and Sharma 1957). We examined two spe- cies cies and found the chromosome numbers of 3. 3. Menispermum L. - Two to four species both S. yunnanensis Luo (Fig. 1， 7) and S. of of Menispermum in Eurasia and North dielsiana Wu (Fig. 1， 8) to be 2n = 26. America America (Kessler 1993) ， and one species M. dahuricum DC. is in China (Luo 1996). General discussion L. was reported The chromosomes of Menispermaceae 紅 e by some authors (Langlet 1928 ， Lindsay very small in size. Diploid numbers 2n = 24 1930 ，Love and Love 1982 ，Tischler 1931) and 26 紅 e the most frequent and 2n = 22 is to to be 2n = 52 ， and M. dauricum from North less common. Polyploidy is relatively rare in China China was reported by Langlet (1 928) to be Menispermaceae (Thanikaimoni 1984). Al- 2n = 52-54 (who also reported M. canadense though our results 紅 e not consistent with to to be 2n = 52-54). previous reports ，we consider x = 13 to be We examined five populations of M. one of the main basic chromosome numbers dahuricum and determined it to be tetraploid for the family. August2004 August2004 Journal of Japanese Botany Vo l. 79 No. 4 245

We thank Professor Li Ze- Xian ， South Joshi A. C. 1934. Chromosome numbers in Menis- China China Institute of Botany ， the Chinese permaceae. Nature(London) 134(3375): 29. Academy of Sciences ， for guidance in the Kessler P. J. A. 1993. Menispermaceae. In: Kubitzki K. K. (ed よ The Families and Genera of Vascular field field and for identification of the species. Plants Plants 2: 402-418. Springer 司 Verlag ，Heidelberg. This This study was financially supported by Langlet O. F. J. 1928. Einige Beobach 加ngen uber die Chinese Chinese Academy of Sciences (Nos. Zytologie der Berberidazeen. Svensk Bo t. Tidskr. KSCX2-SW-104 ，01035108 ，0105110 and 22 (1 -2): 169-184. 05035123). 05035123). Lewis W. H. ，Stripling H. L. and Ross R. G. 1962. Chromosome numbers for some angiosperms of the southern southern United States and Mexico. Rh odora. References References 64(758): 147-16 1. Abraham A. 1942. Chromosome number in Tinospora. Lindsay R. H. 1930. The chromosomes of some Curr. Curr. Sci. 11(7): 282. dioecious Angiosperms. Amer. J. Bo t. 17(2): 152- Baqu 紅 S. R.， Akh t紅 S. and Husain A. 1965. Meiotic 174. chromosome numbers in some vascular plants of Love A. and Love. D. 1982. IOPB chromosome num- Indus Indus Delta. 1. Bo t. Notiser 118(3): 289-298. ber reports LXXIV. Taxon 31: 120-126. Bir Bir S. S. and Thakur. H. 198 1. Chromosome number Luo H. S. 1996. Menispermaceae. Fl ora Reipublicae reports reports LXXII 1. Taxon 30: 842-843. Popularis Sinicae. Tomus 30 (1): 1-8 1. Sciences 一一一 and 一一一 1984. SOCGI chromosome num- Press ，Be 討ing (in Chinese). ber ber reports-I 1. J. Cyto l. Gene t. 19: 11 4- 115. Mathew P. M. 1958. Studies on the Menispermaceae. 一一一，一一一 and Chatha G. S. 1987. Chromosomal stud- Proc. Indian Acad. Sci. ，Sec t. B ，47(5): 27 4- 286. ies ies in certain members of Ranunculaceae and Nakajima G. 1937. Cytological studies on some 恥1ensipermaceae. Proceedings of the Indian dioecious plants. Cytologia ，Fujii Jub. Vo l. 282- Science Science Congress Association 74(3 ，VI): 18 4- 185. 292. Bowden W. M. 1945. A list of chromosome number in Nanda P. C. 1962. Chromosome number of some trees higher higher plants. 11. Menispermaceae to Verbenaceae. and . J. Indian Bo t. Soc. ，41(2): 271-277 Amer. Amer. J. Bo t. 32(4): 191-202. Raven P. H. 1975. The bases of angiosperm Chattopadhyay Chattopadhyay D. and Sharma A. K. 1989. phylogeny: cytology. Ann. Missouri Bo t. Gard. 62: Chromosome studies and unisexuality in Menis- 724 ー764. permaceae. permaceae. Nucleus 32: 173-179. Sandhu P. S. and Mann. S. K. 1988 ，SOCGI plant Darlington Darlington C. D. and Wylie A. P. 1955. Chromosome chromosome number reports-VI 1. J. Cyto l. Gene t. Atlas Atlas of Flowering Plants. Pp. 13-14. G. Allen 23: 219-228. London. London. Sarkar A. K.， Datta N. and Chat 旬rjee U. 1973. Fedorov Fedorov A. 1974. Chromosome Numbers of Fl owering Chromosome survey of cert 泊n angiosperms. 11. Plants. Plants. Pp. 418 -4 19. Otto Koeltz Science Bul l. Bo t. India Surv. 15: 148. Publishers ，Koenigstein. 一一，一一- and 一一一 1980. Chromosome number re- Forman L. L. 1986. 乱1enispermaceae. Fl ora Malesiana ports LXVII. Taxon 29: 36 0- 36 1. 10: 10: 157-253. Sharma A. K. and Bhattacharyya N. K. 1955. Gill Gill B. S， Bir S. S. and Singhal V. K. 198 1. Cytogenetics of some members of Chromosome number reports LXXI. Taxon 30: Menispermaceae. Bul l. Bot. Soc. Benga19(2): 159- 513-514. 513-514. 169. Goldblatt Goldblatt P. 1984. Index to Plant Chromosome 一一一 and Sharma A. 1957. Investigations leading to a Numbers 1979-1981. Missouri Botanical Garden. new theory of differentiation in plant cells. Genetic 一一一 1985. Index to Plant Chromosome Numbers Iberica 9(3): 143-157. 1982-1983. 1982-1983. Missouri Botanical Garden. Sidhu M. ， Bir S. S. and Lata P. 1983. IOPB Guha S. 1979. Cytological studies on Stephania Lou r. Chromosome number reports LXXIX. Taxon 32: Proc. Proc. Indian Sci. Congr. Assoc. (III ， C) 66: 81. 322. Hagerup Hagerup O. 1932. むber Polyploidie in Beziehungzu Thanikaimoni 1984. Menispermacees: palynologie et Klima ， Okologie und Phylogenie (Chromosomen- systematique. Trav. Sec t. Sci. Tech. Ins t. Fr. zahlen zahlen aus Timbuktu). Hereditas 16 (1 -2): 19 -4 0. Pondichery . Hong D.Y. 1990. Plant cytotaxonomy. P. 304. Science Tischler G. 193 1. Pflanzliche Chromosomen-Zahlen. Press ，Beijing (in Chinese). Tabu l. Bio l. Periodicae 7: 109-226. 246 246 植物研究雑誌第79 巻第4号 平成 16 年8月

王恒昌，孟愛平，李建強，何子士山:ツヅ ラフジ科 5 属 8 種の染色体数 中国産ツヅラフジ科 5 属 8 種について染色体数 はいずれも x= 13 の染色体基本数をもっ.既報の を報告した.そのうち ， Tinospora sagitta ω(2n= 3 種ではこれまでの報告に一致するが， Cocculus 52) 52) ， Tinospora hainanensis (2n = 26) ， Diploclisia trilobus では 2n=52 で， Nakajima (1937) が報告し glaucescens (2n glaucescens = 26) ， Stephania yunnanensis (2n = た 2n=50 とは一致しなかった. しかし，この場 26) ，Stephani α dielsiana (2n = 26) の 5 種について 合も染色体基本数は x= 13 であると推定している. は初めての染色体数の報告である.報告した 8 種 (中国科学院武漢植物研究所武漢植物園)