J. Jpn. Bot. 86: (2011)

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1 April 2011 Journal of Japanese Botany Vol. 86 No J. Jpn. Bot. 86: (2011) Su-Juan Lin a, *, Takeshi Suzuki b, Atsushi Ebihara c, Haisheng Lu d, Jianqiu Liu d, Sugong Wu e and Kunio Iwatsuki b : A New Record of Polystichum otomasui Sa. Kurata (Dryopteridaceae) from Fujian Province, China a Department of Biological Science, Faculty of Life and Environmental Science, Shimane University, Nishikawatsu 1060, Matsue, Shimane, JAPAN b Museum of Nature and Human Activities, Hyogo, 6, Yayoigaoka, Sanda, Hyogo, JAPAN c Department of Botany, National Museum of Nature and Science, 4-1-1, Amakubo, Tsukuba, JAPAN d College of Life Science, Fujian Normal University, Fuzhou, Fujian, P. R. CHINA e Kunming Institute of Botany, Chinese Academy of Sciences, 132, Lanheilu, Kunming, Yunnan, P. R. CHINA *Corresponding author: sjlin@life.shimane-u.ac.jp Summary: Polystichum otomasui Sa. Kurata, which had been regarded as an endemic fern to Japan, was discovered in the Wuyi Mountains, Fujian Province, South East China. Cytological investigations confirmed that the new collections were sexual diploids with a chromosome number of 2n = 82 (x = 41) and there were 64 spores per sporangium. These morphological data (including spore perispores) accorded with the characteristics of the Japanese Polystichum otomasui. The evergreen fern Polystichum otomasui Sa. Kurata (Dryopteridaceae) was known as a Japanese endemic, distributed in an extremely restricted area on humus-rich floor of mountain forest in Kyusyu, Japan (Kurata 1963, Nakaike 1992, Iwatsuki 1992, Mitsushima et al. 2009). In our field researches on The comparative study of evergreen forests of Japan and Southern China during , thirteen plants of Polystichum that resembled the Japanese P. otomasui were collected from three populations in Wuyishan National Nature Reserve, Fujian province. One population with only several plants was found on a light slope by a stream under a secondary Lithocarpus forest of Xianfengling at 1500 m altitude, and another two populations with more than ten individuals each were found in an Old Tree Village (1100 m alt.) and Tongmuguan (1700 m alt.) in the shady and humid floors along small streams, under mixed Phyllostachys heterocycla, Lithocarpus and Tsuga evergreen forests. The field research area was further enlarged to Mt. Emeifeng (1714 m alt., N, E), the southwestern part of the Wuyi Mountains (Fig. 1) in 2008, where a big population (more than a hundred individuals) was found on a path slope under the broadleaf evergreen forest. A comparative study was carried out between the Chinese collections and Japanese Polystichum otomasui, and the specimens collected from the Wuyi Mountains were confirmed to be P. otomasui, which is a new record in China (Fig. 2). Morphology Polystichum otomasui is closely related to P. makinoi (Tagawa) Tagawa. It was described as having shining blackish scales on stipes and rachis; sori medial on pinnules of upper pinnae and marginal on auricles of pinnules of lower pinnae or sometimes only one sorus preferentially on each auricle at basal acroscopic side of pinnules; indusia entire (Kurata 1963, Nakaike 1992, Iwatsuki et al. 1995). The morphological characters of the plants from the Wuyi Mountains accord with above mentioned characters of Japanese P. otomasui (Fig. 2).

2 116 植物研究雑誌第 86 巻第 2 号 2011 年 4 月 Fig. 1. Map of Fujian Province, South East China. Dotted circle. Area of Fujian Wuyi Mountains. Solid square ( ). Collection sites in China. Solid triangle ( ). Collection locality of Kumamoto Pref., Japan. The fundamental similarities between the Chinese specimens and Japenese P. otomasui (from Kumamoto Pref., ) were also confirmed by SEM observations of the spores (Fig. 3). The spores of both samples are monolete, about 29 µm 36 µm, ellipsoidal, and echinate fenestrate and more or less cristae in perispore, although there are some more or less discernable difference in the surface appearance which could be explained as intraspecific variation between populations due to geographical isolation. Cytology The basic number of chromosomes in the genus Polystichum was reported to be x = 41 (Manton 1950, Daigobo 1973). Daigobo (1973) reported that the Japanese P. otomasui was a sexual diploid with a chromosome number of n = 41. Two samples from Wuyishan Nature Reserve (Lin & al , ) and another two from Emeifeng mountain (Lin & al , ) were observed cytologically. All of them have the same somatic chromosome number of 2n = 82 (Fig. 4). In addition, two samples (Lin & al , ) were estimated to be diploid based on the genome size estimated by a flow cytometer. All twenty samples collected from the Wuyi Mountains were counted to have 64 spores in each sporangium. Based on these results, the Chinese samples were identified with P. otomasui, a diploid sexual species. It was named Nanbi Erjue as Chinese name. Three related species, P. makinoi (Tagawa) Tagawa, P. acutipinnulum Ching & Shing, and P. wuyishanense Ching & Shing were recorded in the vascular plant checklist of Wuyishan National Nature Reserve (Lin et al. 1993). Polystichum makinoi is a tetraploid species (Kurita 1966, Mitui 1968), morphologically distinguished by lacking black scales on rachis, sori distributed on apical portion of laminae downwardly on auricles of pinnules in lower pinnae (Daigobo 1971, Iwatsuki et al. 1995), and

April 2011 Journal of Japanese Botany Vol. 86 No.2 117 Fig. 2. Polystichum otomasui from Wuyishan National Nature Reserve, Fujian Province, China. spore about 50 µm 60 µm (Lin unpublished).

3 April 2011 Journal of Japanese Botany Vol. 86 No Fig. 2. Polystichum otomasui from Wuyishan National Nature Reserve, Fujian Province, China. spore about 50 µm 60 µm (Lin unpublished). However, P. acutipinnulum is a Chinese endemic species, the type specimens were also collected from Tongmuguan, Wuyishan National Nature Reserve (Ching 1981), is very similar to P. otomasui morphologically, but different in its falcate deltoid pinnules and indusia with irregular projections. Polystichum wuyishanense has not been recognized as a distinct species, which was included in P. acutipinnulum by Kung et al. (2001). Cytological and genetic information of the Chinese Polystichum is unsatisfactory for understanding the relationships among these species. Further systematic researches are expected. Voucher specimens from Fujian Province: Wuyishan National Nature Reserve: on the way from Tongmuguan to an Old Tree Village, 1100 m alt., 2006, Lin & al (FNU, KUN, HYO, TNS); near by the Observation Tower of Xianfengling Peak, 1500 m alt., and on the way to Sangang, on a humid & light slope near a stream under a secondary forest, 2006, Lin & al. 0608Lin-1, 0608Lin-2, 0608Lin-5, 0608Lin-6, 0608Lin- 9 (FNU, KUN, HYO, TNS). Tongmuguan, 1700 m alt., on a humid slope under a bamboo forest, 2007, Lin & al , , , , , , (FNU and Herb. Faculty of Life and Environmental Science, Shimane University, Japan). Taining Prefecture: Emeifeng mountain, 1714 m alt., on the way to Emeifeng Nature Reserve, on a path slope under broad leaf evergreen forest, 2008, Lin & al , , , , , , , , , , , , , (FNU, and Herb. Faculty of Life and Environmental Science, Shimane University, Japan). We would like to express our thanks to curators of Wuyishan National Nature Reserve, Fujian Forestry Bureau for permitting our field investigation on Wuyishan National Nature Reserve. We thank Dr. A. Ohshima and Ms. M. Nagamoto for their technical assistance and help during field work. Thanks are also due to Dr. Mei Sun for her useful comments and help with revising the manuscript.

118 植物研究雑誌第 86 巻第 2 号 2011 年 4 月 Fig. 3. SEM photos of spores, showing perispore sculpture. A, B. Sample from Mt. Emeifeng, Taining, Fujian Prov., China (Lin & al. 080812-13). C, D.

4 118 植物研究雑誌第 86 巻第 2 号 2011 年 4 月 Fig. 3. SEM photos of spores, showing perispore sculpture. A, B. Sample from Mt. Emeifeng, Taining, Fujian Prov., China (Lin & al ). C, D. Sample from Kumamoto Pref., Japan (Nagamoto & al ). The curators of FNU, HUH, HYO, KUN, KYO, TI and TNS are acknowledged for their kind permission to examine specimens during this study, and the Department of Organismic and Evolutionary Biology, Harvard University, is acknowledged for offering S.-J. Lin (the senior author) comfortable research environment during her sabbatical leave period (2010). This study was partly supported by the New Technology Development Foundation, and by Grants-in-Aid of the Ministry of Education, Culture, Sports, Science and Technology of Japan (project no for K. Iwatsuki and no for S.-J. Lin). References Ching R. C., New ferns from Fujian Province (1). Wuyi Sci. J. 1: Daigobo S Taxonomical studies on the fern genus Polystichum in Japan, Ryukyu, and Taiwan. Sci. Rep. Tokyo Kyoiku Daigaku 15:

April 2011 Journal of Japanese Botany Vol. 86 No.2 119 Fig. 4. Somatic chromosomes (2n = 82) of Polystichum otomasui (Lin & al. 070814-1) from Wuyishan National Nature Reserve, Fujian Prov., China.

5 April 2011 Journal of Japanese Botany Vol. 86 No Fig. 4. Somatic chromosomes (2n = 82) of Polystichum otomasui (Lin & al ) from Wuyishan National Nature Reserve, Fujian Prov., China. Bar = 10 µm. Daigobo S Chromosome number of the fern genus Polystichum (1). J. Jpn. Bot. 48: Iwatsuki K Ferns and Fern Allies of Japan. Heibonsha, Tokyo (in Japanese). Iwatsuki K., Yamazaki T., Boufford E. D. and Ohba H Flora of Japan. Volume 1. Pteridophyta and Gymnospermae. Kodansha, Tokyo. Kung H. S., Zhang L. B., Zhu W. M., He Z. R. and Xie Y. T Dryopteridaceae (2). In: Kung H. S. (ed.), Flora Reipublicae Popularis Sinicae. 5(2): Science Press, Beijing (in Chinese). Kurata S Notes on Japanese ferns (32). J. Geobot. 12: Kurita S Chromosome numbers of some Japanese ferns (5). J. Jpn. Bot. 41: Kurata S. and Nakaike T Illustrations of Pteridophytes of Japan Volume 3. Tokyo University Press, Tokyo. Lin L. and Zhang Y Vascular plants checklist of Wuyishan Nature Reserve. In: Zhao X. F., (ed.), The General Scientific Surveys of Wuyishan Nature Reserve. pp Fujian Science Press, Fuzhou. Manton I Problems of Cytology and Evolution in the Pteridophyta. Cambridge University Press, New York. Mitsushima I., Tsuji H. and Takamiya M Morphological, Cytological and molecular genetic studies of Polysticum otomasui Sa. Kurata found in Hita, Ooita Prefecture. Plant of Ooita Prefecture 19: (in Japanese). Mitui K Chromosomes and speciation in fern. Sci Rep. Tokyo Kyoiku Daigaku 13: Nakaike T New Fora of Japan. Pteridophyta. Shibundo, Tokyo (in Japanese). 林蘇娟 a, 鈴木武 b, 海老原淳 c, 盧海声 d, 劉劍秋 d, 武素功 e, 岩槻邦男 b : ナンピイノデの中国福建省新分布記録日本固有種とされてきたナンピイノデ Polystichum otomasui Sa. Kurata が中国の東南部福建省武夷山自然保護区と武夷山脈南部の泰寧県峨嵋峰で発見された. 染色体数 2n = 82(x = 41) であり,1 胞子嚢当り 64 胞子性の有性生殖二倍体であることが確認された. 形態学的な特徴 ( 胞子, 胞子の周皮模様を含む ) は日本の Polystichum otomasui の特徴とよく一致した. ナンピイノデには南碧耳蕨の中国名をつけた. ( a 島根大学生物資源科学部, b 兵庫県立人と自然の博物館, c 国立科学博物館植物研究部, d 中国福建師範大学生命科学学院, e 中国科学院昆明植物研究所 )

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