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Comparison of Karyomorphological Characters in Four Japanese Species of Lycopus and Russian L

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Comparison of Karyomorphological Characters in Four Japanese Species of Lycopus and Russian L Chromosome Botany (2009) 4: 71-77 © Copyright 2009 by the International Society of Chromosome Botany Comparison of karyomorphological characters in four Japanese species of Lycopus and Russian L. europaeus, Lamiaceae Tsuneo Funamoto1,3, Katsuhiko Kondo2 and Tsuyoshi Motohashi2 1Biological Institute, Fundamental Education and Research Center of Pharmaceutical Sciences, Showa Pharmaceutical University, 3-chome, Higashi-Tamagawagakuen, Machida City, Tokyo 194-8543, Japan 2Laboratory of Plant Genetics and Breeding Science, Department of Agriculture, Faculty of Agriculture, Tokyo University of Agriculture, Atsugi City, Kanagawa 234-0034, Japan 3Author for correspondence ([email protected]) Received July 11, 2009; accepted September 15, 2009 ABSTRACT. Resting nuclei, the mitotic prophase and metaphase chromosomes in five species ofLycopus were compared: The resting nuclei were commonly of the complex chromocenter type and the mitotic prophase chromosomes were of the proximal type. Lycopus ramosissimus, L. maackianus, L. unifloru and L. europaeus showed commonly the chromosome number of 2n=22, while L. lucidus showed 2n=66. The chromosome number of 2n=66 in L. lucidus and that of 2n=22 in L. ramosissimus were reported here for the first time, and that of 2n=22 inL. maackianus, L. uniflorus and L. europaeus verified the previous reports. Four species of the genus with 2n=22 could be all diploid and one species with 2n=66 could be hexaploid if the basic chromosome numbers x=11 was accepted. KEYWORDS: Karyomorphology, Japananese Lycopus, Lamiaceae, Russian Lycopus Lycopus L. of the Lamiaceae (Labiatae) consists of cultivated in unglazed plant-pots in shade place in the approximately 14 species (Willis 1982) that are perennial experimental garden of Showa Pharmaceutical University. herbs and grow in wet places, lakeshores and reeds in the Taxonomical treatment followed the system of Murata riverbed in temperate to sub-arctic regions of the northern and Yamazaki (1993) and Doron’kin (2006). The voucher hemisphere. Among the species of the genus, seven are specimens were deposited in Funamoto’s private herbarium found in East Asia; four are distributed in Japan from in Showa Pharmaceutical University. Hokkaido to Kyusyu (Murata and Yamazaki 1993), six are distributed in Russia and adjacent states (Czerepanov Chromosome observations Fleshly growing root-tips 1995), and four are distributed in China (Li and Hedge 5-10 mm long were harvested from their cultivated plants 1994). and pretreated in 2 mM 8-hydroxyquinoline for 4 h at about Cytological studies of this genus have been made in 13 20°C. Then, they were fixed in 45% acetic acid for 10 min species with mostly respect to the chromosome counts of at ca 2°C, and were macerated in the mixture of 45% n=11 and 2n=22, 24 and 30 by many researchers (e.g.; acetic acid and 1N hydrochloric acid (1:1) for 20-23 sec at Rutter 1933; Tischler 1934; Rohweder 1937; Ehrenberg ca 60°C. They were stained with 2% aceto-orcein in a 45% 1945; Gadella and Kliphuis 1963, 1966; Skalinska et al. acetic acid moist chamber for ca 30 min at room temperature. 1971; Strid 1971; Lee 1972; Morton 1973; Löve and Slide preparation was made by the conventional aceto- Kjellqvist 1974;Vachova and Ferakova 1977;Van Den orcein squash method. The slides were faintly heated under Brand et al. 1979; Gill 1979, 1981a,b; Webber and Ball 1980; an alcohol flame for 1-2 sec before observation. Amadei et al. 1982; Löve and Löve 1982a, b; Astanova 1984; Classification of the chromosome types of the resting Dmitrieva 1985; Romano et al. 1986; Sokolovskaya et al. nuclei and the mitotic prophase chromosomes followed 1986; Wentworth et al. 1991; Hollingsworth et al. 1992; Tanaka (1971, 1977). Markova and Goranova 1994; Probatova and Sokolovskaja Measurements of chromosome lengths were made 1995; Dobes and Hahn 1997; Gervais et al. 1997, 1999; in several good metaphase cells. Classifications for the Lövkvist and Hultgård 1999; Shatalova 2000). chromosome types at mitotic metaphase by the centromeric The present study compared chromosome characters in positions followed Levan et al. (1964). the resting nuclei, the mitotic prophase and metaphase chromosomes of four Japanese species and one Russian RESULTS AND DISCUSSION species of Lycopus in Japan and one species in Russia. Lycopus ramosissimus (Makino) Makino, L. lucidus Turcz., L. maackianus (Maxim. ex Heder) Makino, L. uniflorus MATERIALS AND METHODS Michx. and L. europaeus L. studied had common karyo- Plant materials The plants used were collected in 30 morphological characters such as the complex chromocenter localities in Japan and Russia (Fig. 1, Table 1). They were type of the resting nuclei which had numerous chromomeric 72 FUNAMOTO ET AL. Fig. 1. Collection sites of five species of Lycopus in Russia and Japan. (★): L. ramosissimus. (■): L. lucidus. (▲): L. maackianus. (●): L. uniflorus. (☆): L. europaeus. Table 1. Collection sites, sample and chromosome numbers of four species in Japanese Lycopus and L. europaeus in Russia observed in this study Sample Chromosome Species Collection site number number (2n) L. ramosissimus (Makino) Japan, Miyagi Pref., Kurihara-shi, Tsukidate, Utinuma, 38°42’ 65” N, 141°05’ 4 22 Makino 11” E, alt. 15 m Japan, Yamagata Pref., Higashimurayama-gun, Yamabe-machi, Hatadani, 6 22 Hannoki-muma, 38°14’ 08” N, 140°12’ 24” E, alt. 620 m Japan, Yamagata Pref., Nanyo-shi, Hakuryu-ko, 38°03’ 16” N, 140°10’ 85” E, alt. 4 22 210 m Japan, Shizuoka Pref., Fujinomiya-shi, Lake side of Tanuki-ko, 35°20’ 38” N, 4 22 138°33’ 56” E, alt. 670 m Japan, Fukui Pref., Fukui-shi, Busyu-machi, Lake side of Busyu-ike, 36°00’ 79” 4 22 N, 136°01’ 57” E, alt. 270 m Japan, Fukui Pref., Tsuruga-shi, Ikenogouchi, 33°38’ 92” N, 136°08’ 18” E, alt. 3 22 310 m L. lusidus Turcz. Japan, Chiba Pref., Narita-shi, Kitasuga, Lake side of Inba-muma, 35°47’ 50” N, 5 66 140°15’ 66” E, alt. 3 m Japan, Fukui Pref., Tsuruga-shi, Ikenogouchi, 35°39’ 00” N, 136°08’ 19” E, alt. 3 66 300 m Japan, Fukui Pref., Mikatakaminaka-gun, Wakasa-machi, Torihama, 35°33’ 30” 3 66 N, 135°54’ 12” E, alt. 0.5 m KARYOMORPHOLOGICAL CHARACTERS OF LYCOPUS, LAMIACEAE 73 Japan, Hiroshima Pref., Fukuyama-shi, Minomi-machi, 34°27’ 73” N, 133°21’ 3 66 63” E, alt. 5 m Japan, Hiroshima Pref., Mihara-shi, Hongo-machi, Nousyo, 34°24’ 04” N, 4 66 135°00’ 37” E, alt. 20 m Japan, Tokushima Pref., Anan-shi, Gamoda, 33°47’ 29” N, 134°44’ 30” E, alt. 5 m 4 66 L. maackianus (Maxim. ex Japan, Aomori Pref., Kamikita-gun, Rokkasyo-mura, Ichiyanagi-muma, 40°53’ 3 22 Heder) Makino 56” N, 141°22’ 34” E, alt. 10 m Japan, Yamagata Pref., Higashimurayama-gun, Yamabe-machi, Hatadani, 5 22 Hannoki-muma, 38°14’ 08” N, 140°12’ 24” E, alt. 620 m Japan, Yamagata Pref., Nishiokitama-gun, Oguni-machi, Kanomizu, 37°59’ 85” 6 22 N, 139°48’ 50” E, alt. 270 m Japan, Fukushima Pref., Soma-shi, Isobe, 37°47’ 29” N, 140°59’ 30” E, alt. 4 m 5 22 Japan, Fukushima Pref., Fukushima-shi, Tsutiyuonsen, O-numa, 37°41’ 31” N, 5 22 140°19’ 05” E, alt. 660 m Japan, Fukushima Pref., Aizuwakamatsu-shi, Akai-shitsugen, 37°30’ 79” N, 3 22 140°00’ 32” E, alt. 510 m Japan, Niigata Pref., Myoko-shi, Mumanohara, 36°51’ 75” N, 138°17’ 18” E, alt. 5 22 830 m Japan, Nagano Pref., Chikuma-shi, Yawata (Onuma), 36°28’ 93” N, 138°05’ 36” 4 22 E, alt. 820 m Japan, Shizuoka Pref., Fujinomiya-shi, Kodanuki-shitsugen, 35°20’ 92” N, 6 22 138°33’ 55” E, alt. 690 m Japan, Shizuoka Pref., Fujinomiya-shi, Lake side of Tanuki-ko, 35°20’ 42” N, 7 22 138°34’ 08” E, alt. 670 m Japan, Shizuoka Pref., Iwata-shi, Lake side of Tsuruga-ike, 34°44’ 71” N, 137°53’ 5 22 16” E, alt. 10 m Japan, Aichi Pref., Toyohashi-shi, Iwata, 34°44’ 52” N, 137°27’ 29” E, alt. 80 m 3 22 L. uniflorus Michx. Japan, Yamagata Pref., Nishimurayama-gun, Asahi-machi, Oo-muma, 38°19’ 73” 3 22 N, 140°06’ 96” E, alt. 310 m Japan, Yamagata Pref., Higashimurayama-gun, Yamabe-machi, Hatadani, 5 22 Hannoki-muma, 38°14’ 08” N, 140°12’ 24” E, alt. 620 m Japan, Fukushima Pref., Fukushima-shi, Tsutiyuonsen, O-numa, 37°41’ 31” N, 4 22 140°19’ 05” E, alt. 660 m Japan, Nagano Pref., Shimominochi-gun, Sakae-mura, Nonomi, 37°01’ 13” N, 6 22 138°31’ 73” E, alt. 1,040 m Japan, Nagano Pref., Shimotakai-gun, Yamanouchi-machi, Lake side of Biwa- 5 22 ike, 36°42’ 97” N, 138°29’ 15” E, alt. 1,400 m Japan, Nagano Pref., Shimotakai-gun, Yamanouchi-machi, Lake side of 5 22 Mizunashi-ike, 36°43’ 43” N, 138°28’ 67” E, alt. 1,370 m Japan, Nagano Pref., Minamisaku-gun, Koumi-cho, Matsubara, 36°02’ 89” N, 7 22 138°27’ 61” E, alt. 1,130 m Japan, Nagano Pref., Saku-shi, Maeyama, Misasa, 36°11’ 08” N, 138°24’ 04” E, 5 22 alt. 970 m Japan, Nagano Pref., Nagano-shi, Togakushi, Chushya, 36°44’ 50” N, 138°05’ 6 22 13” E, alt. 1,260 m Japan, Nagano Pref., Higashichikuma-gun, Omi-mura, Hijiri-kogen, 36°29’ 04” 6 22 N, 138°04’ 19” E, alt. 970 m Japan, Nagano Pref., Omachi-shi, Yasaka, Aikawa, , 36°29’ 09” N, 137°53’ 58” E, 3 22 alt. 970 m L. europaeus L. Russia, Moscow, Vladenie 171, Dmitrovskoe shosse, Dovue pygue-ozepa, Dolgie 8 22 prubie, 55°56’ 18” N, 037°32’ 08” E, alt.
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