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Page 1 植物研究雜誌 J. Jpn. Bot. 73: 241–247 (1998) Originals Allozyme 植物研究雑誌 J. J. Jpn. Bo t. Originals Originals 73: 241-247 (1998) Allozyme Variation in Six Flowering Plant Species Characterizing Characterizing Ullung Island ，Korea Hye Gi CHUNG ， Jae l¥ lin CHUNG and Myong Gi CHUNG Department Department ofBiology ，Gyeongsang National University ，Chi 吋 u，660-701 KOREA (Received (Received on February 18 ，1998) A prelimin 訂 y allozyme study on six populations of Fagus multinervis (endemic to the the Ullung Island) ， Pinus parviflora ， Hepatica maxima (endmic to the Island) ，Campanula takesimana takesimana (endemic to the Island) ，Tim ・ella polyphylla ， and Thymus quinquecostatus va r. japonicus on the Ullung Island from Korea was made. Populations of F. multinervis and and P. parviflora maintain moderate levels of allozyme variation (mean expected heterozygosity heterozygosity ofO.253 in F. multinervis and 0.149 in P. parviflora) ， whereas those of H. maxima (0.032) ，仁 takesimana (0.021) ，T. polyphylla (0.016) ， and T. quinquecostatus var.japonicus var.japonicus (0.066) maintain low levels of allozyme diversity within populations. For the the latter four taxa ，factors such as founder effect ，genetic drift associated with small population population size ， and probable inbreeding contributed to their situation. The Ullung Island ，75 .4 km 2 area ， is 10- (Yim 1993). catedat catedat 130 0 E and 37 0 N ， ca.141 kmeastofthe Estimates of levels and distribution of ge- Korean Peninsula. The Island had been formed netic diversity within and among rare 'and by volcanic activities from the late Tertiary to endangered species could be useful to provide the the Quaternary. The flora on the Island has a basis on which to build sound programs for been ofbotanical interest because several sub- the conservation of genetic diversity of these tropical tropical plants grow on the Island owing to a plants (Soule 1986). In addition ，genetic diver- warm current ，though the Island is located in sity routinely from allozyme markers can be the the northern temperate region. About 500 vas- used as a yardstick to measure the effective 同 cular cular plants grow on the Island (Oh 1978) ，and ness of in situ and ex situ conservation pro- among them ca. 40 species are endemics. Re- grams (Hamrick et al. 1991). Despite the im- cently ， the number ofvisitors to the Island has portance of know ledge concerning genetic been rapidly increased (several thousands in variation for providing information for con- summer season) because several express pas- ser 、ration purposes ，little is known of the ge- senger senger boats are available. In addition ，several netic variation in plants native to the Island. roads roads on hillsides have been recently enlarged Island plantpopulations generally harborfewer and newly constructed. These activities could polymorphic loci and fewer alleles per locus severel severel y affect the flora and natural habi tats on than mainland populations because they are the the Island. For these reasons ， the Korean gov- probably descended from one or a few found- ernment has designated six sites of natural ers (Hamrick et al. 1979). forests forests as N atural Monuments to protect rare As a part of studies to preserve plant diver- and endemic plants from human disturbances sity on the Island ， the objective ofthis prelimi- 一 241- 242 植物研究雑誌第73 巻第5号 平成10 年10 月 nary nary study is to explore patterns of genetic subshrub ， grows on limestone and andesite variations variations in six plant taxa on the Island. F agus regions in Japan (Hokkaido ，Honshu and multinervis multinervis N akai (Fagaceae) ， an endemic plant Kyushu) ， the northern China ，Mongolia ， and to to the Island ，is an element of cool-temperate China (Ki tamura and Murata 1986). Only one 2 forests forests in East Asia. It is a large ，long-lived ， population (3 ，226 m ) at the Nari Basin is monoecious monoecious tree ， and its wind-bome pollen known on the Ullung Island and has been and and potentially animal-dispersed seeds may designated as the N atural Monument in Korea allow allow fairly long-distance gene dispersal on and is now surrounded by a wire fence because the the Island. Pinus parviflora Siebold & Zucc. of its rarity in South Korea. Flowering indi- (Pinaceae) (Pinaceae) is distributed in J apan (westem viduals of T. quinquecostatus var. japonicus Honshu ， Shikoku and Kyushu) and Ullung are visited by bees (M. Chung pers. obs よ Island. Island. Pinus parviflora is also a component of cool-temperate cool-temperate forests in the region. On the Materials and Methods Ullung Ullung Island ， only a few natural stands in 田 A total of 300 leaf samples was randomly cluding cluding one N atural Monument stand are collected from six populations of the six taxa known (Ahn 1971) ， and the number of indi- on the Ullung Island. Mature leaves (or nee- viduals viduals in each stand are less than hundreds dles) were collected from 50 individuals per (M. (M. Chung pers. obs.). Like other conifers ， population. For Fagus multinervis and Pinus Pinus Pinus parviflora is a large ，long-lived ，wind- parviflora ，individuals (30-60 years old) were 2 pollinated pollinated monoecious tree. The seeds are collected from area covered ca. 5，000 m . Leaf 1.5 1.5 cm long ， 0.8 cm wide with 4 mm wing samples of Hepatica maxima ，Campanula (Ahn 197 1). Hepatica maxima Nakai takesimana ， and Tiarella polyphylla were col- 2 (Ranunculaceae) ，a herbaceous perennial ，is lected from areas covered ca. 500 m . Leaf endemic endemic to the Island. White flowers (1.5 cm samples ，wrapped with a wet paper towel ， wide) wide) are visited by bees (Chung pers. obs.). placed in plastic bags and stored on ice during The achene is 5 mm long. Local populations of transport from the Island to the laboratory. H.m αxima are found in patches ， and isolated Samples were then stored at 4 0 C until proteins from each other under hillside forests on the were extracted. Island. Island. Camp αnula takesimana N akai Leaf samples were cut finely and crushed (Campanulaceae) ， an endemic of the Island ， with a mortar and pestle. A phosphate-polyvi 国 grows grows on the forest margin near coastal areas. nylpy 町 olidone extraction buffer (Mitton et al. Whitish Whitish pu 中 le flowers (3-5 cm long) are 1979) was added to leaf samples to facilitate visited visited by bees and each capsule contains crushing and to aid enzyme stabilization. En- hundreds of seeds. In Korea ， Tiarella zyme extracts were absorbed onto 4x6 mm polyphylla polyphylla D.Don (Saxifragaceae) only oc- wicks cut from Whatman 3 mm chromatogra- curs curs under oak forests near the summit of the phy paper ，which were stored at -70 o C until Ullung Ullung Island ， but the species widely distrib- needed. Electrophoresis was performed using utes utes inJapan (Hokkaido ，Honshu and Shikoku) ， 10.5% starch gels to resolve scorable loci for Taiwan ，China and Himalaya (Kitamura and 13 enzyme systems: acid phosphatase (ACP) ， Murata 1987). Small white flowers are visited triosephosphate isomerase (TPI) ，phosphoglu- by bees (M. Chung pers. obs.) ， and each cap- comutase (PGM) ，fluorescent esterase (FE) ， sule sule contains several small seeds (1 .2-1 .4 mm peroxidase (PER) ， alcohol dehydrogenase long). long). Thymus quinquecostatus Cela k. var. (ADH) ，menadione reductase (MNR) ， japonicus japonicus H.Hara (Lamiaceae) ，a deciduous phosphoglucoisomerase (PGI) ， diaphorase October October 1998 Journal of Japanese Botany Vo l. 73 No. 5 243 (DIA) (DIA) ，isocitrate dehydrogenase (IDH) ，rnalate dures for DIA followed the rnethod described dehydrogenase dehydrogenase (MDH) ，6-phosphogluconate by Cheliak and Pitel (1 984). All other stain dehydrogenase dehydrogenase (PGD) ， and skirnate dehydro- recipes were identical to those described by genase genase (SKDH). Buffer systerns and putative Soltis et al. (1 983). Putative loci were desig- loci loci are listed in Table 1. The staining proce- nated sequentially ， with the rnost anodally Table Table 1. List ofbuffer systerns and resolvable enzyrne loci. FM ，Fagus multinervis; PP ， Pinus P卯α門川ifl μlor α， and and TQ ，Thymus quinquecost α， tus v訂 . japonicus: M indicates a rnonornorphic locus ，P indicates indicates a polyrnorphic locus ， X indicates that the locus was poorly resolved could not be scored. scored. For TQ ，diaphorase (DIA) and 6-phosphogluconate dehydrogenase (PGD) were resolved resolved on a rnorpholine citrate buffer systern Buffer Buffer systern Loci FM PP HM CT TP TQ PMMPMM Acph-1 Acph-1 X P M M 乱4 Acph-2 Acph-2 Acph ・3 Acph-4 Acph-4 MMMX MMMM Tpi-1 Tpi-1 M M M Tpi-2 Tpi-2 M Tpi ・3 ppp MPM Pgm-1 M P Pgm-2 MMMM Fe-1 Fe-1 P M M Fe-2 Fe-2 MMM MMMMPM Per M M 島f A rnodification Adh-1 孔f M 乱f (Hau f1 er 1985) Adh-2 of of Soltis et al. Adh-3 MMMMPMM (1 983) systern 6 Mnr ・-1 p P X M Mnr-2 p M Mnr-3 MPMM MMP MMMMMMMM Pgi-1 Pgi-1 M Pgi ・2 p Dia-1 Dia-1 Dia-2 Dia-2 Dia-3 Dia-3 MMPM MMMMMMM MMMMMMM Idh-1 Idh-1 P M M A rnodification Idh-2 M (Chung and Kang Mdh-1 恥f M 1994) 1994) of Soltis Mdh-2 p eta l. (1 983) Mdh-3 Systern Systern 11 Pgd-1 p M M M Pgd-2 Pgd-2 p 乱4 Ab(attu hvdl H &・・も nfmnYRut-eO mmCch9tvdT ψi30、)戸し vst Skdh-1 x 孔f 恥f X M 町 剖旬 Skdh-2 Skdh-2 x M が日 6.uba ゐ vdkB rl ノ 也、 d パい n .uFら 244 244 植物研究雑誌 第73 巻 第5号 平成10 年 10 月 migrating migrating isozyme designated ‘1' ， the next 0.253) (Table 2) ， though the species is only ‘2' ，and so on.
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