Lilium maculatum var. bukosanense, Miyamasukashi-yuri in Japanese, grows endemically in Saitama Prefecture, Japan, and is listed in the Saitama Prefecture Red Data Book as a critically endangered plant. Articles about its genetic diversity and population genetic structure have not been reported. Before it is lost, population genetic studies can provide information for efficient in situ conservation as well as for possible future use in breeding programs. Therefore, we carried out a population genetic study of natural L. maculatum var. bukosanense using chloroplast DNA (cpDNA) and inter-simple sequence repeat (ISSR) markers. In this study, fresh leaves of 46 L. maculatum var. bukosanense individuals were collected from two populations in Mount Buko. By polymerase chain reaction (PCR)-restriction fragment length polymorphism analysis of cpDNA, we estimated the maternal origin of individuals sampled. Sixteen cpDNA primer pairs were tested, and only four of them produced clear bands. Twenty-four combinations from these PCR fragments combined with six restriction enzymes were assayed; however, only 18 of them could be digested, producing two to four fragments. All individuals of L. maculatum var bukosanense analyzed were virtually identical for all primer-enzyme combinations. These findings suggest that the maternal origin of the 46 individuals analyzed is close. In addition, we analyzed ISSR markers to provide mainly biparental inheritance. PCR products of five anchored simple sequence repeat primers showed polymorphism, indicating that the individuals sampled were not clones, and thus it appears that they could have been reproduced from seeds. The primers yielded a total of 30 polymorphic bands (i.e. an average of six polymorphic bands per primer). The mean genetic distance and genetic identity values between the populations were 0.032 and 0.97, respectively. These findings suggest that all individuals from both populations are genetically similar. The individuals growing at 1000 m have greater genetic variation than those individuals from 650 m and this could possibly be attributed to altitude. A significant correlation between gene diversity and population size is also reported. From the results presented here some strategies for its in situ and ex situ conservation might be considered.

Ovary slice culture, after cut-style poUination, was used to develop interspecific hybrids between Lilium longiflorum and L. concolor. Reciprocal crosses between diploid cultivars (2n = 2x = 24) were carried out. On the days 30, 35,40 and 45th after pollination (DAP), ovaries were sliced and cultured on a modified hormone-free Murashige-Skoog (M-S) medium without NH4NO3, supplemented with 6% sucrose, 50 mg/l yeast extract and 0.25% gelrite at pH 6.3. For the L. longiflorum X L. concolor cross, ovule germination was found to be best at 30 DAP. After transfer to a M-S (half-strength) medium supplemented with 1.5% sucrose and 0.25% gelrite at pH 5.8, diploid and triploid hybrid plants were established. In contrast, ovules from the L. concolor X L. longiflorum cross did not germinate. The hybridity of the plantlets obtained was verified by karyotype and isozyme analysis. The importance ofthe ovary slice culture technique as a tool to develop new hybrids between incompatible lilly plants is discussed.Key words: Lilium spp. -cut-style pollination -interspecific hybrids -isozyme -karyotype -ovary slice culture Lilium longiflorum is one of the most popular lihes in horticulture and many breeders have attempted to raise new hybrid lilies by interspecific crosses between it and other colourful lilies (Kanoh et al. 1988) such as L. coneolor, since the flower of L. longiflorum is always white. It has not so far been possible to obtain interspecific hybrids between L. longiflorum and L. concolor in vivo.The cut-style pollination method has been developed and used in lily plants to overcome cross incompatibility Myodo 1977a, van Tuyl et al. 1991). After pollination, the ovary slice culture technique has also been successfully applied to develop hybrids in the genus Lilium by several researchers (Hayashi et al. 1986, Kanoh et al. 1988, Morimoto 1990, who suggested that this technique might enable the production of interspecific hybrids in lily plants more efficiently and easily than embryo culture.In some lily crosses, fertilization occurs but hybrid embryo growth is restricted by post-fertilization barriers. Some of these irregularities are associated with various types of abnormahty in endosperm development, and the failure of endosperm development occurs at various times after fertilization (Stout 1922, Myodo 1962, Dowrick and Brandram 1970, Asano 1982.In this study, an attempt was made to improve interspecific crosses between L. longiflorum and L. concolor by using ovary slice culture at four different times after cut-style pollination. The hybridity of the plants obtained was evaluated through karyotype and isozyme analysis. Materials and MethodsPollination method: Reciprocal crosses were made between L. longiflorum cv. Wase Teppo Yuri and L. concolor cv. Akahime Yuri from the Takii Company, Japan. In this experiment, cut-style pollination was used according to the method proposed by Asano and Myodo (1977a), and van Tuyl et al. (1991). The poUinations were carried out in a greenhouse.In vitro culture procedure: On days 30...

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Isozyme banding patterns (IBPs) were studied for cultivars of lily (Lilium spp.) by means of horizontal starch-gel electrophoresis (SGE). An array of continuous histidine-citrate buffer systems at eight ranges of pH and four extraction buffers were tested. On the basis of this survey, the extraction buffer two (Eb-2) and the buffer system E at pH 7.7 were found to be suitable for detection of lily isozymes. EC 5.3.1.9) and 6-phosphogluconate dehydrogenase (PGD; EC 1.1.1.44) were assayed. In total 29 cultivars were tested in this study: nine were analysed for all eight enzyme systems, 16 cultivars for seven systems, three for six, and one for five enzyme systems. Some IBP were identified as section-specific biochemical markers. Eight enzymes systems were analysed by constructing a dendogram using the unweighted pair group method, arithmetic average (UPGMA) cluster analysis. The analysis indicated that the lily cultivars could be separated from other Lilium species, except for two L. x formonlogi cultivars: ÔHakubaÕ and ÔHakukoÕ which could not be distinguished from each other by the isozyme patterns assayed here. This study shows that isozymes can provide useful biochemical markers for lily cultivar identification and to estimate the phylogenetic relationships among those cultivars.