The karyotypes of 50 plants of Paris tetraphylla were analyzed by observing the Cbanded segment patterns of chromosomes in Miike (Mi), Kunimi (Kn) and Kikuchi (Kk) populations in Kumamoto Prefecture of Kyushu. Various karyotypes were observed in each population, i.e., 49 different karyotypes in Mi, 50 in Kn and 38 in Kk. The karyotypes tended to be heterozygous for most of the homologous chromosome pairs. This indicated that the karyotypic structure of the populations may be maintained mainly by random crossing. Nevertheless, the frequencies of karyotypes homozygous for all chromosome pairs were higher than the expected values, which were calculated using random combinations of homo-and heterozygous pairs in respective populations. Moreover, in most of the chromosomes composing the genome, the observed frequencies of homozygous pairs were significantly higher than the expected ones, which were calculated randomly based on the observed frequencies of different chromosome types in each population. The inbreeding coefficients (f) indicated that a low degree of inbreeding occurred in the natural conditions. The pollination tests indicated that the species was both self-and cross-compatible and fertilized by predominant crossand inferior self-pollination. This was supported by the collection of air-born pollen grains in the population. In the conclusion, the populations of P tetraphylla were maintained predominantly by random breeding with a low degree of simultaneous inbreeding.