Isoenzyme patterns of 226 accessions of Hydrilla verticillata collected in Japan were compared. 17 and 23 electrophoretic phenotypes were identifiable in diploid and triploid accessions, respectively, in dioecious ones. To the contrery, monoecious plants showed no variation of banding patterns and were assumed to be rametes of the same clone. The cytological change from diploid to triploid was suggested to occur many times in dioecious plants.
Key words: Dioecy--Genetic diversity--Hydrilla verticillata m Multi-enzyme phenotype (MEP)--Monoecy--PolyploidyHydrilla verticillata (L.f.) Royle (Hydrocharitaceae)is a submerged aquatic plant distributed in the Old World and has recently been introduced to the American continent (Cook and LL]6nd 1982 ). Intraspecific variation of H. verticillata has been reported in morphology, sex expression and chromosome number, and occurrence of large genetic variations has been shown by comparative isoenzyme studies (Verkleij et al. 1983a, Pieterse et al. 1984,1985 and random amplified polymorphic DNA (RAPD) (Madeira et al. 1997). Comparative studies have also been conducted to know the differentiation of ecological and physiological traits of various biotypes of H. verticillata (Spencer and Anderson 1986, MacFarland and Barko 1987, Steward and Van 1987, Van 1989, Steward 1991,1992.In our previous study (Nakamura and Kadono 1993), the occurrence of monoecious and dioecious forms of H. verticillata was confirmed in Japan. Dioecious plants were diploid (2n=16) or triploid (2n=24) and monoecious ones were all triploid in chromosome number. In the present study, we tried to reveal further genetic differentiation of H. verticillata in Japan on the basis of isoenzyme study.226 accessions collected from various regions of Japan were used for the electrophoretic analyses of isoenzymes. For details of collection sites, see Nakamura and Kadono (1993). They included 19 monoecious (all triploid) and 207 dioecious accessions. Among dioecious ones, 71 diploid * Present address: Wetland Laboratory, Tatsumi-minami 5-6-29, Ikuno, Osaka, 544-0015 Japan , 9 corresponding author, E-mail kadono@kobe u.ac.jp (female 34, male 33, sex unknown 4) and 136 triploid (female 63, male 67, sex unknown 6) accessions were included.Horizontal starch gel electrophoresis was conducted with the following five enzyme systems: phosphoglucomutase (PGM), phosphogluconate dehydrogenase (PGD), aconitase (ACO), isocitrate dehydrogenase (IDH) and phosphoglucoisomerase (PGI; this enzyme was tested only for dioecious plants).Additional four enzyme systems, that is, shikimate dehydrogenase (SkDH), malate dehydrogenase (MDH), hexokinase (HK), glyceraldehyde 3 phosphate dehydrogenase (G3PD), were also tested preliminarily. However, they were not used because of low activity and poor resolution in the present method.Apical part of the shoots or axillary turions (only in some cases of dioecious plants) were used. They were ground in ice-cold Tris-HCI-PVP buffer and the extracts were centrifuged for 3 min at 10,000 rpm. The supernatant...