Isozyme Genotypes Research Articles

Life cycle expression of inbreeding depression in Eucalyptus regnans and inter-generational stability of its mixed mating system.

Many plants exhibit a mixed mating system. Published models suggest that this might be an evolutionarily stable rather than a transitional state despite the presence of inbreeding depression, but there is little empirical evidence. Through field experimentation, we studied the role of inbreeding depression in eliminating inbred progeny from the reproductive cohort of the forest tree Eucalyptus regnans, and demonstrate a stable mixed primary mating system over two successive generations. Two field experiments were conducted using seed from natural populations. We sowed open-pollinated seeds to simulate a natural regeneration event and determined isozyme genotypes of dominant and suppressed individuals over 10 years. We also planted a mixture of open-pollinated, outcross and selfed families with common maternal parentage; monitored survival of cross types over 29 years; and determined the percentage of outcrosses in open-pollinated seed from a sample of reproductively mature trees using microsatellite analysis. Both experiments demonstrated progressive competitive elimination of inbred plants. By 29 years, the reproductive cohort in the planted experiment consisted only of outcrosses which produced seed which averaged 66 % outcrosses, similar to the estimate for the parental natural population (74 %). Selective elimination of inbred genotypes during the intense intra-specific competition characteristic of the pre-reproductive phase of the life cycle of E. regnans results in a fully outcrossed reproductive population, in which self-fertility is comparable with that of its parental generation. The mixed mating system may be viewed as an unavoidable consequence of the species' reproductive ecology, which includes the demonstrated effects of inbreeding depression, rather than a strategy which is actively favoured by natural selection.

Isozyme variation and differentiation of morphologically cryptic species in the Brachypodium distachyon complex

Isozyme variation of nine enzymes among three morphologically cryptic species of the Brachypodium distachyon complex was studied with PAGE. Aspartate aminotransferase isozymes AAT-A and AAT-C, aryl alcohol dehydrogenase AAD-B, malate dehydrogenase MDH-A and leucine aminopeptidase LAP-A in the allotetraploid B. hybridum display fixed heterozygosities with codominant expression of homoeozymes corresponding to divergent orthozymes of diploids B. distachyon and B. stacei. The multilocus isozyme genotype inherent to most accessions of B. hybridum combines two divergent multilocus isozyme genotypes characteristic of most accessions of B. distachyon and B. stacei. The maternally inherited chloroplast-encoded RUBISCO isozymes show that B. stacei is a maternal progenitor of B. hybridum, whereas the nuclear-encoded isozymes AAD-B, AAT-A, AAT-C, LAP-A, and MDH-A confirm that B. distachyon is the paternal progenitor. These data provide a new isozyme-based support to the karyological and DNA marker data on the allotetraploid nature of B. hybridum and its origin from diploids B. distachyon and B. stacei.

Relationship of Isozyme Genotypes of AGP with Activity of AGP and Starch Content in Wheat Grain

ADP-Glucose pyrophosphorylase(AGP) controls a rate-limiting step in the starch biosynthesis in developing wheat(Triticum aestivum L.) seeds,therefore,determinately affecting seed sink strength. Evidence is presented for the existence of AGP isozymes in common wheat endosperm. Following an approach already shown to be successful in maize(Zea mays L.),native polyacrylamide gel electrophoresis(native PAGE) was designed to identify AGP isozyme patterns from crude enzyme prepara-tions. To explore the relationship of isozyme genotypes with the starch content in wheat grains,activity of AGP and starch content were measured in 60 cultivars from 17 provinces in China. The results showed that there were four alleles,namely AGPa,AGPb,AGPc,and AGPd,with frequencies of 96.7%,80.0%,86.7%,and 16.7%,respectively. Five genotypes were determined in which AGPabc took on the highest frequency of 46.7% and AGPbc took on the lowest frequency of 3.3%. The activity of AGP and starch content in the cultivars with distinct genotypes were significantly different(P0.05). The cultivars with genotype AGPabcd and AGPabc showed a high value of AGP activity and starch content. The lack of AGPa,AGPb,or AGPc in cultivars resulted in a low value of AGP activity and starch content. It indicated that different genotypes of AGP isozyme show distinct genetic impact on enzyme activity and starch content in common wheat.

Hybridization and Introgression Between the Endemic Phoenix canariensis and the Introduced P. dactylifera in the Canary Islands

The proportion of hybridization and introgression between the endemic Phoenix canariensis (Canarian date palm) and the introduced P. dactylifera (date palm), were characterized using a multilocus isozyme genotype, based on 17 isozyme loci. In order to evaluate the multilocus isozyme genotype's ability to detect hybridization and introgression between the endemic Phoenix canariensis and the introduced P. dactylifera, multilocus genotype data from pure and putative hybrid populations were analyzed with a Bayesian-based method, implemented in the STRUCTURE software. The identity of all Phoenix canariensis plants was confirmed. However, in the putative P. dactylifera individuals examined, some of them were designated as hybrid, and others as Canarian date palms. Individuals from each species were clustered separately, while putative hybrid individuals shared ancestors from clusters where both Phoenix species were assigned. In agreement with this, FCA showed that the hybrids were localized between P. canariensis and P. dactylifera individuals' clouds, but with a closer proximity to the former, suggesting an introgression from Phoenix dactylifera to P. canariensis. In addition, the Bayesian cluster revealed a geographical structure within P. canariensis relative to island origin, so populations from the western islands of the archipelago were clustered together and separately from eastern islands.

Genotype-Species Interactions in Neighbourhoods of Forest Tree Communities

Abstract Studies on plant communities of various annual species suggest that there are particular biotic interactions among individuals from different species which could be the basis for long-term species coexistence. In the course of a large survey on species-genetic diversity relationships in several forest tree communities, it was found that statistically significant differences exist among isozyme genotype frequencies of conspecific tree groups, which differ only by species identity of their neighbours. Based on a specific measure, the association of the neighbouring species with the genotypes of the target species or that of the genotypes with the neighbouring species was quantified. Since only AAT and HEK of the five analysed enzyme systems differed in their genotype frequencies among several tree groups of the same target species, a potential involvement of their enzymatic function in the observed differences was discussed. The results of this study demonstrate a fine-scale genetic differentiation within single tree species of forest communities, which may be the result of biotic interactions between the genetic structure of a species and the species composition of its community. This observation also suggests the importance of intraspecific genetic variation for interspecific adaptation.

First Report of Haplotype I-b of Phytophthora infestans in Central Mexico.

Central Mexico is considered a center of genetic diversity for Phytophthora infestans on the basis of a range of genotypic and phenotypic characteristics (3). Surprisingly, while mitochondrial DNA (mtDNA) haplotypes I-a, II-a, and II-b have been reported from central Mexico, haplotype I-b has not been found in central Mexico (1). Therefore, a more extensive search for haplotypes was conducted in areas where sexual reproduction occurs. During the summer of 2003, leaflets of cvs. Rosita and Tollocan with a single lesion of late blight were collected in the area of Villarreal, located in Terrenate County in Tlaxcala, Mexico (170 km northeast of Mexico City). Fourteen P. infestans isolates were characterized for mtDNA haplotype, isozyme genotype (glucose 6- phosphate isomerase [Gpi] and peptidase [Pep]), and mating type. Isolation, mating type, and isozyme genotype were characterized following reported protocols (1,4). MtDNA haplotype was determined by amplifying and digesting the P2 and P4 regions and comparing amplicons to those of reference strains of known haplotype (1,2). Twelve isolates were mtDNA haplotype I-a and two were I-b. While the mtDNA I-b has been associated with the US-1 lineage (mating type: A1, Gpi: 86/100, Pep: 92/100), the genotypes for the Mexican isolates were A2, 86/100 Gpi, 100/100 Pep from cv. Rosita and A2, 86/100 Gpi, 92/100 Pep from cv. Tollocan. To our knowledge, this is the first report of the I-b mtDNA haplotype of P. infestans from central Mexico and it is now clear that all four haplotypes exist in Mexico. This finding therefore, stresses the importance of including a representative regional sampling of Mexican and Andean isolates in studies inferring the origin of this species.

Genetic Diversity of the Population of Phytophthora infestans in Hungary

A total of 123 isolates of Phytophthora infestans were recovered from 43 commercial fields and allotments of potato and tomato in Hungary. Isolates were characterised for mating type, response to metalaxyl, isozyme genotype at glucose-6-phosphate isomerase (Gpi) and peptidase A (Pep) loci, nuclear DNA fingerprint with probe RG57 and mitochondrial haplotype. Both mating types were detected in 8 potato and 3 tomato fields. The frequency of A1 and A2 were 47.5% and 52.5% on potato and 60.5% and 39.5% on tomato. More than sixty per cent of the whole population were resistant (31.1%) or responded intermediately (30.2%) to the fungicide metalaxyl in vitro. All isolates were monomorphic at the Gpi locus (100/100), but four allele combinations 100/100 (33%), 96/100 (28%), 96/96 (31%) and 83/96 (8%) were found at the Pep locus. Genotype Pep 83/96 is rare in Europe and all isolates with this allele combination were of A1 mating type and Ia mitochondrial haplotype. As in other European populations, only two of the f...

Isozyme Variation and Phylogenetic Relationships in Vicia subgenus Cracca (Fabaceae)

The phylogenetic relationships among 27 vetch species belonging to the subgenus Cracca of the genus Vicia were studied in comparison with three species of Lathyrus section Lathyrus on the basis of isozyme variation. Isozymes encoded by 15 putative loci of ten enzymes were resolved by polyacrylamide gel electrophoresis and isozyme variation was analysed by using parsimony and neighbour-joining methods. The analyses revealed 63 parsimony-informative and 36 species-specific orthozymes. Of the latter, 23 are monomophic and are suitable for identification of V. benghalensis, V. palaestina, V. dumetorum, V. pisiformis, V. sylvatica, V. onobrychioides, V. cappadocica, V. cretica, V. articulata, V. tetrasperma, V. ervilia, V. hirsuta and V. loiseleurii. Polymorphism with heterozygous and homozygous isozyme genotypes was found for V. cracca, V. tenuifolia, V. ochroleuca, V. villosa, V. sylvatica, V. cassubica, V. sparsiflora, V. megalotropis, V. altissima, V. onobrychioides, V. cassia, V. cretica and L. heterophyllus, reflecting outcrossing in these species. By contrast, V. benghalensis, V. palaestina, V. disperma, V. dumetorum, V. pisiformis, V. orobus, V. pauciflora, V. tetrasperma and V. loiseleurii had only homozygous isozyme genotypes at polymorphic loci. Isozyme-based phylogenetic trees are presented. Sections Cracca, Ervum, Pedunculatae and Lenticula of traditional taxonomy are monophyletic groups, whereas sections Oroboideae (= Vicilla) and Panduratae appear polyphyletic and section Cassubicae is split into two species-couples linked at a low level of support. Treatment of ervoid species in a separate subgenus Ervum is not supported because of its polyphyly.

Origin of Weedy Rice Grown in Bhutan and the Force of Genetic Diversity

In Bhutan, weedy rice (Oryza sativa L.) was grown together with cultivated rice on terraced paddy fields lower than 2620 m above sea level. Seeds of cultivars and weedy strains were collected at 22 collection sites located from 1000 to 2620 m above sea level. Cultivars with round seeds were frequently found in fields higher than 2250 m, and those with slender seeds in fields lower than 1630 m. All cultivars and weedy strains were divided into indica or japonica types by isozyme (multi-locus) and morpho-physiological (multicharacter) analyses. Japonica cultivars predominated in highland; Indica cultivars predominated in lowland. Plastid type was confirmed by the length polymorphism for the ORF100 region. The japonica cultivars carried non-deletion type ORF100. The indica cultivars carried deletion type ORF100. In contrast, weedy strains showed discrepancy in the combination of the nuclear and cytoplasm types. An intermediate type was found in weedy strains for isozyme genotypes. A recombinant type, which has indica genotypes for isozyme analysis with japonica cytoplasm, and vice versa, was frequently found in weedy strains. These findings suggested that weedy strains would be generated by natural hybridization between indica and japonica. Further, they might fail to recombine their prior genotypes sets for isozyme and cytoplasm. Morphophysiological characters did not show such a tendency. Morpho-physiological analysis of highland plants, in particular, showed indica cultivars and weedy strains with japonica cytoplasm. Hypervariable nuclear microsatellite analysis was then used to compare cultivars and weedy strains. Identical alleles were shared between indica and japonica, and also between cultivars and weedy strains. This suggested that there was gene-flow resulting from natural hybridization.

Restricted seed contribution and clonal dominance in a free‐floating aquatic plant Utricularia australis R. Br. in southwestern Japan

Utricularia australis R. Br. is an aquatic angiosperm species common in natural and irrigation ponds in temperate regions. This species reproduces both sexually and vegetatively, but in southwestern Japan the occurrence of male‐sterile populations, in which plants produce no pollen and propagate only vegetatively, has been recorded. We studied the reproductive contribution of seeds in normal pollen‐producing populations using isozyme analyses, a pollination experiment under culture and field observations. Although seedlings obtained from controlled mating indicated segregation of isozyme, polymorphism of the isozyme genotype was detected mainly among populations, but rarely within each pond population. This suggested clonal dominance and rarity of seed or seedling survival in natural populations. In the pollination experiment, the mean seed set ratio in cross‐pollination between plants of the same isozyme genotype (7.6%) did not differ significantly from self‐pollination (7.6%), but was lower than cross‐pollination between plants of different genotypes (45.7%). The low ratio in crossing between the same genotype plants was ascribed to the clonality of the parents. In general, these results corresponded with the low ratios in seed setting observed in natural populations (7.9–13.7%). All the male‐sterile populations we surveyed showed the same genotype, thus male sterility in the study area was considered to have the same origin.

Novel Genotypes in Phytophthora infestans Populations in Hungary

Twenty-seven isolates of Phytophthora infestans collected in Hungary in 2001 were tested for mating type, response to metalaxyl, isozyme genotype at glucose-6-phosphate isomerase (Gpi) and peptidase A (Pep) loci and nuclear DNA fingerprints with probe RG57. The ratios of the mating types A1 to A2 were 5:6 and 9:7 among isolates from potato and tomato, respectively. Seventeen isolates were sensitive to metalaxyl, 1 isolate responded intermediately and 9 isolates were resistant. No novel combinations of isozyme alleles were found; all isolates were Gpi 100/100, and genotypes at the Pep locus were 96/96 (63%), 83/96 (11%) and 100/100 (26%). In contrast, all of the 22 RG57 fingerprints exhibited patterns that have not been reported in Hungary before. On the basis of combined traits, 22 multilocus genotypes, unnoted elsewhere in Europe, were constructed among the 27 isolates analysed. These results indicate that variability in the Hungarian P. infestans populations is likely due to local events (asexual and sexual interactions) rather than migration from other countries.

Origin of cytoplasm substituted rice cultivars found in Japan.

Genetic variation of Japanese rice cultivars were examined. Five of 450 lowland cultivars and another five of 200 upland cultivars were determined as the indica type by using isozyme genotypes and the remainder were of the japonica type. The major characteristics of these indica cultivars, revealed a slender shape of grains, a short apiculus hair length, a positive allele for Ph reaction, and allele-3 for the Pgd1 locus. Three of these indica cultivars showed a non-deletion ORF100, which is essential to the japonica-type plastid. The plastid subtype identity (PS-ID) sequences of these plastids is 6C7A, which is also a japonica-specific repeat unit. Thus, these cultivars were concluded to be naturally generated cytoplasm substituted lines. These plastids were introduced into a indica genetic background from japonica cultivars grown elsewhere. The rest of the indica cultivars revealed a deletion-type ORF100 and plastid subtype 8C8A, both of which are indica-specific. These cultivars carried indica-type allelic constitutions for diagnostic isozyme loci. However, other characters were identical to the cytoplasm-substituted cultivars in Japan. In East and Southeast Asia, cultivars carrying a indica-type nuclear genotype with a japonica-type plastid are restricted to Aus cultivars in the Bengal region. Genetic and historical records suggest that Japanese indica cultivars and the Aus cultivars are closely related. The Aus cultivars acquire necessary genetic constitutions from both indica and japonica cultivars through naturally occurring out-crossing to adapt to a particular cultivation condition in the region. The wide adaptability enabled them to be introduced into a northern region like Japan.

Importance of genetic characterization and conservation of plant genetic resources: The breeding system and genetic diversity of wild soybean (Glycine soja)

Abstract Plant genetic resources play an important role in the improvement of cultivated plants. To characterize and evaluate the ecological and reproductive features of wild soybean (Glycine soja Sieb. et Zucc.), which is the most probable ancestor of cultivated soybean (G. max (L) Merr.), the breeding system and genetic diversity of G. soja were investigated. The extent of natural cross‐pollination of G. soja was estimated in four populations along the Omono River in Akita Prefecture, Japan by examining allozyme variation. Although it has been previously believed that G. soja is autogamous, as is cultivated soybean, the mean multilocus outcrossing rate (tm) estimate was 13%. These values are much higher than the outcrossing rate previously reported for both G. soja and G. max. Frequent visits by honeybees and carpenter bees to flowers were also observed, which supported this conjecture. Furthermore, to evaluate the genetic variation of G. soja as a genetic resource, the genetic structure of 447 populations over Japan were analyzed. Wild soybean populations had a higher degree of variation of isozyme loci. The GST coefficient of gene differentian values among the sites within the district were particularly high, revealing that the isozyme genotype was greatly different among site populations and homogeneous within the sites. The genetic differentiation among nine districts was observed in the allele frequencies of a few loci, indicating that geographic isolation in the wild soybean population was effectively created through the distance between the districts. The difference in the allele frequency among the districts may be produced under genetic drift. Finally, the importance of the preservation of natural plant populations and the habitats of wild progenitors (i.e. the in situ conservation of plant genetic resources) was emphasized.

Genetic Resources of Primitive Upland Rice in Laos1

Primitive upland cultivars (Oryza sativa L.) were collected in northern Laos. One-hundred-thirty-two cultivars were collected in upland fields at 27 sites. Morphological and physiological traits were recorded. The materials were classified into indica and japonica types based on isozyme genotypes. We classified 106 japonica, 16 indica, two intermediate, and eight heterozygous cultivars. The japonica cultivars were characterized by glabrous hulls and sticky grains. Only two out of 16 indica cultivars were glabrous. The heterozygotes were estimated to be generated by out-crosses between japonica and indica cultivars in upland fields. The intermediate type would be the progeny of such heterozygotes. Higher polymorphism was found at two isozyme loci—Amp1 and Est2—among the japonica cultivars. Genotypic frequencies differed between populations collected from upland fields along roads and along a branch of the Mekong river. Such differences would be caused by different origins of these two populations. In this report, isozymes were indicated as valuable markers to recognize the cultivars to be of independent stock.

Different maternal origins of Japanese lowland and upland rice populations.

Plastid subtype ID (PS-ID) sequences were determined from sequence data based on CA repeats between genes rpl16 and rpl14 in Japanese lowland and upland cultivars. The PS-ID sequences of Japanese rice cultivars showed that there are different maternal origins between lowland and upland cultivars. One subtype, 6C7A, of PS-ID sequences was predominant in all but one Japanese lowland cultivar and carried a combination of the indica-specific subtype 8C8A and japonica-specific nuclear markers for the isozyme genotype. It is probably a nuclear-cytoplasmic recombinant resulting from natural out-crossing and succeeding self-pollination. The origin of the plastid was re-confirmed by the existence of an indica-specific deletion in the plastid genome. In contrast, the Japanese upland cultivars showed two subtypes, 7C6A and 6C7A, of PS-ID sequences. An upland-specific isozyme allele as a nuclear marker was equally predominant in cultivars carrying each subtype. The existence of these particular upland-specific nuclear and cytoplasmic genotypes suggests that the origin of Japanese upland cultivars is different from that of Japanese lowland cultivars. Cultivars carrying the upland-specific nuclear genotype are common in Southeast Asia, but the combination of the upland-specific nuclear and cytoplasmic genotypes which is the same as the Japanese upland predominant type was found in cultivars only in Taiwan and Indonesia. Japanese upland cultivars are closely related to those cultivars.

A tetraploid hybrid plant from 4x × 2x crosses in Vitis and its origin

To study the origin of unreduced (2n) gametes in diploid Vitis cultivars, we surveyed the occurrence of tetraploid hybrid seedlings from 40 interploid crosses with five tetraploid and seven diploid cultivars. A total of 250 seedlings from the interploid crosses were established through embryo culture and by seed sowing. In 20 2x × 4x crosses, no tetraploid hybrid seedlings were derived from 8,902pollinations. In 20 4x × 2x crosses, two tetraploid hybrid seedlings were obtained from 8,057 pollinations. Investigation of isozyme genotypes of the two tetraploid seedlings using three variable enzyme systems indicated that one of the two seedlings resulted from the union of a diploid egg with a 2n male gamete and that failure of second meiotic division resulted in the formation of the 2n male gamete. The percentage of giant pollen grains in `Muscat Bailey A', a pollen parent of the tetraploid seedling, was relatively high(about 5.9%) and 10.9% of the giant pollen grains germinated on agar medium. These results suggested that 2n pollen of diploid cultivars are useful for breeding tetraploid grape.

Glutathione transferase isozyme genotypes in patients with prostate and bladder carcinoma.

Genotype distributions for GSTP1, GSTM1, and GSTT1 were determined in 91 patients with prostatic carcinoma and 135 patients with bladder carcinoma and compared with those in 127 abdominal surgery patients without malignancies. None of the genotypes differed significantly with respect to age or sex among controls or cancer patients. In the group of prostatic carcinoma patients, GSTT1 null allele homozygotes were more prevalent (25% in carcinoma patients vs. 13% in controls, Fisher P =0.02, chi2 P=0.02, OR=2.31, CI = 1.17-4.59) and the combined M1-/T1 -null genotype was also more frequent (9% vs. 3%, chi2 P=0.02, Fisher P = 0.03). Homozygosity for the GSTM1 null allele was more frequent among bladder carcinoma patients (59% in bladder carcinoma patients vs 45% in controls, Fisher P=0.03, chi2 P=0.02, OR=1.76, CI=1.08-2.88). In contrast to a previous report, no significant increase in the frequency of the GSTP1b allele was found in the tumor patients. Except for the combined GSTM1-/ T1-null genotype in prostatic carcinoma, none of the combined genotypes showed a significant association with either of the cancers. These findings suggest that specific single polymorphic GST genes, that is GSTM1 in the case of bladder cancer and GSTT1 in the case of prostatic carcinoma, are most relevant for the development of these urological malignancies among the general population in Central Europe.

Clonal diversity within and among introduced populations of the apomictic vine Bryonia alba (Cucurbitaceae)

Bryonia alba L. (Cucurbitaceae) is a herbaceous Eurasian vine that predominantly reproduces clonally (asexually) through apomixis. We assessed the magnitude and distribution of clonal diversity within and among 23 recently established populations of B. alba in its new range in the western United States, based on the distribution of multilocus isozyme genotypes. Fifty-two unique clones were detected: 30 in the nine populations from eastern Washington and northern Idaho, and the remaining 22 in 14 populations from western Montana, northern Utah, and southern Idaho. On average, each population of B. alba was composed of 6.4 clones, and the proportion of distinguishable clones was 0.275. Multilocus diversity (D) was 0.632 and multilocus evenness (E) was 0.556. Twenty-six of 52 clones (50%) were restricted to a single population, and, on average, each clone occurred in 2.83 populations. Compared with other clonally reproducing plant species, this vine possesses moderate to high levels of clonal diversity in its new range in the western United States. This diversity appears to be a consequence of the events associated with its introduction (including multiple introductions), founder effects, and the proportion of sexual to apomictic reproduction within populations.Key words: invasive vine, apomixis, multilocus genotypes, clonal diversity and evenness, Bryonia alba, Cucurbitaceae.

Isozymes of the Isoetes riparia Complex, II. Ancestry and Relationships of Polyploids

Abstract Isoetes riparia is a complex tetraploid taxon of eastern North America that in its widest sense includes a diversity of morphotypes variously segregated, or included in a single polymorphic species. Although the diploid ancestry of I. riparia has been proposed as I. echinospora × engelmannii, it is uncertain whether this ancestry applies to all elements of the complex, here broadly defined to encompass three segregates of I. riparia and the recently described I. hyemalis (referred to informally as riparia, canadensis, saccharata and hyemalis). To help clarify relationships among these tetraploids, isozyme genotypes for 13 loci coding eight enzymes, were determined for 16 populations representing all four segregates. Duplicate gene expression, evidenced as fixed heterozygosity consistent with allopolyploidy, characterized the tetraploid genotypes of all taxa. Although exhibiting modest to no variation within populations, populations were highly differentiated, most showing a unique combination of ...

Segregation of isozymes in selfed progenies of a synthetic amphidiploid between Solanum integrifolium and S. melongena.

Isozyme and cytogenetic analyses were performed on selfed progenies of a synthetic amphidiploid between scarlet eggplant, Solanum integrifolium (= S. aethiopicum),and eggplant, Solanum melongena `DMP', for estimating genetic uniformity. Isozymes in the 379 examined seedlings segregated into five genotypes (phenotypes) each at the four loci examined, Pgd-2 of phosphogluconate dehydrogenase (E.C.1.1.1.43), Idh-2 of isocitrate dehydrogenase (E.C.1.1.1.41), Pgm-2 of phosphoglucomutase (E.C.2.7.5.1)and Skdh-1 of shikimate dehydrogenase (E.C.1.1.1.25), indicating that the selfed seedlings were not genetically uniform. Most of the examined 15 selfed seedlings exhibited a somatic chromosome number of 48, that is the same number of the synthetic amphidiploid, whereas isozyme genotypes among them were variable. It is suggested that the segregation of isozymes was not caused by variation of chromosome number but by genetic segregation of isozyme genes. The genome of the synthetic amphidiploid was indicated to be unstable.