A comparative cytogenetic study of the tetraploid oat species with the A and C genomes: <Emphasis Type="Italic">Avena insularis,A. magna</Emphasis>, and <Emphasis Type="Italic">A. murphyi</Emphasis>

C-banding of chromosomes and in situ hybridization with the probes pTa71 and pTa794 were used for a comparative cytogenetic study of the three tetraploid oat species with the A and C genomes: Avena insularis, A. magna, and A. murphyi. These species were similar in the structure and C-banding patterns of several chromosomes as well as in the location of the loci 5S rRNA genes and major NOR sites; however, they differed in the number and localization of minor 45S rDNA loci as well as in the morphology and distribution of heterochromatin in some chromosomes. According to the data obtained, A. insularis is closer to A. magna, whereas A. murphyi is somewhat separated from these two species. Presumably, all the three studied species originated from the same tetraploid ancestor, and their divergence is connected with various species-specific chromosome rearrangements. The evolution of A. murphyi is likely to have occurred independently of the other two species.     

A CYTOGENETIC ANALYSIS OF CERTAIN POLYPLOIDS IN GRINDELIA (COMPOSITAE)

Two diploid taxa, Grindelia procera and G. camporum, and 3 tetraploid ones, G. camporum, G. hirsutula, and G. stricta, have been studied to ascertain their interrelationships. Meiosis in diploid parental strains was regular, the common chromosome configuration being 5 rod bivalents and 1 ring bivalent. The average chiasmata frequency per chromosome was 0.60. Pollen fertility was about 90% in all strains examined. Diploid interspecific hybrids had normal meiosis with an average chiasmata frequency of 0.56 per chromosome. No heterozygosity for inversions or interchanges was detected, and pollen fertility was above 85%. Meiosis in parental tetraploid strains was characterized by the presence of quadrivalents in addition to a complementary number of bivalents. The average chiasmata frequency per chromosome was 0.59 and pollen fertility was generally about 80%. Tetraploid interspecific hybrids also had quadrivalents, normal meiosis, and high pollen fertility. Close genetic relationships between the diploids and between the tetraploids are indicated, and geographical, ecological, and seasonal barriers to gene exchange exist. Attempts to obtain hybrids between diploids and tetraploids were successful in a few cases. The hybrids were tetraploid and had normal meiosis and fertility similar to parental and F₁ tetraploids. Their origin was by the union of unreduced gametes of the diploid female parent and normal pollen from the tetraploid parent. On the basis of chromosome homology, normal meiosis, plus high fertility exhibited in the diploid, tetraploid, and diploid X tetraploid interspecific hybrids, these species of Grindelia are considered to be a part of an autopolyploid complex. Gene exchange between diploids and diploids, tetraploids and tetraploids, and diploids and tetraploids is possible. Tetraploid G. camporum may have originated by hybridization between G. procera and diploid G. camporum with subsequent doubling of chromosomes and selection for the combined characteristics of the diploids.     

Habitat differentiation, hybridization and gene flow patterns in mixed populations of diploid and autotetraploid Dactylorhiza maculata s.l. (Orchidaceae)

Detailed ecological, morphological and molecular analyses were performed in mixed populations of diploid and autotetraploid Dactylorhiza maculata s.l. in Scandinavia. Comparisons were made with pure populations of either diploid ssp. fuchsii or tetraploid ssp. maculata. It was shown that mixed populations are the result of secondary contact between ssp. fuchsii and ssp. maculata. No patterns of recent and local autopolyploidization were found. Morphology and nuclear DNA markers (internal transcribed spacers of nuclear ribosomal DNA) showed that diploids and tetraploids from mixed populations have similar levels of differentiation to diploids and tetraploids from pure populations. Vegetation analyses, as well as analyses of environmental variables, revealed that diploid and tetraploid individuals in mixed populations are ecologically well differentiated on a microhabitat level. Diploids and tetraploids in pure populations have wider ecological amplitudes than they do in mixed populations. Triploid hybrids grew in intermediate microhabitats between diploids and tetraploids in the mixed populations. Plastid DNA markers indicated that both diploids and tetraploids may act as the maternal parent. Based on morphology and nuclear markers triploids are more similar to tetraploids than to diploids. There were indications of introgressive gene flow between ploidy levels. Plastid markers indicated that gene flow from diploid to tetraploid level is most common, but nuclear markers suggested that gene flow in opposite direction also may occur. Similar patterns of differentiation and gene flow appeared in localities that represented contrasting biogeographic regions. Disturbance and topography may explain why hybridization was slightly more common and the differentiation patterns somewhat less clear in the Scandinavian mountains than in the coastal lowland. An erratum to this article can be found at     

NOVEL FLAVONOIDS AND RETICULATE EVOLUTION IN THE PHLOX PILOSA–P. DRUMMONDII COMPLEX

The nonanthocyanin flavonoids of the diploids Phlox pilosa ssp. pilosa and P. drummondii ssp. mcallisteri and their putative allotetraploid derivatives P. aspera and P. villosissima were characterized by chromatographic and spectrophotometric analyses. Synthetic F₁ diploid hybrids were also examined. All but one of the compounds encountered were glycoflavone (C-glycosyl) derivatives of either apigenin or luteolin. Although some compounds were present in all four taxa, most were species-specific. Together, the diploids exhibited 15 different flavonoids. Half of each tetraploid ensemble was composed of novel flavonoids, compounds not manifest in either diploid ancestor. In total, the tetraploids produced seven different novel flavonoids. Five novel compounds differed from particular diploid compounds only in number or kind of oxygen-linked sugar substituents. Ostensibly, these five are either identical to diploid biosynthetic intermediates or share common precursors with related diploid end products. The mode of origin of all novel compounds is discussed.     

AUTOTETRAPLOIDY IN HAPLOPAPPUS SPINULOSUS HYBRIDS: EVIDENCE FROM NATURAL AND SYNTHETIC TETRAPLOIDS

Haplopappus spinulosus (Asteraceae) is a herbaceous, perennial weed common throughout the western Great Plains of North America and includes both diploid and tetraploid populations. A number of populations in southeastern Colorado were analyzed cytogenetically and morphologically for two reasons. First, initial observations during a routine population survey showed they were morphologically intermediate between the diploid subspecies glaberrimus and spinulosus, suggesting they might have arisen via hybridization. Second, cytological examination revealed that they were tetraploid. Because there was indication of hybrid origin, it was of interest to determine whether the populations were behaving as autopolyploids or segmental allopolyploids. The distinction between these two polyploid types is not easily made since both are likely to form multivalents at meiosis, but equations derived from a model proposed by Jackson and Hauber (1982) have made it possible to determine statistically whether a tetraploid individual is behaving meiotically as an autotetraploid. Meiotic configuration frequencies at diakinesis were determined for each tetraploid plant sampled, and observed frequencies were compared to those expected for an autotetraploid having the same maximum number of chiasmata per bivalent, chiasma frequency and chromosome number. In general, the meiotic behavior of the tetraploids was no different from that expected for autoploids. The initial hypothesis that the populations were derived from hybridization was tested by a detailed cytogenetic and morphological study of the presumed parental subspecies, F, hybrids, and natural putative hybrids. The evidence supports the hypothesis that the natural autotetraploids arose from the hybridization of ssp. glaberrimus and ssp. spinulosus.     

Cytogenetics of the hybrid Gilia millefoliata × achilleaefolia

Summary Gilia millefoliata andG. achilleaefolia, two annual diploid (n=9) species ofPolemoniaceae, crossed readily in certain combinations but not in others. The F₁ hybrids were vigorous but sterile. They gave rise, apparently by the union of unreduced gametes, to an F₂ generation of tetraploids, which were mostly fertile.Chromosome pairing in the hybrids varied markedly according to the state of nutrition of the plants. The F₁ hybrids formed fewer clear diakinesis figures, fewer bivalents, fewer chiasmata per bivalent, and more attenuated or stretched bivalents when grown in 2 pots of sand than when grown in rich soil (Table 3). A pot-bound allotetraploid individual derived from this hybrid showed the same meiotic irregularities as the starved F₁s until irrigated with a solution of mineral nutrients, after which its chromosomes paired regularly in bivalents (Table 2, Fig. 38).The capacity of the F₁ hybrids to produce polyploids also differed strikingly in the two cultures. The rate of polyploidy of the stunted sand-grown hybrids was 2381 viable tetraploid zygotes per million flowers, while the corresponding figure for the luxuriant field hybrids was only 2.7 per million flowers.For the production of polyploid progeny by diploid parents — a process which should be clearly distinguished from normal fertility — the termpolyploidy rate is proposed. It is suggested that starvation of a structural hybrid may sometimes increase its polyploidy rate by reducing chromosome pairing to the point where restitution nuclei and hence unreduced gametes can be formed.     

Inflorescence culture of wheat-Agropyron hybrids: Callus induction,plant regeneration,and potential in overcoming sterility barriers

Segments of young inflorescences of Triticum aestivum cv. Chinese Spring (CS), its F₁ hybrids with Agropyron trachycaulum and A. scirpeum and backcross derivatives with A. yezoense, A. intermedium and A. junceum, and of a A. yezoense x T. aestivum cv. Wichita hybrid were cultured. Different parts of young spikelets of A. trachycaulum x CS F₁ and A. yezoense x Wichita F₁ 's were also cultured. Percent callus induction was lower in wheat than in the wheat-Agropyron hybrids or backcross derivatives. Percent callus induction from different organs in both hybrids was in the descending order of whole spikelet, spikelet without glumes, rachis, and glumes. No plants could be regenerated from calli of wheat and backcross derivatives except those of CS x A. intermedium combination. Callus induction in hybrids varied from 54 to 84% and plant regeneration from 14 to 31%. The regenerants required no vernalization. Variants including one with top-dense spikes and another with elongated spikelets were recovered. Out of eight A. trachycaulm x CS hybrid regenerants, one had anthers and stigma as opposed to neutral flowers of the original hybrid.     

Chloroplast DNA variation between the common cultivated potato (Solanum tuberosum ssp. tuberosum) and several South American relatives

Summary Chloroplast DNA (ctDNA) from the tuberbearing Solanum species tuberosum, vernei, phureja, and chacoense has been compared by restriction endonuclease analysis. Digestion by Hind III or Xba I reveal no differences, but digestion with Bam HI and Eco RI reveals minor differences in the ctDNA among these species. The ctDNA restriction patterns of the tetraploid common cultivated potato of North America and Europe, S. tuberosum ssp. tuberosum and the South American tetraploid, S. tuberosum ssp. andigena are identical for all four restriction endonucleases. These data suggest that ssp. tuberosum and ssp. andigena contain similar ctDNA and therefore may share a common ancestor, or direct lineage. The ctDNA restriction patterns of S. vernei and S. chacoense are identical for all four restriction endonucleases, and S. phureja ctDNA, can be distinguished from the other diploid ctDNAs by digestion with Bam HI. None of the diploids analyzed contain ctDNA identical to the tetraploids and therefore either did not contribute their chloroplast genomes to the evolution of the tetraploids, or the ctDNA has diverged since this evolutionary event. The ctDNAs studied did not contain restriction polymorphisms which could be correlated to cytoplasmic male sterility in Solanum. This is the first demonstration of ctDNA diversity in the tuber-bearing Solanum species.     

Hybridization between Triticum aestivum L. and Agropyron michnoi Roshev.

Summary Intergeneric hybrids between Triticum aestivum cv Chinese Spring (2n=6x=42, AABBDD) and Agropyron michnoi Roshev. (2n=4x=28, PPPP) were obtained by embryo culture. Their spike characteristics were similar to those of common wheat but, unlike their parents, they were long-awned. The average meiotic chromosome pairing at MI of F₁ hybrids was: 6.39 I +3.75 rodII+8.64 ringII+0.81 III+0.30 IV+0.04 V, the bivalent and multivalent formation of which was much higher than expected from the genomic formulae. It is especially worthwhile to note that the F₁ hybrids were self-fertile, self set being 0.15%, and seeds were easily obtained from the backcross of f₁ plants with hexaploid and tetraploid wheats; here the seed set was more than 20.0%. The polyploid taxa and the position of A. Michnoi in Agropyron are discussed.     

MORPHOLOGY OF SOME MAIZE-TRIPSACUM HYBRIDS

Diploid (2n = 20) and tetraploid (2n = 40) Zea mays L. were crossed with diploid (2n = 36) and tetraploid (2n = 72) Tripsacum dactyloides (L.) L. to produce a series of hybrids combining different numbers of haploid genomes from each parent. Eight hybrid groups and three parental groups were studied morphologically. Twenty-nine quantitative characters were recorded for each sample. Data were analyzed by univariate analysis of variance, multivariate analysis of variance, and discriminant function analysis, in an attempt to evaluate hybrid differences objectively and determine which morphological characters contribute statistically to group separation. The overall MANOVA F test was significant, establishing the presence of real differences between the hybrids; discriminant function analysis indicated that the percent of paired pistillate spikelets/cupule in the lateral inflorescence was the main variable which differentiated hybrids. Duncan's Multiple Range Tests for significant differences between means were applied to five variables contributing maximally to group discrimination, using the appropriate univariate ANOVAs. Pronounced maize-like attributes of backcross hybrids, as compared with corresponding F₁'s possessing similar genome constitutions, gave possible evidence of gene transfer between Zea mays and Tripsacum during backcrossing to maize.     

Species relationships in the avenae

In order to assess the genome homologies of a number of diploid and tetraploid species of Avena, two meiotic characters — mean chiasmata per cell and frequency of types of pairing configurations — have been studied in the species and in a number of diploid, triploid and tetraploid hybrids. The results indicate extensive structural differentiation of the genome of A. longiglumis from that which is common to the other diploids A. strigosa, A. brevis, A. hirtula, A. glabrata and A. wiestii. Structural differentiation is found also between the genomes of the three tetraploids A. vaviloviana, A. abyssinica and A. barbata. Chromosome pairing in triploid hybrids indicates the similarity of the genome in the A. strigosa group to one of those in the tetraploids and a partial but significant affinity with the other. These data, though derived from a very limited range of genotypes, lead to the conclusions that (a) structural differentiation of chromosomes may be common in the genus and important in its evolution, and (b) that current ideas on evolution of the polyploid species through simple allopolyploidy are unlikely to be true. The polyploids probably have a more complex origin in which autopolyploids or near autopolyploids and structural change of chromosomes have played a part.     

Successive domestication and evolution of the Andean potatoes as revealed by chloroplast DNA restriction endonuclease analysis

Five chloroplast DNA (ctDNA) types (W, T, C, S, and A) have previously been identified in the Andean tetraploid cultivated potatoes (Solanum tuberosum ssp. andigena) and three types (C, S, and A) in diploid cultivated potatoes (S. stenotomum). In this study, ctDNA types were determined for an additional 35 accessions of S. stenotomum and 97 accessions of putative ancestral wild species (15 of S. brevicaule, 26 of S. bukasovii, 4 of S. candolleanum, 25 of S. canasense, 17 of S. leptophyes, and 10 of S. multidissectum). The first five ctDNA types were also identified in S. stenotomum. The wild species were also polymorphic for ctDNA types except for S. brevicaule, which had only W-type ctDNA. T-type ctDNA was not found in any of the wild species and could have originated from W-type ctDNA after S. stenotomum arose. The other types of ctDNA evolved in wild species. The geographical distribution of each ctDNA type indicated that A-type ctDNA arose in central Peru and T-type ctDNA in the Bolivia-Argentine boundary. It is implied that potatoes were successively domesticated and that, in parallel, several wild species were differentiated from time to time and place to place from the ancestral species complex. Subsequent sexual polyploidization formed a wide ctDNA diversity among the Andean tetraploid potatoes, and selection from them formed the limited ctDNA diversity found in Chilean tetraploid potatoes (ssp. tuberosum).Hawkes' (1990) classification system is tentatively adopted throughout this text. Synonyms indicated by Hawkes (1990) for the species names described by various authors are presented in parentheses.     

The origin and meiotic behaviour of hexaploid northern wheatgrass (Agropyron dasystachyum)

The occurrence of hexaploid (2n = 6x = 42) forms in some otherwise natural tetraploid populations of Agropyron dasystachyum (2n = 4x = 28) was cytologically detected and studied. The hexaploid plants are morphologically similar to the tetraploids except for a small reduction in the anther size. The general survey of chromosome numbers of natural Northern Wheatgrass (A. dasystachyum 2n = 4x = 28) populations derived from eight different regions of Alberta indicated that the occurrence of hexaploid variants was not restricted to a single locality. A comparative study of chromosome pairing in the natural and the synthetic hexaploids revealed that the naturally occurring 42-chromosomed plants of A. dasystachyum originated as a result of fertilization between unreduced (SSHH) and the natural (SH) gametes, both coming from the tetraploid form of A. dasystachyum. Based on chromosome pairing, the genomes of the natural hexaploid A. dasystachyum have been designated as SSSHHH. The natural hexaploids appear to intercross among themselves and also with tetraploids producing euploid and aneuploid hybrids. The possible evolutionary significance of these findings is briefly discussed.     

Allozyme variation in Japanese species ofChionographis (Liliaceae)

Allozyme variation was examined in three diploid taxaChionographis japonica var.japonica, var.kurokamiana, andC. koidzumiana and three tetraploid taxaC. japonica var.kurohimensis, ssp.hisauchiana, and ssp.minoensis. Results show thatC. japonica var.kurokamiana is genetically closer toC. koidzumiana than to var.japonica. In the tetraploid taxa, fixed heterozygosities were found at several loci, and this supports the hypothesis that these taxa are allotetraploids. Furthermore, the tetraploid taxa have many unique alleles not found in the diploid taxa. This suggests that sufficient time has passed since the origin of tetraploids for new mutations to have been fixed.     

Autopolyploidy in Dactylis glomerata L.: further evidence from studies of chloroplast DNA variation

Summary Chloroplast DNA variation has been used to examine some of the maternal lineages involved in the evolution of the intraspecific polyploid complex, Dactylis glomerata L. Diploid (2x) and tetraploid (4x) individuals were collected from natural populations of the subspecies glomerata (4x), marina (4x) and lusitanica (2x), as well as from sympatric 2x/4x populations of the Galician type. Digestion of their ctDNA with 11 restriction endonucleases revealed enough variation to characterise three ctDNA variants, designated MBMK, MBmK and mBMK. The distribution of these ctDNA variants reflects different stages in their spread among the populations. The MBMK ctDNA variant predominated at both ploidy levels in subspecies glomerata, lusitanica and marina, and in recent tetraploid Galician/glomerata hybrids. The MBmK variant was detected in a single tetraploid individual and probably results from a relatively recent mutation. Fixation of the mBMK minority variant in the diploid and tetraploid Galician populations adds to the evidence concerning the possible origin of the Galician tetraploids. It means that the Galician diploids were maternal ancestors of the tetraploids. This result complements evidence from earlier studies based on morphology or biochemical markers, and reduces the likelihood that the tetraploids arose by hybridisation between an ancient Galician diploid and an alien tetraploid. It is, however, consistent with a true autopolyploid origin of the tetraploids.     

Evidence for autotetraploidy associated with reproductive isolation in Saccharomyces cerevisiae: towards a new domesticated species

Partial or whole‐genome duplications have played a major role in the evolution of new species. We have investigated the variation of ploidy level in a panel of domesticated strains of Saccharomyces cerevisiae coming from different geographical origins. Segregation studies and crosses with tester strains of different ploidy levels showed that part of the strains were well‐balanced autotetraploids displaying tetrasomic inheritance. The presence of up to four different alleles for various loci is consistent with a polyploidization mechanism relying on the fusion of two nonreduced meiospores coming from two S. cerevisiae strains. Autotetraploidy was also in accordance with karyotype and flow cytometry analyses. Interestingly, most bakery strains were tetraploids, suggesting a link between ploidy level and human use. The null or drastically reduced fertility of the hybrids between tetraploid and diploid strains indicated that domesticated S. cerevisiae strains are composed of two groups isolated by post‐zygotic reproductive barriers.     

Analysis of meiosis in triticale (XTriticosecale Wittmack) X rye (Secale cereale L.) F1 hybrids at three ploidy levels

Summary Triticales (XTriticosecale Wittmack) at three ploidy levels (8x, 6x, 4x, x=7) were crossed with diploid rye (Secale cereale L.) to produce a solitary hypopentaploid hybrid (2n=32), and a number of tetraploid (2n=4x=28) and triploid (2n=3x=21) hybrids. The hybrids exhibited a morphology which was intermediate between the parents. The number of bivalents ranged from 1–7 (4.65 per cell) in hypopentaploid, from 2–12 (7.13 per cell) in tetraploid and from 4–9 (6.84 per cell) in triploid hybrids. In 4x and 3x hybrids, trivalents and quadrivalents were also observed at low frequencies (range 0–1; mean 0.01–0.03 per cell). Chiasmata frequency was highest in triploid hybrids (12.44 per cell), lowest in hypopentaploid (5.37 per cell) and intermediate in tetraploids (10.54 per cell). More than 7¹¹ were found in 39.7% pollen mother cells (PMC's) in the 4x hybrids and in 5.0% PMCs in 3x hybrids. It is concluded that an increase in the relative proportion of wheat chromosomes in the hybrids had a slight suppression effect on homologous as well as homoeologous pairing of rye chromosomes. Contrary to this, the relative increase in rye complement promoted homoeologous pairing between wheat chromosomes. In triploid hybrids, the chiasmata frequency as well as the c value were the highest, suggesting that in tetraploid hybrids rye chromosomes had a reduced pairing (low frequency of ring bivalents).     

Cytogenetic studies in the genus Zea

Summary New cytological evidence supporting x = 5 as the basic chromosome number of the genus Zea has been obtained as a consequence of our analysis of the meiotic configurations of Zea mays ssp. mays, Z. diploperennis, Z. perennis and of four F₁ artificial interspecific hybrids. Z. mays ssp. mays (2n = 20) presents regular meiosis with 10 bivalents (II) and is considered here as a typical allotetraploid (A₂A₂B₂B₂). In Z. diploperennis (2n = 20) 10II are formed in the majority of the cells, but the formation of 1III + 8II + 1I or 1III + 711 + 3I in 4% of the cells would indicate its segmental allotetraploid nature (A₁A₁B₁B₁). Z. perennis (2n = 40) had 5IV + 10II in 55% of the cells and would be considered as an auto-allooctoploid (A₁A₁A'₁A'₁C₁C₁C₂C₂). Z. diploperennis x Z. mays ssp. mays (2n = 20) presents 10II in ca. 70% of the cells and no multivalents are formed. In the two 2n = 30 hybrids (Z. mays ssp. mays x Z. perennis and Z. diploperennis x Z. perennis) the most frequent meiotic configuration was 5III + 5II + 5I and in 2n = 40 hybrid (Z. diploperennis x Z. perennis) was 5IV + 10II. Moreover, secondary association was observed in the three abovementioned tetraploid taxa (2n = 20) where one to five groups of two bivalents each at diakinesis-metaphase I was formed showing the affinities between homoeologous genomes. The results, as a whole, can be interpreed by assuming a basic x = 5 in this polyploid complex. The main previous contributions that support this working hypothesis are reviewed and its phylogenetic implications studied are discussed.     

Meiotic chromosome pairing behaviour of natural tetraploids and induced autotetraploids of Actinidia chinensis

Key message

Non-preferential chromosome pairing was identified in tetraploid Actinidia chinensis and a higher mean multivalent frequency in pollen mother cells was found in colchine-induced tetraploids of A. chinensis compared with naturally occurring tetraploids.

Abstract

Diploid and tetraploid Actinidia chinensis are used for the development of kiwifruit cultivars. Diploid germplasm can be exploited in a tetraploid breeding programme via unreduced (2n) gametes and chemical-induced chromosome doubling of diploid cultivars and selections. Meiotic chromosome behaviour in diploid A. chinensis ‘Hort16A’ and colchicine-induced tetraploids from ‘Hort16A’ was analysed and compared with that in a diploid male and tetraploid males of A. chinensis raised from seeds sourced from the wild in China. Both naturally occurring and induced tetraploids formed multivalents, but colchicine-induced tetraploids showed a higher mean multivalent frequency in the pollen mother cells. Lagging chromosomes at anaphase I and II were observed at low frequencies in the colchicine-induced tetraploids. To investigate whether preferential or non-preferential chromosome pairing occurs in tetraploid A. chinensis, the inheritance of microsatellite alleles was analysed in the tetraploid progeny of crosses between A. chinensis (4x) and A. arguta (4x). The frequencies of inherited microsatellite allelic combinations in the hybrids suggested that non-preferential chromosome pairing had occurred in the tetraploid A. chinensis parent.     

Chromosome doubling of 2x Solanum species by oryzalin: method development and comparison with spontaneous chromosome doubling in vitro

A potato breeding scheme implies the possibility of ploidy level manipulation either by reducing the chromosome number of cultivars from 48 to 24 to be able to cross them with diploid related species or by doubling diploid material to reach the generally optimal tetraploid level. In vitro spontaneous chromosome doubling is widely used but can lead to somaclonal variation. Since oryzalin has proven to be efficient as a chromosome doubling agent on potato cell suspension cultures, we tried this herbicide on various Solanum species and interspecific diploid hybrids. A 24 h dip in a 28.8 M aqueous oryzalin solution applied on apical buds was the most efficient treatment in terms of tetraploid plant production (mean = 4.1 tetraploid plants for 10 treated buds over 4 genotypes). However 50–100% of the regenerated tetraploid plants acclimatized after in vitro treatment proved to be chimaeric. Consequently, a selection procedure in the progeny was necessary to obtain real and stable doubled clones and final yields were low. This technique is easy to apply and could be a good alternative to chromosome doubling by spontaneous in vitro regeneration in the case of refractory genotypes especially where somaclonal variation is problematic. Percentage of tetraploids among the regenerated plants varied from 6 to 29% with the oryzalin doubling technique while it varied from 20 to 78% by in vitro spontaneous doubling for five diploid genotypes. An observation of the progeny indicated that chimaeras were more frequent using oryzalin (50–100% of the initially supposed tetraploid plants) than when chromosomes doubled spontaneously (4–67% of the initially supposed tetraploid plants).