Tetraploid Clones Research Articles

In liquid medium colchicine treatment induces non chimerical doubled-diploids in a wide range of mono- and interspecific diploid banana clones

Introduction . The worldwide production of banana and plantain, which is estimated to 106 Mt·year–1 , relies on a narrow genetic basis. Thus, banana production is fragile regarding emergent diseases, and creation of new varieties of banana stands out as a real necessity for its sustainability. In banana, various improvement strategies aim to create triploid hybrids. The objective of this report is to present a simple in vitro methodology to induce stable tetraploid plants by colchicine treatment, usable as parent for obtaining triploid varieties. Materials and methods . Twenty-one diploid M. acuminata clones and three interspecific M. acuminata / M. balbisiana diploid clones were treated as proliferating culture in liquid medium with 1.25 mM of colchicine for 48 h. Plant screening has been performed by morphological identification in greenhouse, chromosome counts on root tips and flow cytometry on leave blades on vitroplants using propidium iodide staining. Results . Chromosome counts led to distinguish diploid and tetraploid plants but did not afford to detect chimeras. Flow cytometry allowed an early screening of a larger number of plants leading to detect rapidly chimerical plants. It was observed that some stable 2x / 4x cytochimeras are periclinal. Tetraploid clones were obtained with all diploid genotypes. General behaviour of tetraploids in the field was globally weaker than the corresponding diploids. Nevertheless, all the doubled-diploids flowered and crossed with diploid plants to obtain triploid progenies. Conclusion . This study has clearly shown that induction of stable doubled-diploid plants can be obtained from a wide range of genetically different bananas. These results open the way to the systematic use of doubled-diploids by banana breeding programs for the release of enhanced triploid varieties.

Selective Resistance of Tetraploid Cancer Cells against DNA Damage‐Induced Apoptosis

Aneuploidy and chromosomal instability, which are frequent in cancer, can result from the asymmetric division of tetraploid precursors. Genomic instability may favor the generation of more aggressive tumor cells with a reduced propensity for undergoing apoptosis. To assess the impact of tetraploidization on apoptosis regulation, we generated a series of stable tetraploid HCT116 and RKO colon carcinoma cell lines. When comparing diploid parental cells with tetraploid clones, we found that such cells were equally sensitive to a series of cytotoxic agents (staurosporine [STS], hydroxyurea, etoposide), as well as to the lysis by natural killer cells. In strict contrast, tetraploid cells were found to be relatively resistant against a series of DNA-damaging agents, namely cisplatin, oxaliplatin, camptothecin, and gamma- and UVC-irradiation. This increased resistance correlated with a reduced manifestation of apoptotic parameters (such as the dissipation of the mitochondrial transmembrane potential and the degradation of nuclear DNA) in tetraploid as compared to diploid cells subjected to DNA damage. Moreover, tetraploid cells manifested an enhanced baseline level of p53 activation. Inhibition of p53 abolished the difference in the susceptibility of diploid and tetraploid cancer cells to DNA damage-induced apoptosis. These data point to an intrinsic resistance of tetraploid cells against radiotherapy and DNA-targeted chemotherapy that may be linked to the status of the p53 system.

Clone-specific differences in Phragmites australis: Effects of ploidy level and geographic origin

Phragmites australis (Cav.) Trin. ex Steud. is virtually cosmopolitan and shows substantial variation in euploidy level and morphology. The aim of this study was to assess clone-specific differences in morphological, anatomical, physiological and biochemical traits of P. australis as affected by the geographic origin, the euploidy level (4 x, 6 x, 8 x and 12 x), and to assess differences between native and introduced clones in North America. Growth, morphology, photosynthetic characteristics, photosynthetic pigments and enzymes were measured on 11 geographically distinct clones propagated in a common environment in Denmark. Any differences between the measured parameters were caused by genetic differences between clones. Overall, the largest differences between clones were found in ontogeny, shoot morphology and leaf anatomy. The North Swedish clone was adapted to short growing seasons and sprouted very early in the spring but senesced early in July. In contrast, clones from southern regions were adapted to warmer and longer growing seasons and failed to complete the whole growth-cycle in Denmark. Some clones from oceanic habitats with climatic conditions that do not differ much from conditions at the Danish growth site did flower in the common environment. The octoploid genotype in general had larger dimensions of leaves, taller and thicker shoots and larger cell sizes than did the hexaploid and tetraploid clones. The dodecaploid clone was neither bigger than the octoploid, nor significantly different from tetraploid and hexaploid clones in most of the morphological characters observed. Stomatal density decreased with increasing ploidy level, while length of guard cells increased. Tetraploid clones generally had morphometric dimensions, similar to hexaploids. Hence, polyploidy did not always result in an increase in plant size, probably because the number of cell divisions during development is reduced. Four North American clones were included in the study. The clone from the Atlantic Coast and the supposed invasive European clone resembled each other. The Gulf Coast clone differed from the rest of the clones in having leaf characters resembling Phragmites mauritianus Kunth. Thus, morphological characters are not unmistakable parameters that can be used to discriminate between introduced and native clones. The physiological and biochemical processes also differed between clones, but these processes showed considerable phenotypic plasticity and were therefore very difficult to evaluate conclusively. It is concluded that P. australis is a species with very high genetic variability which is augmented by its cosmopolitan distribution, clonal growth form and the large variation in chromosome numbers. It is therefore not surprising that large genetically determined differences in ontogeny, shoot morphology and leaf anatomy occur between clones.

Essential oil composition of diploid and tetraploid clones of ginger (Zingiber officinale Roscoe) grown in Australia.

Ginger oil, obtained by steam distillation of the rhizome of Zingiber officinale Roscoe, is used in the beverage and fragrance industries. Ginger oil displays considerable compositional diversity, but is typically characterized by a high content of sesquiterpene hydrocarbons, including zingiberene, ar-curcumene, beta-bisabolene, and beta-sesquiphellandrene. Australian ginger oil has a reputation for possessing a particular "lemony" aroma, due to its high content of the isomers neral and geranial, often collectively referred to as citral. Fresh rhizomes of 17 clones of Australian ginger, including commercial cultivars and experimental tetraploid clones, were steam distilled 7 weeks post-harvest, and the resulting oils were analyzed by GC-MS. The essential oils of 16 of the 17 clones, including the tetraploid clones and their parent cultivar, were found to be of substantially similar composition. These oils were characterized by very high citral levels (51-71%) and relatively low levels of the sesquiterpene hydrocarbons typical of ginger oil. The citral levels of most of these oils exceeded those previously reported for ginger oils. The neral-to-geranial ratio was shown to be remarkably constant (0.61 +/- 0.01) across all 17 clones. One clone, the cultivar "Jamaican", yielded oil with a substantially different composition, lower citral content and higher levels of sesquiterpene hydrocarbons. Because this cultivar also contains significantly higher concentrations of pungent gingerols, it possesses unique aroma and flavor characteristics, which should be of commercial interest.

Mapping the R10 and R11 genes for resistance to late blight (Phytophthora infestans) present in the potato (Solanum tuberosum) R-gene differentials of Black

The R10 and R11 late blight differentials of Black (tetraploid clones 3681ad1 and 5008ab6) were crossed with the susceptible potato (Solanum tuberosum) cultivar Maris Piper and the progeny were assessed for blight resistance in a whole plant glasshouse test using race 1,2,3,4,6,7 of Phytophthora infestans. The disease scores for the R10 population displayed a continuous distribution whereas the progeny in the R11 population could be categorised as resistant or susceptible. A bulk segregant analysis using amplified fragment length polymorphism assays was done on the ten most resistant and ten most susceptible progeny in each population and two closely linked markers were found to be associated with resistance. R11 mapped to 8.5 cM from marker PAG/MAAG_172.3 and R10 mapped as a quantitative trait locus in which marker PAC/MATC_264.1 explained 56.9% of the variation in disease scores. The results were consistent with R10 and R11 being allelic versions of genes at the R3 locus on chromosome 11. The implications are discussed for mapping R-genes which fail to give complete immunity to a pathogen.

Gingerol content of diploid and tetraploid clones of ginger (Zingiber officinale Roscoe).

Ginger (Zingiber officinale Roscoe), a monocotyledonous, sterile cultigen, is widely used as a spice, flavoring agent, and herbal medicine. The pungency of fresh ginger is due to a series of homologous phenolic ketones of which [6]-gingerol is the major one. The gingerols are thermally unstable and can be converted to their corresponding shogaols, which are present in dried ginger. Fresh rhizomes of 17 clones of Australian ginger, including commercial cultivars and experimental tetraploid clones, were assayed by HPLC for gingerols and shogaols. [6]-Gingerol was identified as the major pungent phenolic compound in all samples, while [8]- and [10]-gingerol occurred in lower concentrations. One cultivar known as "Jamaican" contained the highest concentrations of all three gingerols and was the most pungent of the clones analyzed. Gingerols were stable in ethanolic solution over a 5-month period when stored at 4 degrees C. Shogaols were not identified in the extracts prepared from fresh rhizomes at ambient temperature, confirming that these compounds are not native constituents of fresh ginger. In contrast to previous findings, this study did not find significant differences in gingerol concentrations between the tetraploid clones and their parent diploid cultivar.

Resistance to soft rot (Erwinia carotovora subsp.atroseptica) in tetraploid potato families obtained from 4x-2x crosses

Diploid potato clones, interspecific hybrids ofSolanum species, having in their originS. tuberosum,S. chacoense, S. yungasense, S. phureja, S. gourlayi, andS. demissum, with resistance to soft rot, were crossed to tetraploid potato clones in 4x-2x crosses. The 24 tetraploid families obtained in a North Carolina II design were examined for tuber resistance to soft rot in a laboratory test and for basic agronomic traits in field trials conducted for two consecutive years. In addition, one family originating from a 4x-2x cross of two susceptible parents was tested. Statistical analysis of the data revealed significant general (GCA) and specific combining ability (SCA) effects, year, GCA (female) × year, GCA (male) × year, and SCA × year upon the inheritance of resistance to soft rot. About 35% of the progeny was selected as resistant to tuber soft rot, and of these 11% showed high resistance combined with good tuber yield, tuber weight, and tuber appearance. The relationships between resistance to soft rot and chosen agronomic traits were not noted or were weakly significant and sporadic. The resistance to tuber soft rot found in diploid potato hybrids can be transferred to the cultivated tetraploid pool through 4x-2x crosses, and a high frequency of offspring posses resistance.

Net Nitrate Uptake by the Roots of Different Potato Haploids

ABSTRACT Seventy haploids were obtained by crossing the pollinator Solanum tuberosumL. Group Phureja (P.I.1.22) (2n = 2x) with tetraploid clones of Solanum tuberosumL., “Spunta” and `Desirée,' two adapted Italian varieties. By tuber morphology screening, they were grouped into six classes and six haploids were selected as representative for each class. Esterase isozyme analysis on leaves of the selected haploids indicated the presence of a considerable degree of genetic variability. Six potato haploids were evaluated for differences in nitrate acquisition efficiency. A wide variability of net nitrate uptake rate, ranging from 2.36 to 4.23 μ moles/h/g FW, was observed among the potato genotypes. Furthermore, the time needed for the expression of the nitrate uptake induction showed large differences (from 6.93 to 46.2 h). Relative growth rate of potato haploids correlated with the accumulated nitrate (At) while any correlation with the parameters relative to the induction phase of nitrate uptake was observed. Results indicated that, besides morphological variability, potato haploid populations showed a high variability in nitrate acquisition efficiency. This variability must be gauged in order to select potato varieties with improved capacity for nitrate uptake.

In vitro chromosome-doubling in tulip (Tulipa gesneriana L.)

SummaryMost current tulip cultivars are diploid (2n = 24), some are triploid (Darwin Hybrids) and there are some rare tetraploid cultivars. Crosses between diploids are generally successful, but crosses between diploid cultivars and tetraploid selected species often fail, which constitutes a major obstacle to broadening of the genetic base. Our objective was thus to develop a method to obtain tetraploid material from diploid genotypes using a stem-disc regeneration process and the chromosome-doubling agent oryzalin, applied in different ways. Tetraploid clones were obtained from all treatments and from all cultivars tested. The final yield of tetraploids was low (0 – 3.75 tetraploids for one treated flower stem), but was sufficient to initiate crossing experiments between diploid and tetraploid progenitors to obtain triploid offspring.A genotypic effect was observed during the regeneration process.The cultivars ‘Lucky Strike’ and ‘Gander’ were more responsive in vitro than cvs. ‘Lustige Witwe’ and ‘Don Quichotte’. However, the percentage of tetraploid clones finally obtained was not influenced by genotype or by the way in which oryzalin was applied.

Distribution, mechanisms and evolutionary significance of clonality and polyploidy in weevils

Abstract 1 Genetical mtDNA relationships of 41 taxa of weevils were examined using cladistics. Ingroup taxa belong to Otiorhynchus scaber and O. nodosus and outgroup comparison was made with O. singularis. All three species are minor forest pests.2 Otiorhynchus scaber specimens are either diploid sexuals or diploid, triploid and tetraploid clones, from two different populations (Slovenia and Austria) that belong to two different evolutionary lineages. Otiorhynchus nodosus specimens are tetraploid clones. Both species show geographical parthenogenesis, as do many other Otiorhynchus species.3 Mitochondrial data indicate that O. nodosus clones are more closely related to Slovenian sexuals of O. scaber than these are to sexuals from Austria. It also shows that almost all clones of O. scaber collected in one of the two regions where sexuals are found are more closely related to sexuals from the other region.4 Three different hypotheses that may explain the distribution of O. scaber, mechanisms important for the evolution of the clones and implications of the presence of Wolbachia are discussed.5 We conclude that parthenogenesis is likely to be linked to hybridization in O. scaber and that hybridization events between ancestors of O. nodosus and O. scaber are the probable cause of the presence of O. nodosus in the ingroup. We also find that polyploid clones are superior colonizers compared to sexuals and diploid clones, in O. scaber.6 The results suggest that systems where both sexuals and clones exist are more complex than previously suggested. The mapping of genetic variation in clonal complexes and the tracing of clonal origins may be useful in pest management.

Genetic diversity in sexual diploid and apomictic tetraploid populations of Paspalum notatum situated in sympatry or allopatry

Paspalum notatum is a subtropical grass widely distributed in the temperate areas of America. Diploids are sexual while polyploids give rise to clonal seeds through aposporous apomixis. RAPD markers were used to analyze the genetic structure of three natural populations: i) diploids reproducing sexually (R2X); ii) sympatric apomictic tetraploids collected in the vicinity of the diploids (R4X); iii) allopatric apomictic tetraploids growing in isolation (C4X). The apomictic reproduction rate was evaluated by the use of molecular markers in progeny tests, while chromosome-counting allowed the verification of ploidy levels. Data revealed that the R4X group presented a variation considerably higher than that observed for C4X. Jaccard’s coefficients were used to produce a cluster diagram using the UPGMA method. All but one tetraploid genotypes grouped together and were associated to diploid genotype A21. The possibility of occasional generation of novel tetraploid clones from the interaction between tetraploid and diploid individuals is discussed.

Characterisation of Tetraploid and Diploid Clones of Spodoptera frugiperda Cell Line

We have isolated and characterised diploid and tetraploid clones from the normally heterologous Spodoptera frugiperda (Sf-9)cell line by dilution cloning technique. Tetraploid clones were found to have cell sizes in excess of 35% larger than that of the diploid clones. In contrast, the maximum cell numbers achieved in batch cultures of diploid clones were on average 185% higher than the tetraploid cell numbers. Growth rates and metabolic quotients during the exponential phase were similar for both clones. Tetraploid cells infected with wild-type and recombinant green fluorescent protein (GFP) baculovirus, resulted in more polyhedra or GFP product per cell. Importantly, the difference between the clones either completely diminished or reduced to 50% when the yield was assessed in terms of the amount of polyhedra or GFP per mL of medium, respectively. These results indicate that the existing heterogeneity in insect cell populations with respect to ploidy level, are correlated to cell growth and product yield.

Genetic heterogeneity and ploidy level analysis among different gynogenetic clones of the polyploid gibel carp.

Some triploid and tetraploid clones have been identified in the gynogenetic gibel carp, Carassius auratus gibelio Bloch, by karyotypic and cytologic analyses over many years. Further, 5-20% males and karyotypic diversity have been found among their natural and artificial populations. However, the DNA contents and the relation to their ploidy level and chromosome numbers have not been ascertained, and whether normal meiosis occurs in spermatogenesis needs to be determined in the different clones. The sampled blood cells or sperms were mixed with blood cells from chicken or individual gibel carp and fixed in 70% pre-cooled ethanol overnight at 4 degrees C. The mixed cell pellets were washed 2-3 times in 1x phosphate buffered saline and then resuspended in the solution containing 0.5% pepsin and 0.1 M HCl. DNA was stained with propidium iodide solution (40 microg/mL) containing 4 kU/ml RNase. The measurements of DNA contents were performed with Phoenix Flow Systems. Triploid clones A, E, F, and P had almost equal DNA content, but triploid clone D had greater DNA content than did the other four triploid clones. DNA content of clone M (7.01 +/- 0.15 pg/nucleus) was almost equal to the DNA content of clone D (5.38 +/- 0.06 pg/nucleus) plus the DNA content of common carp sperm (1.64 +/- 0.02 pg/nucleus). The DNA contents of sperms from clones A, P, and D were half of their blood cells, suggesting that normal meiosis occurs in spermatogenesis. Flow cytometry is a powerful method to analyze genetic heterogeneity and ploidy level among different gynogenetic clones of polyploid gibel carp. Through this study, four questions have been answered. (a) The DNA content correlation among the five triploid clones and one multiple tetraploid clone was revealed in the gibel carp, and the contents increased with not only the ploidy level but also the chromosome number. (b) Mean DNA content was 0.052 pg in six extra chromosomes of clone D, which was higher than that of each chromosome in clones A, E, F, and P (about 0.032 pg/chromosome). This means that the six extra chromosomes are larger chromosomes. (c) Normal meiosis occurred during spermatogenesis of the gibel carp, because DNA contents of the sperms from clones A, P, and D were almost half of that in their blood cells. (d) Multiple tetraploid clone M (7.01 +/- 0.15 pg/nucleus) contained the complete genome of clone D (5.38 +/- 0.06 pg/nucleus) and the genome of common carp sperm (1.64 +/- 0.02 pg/nucleus).

A fluorogenic 5' nuclease (TaqMan) assay to assess dosage of a marker tightly linked to red skin color in autotetraploid potato.

We have recently identified an allele of dihydroflavonol 4-reductase ( dfr) that cosegregates with the ability of potato ( Solanum tuberosum L) to produce red pelargonidin-based anthocyanin pigments. A rapid assay to assess dosage of this allele in cultivated potato, an autotetraploid, would be useful for breeding programs that develop red-skinned cultivars. To identify regions of dfr that are conserved between alleles, as well as regions that are variable, a portion of the gene was sequenced from several cultivated and wild potato clones. In one region the sequence of the 'red' dfr allele differed at two nucleotide positions from the three other sequence classes observed. A fluorogenic oligonucleotide probe labeled with 6-FAM was designed to anneal specifically to the red allele in this region, while a second probe labeled with VIC was designed to anneal to the 'not-red' dfr alleles. PCR primers that annealed to conserved sequences flanking the variable region were also developed. When subjected to a fluorogenic 5' nuclease (TaqMan) allelic discrimination assay all diploid clones tested clustered into three distinct groups based on the relative amounts of FAM and VIC released. These three groups represented clones homozygous for the red allele, heterozygous for the red allele, and homozygous for the not-red allele(s). When tetraploid clones were tested they separated into five distinct clusters, three of which were shared with diploid clones. The five clusters were interpreted to represent clones quadruplex, triplex, duplex, simplex and nulliplex for the red dfr allele. This interpretation was supported by monitoring the segregation of red allele dosage in several tetraploid crosses. To the best of our knowledge this is the first report of a fluorogenic 5' nuclease assay being used for allelic discrimination in an autopolyploid.

Hypocellular acute promyelocytic leukemia with a tetraploid clone characterized by two t(15;17)

We describe a case of hypocellular acute promyelocytic leukemia with a tetraploid clone characterized by two t(15;17). The large leukemia cells had a bizarre nuclear configuration and multiple Auer rods. A bone marrow biopsy specimen revealed a markedly hypocellular marrow (<10% cellularity) in the absence of myelofibrosis. Myelodysplastic features were not detected. Chromosome analysis of marrow cells revealed a karyotype of 92,XXYY,del(2)(q?),t(15;17)(q22;q21)×2. Interphase fluorescence in situ hybridization revealed that the marrow cells were composed of a tetraploid clone carrying double t(15;17) and normal diploid cells. The leukemia responded well to all- trans retinoic acid. We think that the tetraploidy could be caused by endoreduplication or endomitosis of the diploid clone with single t(15;17). The unique karyotype largely contributed to the cell morphology and marrow hypoplasia, while it may not have affected on the prognosis of the acute promyelocytic leukemia.

Tetraploid acute promyelocytic leukemia with double t(15;17) and PML/ RARA rearrangements detected by fluorescence in situ hybridization analysis

Acute promyelocytic leukemia (APL) is characterized by a serious hemorrhagic syndrome, unique morphologic findings, and its response to retinoids. Tetraploidy is a very rare chromosomal abnormality in acute myelocytic leukemia. This report presents a unique case of APL with a tetraploid clone characterized by two t(15;17) without other chromosomal changes, as well as PML/ RARA rearrangements confirmed fluorescence in situ hybridization. The morphology of the blast cells was that of the classic M3 subtype, but the mean blast size exceeded that of control APL cases with diploidy. A chromosomal study revealed a 92,XXXX,t(15;17)(q22;q21)×2 karyotype in all 20 metaphase spreads. Despite all- trans-retinoic acid (ATRA) treatment and chemotherapy, leukemic cells persisted in the blood, and the patient died of an intracranial hemorrhage on the 16th day after admission.

Regeneration of a Tetraploid Clone from Callus Culture of Asparagus officinalis L. through Somatic Embryogenesis

A tetraploid callus line of Asparagus officinalis L. was identified from 4 regenerated callus lines. Plants were regenerated from these callus lines following somatic embryogenesis and the tetraploid clone of this species was established in the field with 80% survival rate. The embryogenic callus was induced in Murashige and Skoog's (MS) medium in presence of α-naphthaleneacetic acid (NAA) (0.2 mg/l) and kinetin (0.02 mg/l) and proliferated as well as maintained in 2,4-dichlorophenoxyacetic acid (2,4-D) (1.0 mg/l) containing MS medium. Somatic embryos were initiated in Gelrite-solidified MS medium with variable levels of NAA and 6-(γ-γ-dimethylallylamino) purine (2ip). A higher level of carbohydrate enhanced embryo conversion efficiency. The embryos induced in presence of 10% glucose for 2 weeks and subsequently transferred to 2% sucrose level showed higher conversion rate than those maintained in 3% sucrose concentration. Karyotype analysis of diploid and tetraploid clones revealed exact duplication of the diploid set in tetraploid plants.

A Patient with Congenital Ichthyosis Hystrix (Disseminated Congenital Naevus) and Acute Lymphoblastic Leukaemia

Congenital ichthyosis is an uncommon spectrum of hereditary and sporadic hyperkeratotic epidermolytic disorders that may occur in isolation or in association with other clinical syndromes. The pattern of distribution of the hyperkeratotic lesions and family history form the basis of diagnosis. Ichthyosis may be a marker for immune deficiency and cancer prone conditions. We report a 16-year old Chinese patient with a diagnosis of ichthyosis hystrix hystrix (disseminated congenital naevus) since birth who presented with T cell acute lymphoblastic leukaemia with tetraploid clones. There are four reports of acute leukaemia complicating ichthyosis, all involving paediatric acute lymphoblastic leukaemia. The possible mechanisms involved are discussed.

Variations in leaf anatomy and gas exchange in tea clones with different ploidy

SummaryExperiments were conducted in Kenya to evaluate the variation in leaf anatomy and gas exchange in 25 diploid, triploid and tetraploid clones of tea ( Camellia sinensis L.). There were significant variations in the different attributes within and between the different groups. The number of stomata and epidermal cells per unit leaf area decreasedwith increasing ploidy, reflecting larger cells in the tetraploids than in the diploids and triploids. Specific leaf mass (SLM) was also higher in the tetraploids. Net CO2 assimilation (A) and stomatal conductance (gs) were lower in the tetraploids than in either diploids or triploids and lower in triploids than diploids. These changeswere related to the lower number of stomata and epidermal cells with a correlation between A and number of stomata (R2.=.0.58) and number of epidermal cells (R2.=.0.85). Net CO2 assimilation and stomatal conductance were also strongly correlated (R2.=.0.73). Gas exchange was not related to specific leaf mass (P>0.05). It is concluded that gas exchange is strongly influenced by leaf anatomy and ploidy level.

Molecular marker analysis of 24- and 25-chromosome plants obtained from Solanum tuberosum L. subsp. andigena (2n = 4x = 48) pollinated with a Solanum phureja haploid inducer.

Clones with 24 or 25 chromosomes were obtained by pollinating an Andean cultivated tetraploid potato (Solanum tuberosum subsp. andigena clone 94H94, 2n = 4x = 48) with the Solanum phureja haploid-inducer clone 1.22. Their genetic composition was analyzed in an RAPD assay using 135 decamer primers and in an RFLP assay using 45 single-copy DNA probes. In total, 22 RAPD and 20 RFLP markers were found to be specific to S. phureja. None of these markers were found in the 24- and 25-chromosome clones. RFLP genotypes for the 45 RFLP loci were further determined for each clone. Genotypes of the 24-chromosome clones were characterized using two alleles randomly selected from four alleles of the parental tetraploid clone for almost all RFLP loci. Five 25-chromosome clones had extra alleles for all of the RFLP loci of chromosomes 4, 8, 10, 11, and 12, respectively, suggesting primary trisomy for one of these chromosomes. Clones with genotypes showing double reduction were also identified. Therefore, the obtained clones likely originated from random samples of female gametes, and hence are euhaploids or aneuhaploids of S. tuberosum subsp. andigena, strongly supporting parthenogenesis to be a primary mechanism for haploid induction in potato.