Tuberosum Haploids Research Articles

Hybridization barriers between diploid Solanum tuberosum and wild Solanum raphanifolium

Wild potato germplasm represents a unique, diverse and accessible resource for disease and pest resistance, along with useful agronomic traits that may be introgressed into the cultivated potato (Solanum tuberosum L.). Hybridization of diploid wild Solanum species with haploids (2×) of cultivated potato (4×) is generally an effective technique for introducing genetic diversity and desirable traits into potato. However, in this study, hybridization barriers were found in crosses between S. tuberosum haploids and the wild species S. raphanifolium. Male sterility, likely due to nuclear-cytoplasmic interactions, was observed in some haploid tbr × S. raphanifolium hybrids. In addition, pollen-pistil incompatibilities were observed in backcross, F2, and reciprocal cross hybridization attempts that failed to produce seed. More crosses were successful when F1 clones were crossed as females to wild clones than to cultivated clones. When crosses were made in the other direction, with F1 hybrids used as male parents, seeds were almost never produced.

Interspecific somatic hybrids between Solanum bulbocastanum and S. tuberosum and their haploidization for potato breeding

Protoplast fusion between incongruent Solanum bulbocastanum and S. tuberosum haploids was accomplished to produce hybrids combining elite traits from both parents. We identified 11 somatic hybrids out of 42 regenerants analyzed through ISSR markers. Some hybrids had loss or gain of fragments compared to the parents, likely due to rearrangements and deletions of chromosome segments after fusion, and/or to somaclonal variation during hybrid regeneration. Increased heterotic vigor for some traits as well as high diversity was observed as the effect of both ploidy and fusion combination. Microsporogenesis analysis indicated the occurrence of multivalent configurations and several meiotic abnormalities, such as chromosomes bridges and various spindle orientations. Since all hybrids were sterile, in vitro anther culture was employed for haploidization as a possible strategy to overcome barriers to hybridizations. Haploids were obtained from all the tetraploid S. bulbocastanum (+) S. tuberosum somatic hybrids tested, although with differences in both the number of embryos per 100 anthers cultured and the number of differentiated green plantlets. This is the first report on the successful production of haploid plants from S. bulbocastanum (+) S. tuberosum hybrids.

Interspecific somatic hybrids between Solanum bulbocastanum and S. tuberosum and their haploidization for potato breeding

Protoplast fusion between incongruent Solanum bulbocastanum and S. tuberosum haploids was accomplished to produce hybrids combining elite traits from both parents. We identified 11 somatic hybrids out of 42 regenerants analyzed through ISSR markers. Some hybrids had loss or gain of fragments compared to the parents, likely due to rearrangements and deletions of chromosome segments after fusion, and/or to somaclonal variation during hybrid regeneration. Increased heterotic vigor for some traits as well as high diversity was observed as the effect of both ploidy and fusion combination. Microsporogenesis analysis indicated the occurrence of multivalent configurations and several meiotic abnormalities, such as chromosomes bridges and various spindle orientations. Since all hybrids were sterile, in vitro anther culture was employed for haploidization as a possible strategy to overcome barriers to hybridizations. Haploids were obtained from all the tetraploid S. bulbocastanum (+) S. tuberosum somatic hybrids tested, although with differences in both the number of embryos per 100 anthers cultured and the number of differentiated green plantlets. This is the first report on the successful production of haploid plants from S. bulbocastanum (+) S. tuberosum hybrids.

Glycoalkaloids as Biomarkers for Recognition of Cultivated, Wild, and Somatic Hybrids of Potato

Cultivated and wild potato species synthesize a wide variety of steroidal glycoalkaloids (GAs). During breeding programs, species genomes are often put together through either sexual or somatic hybridization. Therefore, the determination of the GA composition of hybrids is very important in that it may affect either human consumption, or resistance to pathogen and pests. Here, we report the results of GA analysis performed on wild Solanum bulbocastanum, haploids of cultivated potato S. tuberosum and their interspecific somatic hybrids. GAs were extracted from tubers and analyzed by HPLC. HPLC Profile of S. tuberosum haploids showed, as expected, the presence of alpha-solanine and alpha-chaconine. The profile of S. bulbocastanum extract showed lack of alpha-solanine and alpha-chaconine, and the presence of four GAs. The GA pattern of the somatic hybrids was the sum of their parents' profile. This represents a noteworthy tool for their unequivocal recognition. Interestingly, two hybrids produced not only GAs of both parents but also new compounds to be further investigated. This provided evidence that somatic hybridization induced the synthesis of new metabolites. The nature of the probable unidentified GAs associated to S. bulbocastanum and its somatic hybrids was ascertained by chemical degradation and spectroscopic analysis of their aglycones and sugar moieties. Our results suggest their close relation with GAs of both wild and cultivated potato species.

Nuclear and cytoplasmic genome composition of Solanum bulbocastanum (+) S. tuberosum somatic hybrids

Somatic hybrids between the wild incongruent species Solanum bulbocastanum (2n = 2x = 24) and S. tuberosum haploids (2n = 2x = 24) have been characterized for their nuclear and cytoplasmic genome composition. Cytologic observations revealed the recovery of 8 (near-)tetraploid and 3 hexaploid somatic hybrids. Multicolor genomic in situ hybridization (GISH) analysis was carried out to study the genomic dosage of the parental species in 5 somatic hybrids with different ploidy. The GISH procedure used was effective in discriminating parental genomes in the hybrids; most chromosomes were unambiguously colored. Two (near-)tetraploid somatic hybrids showed the expected 2:2 cultivated-to-wild genomic dosage; 2 hexaploids revealed a 4:2 cultivated-to-wild genomic dosage, and 1 hexaploid had a 2:4 cultivated-to-wild genomic dosage. Characterization of hybrid cytoplasmic genomes was performed using gene-specific primers that detected polymorphisms between the fusion parents in the intergenic regions. The analysis showed that most of the somatic hybrids inherited the plastidial and mitochondrial DNA of the cultivated parent. A few hybrids, with a rearranged mitochondrial genome (showing fragments derived from both parents), were also identified. These results confirmed the potential of somatic hybridization in producing new variability for genetic studies and breeding.

Molecular characterization of the progeny of Solanum etuberosum identifies a genomic region associated with resistance to potato leafroll virus

Potato leafroll virus (PLRV; Genus Polerovirus; Family Luteoviridae) is one of the most important virus pathogens of potato worldwide and breeders are looking for new sources of resistance. Solanum etuberosum Lindl., a wild potato species native to Chile, was identified as having resistances to PLRV, potato virus Y, potato virus X, and green peach aphid. Barriers to sexual hybridization between S. etuberosum and cultivated potato were overcome through somatic hybridization. Resistance to PLRV has been identified in the BC1, BC2 and BC3 progeny of the somatic hybrids of S. etuberosum (+) S. tuberosum haploid × S. berthaultii Hawkes. In this study, RFLP markers previously mapped in potato, tomato or populations derived from S. palustre (syn S. brevidens) × S. etuberosum and simple sequence repeat (SSR) markers developed from tomato and potato EST sequences were used to characterize S. etuberosum genomic regions associated with resistance to PLRV. The RFLP marker TG443 from tomato linkage group 4 was found to segregate with PLRV resistance. This chromosome region has not previously been associated with PLRV resistance and therefore suggests a unique source of resistance. Synteny groups of molecular markers were constructed using information from published genetic linkage maps of potato, tomato and S. palustre (syn. S. brevidens) × S. etuberosum. Analysis of synteny group transmission over generations confirmed the sequential loss of S. etuberosum chromosomes with each backcross to potato. Marker analyses provided evidence of recombination between the potato and S. etuberosum genomes and/or fragmentation of the S. etuberosum chromosomes.

Advantages of Wild Diploid Solanum Species Over Cultivated Diploid Relatives in Potato Breeding Programs

For breeding programs of the tetraploid potato (Solanum tuberosum), both wild and cultivated diploid relatives are valuable sources of genetic diversity. While both types of germplasm are used in breeding programs, there are several advantages to using wild relatives. Diploid relatives are typically crossed with haploids (2n = 2x = 24) from tetraploid S. tuberosum to improve daylength adaptation. Most haploids are male sterile, so they are typically used as female parents. Cultivated diploids, such as members of the Phureja Group, produce male sterile hybrids when crossed as females to haploids; wild relatives, such as S. tarijense, often produce male fertile hybrids. Tuber yield following crosses of haploids to cultivated or wild relatives is often high. However, cultivated relatives generally produce hybrids with a high set of small tubers; hybrids from wild relatives are variable, but many are similar to cultivars in tuber size and set. While tubers of hybrids from cultivated relatives are typically rough, with deep eyes and raised internodes, those from wild relatives are often smooth. Tuber dormancy in hybrids with cultivated relatives is generally short, while that in hybrids with wild species is longer, allowing for storage over winter. Finally, resistance to several major diseases and stresses has been found in wild species and their hybrids with S. tuberosum haploids. The desirable traits in hybrids are transmitted to tetraploids via unilateral sexual polyploidization (4x × 2x or 2x × 4x crosses in which the diploid parent produces 2n gametes). Wild Solanum species are recommended for use in potato breeding programs as sources of genetic diversity that can be adapted easily following hybridization with S. tuberosum haploids.

Unilateral and bilateral hybridization barriers in inter-series crosses of 4x 2EBN Solanum stoloniferum, S. pinnatisectum, S. cardiophyllum, and 2x 2EBN S. tuberosum haploids and haploid-species hybrids

Wild Mexican potato species are an important untapped source of useful variation for potato improvement. Introgression methods such as 2n gametes, chromosome doubling, and crossing with disomic 4x 2 endosperm balance number (EBN) bridge species have been used to overcome post-zygotic endosperm failure according to the EBN hypothesis. Stylar barriers can prevent zygote formation, bilaterally when zygote formation is blocked in both directions of the cross or unilaterally when zygote formation is blocked in self incompatible (SI) × self compatible (SC) crosses. In several Solanaceae species, the S-locus for SI has been implicated in interspecific incompatibility. The objectives of this research were to determine if: (1) disomic 4x 2EBN Solanum stoloniferum can be used as a bridge species for introgression of the Mexican 2x 1EBN species Solanum cardiophyllum and Solanum pinnatisectum, (2) pre- and/or post-zygotic barriers limit hybridization among EBN compatible Solanum inter-series crosses, and (3) reproductive barriers act unilaterally or bilaterally. Fruit formation and seed set was recorded for inter-pollinations of S. stoloniferum, 4x 2EBN chromosome doubled S. cardiophyllum and S. pinnatisectum, and 2x 2EBN S. tuberosum haploids (HAP) or haploid-species hybrids (H-S). In vivo pollen tube growth was analyzed for each cross combination with fluorescence microscopy. Attempts to create bridge hybrids between S. stoloniferum, and S. cardiophyllum or S. pinnatisectum were not successful. Pre- and post-zygotic barriers prevented seed formation in crosses involving S. cardiophyllum and S. pinnatisectum. Self compatibility in S. stoloniferum and S. pinnatisectum suggests that the S-locus does not contribute to the stylar barriers observed with these species. Alternatively, the presence of functional and nonfunctional (SC) S-alleles may explain interspecific incompatibility in intra- and inter-ploidy crosses. A non-stylar unilateral incongruity was discovered in H-S/HAP × S. stoloniferum crosses, indicating either a post-zygotic barrier, or a pre-zygotic barrier acting at or within the ovary. Furthermore, lack of S. stoloniferum pollen rejection may occur through absence of S. stoloniferum pollen-active genes needed to initiate pollen rejection, or through competitive interaction in S-locus heterozygous S. stoloniferum pollen. Introgression strategies using these species would benefit potato breeding by introducing genetic diversity for several traits simultaneously through co-current introgression.

Introgression of resistance to late blight (Phytophthora infestans) from Solanum pinnatisectum into S. tuberosum using embryo rescue and double pollination

Efforts to generate hybrids from crosses of Mexican diploid 2x(1EBN) species × 2x(2EBN) Solanum tuberosum haploids via embryo rescue and double pollination were conducted. The species included were S. bulbocastanum, S. pinnatisectum and S. trifidum. The species S. phureja was used as a second pollinator. After 3,077 pollinations were made during the spring and summer of 1995, 31 potential hybrids were obtained; however, they were discarded because of their S. phureja second pollinator characteristics. The length of time before conducting embryo rescue [20–25 days after pollination (DAP)] was presumed responsible for this failure. In 1996, 3,288 pollinations were made, resulting in 184 fruit. Embryo rescue was performed between 15 and 20 DAP and resulted in 3,867 embryos of which 530 developed into seedlings. In January of 1997, a single unique hybrid was discovered in this population and was almost discarded because of its S. phureja anthocyanin nodal banding marker. This diploid hybrid arose from a cross between the haploid US-W 13,089 (Sebago) × S. pinnatisectum (PI275233). The hybrid was hypoploid (22 chromosomes). Its hybridity was confirmed through morphology, chromosome number and RAPD analysis. Its resistance to late blight was tested in an unsprayed replicated experiment at the UW Agricultural Research Station at Hancock, WI during the summer of 1997, where the results indicated a high level of field resistance. The hybrid exhibited a low degree of male and female fertility. From meiotic analysis, it was evident that normal pairing between the two genomes occurred, although the presence of univalents was observed. No hybrids with S. bulbocastanum and S. trifidum were obtained. These results support the view that transfer of valuable characters from S. pinnatisectum to S. tuberosum is feasible; however, aneuploidy and the presence of univalents may inhibit this effort.

2n gametes in the potato: essential ingredients for breeding and germplasm transfer

2n gametes are the result of meiotic mutations occurring during micro – and mega-sporogenesis. They have been identified in several plant species of different taxa. The potato is probably the crop plant where they have been most intensively studied and also more appropriately used for the genetic improvement of cultivated genotypes. This paper reviews how 2n gametes allow potato breeders to broaden the genetic basis of the cultivated Solanum tuberosum, introducing both new genes for the improvement of traits of interest and allelic diversity to maximize heterozygosity. We provide molecular and breeding evidence that, in the potato, 2n gametes represent a unique tool to transfer target genes from wild forms to the cultivated tetraploid gene pool. In fact, species directly crossable to S. tuberosum haploids can be exploited through sexual polyploidization crossing schemes. For those which have developed crossability barriers, specific crossing schemes based on ploidy bridges can be designed. In this paper we also give possible hypotheses to explain conflicting results on the genetic control and meiotic mutations responsible for 2n-gamete formation in the potato.

Effect of functional female and male fertilities on crossability in diploid potato breeding

Functional female fertility (FFF) and functional male fertility (FMF) were studied in diploid potato clones, includingS. tuberosum haploids, F1 hybrids between haploids and otherSolanum species, and in progenies from subsequent hybridisations. The results of 402 crosses between 91 diploid potato clones revealed that 60% of all genotypes had low levels of FFF as well as low levels of FMF regardless of their origin and degree of hybridity. Failed crosses (without any fruits at all) and those with a low level of success (with less than 3 seeds per pollinated flower) constituted 69% of all studied crosses. Most of these had resulted from parents with low levels of FFF and FMF, and these two characteristics were expressed independently. Functional female fertility affected crossability behaviour no less than FMF, therefore both FFF and FMF should be controlled in diploid potato clones involved in breeding. Reliable FFF and FMF estimates can be based only on crossing results. The coefficient of multiple correlation between seeds per pollinated flower in crosses and FFF and FMF estimates of the parental clones was 0.80 (P<0.01). Hence the compilation and analysis of the results of diploid potato crosses allow a reliable assessment of FFF and FMF and consequently the prediction of crossability with a degree of sufficient accuracy.

Resistance to blackleg and tuber soft rot in sexual and somatic interspecific hybrids with different genetic background

Diploid interspecific hybrids from crosses between Solanum tuberosum haploids and S. canasense, S. multidissectum, and S. taTijense, along with tetraploid and hexaploid somatic hybrids S. tuberosum (+) S. commersonii, were screened for resistance to blackleg and tuber soft rot by Erwinia carotovora subsp, atroseptica and E. carotovora subsp, carotovora. Among the diploid hybrids, those involving clone mlt la of S. multidissectum and tar 2b of S. tarijense yielded the highest number of genotypes resistant to both blackleg and tuber soft rot. By contrast, all the hybrids involving clone tar 1 lb of S. tarijense were susceptible to both bacterial diseases. As far as resistance of somatic hybrids is concerned, the most interesting genotype was the tetraploid one which showed resistance to both blackleg and tuber soft rot. Significant correlations were found between E. carot0vora subsp, atroseptica and E. carotovora subsp, carotovora both in resistance to blackleg and in resistance to tuber soft rot, suggesting that resistance to the two subspecies is controlled by the same gene/s or that the relevant genes are linked. On the other hand, correlations between blackleg and tuber soft rot were never significant, indicating that different mechanisms may control resistance in tuber and stems. Alternatively, the resistance could be controlled by the same gene/s under a different spatial/temporal expression pattern. Twelve diploid sexual hybrids and one tetraploid somatic hybrid were selected for resistance to blackleg and/or tuber soft rot and for tuber characteristics.

Male fertility and freezing tolerance of hybrids involvingSolanum tuberosum haploids and diploidSolanum species

Male sterility of (haploid x species) hybrids often hampers the exploitation of wildSolanum species in breeding for cold tolerance. In a study aimed at finding ways round these problems, we found that one accession of the cold tolerant wild speciesS. sanctae-rosae gave progenies segregating for male fertility in crosses withtuberosum haploids. Moreover, a source of male fertility restorer gene(s) was hypothesized in the (haploid x species) hybrid UP88-C80 (tbr x chc). A relatively high freezing tolerance was found in the progenies coming fromtuberosum haploid xsanctae-rosae crosses and some hybrids showed a freezing killing temperature as low as −5.4°C. Although the most tolerant clones did not shed pollen, at least one clone (Sr6,tbr x sct) combined male fertility, 2n pollen production and good freezing tolerance (−3.9°C) and might be directly used in crosses with tetraploidS. tuberosum in order to introgress resistance to low temperature intuberosum form.

Variation for agronomic and processing traits in Solanum tuberosum haploids × wild species hybrids

The broad range of wild and cultivated species relatives of the commerical potato (Solanum tuberosum L.), guarantees the existence of a gene reservoir to be screened for traits that are absent or present only in low frequencies in the cultivated germplasm. Haploids (2n=2x=24) extracted from the cultivated tetraploid (2n=4x=48) germplasm cross easily with diploid (2n=2x=24) potato species and produce fertile progeny. As most haploid × species hybrids tuberize under long day conditions they can be maintained clonally and evaluated for the traits of interest. Three populations involving Tuberosum haploids × Solanum bukasovii (tub x buk), Tuberosum haploids × Solanum sparsipilum, (tub × spl) and Tuberosum haploids × Solanum berthaultii (tub x ber) clones, were planted at two locations in Wisconsin. The haploids were derived from W231, a selected clone from the University of Wisconsin Potato Breeding Program. The objectives were: to evaluate the H-S populations for agronomic and processing traits, and to determine the phenotypic associations between them. Data were recorded on haulm maturity (HM), tuber weight (TW) and tuber number (NT) per plant, specific gravity (SG), chip color after harvest (CH), and chip color after storage at 4° C and one week of reconditioning at room temperature (CH1). Results for SG and CH indicated good potential of the germplasm evaluated to introgress these traits into the commercial potatoes. For instance, the SG mean for tub x buk, tub x spl and tub x ber were 1.079, 1.086 and 1.082, respectively, and their means for CH were 4.8, 3.9 and 3.5. Chip color after storage and reconditioning was found in low frequency in the populations. Four clones in the tub x spl population, and three clones in the tub x ber population had CH1 ≤ 4.0, the commercially acceptable score for this trait. Significant (p ≤ 0.01) phenotypic correlations (r) were found between HM and SG (r=−0.46 in tub x buk, r=−0.61 in tub x spl, and r=−0.34 in tub x ber), NT and TW (r=0.79 in tub x buk, r=0.88 in tub x spl, and r=0.71 in tub x ber), and TW and SG (r=0.40 in tub x buk, and r=0.36 in tub x spl). The correlation coefficients between processing traits were not significant, which may indicate the presence of separate genetic mechanisms governing the inheritance of these traits.

Male fertility and cytology of triploid hybrids between tetraploidSolanum commersonii (2n=4x=48, 2EBN) and Phureja-Tuberosum haploid hybrids (2n=2x=24, 2EBN)

Solanum commersonii Dun. is a diploid (2n=2x=24, 1EBN) wild species of potential value for potato breeding. It is a reproductively isolated species and cannot be crossed with Tuberosum haploids (2n=2x=24, 2EBN) or other diploid 2EBNSolanum species. In order to overcome the EBN barriers, triploid hybrids were produced between Phureja-Tuberosum haploid hybrids, which form 2n pollen grains by parallel spindles, and tetraploidS. commersonii. Microsporogenesis analysis of the triploids indicated a trend towards low values of chromosome distribution at Anaphase I; lagging chromosomes were often observed as well. Despite these abnormalities, the percentage of stainable pollen was very high, ranging from 5.0% to 74.3%. A high variation in pollen grain diameter was also evident. Parallel and tripolar orientation of spindles at Metaphase II of microsporogenesis was a common feature of all the triploids analyzed, but dyads and triads were observed at a very low frequency. Therefore, also the frequency of 2n pollen was very low; the different size of stainable pollen appears to represent the ploidy levels which are possible according to the distribution of chromosomes in Anaphase I. The results obtained also suggest thatS. commersonii could have minor genes acting at the end of meiosis in such a way that, despite the presence of parallel/tripolar spindles, dyads/triads are not formed.

Somatic hybrids between Solanum etuberosum and diploid, tuber bearing Solanum clones

Electrofusion was used to obtain somatic hybrids between Solanum etuberosum (2n=2x=24) and two diploid potato lines. These hybridizations were conducted to determine if haploidxwild species hybrids are better fusion partners than conventional S. tuberosumGp. Tuberosum haploids. Restriction fragment length polymerase (RFLP) analyses of the putative somatic hybrids confirmed that each parental genome was present. The somatic hybrids between S. etuberosum and a haploid S. tuberosum clone, US-W730, were stunted and had curled, purple leaves. In contrast, somatic hybrids between S. etuberosum and a haploidxwild species hybrid (US-W 730 haploidx S. berthaultii), were vigorous and generally tuberized under field conditions. These hybrids were designated as E+BT somatic hybrids. Analyses of 23 E+BT somatic hybrids revealed a statistically significant bias towards the retention of S. etuberosum chloroplasts. Stylar incompatibilities were observed when the E+BT somatic hybrids were used as pollen donors in crosses with S. tuberosum cultivars. Reciprocal crosses did not show this incompatibility. The progeny were vigorous and had improved tuber traits when compared to the maternal E+BT parent. RFLP analyses of three sexual progeny lines confirmed the presence of all 12 S. etuberosum chromosomes. In two of these lines, RFLPs that marked each of the 24 chromosome arms of S. etuberosum were present. However, RFLP markers specific for regions on chromosomes 2, 7, and 11 were missing from the third clone. Because other markers for these chromosomes were present in the progeny line, these results indicated the likelihood of pairing and recombination between S. etuberosum and S. tuberosum chromosomes.

Resistance to potato virus Y in somatic hybrids between Solanum etuberosum and S. tuberosum x S. berthaultii hybrid

Somatic hybrids between a potato virus Y (PVY) resistant Solanum etuberosum clone and a susceptible diploid potato clone derived from a cross between S. tuberosum Gp. Tuberosum haploid US-W 730 and S. berthaultii were evaluated for resistance to PVY. All but one of the tested somatic hybrids were significantly more resistant than cultivars 'Atlantic' and 'Katahdin'. However, none was as resistant as the S. etuberosum parent. One hexaploid somatic hybrid, possibly the product of a triple-cell fusion involving one S. etuberosum protoplast and two haploid x S. berthaultii protoplasts, was as susceptible to PVY infection as the cultivars. Tetraploid progeny of the somatic hybrids, obtained from crosses with Gp. Tuberosum cultivars, were neither as resistant as the maternal somatic hybrid parent, nor as susceptible as the paternal cultivar parent. It appears that the introgression of PVY resistance from (1EBN) S. etuberosum into (4EBN) S. tuberosum (EBN-endosperm balance number) will be successful through the use of somatic hybridization and subsequent crosses of the somatic hybrids back to S. tuberosum.

Inheritance of potato chip color at the 24-chromosome level

Potato chip color is an important trait in determining quality and acceptability of cultivars for processing. Therefore, the genetics of potato chip color is important in varietal breeding. The use of 2xS. tuberosum Group Tuberosum haploid × 2x wild species hybrids provides disomic inheritance for genetic analysis of chip color. Two hundred forty haploid-species hybrids representing crosses between 15 haploids and 10 species were evaluated for potato chip color after 21 days reconditioning (18 to 20C) following 6 months at 4C. Phenotypic variation for chip color was found among the haploid-species hybrids. Crosses between good, medium, and poor performing clones were made in all combinations and 136 families were obtained. Progeny were evaluated for reversion resistance (chipping direct from 10C storage) and reconditioning (chipping from 4C after tuber warming) chip color. Variation for chip color was found among the progeny for each trait. Significant correlations between reversion resistance and reconditioning were found. Yet, the use of at least two chip tests (one for reversion resistance and one for reconditioning) is more informative than the use of one test and subsequent predictions made, regarding the other trait. A three locus hypothesis for both reversion resistance and reconditioning is proposed for the inheritance of each trait. One dominant allele at each of the three loci is necessary for good chipping. One or two loci may be common to both characteristics. Good chipping 2x hybrids that produce 2n pollen can be used to transfer this trait to the 4x level through use of 4x × 2x crosses.

Identification of genetic factors influencing chip color in diploid potato (Solanum spp.)

A genetic map was constructed with a combination of isozymes, restriction fragment length polymorphisms (RFLPs) and randomly amplified polymorphic DNA (RAPDs) to apply quantitative trait loci (QTL) analysis to identify genetic factors that contribute to chip color in potato. The diploid population used was a cross between aSolanum tuberosum haploid andS. chacoense hybrid used as female parent and aS. phureja clone used as male. Chip color was determined visually on samples fried from tubers stored at 10C. On a scale of 1 (light color) to 10 (dark color), the population ranged from 2 to 8 while the parents average chip color was 3.5. Based upon one-way ANOVAs (P < 0.05), 13 genetic markers showed significant associations which represent a total of six QTLs. A multiple locus model based upon the markers that have the largest effect per QTL explained 43.5% of the phenotypic variation for chip color in the population and increased to 50.5% when one significant epistatic interaction was included in the model. All the significant marker associations were identifed in theS. tuberosum-S. chacoense hybrid. Through preliminary data, the results of this study suggest that additive effects contribute a significant portion of the genetic variation for chip color. The identification of these QTLs for chip color variation provides the means to apply marker-assisted selection to introgress these genes into the cultivated potato germplasm.

Breeding for 2n egg production in haploid × species 2x potato hybrids

The identification of diploid(2x), Tuberosum haploid × wild species hybrids (HS) which produce high frequencies of 2n eggs and have good tuber appearance is important for obtaining 4x progeny from 2x × 2x crosses in potato. More than 1700 HS clones were screened for the occurrence and frequency of 2n eggs through 2x × 4x crosses during two pollination periods. The resulting seed/fruit provides an estimate of 2n egg frequency. About 650 HS clones produced 2n eggs; 63 had 10–114 seeds/fruit; and 27 were selected for good tuber type. There was a large variation in seed set after 2x × 4x crosses, both within and between clones. Comparison of fruit and seed set between the two pollination periods using the same 595 clones indicated that environmental factor significandy affected 2n egg frequency. The mode of 2n egg formation was mainly due to omission of the second meiotic division controlled by a recessive gene (os). The gene frequency foros varied from 0.28 to 0.76 among six, 2x taxa. Both additive and dominance variance were significant for seed set after 2x × 4x crosses. Narrow sense heritability was estimated as 0.24. The average degree of dominance was equal to 1.66 which could indicate overdominance. Alternatively, this could be due to linkage disequilibrium,i.e. pseudooverdominance. However, dominance variance was greater than the additive variance indicating recurrent selection can be useful for improving the frequency of 2n egg production in the 2x population. The selected individuals, based on their phenotypic performance, should be progeny tested to identify the best parents for use in the next cycle of recombination.