Utilizing the frost resistance of diploid solanum species

Introductions of 13 frost-resistant diploid (2n=2x=24)Solanum species (2 non-tuberous) were inter-crossed to produce F₁ hybrids involving two frost resistance sources. Successful F₁ combinations were: (i) inter-crossed to produce hybrids involving 3 to 4 frost resistance sources, (F₁×F₁); and (ii) crossed with haploids (2n=24) of frost-susceptible tetraploid (2n=4x=48)S. tuberosum subsp.tuberosum, (F₁×tbr). Thirty-five of the 75 crosses between the 13 species produced viable seed. The F₁ and F₁×F₁ progenies segregated to yield plants more resistant to freezing temperatures than the parental species. All attempts to cross these hybrids with 24-chromosometbr were successful. Although tuber production improved following the first cross totbr, the frequency of resistant plants decreased approximately 50%. Thus far, all first back-cross attempts, (F₁×tbr)×tbr, have succeeded. Utilizing hybrids synthesized from several sources of frost resistance as the donor parents, and clonal selections of 24-chromosometbr as the recurring parents, may provide a high order of resistance throughout the backcrossing program and implement selection of frost-resistant, long-day tuberizing parental stocks acceptable to the potato breeder.     

Cytoplasmic substitution inSolanum. I. Seed production,germination, and sterilities of reciprocal backcross generations

Summary Genes ofSolanum tuberosum L. ssp.tuberosum were introduced into cytoplasm ofS. tuberosum ssp.andigena (Juz. & Buk.) Hawkes andS. phureja Juz. & Buk. to reduce cytoplasmic sterilities. Alleles that influenced berry set and seed content produced differences between reciprocal progenies. Their expressions were interpreted in terms of genes of the maternal and/or paternal parents. When the ssp.andigena genotype was introduced from the maternal parent if favoured high berry set and low seed number. Maternal ssp.tuberosum favoured lower berry set and higher seed number. The source ofS. phureja cytoplasm, aS. phureja×S. chacoense F₁, used as maternal parent favoured lower berry and seed set. During successive backcrosses progeny expressions approached those of the recurrent ssp.tuberosum pollen parents. A low level of pollen sterility occurred in BC₂ plants, unrelated to direction of cross or cytoplasmic factors. By BC₄, seed set and fertility were as good as ssp.tuberosum, and recurrent backcrossing could be terminated without loss of seed production or fertility. Pennsylvania Agricultural Experiment Station Journal Series No. 7493.     

Acaphu: A tetraploid,fertile breeding line,selected from anS. acaule ×S. phureja cross

An artificial, fertile breeding line was obtained by crossingS. acaule, Bitter toS. phureja Juz et Buk. It has 2n = 48 chromosomes and was formed by fertilization ofS. acaule eggs (n = 24) andS. phureja pollen grains (2n = 24). It possesses valuable characters ofS. acaule andS. phureja combined, and is highly male and female fertile, producing abundant seed by selfing or by crossing to tetraploid, cultivated potato clones. Since it is tetraploid and easy to cross, it is expected to be more efficient for transferring valuable genes fromS. acaule to cultivars than the natural, well known, hybridogenic speciesS. ×juzepczukii (2n = 36) and S. ×curtilobum (2n = 60).     

The effect of direction of hybridization (4x × 2x vs. 2x × 4x) on yield of cultivated potatoes

The types of reciprocal differences found in higher plants are generally limited to those characters which are controlled by one or two major genes. There are, however, a few examples where the progeny from reciprocal crosses differ in yield or other quantitative characters. Both have been ascribed mainly to cytoplasmic inheritance. In cultivated tetraploid potatoes (Solatium tuberosum Group Tuberosum) differences in yield of the progeny from reciprocal crosses have been found. These reciprocal differences are attributed to the mode of 2n gamete formation rather than cytoplasmic differences. Preliminary experiments involving reciprocal crosses between Tuberosum cultivars (2n=4x=48) and Phurejahaploid Tuberosum hybrids (2n=2x=24) indicated that some progenies from 4x x 2x crosses yield more than progenies from reciprocal crosses. It was hypothesized that the differences in tuber yield may be due to one or more of the following factors: (i) the effect of high fruit set on yield —i.e. almost 100% of the progenies from 2x X 4x crosses are male fertile and thus higher fruit set is expected than in the 4x X 2x progenies where only about 57% of the F₁ clones are male fertile; (ii) the advantage of being in 4x cytoplasm if the nucleus is 4x; and (iii) the difference in mode of 2n gamete formation-first division restitution (FDR) gives highly heterozygous gametes and second division restitution (SDR) results in highly homozygous gametes. Six families of reciprocal crosses between three 4x cultivars (Merrimack, Superior, and Wis 639) and three 2x Phureja-haploid clones (W5293.3, W5295.7, and W5845.1) were planted in 1970 in Rhinelander in a split-plot design with two tubers of each clone in each of two replicates. Flowers were removed from one of the paired plants in each family. Even though the fruit yields were higher in 2x X 4x progenies as expected, tuber yields were not affected by fruit formation. Therefore, factor (i) is apparently not involved in the reciprocal differences observed. Significantly higher tuber yields were obtained from 4x X 2x families only with the 2x clone W5295.7. This eliminated factor (ii), since no difference in reciprocals was obtained with the other 2x clones. Factor (iii) could thus be responsible for differences in reciprocal crosses, since cytological observations indicate only W5295.7 of the 2x clones used forms 2n pollen by FDR.     

Glycoalkaloid content in tubers of hybrid and backcross populations from aSolanum tuberosumx s. chacoense cross

The glycoalkaloid contents of tubers were measured in hybrid and back-cross populations derived from a cross ofSolarium chacoense Bitter (PI 472810, 2n=2x=24) withSolanum tuberosum L. (2n=4x=48). In tubers sampled from near the median size in each population and parent, the mean total glycoalkaloid (TGA) contents of theS. chacoense andS. tuberosum parent genotypes, and the F₂ and F₄ hybrid genotypes were 245, 4, 39, and 35 mg/ 100 g fresh weight (fw), respectively. Backcrossing the F₂ genotypes toS. tuberosum reduced the mean TGA content to 15 mg/100 g fw. A second backcross further reduced the content to 9 mg/100 g fw. However, some genotypes in both backcross populations (25% and 10%) had TGA contents above the generally accepted upper limit of 20 mg/100 g fw. Tuber size was significantly smaller in theS. chacoense parent and the hybrid populations compared with the size in theS. tuberosum parent and backcross populations. In a second sampling of tubers from each population, intentionally selected to equalize tuber size among populations, the mean tuber size for populations was reduced by 79%. The mean population TGA content increased by 44%. However, the differences and rankings among parent, hybrid, and backcross populations were the same for both tuber samplings.     

Male sterility and 2n pollen in 4x progenies derived from 4x×2x and 4x×4x crosses in potatoes

Summary The objectives of these experiments were to evaluate male sterility (MS) and 2n pollen frequency in 77 4x×2x families and 26 4x×4x families. The 2x parents were haploid-species hybrids and the 4x parents mainly clones withS. tuberosum ssp.tuberosum (tbr) andS. tuberosum ssp.andigena (adg) in their genetic background. The female parents had different cytoplasms: adg, tbr,S. demissum (dms), andS. stoloniferum (sto). Families from female parents with adg or dms cytoplasm did not have MS plants. However, families derived from crosses of many tbr females with 2x hybrids and a 4x adg clone had a high percentage of MS plants; all the progenies from cv. Serrana (sto cytoplasm) possessed tetrad sterility. These results can be explained as due to an interaction between a dominant male sterility (Ms) gene with tbr and sto cytoplasms. Some MS 4x hybrids (tbr×adg) used as females had the Ms gene, but male fertility was restored in some plants of their progenies when they were crossed with 4x tbr clones. This indicates that some tbr clones have a fertility restorer gene (Rt). The results from both 4x×2x and 4x×4x families fit the expected ratios assuming chromatid segregation for both the Ms and Rt loci. The gene frequency of parallel spindles was estimated in the 4x population as 0.74.     

Variation for 2n pollen production and male fertility in wildSolanum germplasm resistant toPhytophthora infestans (Mont.) de Bary (US-8)

Wild potato species possess genetic variability for valuable traits including resistance toPhytophthora infestans, the causal agent for potato late blight disease. Breeding schemes using 2n gametes are a powerful tool for introgressing these traits. Forty-five plant introductions (PIs) across 12Solanum species representing various Endosperm Balance Numbers (EBN) and having resistance toP. infestans (US-8, isolate ND 95-2) were screened for 2n pollen production and male fertility. Species evaluated were 2x(lEBN)S. bulbocastanum, S. cardiophyllum, S. pinnatisectum, andS. trifidum; 2x(2EBN)S. berthaultii, S. megistacrolobum, S. microdontum, andS. verrucosum; 4x(2EBN)S. fendleri andS. stoloniferum; 6x(4EBN)S. guerreroense; and 2x(unknown EBN)S. polyadenium. Acetocarmine staining and cytological analyses were used to determine 2n pollen production and pollen stainability from genotypes grown across three locations. Based on frequency, genotypes were assigned to 2n pollen production categories (0≤1%, Low=1%–10%, Medium=11%–60%, and High≥60%) and pollen stainability categories (0≤1%, 1=196–5%, 2=6%–10%, 3=11%–20%, 4=21%–50%, and 5≥50%). Eleven of the 12 species and 37 of the 45 PIs examined contained genotypes producing ≥1% 2n pollen with ranges of 0%–63.6% and 0%–81.8%, respectively. Dependence was found between location and pollen stainability, EBN and pollen stainability, but not for location and frequency of genotypes with 2n pollen. Stability for 2n pollen production was observed across environments. Among the 1EBN germplasm, dependence was found between 2n pollen and pollen stainability. Resistance toP. infestans was found to be independent of 2n pollen production; therefore, identifying genotypes combining 2n pollen production, fertility, and resistance toP. infestans was possible.     

Evolutionary pathway of T-type Chloroplast DNA in potato

Potato was domesticated in the Andes of South America. However, the presently worldwide-grown potato (Solanum tuberosum L. ssp.tuberosum, 2n=4x=48) has characteristic T-type chloroplast DNA that was introduced after late blight epidemics in the mid-19th century from the Chilean potato (2n=4x=48) grown in the southern coastal regions in Chile. Among many wild potato species, the same chloroplast DNA was found only in some populations of a diploid speciesS. tarijense Hawkes (2n=2x=24), which ranges from central Bolivia to northwest Argentina. To elucidate an evolutionary pathway of T-type chloroplast DNA fromS. tarijense to Chilean potato, 215 accessions ofS. stenotomum Juz. et Buk., considered to be the most primitive, diploid cultivated potato species, from which all the Andean cultivated species evolved, and 286 accessions of the most widely grown, Andean tetraploid cultivated speciesS. tuberosum L. ssp.andigena Hawkes (2n=4x=48) were examined in this study. No accession ofS. stenotomum had T-type chloroplast DNA, while nine accessions, mostly from northwest Argentina, ofS. tuberosum ssp.andigena had T-type chloroplast DNA. Therefore, I conclude that some populations ofS. tarijense having T-type chloroplast DNA were naturally crossed as female withS. tuberosum ssp.andigena from which the Chilean potato was selected.     

Evaluation for chipping and tuber soft rot (Erwinia carotovora) resistance in potato clones from unilateral sexual polyploidization (2x × 4x)

The aim of this work is the evaluation of tetraploid potato clones with germplasm fromS. chacoense andS. phureja for chipping and resistance to tuber soft rot (Erwinia carotovora subsp.carotovora). Tetraploid potato clones were obtained via 2n eggs through 2x–4x crosses. Cytological mechanisms were investigated in the diploid hybrid 2n-egg producer used as parent in the unilateral sexual polyploidization scheme; the omission of the second meiotic division and cytokinesis failure at the end of meiosis were identified with a genetic consequence equivalent to second division restitution (SDR). Tuber marketable yield, specific gravity and chip-processing ability evaluated at harvest and after cold storage with and without reconditioning were higher in some clones as compared to theS. tuberosum tetraploid parent. Tuber soft rot resistance, contributed byS. chacoense, was successfully transferred to the4x level. After inoculation withErwinia carotovora subsp.carotovora and estimation of the rotted area, one tetraploid clone proved to be resistant and three partially resistant. The genetic value of exotic germplasm and sexual polyploidization strategy are discussed in relation to potato improvement.     

Cytoplasmic substitution inSolanum. II. Tuber characteristics of reciprocal backcross progeny

Summary Recurrent backcrosses (BC) were used to introduce nuclear factors ofS. tuberosum L. ssp.tuberosum into the cytoplasm of Andean potatoesS. tuberosum ssp.andigena (Juz. and Buk.) Hawkes andS. phureja Juz. and Buk. Each generation BC was produced in reciprocal directions to test the effect of cytoplasmic substitution on yield. This report analyses tuber number, tuber weight, and tuber length of BC₂ and BC₃ substitution into ssp.andigena cytoplasm and BC₂ and BC₂ substitutions intoS. phureja cytoplasm. Direction of cross had no consistent significant effect on yield components of ssp.andigena substitution lines. InS. phureja reciprocal progenies direction of cross was responsible for significant differences of some reciprocal sets. When there were significant differences the higher yield usually occurred when ssp.tuberosum was the pistillate parent, but in some progenies when ssp.tuberosum was the pistillate parent the yield was significantly lower than in the reciprocals. It is suggested that differences between reciprocals resulted from chromosomally encoded gene action, resulting from maternal and/or paternal effects, rather than cytoplasmic factors. Authorized for publication as paper No. 7494 in the Journal Series of The Pennsylvania Agriculture Experiment Station.     

Selection for 2n gametes and tuberization inSolanum chacoense

A breeding scheme is outlined which broadens the genetic base of the potato by incorporation of germplasm fromS. chacoense (Argentina) with Group Phureja and haploids of Group Tuberosum. Selection for adaptation to cultivation and production of 2n gametes is carried out at the diploid level, with the benefits of disomic ratios.S. chacoense provides increased genetic diversity, vegetative vigor and profuse flowering. About one-fifth of the 654 plants examined produced 2n eggs; clones with high 2n egg production may be obtained either by screening within particular introductions or by intercrossing known 2n egg producers. Only 16 of 1,212 plants screened produced 2n pollen —one by first division restitution. There is great variation among introductions and individuals for tuberization in the greenhouse. Selection for early tuberization in the field may be difficult due to extremely long stolons, but it would be facilitated by incorporation of the short stolon characteristic from the topiary mutant typeS. infundibuliforme. Male sterile Tuberosum haploids hybridize withS. chacoense to give male fertile interspecific F₁ progeny, some of which produce 2n eggs or 2n pollen.     

A restorer gene for genetic-cytoplasmic male sterility in cultivated potatoes

Fifty-nine clones (cultivars and advanced selections) ofSolanum tuberosum Group Tuberosum were pollinated with two Group Phureja-haploid Tuberosum hybrids, which produce FDR 2n pollen (ps/ps) and are heterozygous for a dominant gene (Ms/ms) which interacts with Tuberosum cytoplasm to result in male sterility. Twenty-eight and thirty-four plants from each family were transplanted to the field in two different locations, respectively. Pollen samples were collected from each plant and stained with acetocarmine glycerol jelly. Plants with over 95% unstainable pollen were considered male sterile. The families could be put into distinct classes according to the ratio of male sterile to male fertile plants; 3 sterile: 1 fertile (23 clones), 2 sterile: 3 fertile (24 clones), 1 sterile: 5 fertile (3 clones), and 0 sterile: 1 fertile (3 clones). Four families deviated significantly from any expected ratio. The variation for male fertility among families resides in the tetraploid parent, since the male parents have the same genotype (Ms/ms) for theMs locus. The locus of this gene is very distal to the centromere such the 2n gametic output is 1 Ms/Ms : 2 Ms/ms : 1 ms/ms. The variation in male sterility is explained by the presence or absence of a dominant allele of a restorer gene (Rt) in tetraploid Tuberosum. This locus is also very distal to the centromere. The frequency of theRt gene was estimated to be 0.20 in Tuberosum. The finding ofRt gene explains some of the conflicting results in male fertility of haploid Tuberosum x 2x cultivated and wild species hybrids. It also provides an opportunity to partially circumvent this male sterility. Haploids (2n = 2x = 24) extracted from 4x clones withRt genes would have male fertile progeny in crosses with species carrying theMs gene.     

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

Summary 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. Contribution no. 137 from the Research Centre for Vegetable Breeding-CNR. Research supported by National Research Council of Italy, special project RAISA, subproject no. 2, paper no. 2776.     

Breeding Potential of Potato Somatic Hybrids: Evaluations for Adaptability,Tuber Traits,Late Blight Resistance,Keeping Quality and Backcross (BC<Subscript>1</Subscript>) Progenies

Ten tetraploid interspecific potato somatic hybrids developed earlier through protoplast fusion between the dihaploid Solanum tuberosum L. ‘C-13’ and the diploid wild species Solanum pinnatisectum Dun. were used in this study. Somatic hybrids and standard control cultivars were evaluated for adaptability, tuber traits, late blight resistance and keeping quality attributes (dormancy, sprouting, weight loss and appearance after 75 days of storage) during two successive winter crop seasons (short-days) in the sub-tropical plains of India, where nearly 90% of India’s potatoes are grown. Somatic hybrids showed medium to good plant vigour and late to very late plant maturity. Though none of the somatic hybrids produced equal or higher tuber yield than the control cultivars, except clone P7, all other somatic hybrids produced significantly higher marketable and total tuber yield after a 90-day growth than the parent C-13. Most of the somatic hybrids possessed higher tuber dry matter concentration, resistance to late blight and better keeping quality attributes than the control cultivars. Notably, the tubers of somatic hybrids showed a tendency for colour change from white to purple on exposure to sunlight. Breeding potential of somatic hybrids was further assessed by crossing them with commonly grown local potato cultivars for the development of backcross (BC₁) progenies. In the hybridization programme, berries were formed while somatic hybrids were used either as male or female parents with common cultivars. Our results revealed that the use of bulk pollen of somatic hybrids not only resulted in higher pollination success but also helped to achieve higher numbers of true potato seeds (TPS) per berry. Despite the formation of berries, TPS was absent in 22 crosses using somatic hybrids as a female or male parent which may be due to ovule or embryo abortions. Segregating BC₁ generations were evaluated under the same sub-tropical conditions for further exploitation in potato breeding. Based on our study, the somatic hybrids P4, P8 and P10 can be utilized for the introgression of important characters such as high tuber dry matter concontration, resistance to late blight and excellent keeping quality attributes into the cultivated potato via conventional breeding methods for cultivar development in the sub-tropical plains of India.     

Cytoplasmic evaluations during substitution backcrossing inSolanum

Summary Sterility-resistant cytoplasmic factors ofSolanum tuberosum spp.andigena (Juz. & Buk.) Hawkes and ofSolanum phureja (Juz. & Buk.) were combined with chromosomal genes ofS. tuberosum ssp.tuberosum. Eleven morphological characters of reciprocal F₁ and BC₁ progenies were monitored to evaluate rate of progression towards characteristics occurring in the recurrent ssp.tuberosum parent. Female and male fertilities of F₁ and BC₁ populations were also evaluated. Cytoplasmic factors did not influence morphological parameters consistently, except those relating directly to fertility. Differences between reciprocal progeny depended more upon the genes of individual parents used in crosses than on cytoplasm source. No consistent reciprocal differences occurred in tuber characters and those differences which arose seem to reflect a maternal rather than a cytoplasmic influence. F₁ and BC₁ progenies containing the sterility-resistant cytoplasmic factors of ssp.andigena or ofS. phureja had higher fertility than their respective reciprocal progenies with cytoplasm of ssp.tuberosum. Pennsylvania Agricultural Experiment Station Journal Series No 6212.     

Performance of populations derived by selecting for resistance to potato leafhopper in A 4XSolanum Tuberosum X 2XSolanum Chacoense cross

Intercross and backcross populations derived from aSolanum tuberosum L. (2n = 2x = 48) ×Solanum chacoense Bitt. (2n = 2x = 48) cross by selecting for resistance to potato leafhopper (PLH) (Empoasca fabae Harris), were tested in the field. In F2 and F4 populations, the levels of nymphal infestation and leaf necrosis from PLH feeding were considerably lower and tuber yields were much lower compared with those measured on the susceptible cultivar Kennebec. No differences were found between the two populations. Backcrosses of the F2 and F4 populations toS. tuberosum decreased resistance to PLH. Resistance to infestation was still greater than that of Kennebec; severity of leaf necrosis equaled that of Kennebec. Tuber yields increased, but were less than Kennebec’s yield. Foliage types shifted closer to those ofS. tuberosum. Three generations of selection and intercrossing within the backcross populations did not change the level of resistance to PLH but did tend to lower tuber yields. Foliage types remained primarilyS. tuberosum in appearance although distinctive types segregated with increased frequency. A second backcross toS. tuberosum did not change the level of resistance to PLH or the level of tuber yield relative to those of the first backcross or Kennebec.     

Haploid extraction inSolanum tuberosum group Andigena

Haploids were extracted from 66 differentSolanum tuberosum Group Andigena introductions usingS. tuberosum Group Phureja clone 1.22 as pollinator. The overall frequency of haploids was 13 per 100 fruit, with a high of 106. The overall average number of seeds per fruit was 1.4. Some Group Andigena clones produced primarily haploid, others primarily triploid, and still others primarily tetraploid progeny. The “triploid block” is still evident in these materials even though some clones produce primarily triploid progeny. It is suggested that the ploidy level of progeny produced in 4x - 2x crosses may be under relatively simple genetic control.     

Production and characterization of “second generation” somatic hybrids derived from protoplast fusion between interspecific somatohaploid and dihaploidSolanum tuberosum L.

Protoplasts were fused to produce somatic hybrids between a triploid (2n=3x=32-34) interspecific somatohaploid betweenSolanum brevidens Phil. andS. tuberosum L., and a dihaploid (2n=2x=24) anther-derived line ofS. tuberosum cv. Van Gogh. A total of 265 plants were regenerated from protoplast fusion derived calli and their hybridity was verified using fusion partner specific RAPD markers. These “second generation” somatic hybrids were aneuploid pentaploids (2n=5x=51-65) with a 2C DNA content ranging from 3.36 to 4.43 pg, which corresponded to the sum of the 2C values of each of the fusion partners (somatohaploid: 2.22 pg; and the dihaploid line of cv. Van Gogh: 1.87 pg). Most of the “second generation” somatic hybrids were vigorous, but variable in morphology. They were extremely resistant to PLRV and they had tolerance to PVY infection derived from the somatohaploid fusion partner. Even though most of the “second generation” hybrids tuberized, the tuber morphology was variable and most were poorly shaped. InErwinia soft rot resistance tests, the tubers showed higher level of resistance than the tetraploidS. tuberosum cultivars, the dihaploidS. tuberosum fusion partners and the hexaploid somatic hybrids betweenS. brevidens andS. tuberosum. The “second generation” somatic hybrids were all male sterile and failed to produce berries or seeds.     

Intermating of potato haploids and spontaneous sexual polyploidization—Effects on heterozygosity

Solanum tuberosum Group Tuberosum haploids (2n = 2x = 24) were inter-mated using bulk pollen. Nearly half of the resulting progeny were tetraploid, arising from bilateral sexual polyploidization. Subsequent yield trials indicated that the diploid progeny did not yield significantly more than the original haploids, but the tetraploid progeny did yield significantly more than their diploid sibs and the original haploids. Tetraploid cultivars and conventional tetraploid breeding families significantly out-yielded the haploid-derived tetraploids. These results and the theoretical considerations which are presented indicate that inter-mating haploidsper se does not increase heterozygosity. More importantly, these results indicate that without the introduction of new germplasm, haploidization followed by repolyploidization can only be expected to result in tetraploids with heterozygosity equal to or below that of their parents.     

2n pollen formation via parallel spindles in the potato cultivar sebago

Parallel spindles occur during microsporogenesis in the cultivar Sebago (2n=4x =48) and are the basis of 2n pollen production in this clone. The relations between the frequencies of parallel spindles, 2n microspores, and 2n pollen are striking. The predicted frequency of 2n pollen based on the percentage of microsporocytes with parallel spindles is highly correlated with the frequency of 2n pollen observed. The percentage of 2n microspores at the sporad stage also corresponds very closely with the observed percentage of 2n pollen. Sebago is thus nulliplex (psp sp sps) for the meiotic mutant which governs spindle orientation at the second meiotic division in microsporocytes. This finding agrees with the prediction that if parallel spindles were involved in the origin of cultivated tetraploid potatoes, the genesps should be present in some 4x cultivars. It adds further support to the concept that 2n gametes have been a creative factor in the origin and evolution of cultivated tetraploids.