Solanum Incamayoense: A new tuberous species of Argentina

Solarium incamayoense is described as a new wild tuber-bearing species from dept. Rosario de Lerma, prov. Salta, Argentina. This species is morphologically related toS. gourlayi Hawkes from which it differs by the shape, size and colour of the leaflets and by the habit of growth. The species is diploid (2n = 24).     

Lack of association between genetic and geographic origin characteristics for the wild potatoSolarium sucrense Hawkes

Collecting germplasm to broaden breeding resources is an essential activity of genebanks. Research to understand how genetic diversity is partitioned in nature might help to identify collections rich in diversity. Previous studies among wild populations ofSolanum fendleri (a disomic polyploid selfer) andS. jamesii (a diploid outcrosser) revealed no significant associations between genetic and ecogeographic variation. Even physical separation did not predict genetic differences. In this study, 28 populations ofS. sucrense Hawkes (2n=4x=48), a Bolivian species with another breeding system (polysomic polyploid oucrosser), were evaluated. The objective was to assess whether genetic differences between populations are predicted by differences in geographic parameters at the natural site of origin. Genetic differentiation was estimated by using 216 RAPD markers. The average genetic distance (GD) found between pairs of populations was 31% (ranging from 8% to 44%). Correlations of GD with latitude, longitude, altitude and distance were not significant. Multiple regression analysis also confirmed that GD was not explained by the geographic parameters used. We conclude that geographic origin data is not very useful in gauging interpopulation genetic diversity in the genebank.     

Genetic heterogeneity estimated by RAPD polymorphism of four tuber-bearing potato species differing by breeding system

Most wild potato germplasm in genebanks is collected, preserved, and evaluated as botanical seed populations that may be highly heterozygous and heterogeneous. However, some species are selfers so potentially very homozygous, perhaps also homogeneous. Intrapopulation heterogeneity increases sampling error that can undermine consistency in seed regeneration in the genebank, screening results, germplasm collecting, and estimates of taxonomic relationships. Thus, knowledge of genetic heterogeneity (GH) can predict the need to commit more resources for larger sample sizes or replication when populations of a given species are being regenerated, evaluated, collected, and classified. This study investigated withinpopulation GH in 32 potato populations comprising four different breeding systems observed inSolatium species:S. fendleri (2n=4x=48, disomic selfer),S. jamesii (2n=2x=24, outcrosser),S. sucrense (2n=4x=48, tetrasomic outcrosser), andS. verrucosum (2n=2x=24, selfer). RAPD markers were used to estimate heterogeneity among 24 individuals per population. Populations ofS. verrucosum were quite homogeneous with an average GH of 6.0%. Similarly low heterogeneity was detected among the eight populations ofS. fendleri (average GH=7.1%). In contrast,S. jamesii andS. sucrense had a much higher GH of 29.4% and 44.1%, respectively. These results demonstrate and quantify the great difference in intrapopulation heterogeneity among wild potato species. Calculations based on intrapopulation heterogeneity indicate that samples should be composed of 25 to 30 random plants for low sample variation that is uniform for all species.     

Fine screening potato (Solanum) species germplasm for tuber calcium

A representative sample of the US potato germplasm collection was screened in the greenhouse for ability to accumulate tuber calcium. Germplasm with very high and very low calcium accumulation capacity has been identified by progressively screening for extremes among species, among populations (accessions), and finally among genotypes (i.e., fine screening). Among accessions, the bestS. gourlayi andmicrodontum populations averaged more than six-fold greater tuber calcium than the poorestS. kurtzianum population. When screening was performed at the genotype level, the best individuals ofgourlayi andmicrodontum had over eight-fold greater calcium than the poorestkurtzianum individual. When grown in the field, calcium levels ofS. microdontum selections were confirmed to be significantly higher thantuberosum cultivars. When selected genotypes were intercrossed and their progeny were compared to their original populations, mean tuber calcium was significantly changed in the intended direction of selection forS. kurtzianum (made lower) andS. microdontum (made higher). These stocks are expected to be useful tools for study of the genetic and physiological bases of tuber calcium accumulation, refining screening methods, and breeding improved cultivars.     

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).     

Male fertility and 2n pollen production in haploid-wild species hybrids

Hybrids (2n = 2x = 24) from crosses between Group Tuberosum haploids (2n = 2x = 24) and 24-chromosome, wild species must be fertile and produce 2n gametes if they are to be used in breeding schemes involving sexual polyploidization. Male fertility of the 2x hybrids depends primarily on the species parent. Hybrids involvingS. berthaultii, S. chacoense, S. kurtzianum, S. spegazzinii, andS. tarijense as parents are male fertile. The haploid parents are male sterile, so the species not only contribute genetic diversity and desirable traits, but also genes necessary for male fertility.S. infundibuliforme, S. raphanifolium, andS. sanctae-rosae parents produced male sterile hybrid progeny, while use ofS. boliviense, S. bukasovii, S. canasense, S. microdontum, andS. sparsipilum results in families containing both fertile and sterile plants. Sterility may be due to interactions between Tuberosum cytoplasm and dominant nuclear genes from the species. Genetic male sterility and environmental conditions may also be responsible for low pollen stainability in some hybrids. Many fertile, 2x hybrids produce 2n pollen. Selection for 2n pollen in the species is fast and efficient, but meiotic analysis is required in male sterile haploids. Fortunately, selection in the species parent alone results in a large number of 2n pollen-producing hybrids. These 2x hybrids have been used to generate tetraploids via 4x × 2x crosses.     

Resistance toFusarium sambucinum in wild and cultivatedSolanum species

Several species ofFusarium cause both dry rot in storage and seedpiece decay in the field, which results in severe losses in all major potato-growing regions. In temperate climates twoFusarium species are the principle causes of tuber dry rot-F.sambucinum andF. coeruleum. Resistance toF. sambucinum was identified in cultivated and wildSolanum species. Relatively high levels of resistance exist in the following 2EBN species:S. boliviense (PI#’s 265860, 310975, 310974, 545853, 545889, 545964),S. gourlayi ssp.gourlayi (PI#’s 473344, 545865),S. microdontum (PI#’s 310979, 473171),S. sancta-rosae (PI# 498392),S. kurtzianum (PI# 472930),S. fendleri ssp.fendleri (PI# 262895),S. gandarillasii (PI#’s 265866, 283076),S. oplocense (PI# 498281) andS. vidaurrei (PI# 498332). The resistance is highly heritable but appears, at least in some of the species, to be associated with recessive alleles.     

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.     

Germplasm transfer from the wild tetraploid speciesSolanum acaule bitt. to the cultivated potato,S. tuberosum L. using 2N eggs

Solanum acaule Bitt. (acl ¹, 2n=4x=48 and 2n=6x=72) is a wild potato species in which resistance to adverse biotic and abiotic agents has been found. The tetraploid subspeciesacaule andaemulans cannot be easily crossed with the common potato,S. tuberosum L. Grouptuberosum (tbr, 2n=4x=48) because the development of the endosperm in the hybrid seed is abnormal. Since 4xacl behaves as a diploid in intra and interploidy crosses, it was postulated that the hybridization barrier withtbr could be overcome if the wild species produced 2n gametes. One hundred and ninety-seven plants of 4xacl (22 introductions) were screened for pollen of heterogeneous size; only two plants produced this type of pollen, with 3% and 5% of pollen in the large class, respectively. On the other hand, 427 plants (47 introductions) were screened for 2n eggs through controlled crosses withtbr. Forty-one plants (13 introductions) produced 219 berries of which 46 contained at least one plump seed. Seed from berries with low seed set gave rise to hexaploid hybrids possibly through the functioning of 2n eggs from 4xacl. A sample of these hybrids was successfully backcrossed as female parents totbr, giving rise to 5x BC₁ progenies. These BC₁ progenies could be easily back-crossed totbr, yielding tetraploid or near tetraploid BC₂ plants.     

Reduction of Seed Dormancy in Echinacea pallida (Nutt.) Nutt. by In-dark Seed Selection and Breeding

Strong seed dormancy has been an obstacle for field production of Echinacea species. Previous research on overcoming Echinacea seed dormancy has been extensive and focused on treatment methods, which involve time and expense, and are incompatible with organic production if synthetic chemicals are used. We have attempted to genetically reduce seed dormancy through selection and breeding in Echinacea, by using Echinacea pallida as a model species. Three accessions were used in this study. Nine parent plants of each accession selected from early, in-dark germinated seeds (in-dark plants) or from late, in-light seeds (in-light plants) were planted and grouped by accession and germination treatment method for seed production through a polycross method. Germination tests indicated that these in-dark plants produced seed (in-dark seed) with significantly reduced seed dormancy when tested under light or dark condition in comparison to the seed of the in-light plants (in-light seed). Among the three accessions, the in-dark seed germinated at much higher rates than did the in-light seed, more than 2× at 25 °C under light and up to an 83× increase in darkness, and up to an 8× increase over the corresponding parental seed lots under comparable germination conditions. In addition to these increases in germination, the in-dark seed showed early and synchronized germination as compared to the in-light seed. Since these results were achieved through only one cycle of selection and breeding, they strongly suggest that we have developed a very effective method for modifying seed dormancy in Echinacea.     

Vulnerability of alleles in the US Potato Genebank extrapolated from RAPDs

Crop genebank managers must be concerned with preserving low-frequency alleles that are vulnerable to being lost from the genebank because they are not sampled as seed increase parents. Traits conferred by such alleles are also vulnerable to being overlooked because they are not sampled in plants evaluated for breeding traits. Mathematical models and recommendations that follow from hypothetical vulnerable alleles within single populations have been made by various researchers. This work was undertaken to empirically detect the prevalence of such alleles in the US Potato Genebank by extrapolation from RAPD markers. Two outcrossing species known to be very heterogeneous were selected,Solanum jamesii Torr. (2n=2x=24) andSolanum sucrense Hawkes (2n=4x=48). RAPD band frequencies at a total of 83 marker loci within each of 15 populations were assessed using 24 plants per population. For both species, about 10% of within-population loci had vulnerable alleles (banded plant frequency < 40%), considering just the particular population in question. However, about half of these were fixed in at least one other population (making that marker invulnerable to loss). The highest within-population banded plant frequency for loci not fixed in any population was, with one exception, always > 40%. So, while several alleles may be vulnerable within a given population, these were nearly always fixed or nearly fixed (invulnerable) in another population. This suggests that the safest strategy for maximizing conservation of alleles is to apportion available resources over many separate populations, rather than adopt a preservation protocol so rigorous that it can only be practically applied to a limited number of populations.     

In vitro shoot regeneration and chromosome doubling in 2X and 3X potato clones

The possibility of scaling ploidy levels up and down offers several advantages for genetic studies and breeding work in potato. Shoot regeneration and chromosome doubling in plants regenerated fromin vitro cultures were investigated in 4 diploid (2n=2x=24) and 2 triploid (2n=3x=36) clones. Internode and leaf expiants were taken from plants propagated eitherin vitro as shoot cultures, orin vivo in a greenhouse. Two regeneration procedures were compared. Regeneration frequencies were significantly higher using a two-step regeneration protocol and from leaf explants, while doubling was more efficient starting fromin vivo grown plants. No doubling was observed in triploid clones. Considering altogether the percentage of regeneration and doubling, and the number of shoots regenerated per explant, the most efficient conditions were considered leaf explants taken fromin vivo grown plants, and cultured according to a two-step regeneration protocol.     

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.     

Response of six wild potato species to chloride and sulfate salinity

Eleven accessions belonging to six species ofSolanum (S. bulbocastanum,S. chacoense, S. gourlayi, S. microdontum, S. papita, and S.sparsipilum) were tested for resistance to salt. Resistance was measured by survival and growth (dry weight of tops) of plants treated with solutions of NaCl and Na₂SO₄. The six species could be grouped into four classes based on good to poor resistance to salt solutions (1.S. chacoense, 2.S. gourlayi, 3.S. microdontum, S. sparsipilum, S. bulbocastanum and 4.S. papita). There was a close relationship (r=0.94**) between the survival of plants and growth of surviving plants treated with salt solutions. There was no relationship between known heat or drought resistance of various accessions and salt resistance. Results indicate there is a large genetic base available to improve the salt resistance of the cultivated potato.     

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.     

Seed protein variation within accessions of wild and cultivated potato species and inbredSolanum chacoense

Summary Soluble proteins in seed potato were analyzed by electrophoresis (PAGE). The Index of Variation (IV) was calculated in order to estimate variation within each accession of wild and cultivated potato species andS. chacoense selfed lines. Seeds of 34 accessions representing 14 species from six taxonomic series were analyzed. The IV ofS. chacoense selfed lines decreased from 47.7 in the S₂ to 12.9 in the S₇ generation. The average IV for the self-incompatible diploid species was 41.2, whereas that of the selfcompatible diploid species,S. polyadenium, was 21.3. TetraploidS. tuberosum ssp.andigena andS. sucrense had an average IV of 62.2, which is much higher than that observed in any other diploid species. The naturally selfing speciesS. acaule (4x), and its colchicine-doubled derivative (8x), showed very low variation (IV=7.5). The IV appears to reflect heterozygosity and is affected by the breeding mode and the genetic constitution of diploid parents for derived polyploids.     

Distribution and significance of diplandroids among the diploid solanums

Clones of several diploid species capable of generating diplandroids (2n pollen) were identified by screening for the occurrence of 2n pollen. Diplandroid production was detected inSolatium bukasovi, S. canasense,S. cardiophyllum, S. chacoense, S. leptostigma, S. megistacrolobum, S. tarijense, S. tuberosum Group Phureja and Group Stenotomum. Parallel spindles at Ana II was the mechanism producing 2n pollen inS. chacoense, Group Phureja, and Group Stenotomum. The widespread distribution both taxonomically and geographically suggests a possible role for diplandroids in evolution. It also encourages exploration of a proposed breeding method in which tetraploids are generated from 2x × 2x matings.     

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.     

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.     

Agronomic characterization of diploid hybrid families derived from crosses between haploids of the common potato and three wild Argentinian tuber-bearing species

Five diploid hybrid families derived from crosses between haploids of three Argentinian cultivars and three diploid wild species,Solanum gourlayi, S. chacoense andS. spegazzinii, were evaluated in a field experiment in Balcarce. All plants were androsterile and, thus, could not be screened for 2n pollen production. Data on agronomic tuber traits were statistically analyzed. Differences among families were non-significant for tuber number/plant, but were significant for tuber weight/plant and dry matter content (with values above 21.0% in two of them). The relative frequencies of tubers in the categories established for size, skin texture, and eye depth were family dependent, but they were familiy independent in the categories established for tuber shape and skin color. Great morphological variability was detected for all the traits studied, and plants with high yield and high dry matter content were identified. These families can be used as sources of germplasm in breeding and to explore heterotic responses in 2x × 4x crosses, given the capacity of some plants to produce 2n eggs, as found in a related investigation.