Natural triploid hybrids between Solanum acauleBitter and S. infundibuliformePhilippi in the province of Jujuy,Argentina

Summary The occurrence of natural triploid hybrids between tetraploid Solanum acauleBitter subsp. acaule and diploid S. infundibuliformePhilippi both tuber-bearing Solanums, in the Puna region of Jujuy, is reported. The natural hybrids, which are morphologically intermediate between their putative parents and completely male sterile, have a somatic chromosome number of 36. The artificial hybrids, obtained by crossing both parental species, closely resemble the natural ones thereby confirming their parentage. It is proposed to designate the natural hybrids Solanum x viirsooi hybr. nov.     

A biosystematic study of the origin of the cultivated diploid potato,Solanum x Ajanhuiri Juz. et Buk.

Summary The origin and nature of the diploid cultivated potato species S. ajanhuiri Juz. et Buk. was studied. Several lines of evidence indicate that S. ajanhuiri might be derived from natural crosses between primitive cultivars of the diploid species S. stenotomum and the wild species S. megistacrolobum. Morphological comparisons were made between S. stenotomum x S. megistacrolobum F₁ hybrids and naturally occurring S. ajanhuiri to investigate this hypothesis. Comparisons were also made between S. ajanhuiri x S. stenotomum crosses and the F₂ generation of the first-mentioned cross.Crosses between the two major groups of S. ajanhuiri cultivars, Ajawiri and Yari, showed not only genetic breakdown but also a wide range of phenotypic variation similar to those of artificial F₂ families of S. stenotomum x S. megistacrolobum. Furthermore, there was strong evidence showing that the Yari group of S. ajanhuiri could almost certainly be an F₁ S. stenotomum x S. megistacrolobum hybrid, whereas the Ajawiri group could be a backcross of an F₁ hybrid to S. stenotomum. These results added further support to the hypothesis of a hybrid origin of S. ajanhuiri, as well as indicating its putative parents. It is suggested that this hybridogenic taxon be retained at the species level under the name Solanum x ajanhuiri.     

Meiosis in different F1-hybrids of Solanum acaule Bitt. x S. bulbocastanum Dun. and its bearing on genome relationship,fertility and breeding behaviour

Summary Meiosis was studied in some triploid, tetraploid and hexaploid F₁-hybrids from Solanum acaule x S. bulbocastanum and in a triploid F₁-hybrid from S. acaule x S. tuberosum-haploid.The only anomaly found was stickiness at metaphase I, the degree of which appeared to be related to the proportion of the S. bulbocastanum chromosomes in the hybrids. No stickiness occurred at pre- and post-metaphase stages.A clear allosyndetic pairing between chromosomes of the two S. acaule genomes was observed in all triploid and tetraploid hybrids. These genomes apparently are closely related and little differentiated. The triploids S. acaule x S. bulbocastanum had 0–1 trivalent per cell, whereas 2–5 trivalents per cell were observed in the triploid S. acaule x S. tuberosum-haploid. Therefore S. acaule is more closely related to S. tuberosum L. than to S. bulbocastanum. A small amount of pairing between S. acaule and S. bulbocastanum is apparent from the occurrence of multivalents in all hybrids.Hexaploid F₁'s (2n=72) showed a nearly complete homologous pairing of chromosomes (35.2 bivalents per cell) and generally a normal separation of chromosomes at anaphase: 36-36. This offers an explanation for their high fertility. Triploid F₁'s from S. acaule x S. bulbocastanum have a high frequency of univalents leading to irregular separation of chromosomes at anaphase and consequently to unbalanced gametes and extreme sterility. In the tetraploid F₁'s (2n=48) nearly complete bivalent pairing was observed, 50% expectedly being homologous and 50% homoeologous pairing. Separation of chromosomes at anaphase was generally normal 24-24. In spite of this normal behaviour and allowing for tight stickiness at metaphase the tetraploids are very sterile. A satisfactory explanation cannot yet be given.Selfing and intercrossing hexaploid F₁'s gives normal berry set and many seeds per berry. However crosses with S. tuberosum and even those with the fertile hexaploid hybrid from 8x-S. acaule x 4x-S. tuberosum are little successful: berry set is far below normal and the berries are either parthenocarpic or contain only one or two seeds. These rather unexpected results warrant further investigation. Large-scale selfings and intercrosses of triploid and tetraploid hybrids have not been successful as yet. Among the female gametes of tetraploid hybrids a few appeared to be functional in crosses with hexaploid hybrids and in those with S. bulbocastanum.     

The breeding of the cultivated potato species solanum x juzepczukii and S. x curtilobum II. The resynthesis of S. x juzepczukii and S. x curtilobum

Summary The possibility of combining anew the genomes of wild and cultivated progenitors of triploid S. x juzepczukii and pentaploid S. x curtilobum by following the known evolutionary pathway of these species was investigated.The resynthesis of S. x juzepczukii was easy, and a wide range of synthetic forms was bred. Among these were forms with higher frost resistance (-5°C) than has been found in natural S. x juzepczukii. The total tuber glycoalkaloid content of several synthetic hybrids was lower than or as low as that of natural clones. Most synthetic hybrids were more vigorous than natural S. x juzepczukii and produced about the same types of tubers as are found in the natural range of variation. The best diploid parents were found in the species S. goniocalyx.The attempt to resynthesize pentaploid S. x curtilobum has not been successful but tetraploid plants were obtained in the process. An explanation for the occurrence of tetraploids resulting from triploid x tetraploid and/or diploid crosses is offered.The newly bred tetraploids contain at least one genome from S. acaule (possibly two) and hybridize easily with ssp. andigena. They thus provide a means for the transfer of S. acaule germ plasm into the tetraploid cultivated gene pool which would profit from the frost resistance of S. acaule.     

Breaking the crossability barriers between disomic tetraploid Solanum acaule and tetrasomic tetraploid S. tuberosum

Summary A combination of compatible second pollinations and embryo rescue was applied for systematic production of true tetraploid hybrids from crosses between disomic tetraploid Solanum acaule and tetrasomic tetraploid potato, S. tuberosum. Several genotypes of tetraploid potatoes were pollinated with S. acaule, and the compatible second pollinations were made on the following day, with a genotype of S. phureja, IvP 35 to promote fruit development. Embryo rescue was carried out in 21 families, 14 to 27 days after the first pollination. A total of eight plants were obtained from the embryo rescue and their chromosome numbers were counted in the root tips. Three of the eight plants were identified as tetraploid, and five others as diploid. Morphology, isozyme banding patterns, and pollen stainability, as well as potato spindle tuber viroid (PSTVd) resistance, indicated the hybrid nature of the three plants. This is the first report of successful tetraploid hybrid production between disomic tetraploid S. acaule (4x) and tetrasomic tetraploid potatoes. Seed set from the crosses between one of hybrids and diploid potatoes indicated workable levels of both male and female fertility for introgression of valuable genes from S. acaule into the cultivated potato gene pool. The methodology used may be applied to other disomic tetraploid tuber-bearing Solanum species and with some modifications also to distantly related solanaceous species and genera.     

Immunity to Clavibacter michiganensis subsp. sepedonicus: Screening of exotic Solanum species

Summary Accessions from exotic Solanum species, including diploid and tetraploid species, were screened for immunity to Clavibacter michiganensis subsp. sepedonicus, the causal agent of potato ring rot. The diploid species included S. infundibuliforme, S. lesteri, S. megistacrolobum, S. tuberosum Group Phureja, S. polyadenium, S. pinnatisectum, S. raphanifolium, S. sparsipilum, S. sanctae-rosae, S. tuberosum Group Stenotomum, S. toralapanum, and S. verrucosum. The tetraploid species included S. tuberosum Group Andigena, S. acaule, S. fendleri, S. hjertingii, S. oplocense, S. polytrichon, and S. stoloniferum. Apparent immunity was initially found in several diploid species, but was not present during subsequent retesting. Immunity was found in nine accessions of tetraploid S. acaule. These accessions maintained their immunity during testing over an eight-month period. S. acaule appears to be a good source of immunity for introgression studies.     

Tetrahaploids from colchicine-induced octaploid Solanum acaule subsp. Aemulans

Summary Crosses between colchicine-induced octaploid Solanum acaule ssp. aemulans (2n=8x =96) with diploid S. tuberosum, Group Phureja, carrying a homozygous embryo marker, produced a high percentage of S. acaule tetrahaploids besides pentaploid and hexaploid F₁ hybrids. These results are discussed in relation to the behaviour of autopolyploid and allopolyploid Solanum species, which basically differ in the occurrence of haploids and the ploidy level of F₁ hybrids after pollination of both types of poly-ploids with Phureja.     

Induction of haploids and aneuhaploids in colchicine-induced tetraploid Solanum chacoense Bitt.

Summary The species Solanum chacoense BITT. (2n=2x=24) is a tuber-bearing, self-incompatible species which is important both for breeding and for genetic research. It crosses readily with most other tuber-bearing Solanum species including the common potato S. tuberosum (2n=4x=48). Gametophytic incompatibility hampers research in and utilization of this species. Doubling the chromosome number by colchicine makes it a self-compatible autotetraploid. By crossing selfed progeny of 4x-S. chacoense with a number of haploid-inducing diploid Solanum species a high yield of different dihaploid S. chacoense individuals (2n=24) could be obtained from one originally diploid clone: S. chacoense CPC 1153. More than 160 haploids showing a large variability were identified. The average haploid frequency was 53.7 per 100 berries. Most hybrid plants (70–100%) from four 4x × 2x crosses studied were tetraploid. The frequency of triploid hybrids was low (0–10%). Haploid-inducing capacity of fifteen male parents used in this study varied from 0–141 haploids per 100 berries. Careful examination of 156 haploids revealed 15 viable aneuhaploids (2n=25, 26, and 27), i.e. 9.4%. The potential value of these aneuhaploids is discussed.About 50% of the haploids were sufficiently male fertile to use them in crossing. A few of them set berries after selfing. Five aneuhaploids including the one with 27 chromosomes were successfully crossed as females with a diploid hybrid clone.     

Chloroplast DNA variation in the wild potato species, Solanum acaule and S. albicans

The chloroplast DNA of Solanum acaule (109 accessions) and S. albicans (9 accessions) was investigated by restriction endonuclease analysis. Unexpectedly, all the accessions analyzed had C type chloroplast DNA in common. This suggested that S. acaule originated from a species with C type chloroplast DNA. DraI restriction digestion revealed further differentiation of C type chloroplast DNA into 8 types. The DraI polymorphism indicated the province of Salta in Argentina and the nearby regions to be a center of diversity for S. acaule. Surprisingly, S. albicans as well as S. acaule both ssp. acaule and ssp. punae, from Peru were virtually indistinguishable, although by morphology and/or cytology all three taxa are easily distinguished. This revised version was published online in July 2006 with corrections to the Cover Date.     

Enhancement of diploid Solanum chacoense Bitt. using adapted clones of Solanum phureja Juz. & Buk.

Summary F₁ hybrids between Solanum phureja and S. chacoense were studied in a new approach to enhance wild diploid potato species using adapted S. phureja clones as the female parent. S. chacoense parents had few tubers, whereas S. phureja parents had been previously selected for tuberization under long days. Twenty hybrid families were evaluated for haulm vigor, vine maturity, stolon development, frequency of tuberization, tuber set, tuber yield and average tuber weight under naturally occurring increasing or decreasing photoperiods in spring and fall growing seasons in Morocco. The hybrid plants were more vigorous, later maturing with thicker stolons in the spring season. Frequency of tuberization and tuber set did not vary with season. Tuber yield and average tuber weight were higher in fall; the maximum yield was 2.4 and 4.6 kg per hill under spring and fall conditions, respectively. Remarkable variation was found both between and within hybrid families for all traits in this first cycle of enhancement. The selection of vigorous, fertile clones incorporating the S. chacoense genome is possible by using S. phureja as an adapted germplasm source.     

Identification of somatic hybrids of dihaploid Solanum tuberosum lines and S. brevidens by species specific RAPD patterns and assessment of disease resistance of the hybrids

Summary Symmetric somatic hybrids were produced by electrofusion of protoplasts of two dihaploid tuber-bearing potato (Solanum tuberosum L.) lines and Solanum brevidens Phil., a diploid non-tuber-bearing wild potato species. A total of 985 plants was obtained. Verification of nuclear hybridity of putative hybrids was based on additive RAPD patterns, general morphological characteristics and chromosome counts. 53 (90%) calli regenerated into plants which were identified as somatic hybrids. Most of the hybrids were aneuploids at the tetraploid (4×) or hexaploid (6×) level. The 20 hybrids tested expressed a high level of resistance to potato virus Y (PVY ^N ) characteristic of the S. brevidens parent. Resistance to late blight (Phytophthora infestans (Mont.) de Bary) varied between hybrids, but was on average better than that of the fusion parents. Resistance of hybrids to bacterial stem rot (Erwinia carotovora subsp. atroseptica (van Hall) Dye) was not superior to that of commercial potato cultivars.     

The nature of the Bolivian weed potato species Solanum sucrense Hawkes

Summary Two theories for the origin of the Bolivian weed potato species Solanum sucrense Hawkes have been tested. The first was that it had an allopolyploid origin as a hybrid of the cultivated tetraploid S. tuberosum ssp. andigena (Juz. et Buk.) Hawkes with the wild/weed tetraploid cytotype of S. oplocense Hawkes. The second hypothesis postulated that it was a cross of the weed diploid species S. sparsipilum (Bitt.) Juz. et Buk. with S. oplocense.Synthetic hybrids of S. tuberosum ssp. andigena x S. oplocense and of S. sparsipilum x S. oplocense have been compared morphologically with S. sucrense accessions. Their crossability has also been investigated.The crossability and morphological studies strongly suggested that the former hypothesis was the more likely. This was borne out by observations made on the 1974 University of Birmingham Expedition when segregating populations of S. sucrense and S. oplocense were collected growing with feral S. tuberosum ssp. andigena.The evidence from this study indicates that the S. tuberosum ssp. andigena x S. oplocense hybrid has formed the basis of the S. sucrense gene pool. It is suggested that this hybridogenic taxon be maintained under the name Solanum x sucrense.The clarification of the taxonomic relationships of these three species will be of interest to Globodera resistance breeders, in view of the fact that these taxa are resistant to various pathotypes of the golden nematode.     

Double-bridge hybrids of Solanum bulbocastanum and cultivars of Solanum tuberosum

Summary Solanum bulbocastanum (2n=2x=24) has valuable characters for potato breeding, but cannot be hybridized directly with S. tuberosum cultivars. Both S. acaule (2n=4x) and S. phureja (2n=2x) were used as bridging species. Triploid S. acaule × S. bulbocastanum were doubled with colchicine and the resulting fertile hexaploid F₁'s crossed with S. phureja. The triple hybrids obtained were tetraploid or nearly so. The two genomes of S. acaule in these triple hybrids probably pair preferentially, which may provoke pairing and possibly crossing over between the chromosomes of S. bulbocastanum and S. phureja.More than 20000 pollinations of the triple hybrids with four potato cultivars had to be made to produce 40 quadruple hybrids. These highly vigorous hybrids varied greatly in many morphological characters, resistance to Phytophthora infestans, fertility and crossability. The chromosome numbers are 48 (24 hybrids), 49 and 46, but some higher ploidy levels (65, 66, 72 chromosomes) were found as well. Their origin is to be sought in the fusion of an unreduced egg cell from triple hybrids (either euploid or hypoploid) and a reduced male gamete from the cultivars. This view is corroborated by their extreme resistance to Phytophthora. Also some 48-chromosome hybrids are highly resistant, which may indicate introgression from S. bulbocastanum.Most quadruple hybrids are readily inter-crossable and crossable as females with cultivars; several also as males. Two could be hybridized with S. bulbocastanum, but the few seeds dit not germinate.Studies of pachytene stage of meiosis revealed the presence of a S. bulbocastanum chromosome in at least one tetraploid hybrid, which is highly resistant to Phytophthora. At metaphase I of meiosis chromosome associations higher than quadrivalents were not found. Except in one hybrid, the frequency of quadrivalents did not exceed one per cell and the average proportion of chromosomes associated as bivalents amounted to 90%.The quadruple hybrids (double-bridge hybrids) appear good starting material for breeding programmes aimed at introducing genes from S. bulbocastanum into S. tuberosum cultivars.     

Studies on the use of Solanum acaule as a bridge between Solanum tuberosum and species in the series Bulbocastana,Cardiophylla and Pinnatisecta

Preliminary results have indicated that Solanum acaule can serve as an intermediate for gene transfer from Solanum species in the Mexican series Bulbocastana, Cardiophylla and Pinnatisecta to Solanum tuberosum. Clones of S. acaule vary in their effectiveness as female parents in crosses with the Mexican species. The F1 hybrids obtained were sterile triploids. Fertile hexaploids were produced from the triploids by colchicine treatment. The hexaploid from the cross S. acaule x S. pinnatisectum was successfully crossed with diploid and tetraploid forms of series Tuberosa. Some of the resultant triple hybrids were self-fertile.     

Resistance to viruses in F1 hybrids produced by direct crossing between diploid Solanum series Tuberosa and diploid S. brevidens (series Etuberosa) using S. phureja for rescue pollination

Resistance to potato viruses was examined in the F₁ hybrids (TET) obtained from a cross between a diploid (2n = 24), tuber-bearing interspecific hybrid 87HW13.7 (Solanum tuberosum W231 ×S. multi-dissectum PI 473354) and a diploid (2n = 24), nontuber-bearing wild potato species (S. brevidens CPC 2451) using S. phureja IvP35 (2n = 24) for rescue pollination. The parental plants were susceptible to PVX, whereas two hybrids (TET38.2 and TET38.9) and S. phureja IvP35 reacted with hypersensitivity to PVX. Two hybrids (TET 38.9 and TET 38.12) were extremely resistant to PVY°, which was similar to S. brevidens and S. phureja IvP35, whereas the remaining two hybrids were moderately resistant to PVY°. No resistance to PVA and PLRV was observed in the progenies, in contrast to S. brevidens which was extremely resistant to PVA and PLRV. Hypersensitivity to PVX in two progenies suggested (1) integration by somatic translocation or heterofertilization and expression of genes from the rescue pollinator S. phureja IvP35, or (2) transgressive or complementary gene action.     

Characterization of morphological variation and cold resistance in interspecific somatic hybrids between potato (Solanum tuberosum L.) and S. brevidens Phil.

Summary Somatic hybrids between Solanum tuberosum L. cv. Gracia (2n=4x=48) and Solanum brevidens Phil. (2n=2x=24) were produced via fusion of mesophyll protoplasts. Selection of the protoplast derived putative hybrid calli was based on their vigorous growth. Additive isozyme patterns and chromosome numbers as well as the expression of parental morphological characters have proved the hybrid origin of the selected regenerants. Extensive chromosome loss during the regeneration process resulted in aneuploid hybrids with high frequency. Genomic instability could not be detected in these plants during the period of vegetative propagation. Regenerants from hybrid tissues exhibited wide morphological variation especially in tuber formation. The detailed morphological analysis based on the use of multivariate method (principal component analysis, PCA) enabled to identify morphological groups among the hybrid clones. The positioning of hybrid clones in the PCA space could not be correlated with chromosome numbers. The genomic ratio represented by the tetraploid and diploid parents influenced the morphology of somatic hybrid population according to the applied analytical system. Two selected hybrid clones have exhibited an intermediate degree of frost tolerance compared to the parents, based on the recovery of plants from lower buds after freezing of potted plants.     

Breeding of the cultivated potato species Solanum x juzepczukii Buk. and Solanum x curtilobum Juz. etBuk.

Summary This study investigated the possibility of recombining anew the genomes of the wild and cultivated progenitors of triploid S. juzepczukii and pentaploid S. curtilobum by following the known evolutionary pathway of these two species. Before starting the actual breeding work, the natural variation of S. juzepczukii, S. curtilobum and their wild progenitor S. acaule was studied from the point of view of morphology, quantitative and qualitative tuber glycoalkaloid content and frost resistance. The morphological study was supplemented by a study of the soluble tuber proteins employing polyacrylamide slab-electrophoresis. From 137 accession of S. juzepczukii only 19 morphotypes were identified, 18 of which were also different in their protein spectra. The only red-tubered S. juzepczukii revealed a protein spectrum identical to that of the largest white-tubered group. On phylogenetic grounds, the occurrence of a red-tubered S. juzepczukii cannot be explained. It is concluded that this red clone is a somatic mutant for tuber colour which arose from a whitetubered clone. S. curtilobum was restricted in its variation to just two morphotypes differing only in tuber colour which are, however, identical chemotypes. This would be the case if one of the clones was a somatic mutant for tuber colour from the other one. The glycoalkaloids -solanine, -chaconine, tomatine, demissine and - and -solamarine are shown to be useful taxonomic characters which confirm earlier hypotheses on the origin of S. juzepczukii and S. curtilobum. Laboratory tests showed the two cultivated species to be resistant to about –3°C whereas S. acaule is resistant to temperatures sometimes below–5°C. The diploid progenitor of S. juzepczukii, S. stenotomum, also has forms resistant to –3°C. The results of this study demonstrate that the proposed breeding scheme is possible.     

Resistance to Meloidogyne chitwoodi,M. fallax and M. hapla in wild tuber-bearing Solanum spp.

Summary Over 5000 plants from 64 tuber-bearing wild Solanum spp. have been individually screened for resistance to Meloidogyne chitwoodi, M. fallax and M. hapla. Seedlings were analyzed by means of counting number of egg masses and resistance was verified by retesting low-scoring plants using stem cuttings. Resistance to both M. chitwoodi and M. fallax was observed in S. bulbocastanum, S. cardiophyllum, S. brachistotrichum, S. fendleri and S. hougasii. Only in S. chacoense and to a lesser extent in S. stoloniferum and S. gourlayi differential results between M. chitwoodi and M. fallax were observed. Resistance to M. hapla was found in S. bulbocastanum, S. brachistotrichum, S. cardiophyllum, S. arnezii, S. chacoense, S. tarijense, S. boliviense, S. gourlayi, S. microdontum, S. sparsipilum, S. spegazzinii, S. sucrense, S. acaule and S. hougasii. The occurrence of resistance in wild Solanum species in relation to their taxonomic status and the implications for introgression of resistance into S. tuberosum are discussed.     

Fertile somatic hybrids of Solanum etuberosum (+) dihaploid Solanum tuberosum and their backcrossing progenies: relationships of genome dosage with tuber development and resistance to potato virus Y

The wild non-tuberous species Solanumetuberosum is resistant to biotic andabiotic stresses, but is very difficult tocross with cultivated potato. Therefore,interspecific somatic hybrids between adihaploid clone of potato S.tuberosum (2n=2x=24, AA genome) and thediploid species S. etuberosum(2n=2x=24, EE genome) were produced byprotoplast fusion. Among the 7 fertilefusion hybrids analysed by genomic insitu hybridisation (GISH), three groups ofplants were found with the genomicconstitution of AAEE, AAEEEE and AAAAEE.Four fusion hybrids had exactly theexpected chromosome composition, while eachof the three aneuploid hybrids had lost twochromosomes of S. etuberosum. Twobackcross progenies were developed, andGISH analysis was applied to analysetransmission of the parental chromosomesinto the sexual generations. BC₁hybrids derived from the crosses of thehexaploid somatic hybrids with tetraploidpotato were pentaploid with thetheoretically expected genomic compositionor with slight deviation from thisexpectation. In the three BC₂ hybridsanalysed by GISH seven to 12 chromosomes ofS. etuberosum were detected in thepredominant S. tuberosum background.No recombinant chromosomes in the hybridswere detected. Genome dosage affects tuberformation in hybrids and their progenies,but has less effect on resistance to potatovirus Y (PVY) in fusion hybrids. Severalgenotypes of the fusion hybrids andBC₁ progeny did not show viralinfection even in the graftingexperiments.     

Breeding behavior for tuber protein in Solanum tuberosum and Tuberosum-phureja hybrids

Summary True protein content among tubers within a plant of Solanum tuberosum cv. Oneida was found to be negatively correlated with tuber size. A similar study of S. phureja genotype 148-17 revealed no correlation. Tuber protein was determined for 250 genotypes in each of four hybrid potato populations in a factorial mating design with four potato cultivars as stylar parents mated to four groups of pollen parents (4x cultivars, 4x high protein selections, 2x S. phureja, 2x high protein S. phureja). The hybrid population derived from the 4x high protein selections was significantly higher in protein content than the other populations. High estimates of general combining ability for tuber protein content were found for both stylar and pollen parents. Poor photoperiodic adaptation to growing conditions in northern latitudes was a possible explanation for the variable protein phenotypes typical of S. phureja and the lack of transmission of the high protein character in phureja to 4x-2x hybrids. Although protein content was negatively correlated with total yield, high protein segregates with good yield potential were identified in all four populations.Scientific Journal Series Article 11, 616 of the Minnesota Agricultural Experiment Station.