The importance of wild potato species resistant to the potato cyst nematode,Globodera pallida,pathotypes P4A and P5A,in potato breeding. II. The crossability of resistant species

Summary Over 2400 pollinations were made to investigate the crossability relationships between cultivated potatoes, and wild diploid species from Series Tuberosa and Circaeifolia, as well as wild polyploid species in Series Tuberosa and Longipedicellata resistant to potato cyst nematode, Globodera pallida pathotypes P₄A and P₅A. Wild diploids in Series Tuberosa crossed easily with cultivated diploid species, except with Solanum lignicaule where most pollinations failed, and seed set was extremely low (0.2 seeds per pollination or less). It is suggested that this species is 1EBN. S. capsicibaccatum is clearly isolated from Series Tuberosa, but can form hybrids with S. lignicaule, which can act as a bridging species to S. tuberosum haploids. S. gourlayi and S. oplocense can be crossed with both subspecies of S. tuberosum, but S. papita, Series Longipedicellata is reproductively isolated from the tetraploid cultigens. The crossability data are discussed in the light of germplasm utilisation for breeding potato varieties resistant to potato cyst nematode.     

Resistance to late blight (Phytophthora infestans) in ten wild Solanum species

Nineteen accessions of the tuber-bearing species Solanum berthaultii, S. chacoense, S. leptophyes, S. microdontum, S. sparsipilum, S. sucrense, S. venturii, S. vernei and S. verrucosum were tested for their resistance to late blight in two years of field experiments. Plants were artifically inoculated with zoospores of race 1.2.3.4.5.7.10.11 and the development of the disease was followed. Resistance ratings, calculated as the areas under the disease progress curves (ADPC), demonstrated a high resistance in all accessions except in S. sparsipilum, S. leptophyes and their interspecific hybrid. Segregations suggest that major genes for resistance are present in S. sucrense and S. venturii, and may also play a role in S. verrucosum. It is not yet certain wether the resistance of the other accessions is comparable to the partial and durable resistance of S. tuberosum cultivars like Pimpernel, as inheritance and mechanism have yet to be established. However, segregations suggesting the presence of single major genes with complete dominance were not found in these other accessions. Tuber initiation in the field occurred in only one accession, S. tuberosum ssp. andigena, and maturity of the clones was not related to their resistance. In the other accessions maturity types could not be assessed, as the clones require short day conditions for tuber initiation.     

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.     

Natural hybridization between Solanum acaule Bitt. and S. megistacrolobum Bitt. in the province of jujuy,Argentina

Summary Evidence is provided that hybridization of the tetraploid (2n=4x=48), self-fertile tuber-bearing species Solanum acaule Bitt. with the diploid (2n=2x=24), self-incompatible, tuber-bearing S. megistacrolobum Bitt. takes place in several localities of the province of Jujuy in the high, cold plateaux of the Argentine Puna. The triploid hybrids (2n=3x=36) closely resemble S. acaule in growth habit, leaf morphology and floral structures and for this reason they can be easily overlooked for that species in the field. Experimental data show that S. acaule can be crossed with S. megistacrolobum though the crossability is rather low and variable according to the particular cross considered. The artificial hybrid obtained compares well with the natural hybrid in morphology and chromosome number. The hybrids, though almost completely male sterile, are successful colonizers of disturbed areas around farmers' dwellings, cattle enclosures and other areas where the soil is rich in organic matter.There is some evidence that the Tilcara material of S. acaule subsp. aemulans and the hybrids of S. acaule x S. megistacrolobum have some characters in common, which can be interpreted as having a similar origin.It is postulated that S. acaule subsp. aemulans, in Jujuy at any rate, is not a primitive form of S. acaule as thought by Hawkes and Hjerting, but rather a fertile hybrid derivative of S. acaule x S. megistacrolobum through the functioning of 2n gametes.We also provide evidence that S. bruecheri Correll should not be considered a hybrid of S. acaulle x S. megistacrolobum but a synonym for S. gourlayi Hawkes. The new name, S. x indunii Okada et Clausen, is proposed to designate this hybrid.     

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.     

The nature of Solanum × chaucha Juz. et Buk., a triploid cultivated potato of the South American Andes

Summary The variation in Solanum × chaucha Juz. et Buk. (2n=3x=36) was studied using 114 accessions from the Potato Collection maintained by the International Potato Center. These accession represented less than 5% of the collection originally screened for ploidy. Twenty morphological characters of the tubers, inflorescences and leaves were scored. The accessions were grouped into 20 morphotypes on the basis of the tuber characters. Two-qualitative characters, red-anther tip and sky-blue flowers were diagnostic of 4 morphotypes, but the variation of the quantitative characters overlapped among morphotypes, and were of limited value. Electrophoretic separation of tuber proteins in 7.5% polyacrylamide gels generally confirmed the morphotype groupings. It is concluded that each of the morphotypes represents a single genotype, and these are named according to the International Code of Nomenclature for Cultivated Plants. The variation within S. × chaucha apparently has been restricted by its hybrid origin.     

The effect of seed tuber weight on characters in the first and the second clonal generation of potato populations

Summary Fifty-five populations from crosses of three Solanum tuberosum ssp. tuberosum cultivars and seven selected S. tuberosum ssp. andigena clones adapted to the northern temperate zone have been studied in the seedling and the first and second clonal generation. This paper only deals with the effect of seed tuber weight on various important agronomic characters which are selected for in early clonal generations, independently of the type of crosses.The results showed that a distinction has to be made between the effects of differences in seed tuber weight within and between seedlings, i.e. genotypes. The effects on the first clonal generation of differences in weight of the tubers within seedlings and those of the first heaviest tubers between seedlings were similar and comparable to seed tuber weight effects within cultivars. In contrast to the within seedlings effects the between seedlings effects proved to be heritable. This is concluded from the fact that, on average, independent of seed tuber weight seedlings with larger first heaviest tubers performed better in the second clonal generation than those with smaller ones. Notably plant height, general impression, date of maturity and particularly tuber yield and mean tuber weight in the second clonal generation were related to the weight of the first heaviest tubers of the seedlings. The potentialities of these findings for an early generation selection are discussed.     

Resistance to late blight (<Emphasis Type="Italic">Phytophthora infestans</Emphasis>) in ten wild <Emphasis Type="Italic">Solanum</Emphasis> species

Nineteen accessions of the tuber-bearing species Solanum berthaultii, S. chacoense, S. leptophyes, S. microdontum, S. sparsipilum, S. sucrense, S. venturii, S. vernei and S. verrucosum were tested for their resistance to late blight in two years of field experiments. Plants were artifically inoculated with zoospores of race 1.2.3.4.5.7.10.11 and the development of the disease was followed. Resistance ratings, calculated as the areas under the disease progress curves (ADPC), demonstrated a high resistance in all accessions except in S. sparsipilum, S. leptophyes and their interspecific hybrid. Segregations suggest that major genes for resistance are present in S. sucrense and S. venturii, and may also play a role in S. verrucosum. It is not yet certain wether the resistance of the other accessions is comparable to the partial and durable resistance of S. tuberosum cultivars like Pimpernel, as inheritance and mechanism have yet to be established. However, segregations suggesting the presence of single major genes with complete dominance were not found in these other accessions. Tuber initiation in the field occurred in only one accession, S. tuberosum ssp. andigena, and maturity of the clones was not related to their resistance. In the other accessions maturity types could not be assessed, as the clones require short day conditions for tuber initiation.     

The breeding potential of wild potato species resistant to the potato tuber moth,Phthorimaea operculella (Zeller)

Summary The potato tuber moth, Phthorimaea operculella (Zeller) is an important pest of potatoes in the field and in stores in warm environments throughout the world. In this study genetic resistance to potato tuber moth was identified in clones of Solanum sparsipilum (coded MBN) originally developed for resistance to bacterial wilt and root-knot nematode. Resistance to this pest in S. sucrense and S. tarijense as well as S. sparsipilum was exploited in wide crosses with diploid and tetraploid cultivated potatoes, and haploids derived from S. tuberosum; hybrid progenies were produced. Crosses between resistant S. pinnatisectum or S. commersonii and cultivated potatoes failed completely, although S. commersonii did hybridise with two bridging species S. lignicaule and S. capsicibaccatum which are slightly compatible with cultivated potatoes. Resistance to potato tuber moth was transferred to all progenies except those in which S. tarijense was the resistant parent. The development of potatoes resistant to potato tuber moth is discussed in the context of population breeding for the lowland tropics.     

Cytogenetic studies and the occurrence of triploidy in the wild potato species Solanum commersonii Dun.

Summary The South American wild potato species Solanum commersonii Dun. exists as two subspecies, subsp. commersonii and subsp. malmeanum (Bitter) Hawkes and Hjerting, and as both diploid and triploid cytotypes. The two subspecies were completely cross fertile but there were reciprocal differences in the number of seeds set per berry and the mean seed weight. Crosses between diploids and colchicine-induced autotetraploids yielded only a few seeds per berry, but 16 of the 18 germinated seeds were triploid or nearly so. Such a high proportion of triploids is unusual for such crosses in potatoes. Some triploids has a high percentage of stainable pollen. The origin of naturally occurring triploids is discussed in the context of 2n gametes, a triploid block, and endosperm balance numbers.     

Identification of RAPD markers linked to the Ns locus in potato

Using the RAPD method and bulked segregant analysis we identified four RAPD markers linked to a dominant gene Ns, responsible for a hypersensitive reaction of potato (Solanum tuberosum L.) to potato virus S (PVS) infection. The markers OPE15⁵⁵⁰, OPJ13⁵⁰⁰, OPG17⁴⁵⁰ and OPH19⁹⁰⁰ were found to be closely linked to the Ns gene in diploid potato clones. They are situated at 2.6, 3.3, 4.6 and 6.6 cM from Ns, respectively. As a source of the gene, clone G-LKS 678147/60, which is known to carry Ns transferred from S. tuberosum ssp. andigena was used. These RAPD markers were not amplified in resistant tetraploid clones containing Ns derived from the clone MPl65 118/3, also having an andigenum origin. This suggests that there may be two separate loci of Ns in the sources identified, or different alleles with the same specificity at a single locus, or that the genetic background of tetraploids tested results in different RAPD amphlification patterns.     

Somatic chromosome elimination and meiotic chromosome pairing in the triple hybrid 6x-(Solanum acaule × S. bulbocastanum) × 2×-S. phureja

Summary Two selected hexaploid F₁ clones from the cross Solanum acaule x S. bulbocastanum were intercrossed and the resulting hybrid plants pollinated with the diploid species S. phureja, in order to obtain tetraploid triple hybrids with the same ploidy level as S. tuberosum cultivars (2n=4x=48).Apart from three trihaploids a large population of triple hybrids was obtained, showing chromosome mosaicism in root tip cells (euploid + hypoploid chromosome numbers) and a uniform, mostly hypoploid chromosome number in the pollen mother cells. It is demonstrated that somatic chromosome elimination in the early stages of development is the most probable cause.From detailed pachytene observations as well as from the chromosome associations observed at metaphase I it is evident, that there is normal pairing between the four genomes in the triple hybrids. Although S. bulbocastanum is a quite distinct species with a rigid crossability barrier with S. phureja, the chromosomes of these two species appear to have a high degree of homology. Especially the formation of quadrivalents involving all twelve groups of four homeologous chromosomes, indicated that the four parental genomes (two from S. acaule, one from S. bulbocastanum and one from S. phureja) are not differentiated to the extent of affecting normal pairing and chiasma formation. These results support the view, that the transfer of valuable characters from S. bulbocastanum to S. tuberosum cultivars is feasible even when these characters are polygenically controlled.     

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.     

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.     

Use of Vaccinium octoploids to facilitate 4x-6x gene transfers

The use of an 8x Vaccinium produced through doubling a tetraploid hybrid with colchicine was studied as a means of facilitating gene exchange between V. corymbosum L. and V. ashei Reade. Analysis included meiotic observations and crossability studies of the 8x plant, as well as attempting to develop 6x breeding lines. Meiotic analysis revealed the presence of micronuclei in Telphase II products and polyspory in sporads. In the crossability studies five 4x highbush plants were pollinated with 8x o2 4x pollen. Data gathered included: % fruit set, average weight/fruit, average number of seeds/fruit, average number of seedlings/pollination, and number of 6x seedlings. Octoploid pollinations were significantly lower than 4x pollinations in all parameters. One 6x plant was produced from the 4x-8x crosses but was found to be mitotically unstable, having somatic cells with chromosome numbers ranging from 48 to 168.     

Molecular Analysis of Local Potato Cultivars from Tenerife Island Using Microsatellite Markers

We have used 19 SSR markers to fingerprint 41 local potato cultivars from 10 locations of Tenerife Island. These varieties represent relicts of the early introductions originating from South America and have been characterised previously morphologically and ecophysiologically. The SSR primers generated a varying degree of polymorphisms. A total of 67 alleles were observed, 12 of them were present in all cultivars. Several accession and group specific alleles were detected. Similarity coefficients were computed from the molecular data and cluster analyses were performed. Generally, cultivar groups with identical or related common names showed the same SSR patterns or clustered closely together. According to the molecular patterns misleading or confounded names were evident for four accessions. The dendrogram clusters were generally in good agreement with previous classifications of the accessions as Solanum tuberosum subsp. andigena, S. tuberosum subsp. tuberosum and Solanum chaucha genotypes. However in four cases the molecular patterns showed discrepancies with previous species assignments suggesting the need for a more detailed and comparative study of these accessions.     

Isolation and characterisation of somatic hybrids of diploid Solanum tuberosum and Solanum brevidens and the use of amylose-free starch mutation for detection of introgression

Summary Somatic hybrids of diploid amylose-free (amf) Solanum tuberosum and diploid S. brevidens were made by Poly-Ethylene-Glycol (PEG) or electrofusion methods. For the isolation of interspecific hybrids the use of selection markers (kanamycin and hygromycin resistance) was useful but not essential. In this 2x+2x interspecific combination 4x and 6x somatic hybrids were obtained. Seed set was the best in 4x×4x (S. tuberosum) backcrosses, but seed germination was the best in 6x×4x combinations, using in vitro germination of unripe seeds harvested 25 days after pollination. A high degree of pollen stainability (30–40%) was observed in 7 tetraploid hybrids and very low in all hexaploids. After iodine staining, the recessive amf marker was expressed by a red colour instead of blue, visible in starch-containing cells like columella cells of root tips, (micro)tubers or microspores. As expected, complementation was observed in starch-containing cells of the fusion hybrids. Segregation of the amf marker was clearly observed in microspores of 4x and 6x hybrids. Segregation ratios in the 4x hybrids showed variable recombination frequencies. In the backcross progeny of hexaploid F12-5 with a tetraploid amf mutant one amylose-free recombinant among 67 plants was found, indicating the occurrence of meiotic recombination in the megaspore mother cells.     

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.     

Effects of aminoethoxyvinylglycine (AVG), environment,and genotype in overcoming hybridization barriers between Cucumis species

Summary It was attempted to overcome crossing barriers between the cucumber (Cucumis sativus L.), the melon (C. melo L.) and two wild Cucumis spp. (C. metuliferus Naud. and C. zeyheri 2x Sond.) by application of aminoethoxyvinylglycine (AVG) and by limiting the amount of rooting substrate. The reciprocal crosses between both wild species were used as a model system.Crosses between C. metuliferus and C. zeyheri 2x succeeded well if the maternal plants were grown in containers of 10 or 25 1 instead of in open soil. This treatment also improved crossability in the cross C. metuliferus x C. melo. It strongly enhanced the number of female flowers per plant in C. metuliferus. Application of AVG to pollinated flowers initially seemed also to improve crossability in these cross combinations, but later on this effect was not found again.Significant genetic variation for crossability was found in C. zeyheri 2x. Rare plants yielded numerous fruits with embryos after crosses with C. metuliferus and also with C. sativus. Clones from these plants gave similar results.Excessive temperature and radiation decreased crossability especially in crosses with C. metuliferus and C. sativus as maternal parent.     

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.