Identification of the tetrad-sterility-causing Solanum stoloniferum cytoplasm in interspecific hybrids with S. tuberosum

Tetrad sterility, in which only clumps of four premature pollen grains are released from anthers, has been observed in some modern potato cultivars. It is a form of cytoplasmic male sterility caused by the cytoplasm derived from the Mexican tetraploid species Solanum stoloniferum Schlechtd. et Bché., an important source of resistance to Potato virus Y in potato breeding. However, since S. stoloniferum is highly polymorphic, the source of tetrad-sterility-causing cytoplasm is unknown among diverse S. stoloniferum accessions. In this study, we directly crossed 24 S. stoloniferum accessions with pollen from 4x S. tuberosum and obtained 39 hybrids from 12 accessions. Nineteen hybrids from six accessions showed tetrad sterility, with either D/γ- or W/γ-type cytoplasm, and were triploid, tetraploid, or hexaploid. The W/γ-type cytoplasm was not necessarily associated with tetrad sterility. Sequence comparisons of 17 mitochondrial genes and their intergenic regions revealed a length polymorphism in the intergenic region between rpl5 and rps10, in which an amplified band of 859 bp was associated with tetrad sterility. This specific cytoplasm causing tetrad sterility is named TSCsto. The 859-bp band would be a useful diagnostic marker for identifying TSCsto in potato breeding.

Screening of Wild Potatoes Identifies New Sources of Late Blight Resistance

Late blight (LB) of potato is considered one of the most devastating plant diseases in the world. Most cultivated potatoes are susceptible to this disease. However, wild relatives of potatoes are an excellent source of LB resistance. We screened 384 accessions of 72 different wild potato species available from the U.S. Potato GeneBank against the LB pathogen Phytophthora infestans in a detached leaf assay (DLA). P. infestans isolates US-23 and NL13316 were used in the DLA to screen the accessions. Although all plants in 273 accessions were susceptible, all screened plants in 39 accessions were resistant. Resistant and susceptible plants were found in 33 accessions. All tested plants showed a partial resistance phenotype in two accessions, segregation of resistant and partial resistant plants in nine accessions, segregation of partially resistant and susceptible plants in four accessions, and segregation of resistant, partially resistant, and susceptible individuals in 24 accessions. We found several species that were never before reported to be resistant to LB: Solanum albornozii, S. agrimoniifolium, S. chomatophilum, S. ehrenbergii, S. hypacrarthrum, S. iopetalum, S. palustre, S. piurae, S. morelliforme, S. neocardenasii, S. trifidum, and S. stipuloideum. These new species could provide novel sources of LB resistance. P. infestans clonal lineage-specific screening of selected species was conducted to identify the presence of RB resistance. We found LB resistant accessions in Solanum verrucosum, Solanum stoloniferum, and S. morelliforme that were susceptible to the RB overcoming isolate NL13316, indicating the presence of RB-like resistance in these species.

Development of chromosome-specific markers for a study on introgressive hybridization of potato with the wild Mexican allotetraploid species Solanum stoloniferum Schltdl

In order to involve valuable germplasm of the wild Mexican allotetraploid potato species Solanum stoloniferum Schltdl. (genomic composition ААВВ) into breeding, pentaploid interspecific hybrids (ААAAВ) with cultivated potato S. tuberosum L. (АААА) and their backcross progenies are usually used. Homologous synapsis in meiosis of such hybrids is expected only between chromosomes of the A subgenome, therefore a question arose about a possibility of introgressing genetic material of the subgenome B into the A genome of cultivated potato. In this connection, development of various schemes for the B subgenome introgression into the genome of cultivated potato is considered as a topical issue. The previous research has yielded four schemes of S. stoloniferum involvement into breeding, which imply backcrossing with cultivated potato of the following interspecific hybrids: (1) hexaploids (genomic composition ААААВВ, the conventional introgression scheme), (2) tetraploids (putatively, АААВ), (3) self-pollination progeny of a 4x hybrid and (4) pentaploid hybrids with a putative genome composition of АААВВ. The present paper presents the first results of the development of chromosome-specific DNA markers for the identification of S. stoloniferum chromosomes in interspecific hybrids. An S. stoloniferum accession PI 205522 with a high degree of resistance to late blight and PVY had been found to possess several DNA-markers of the R-genes conferring resistance to these pathogens and was used in hybridization as a promising parent. A set of 23 SSR- and CAPS markers with the known chromosome location in S. tuberosum was generated. These markers detect polymorphism between parent genotypes, i.e., the diploid clone IGC 10/1.21 of cultivated potatoes S. tuberosum, and accession PI 205522 of S. stoloniferum. All the markers specific for the wild species were found in triploid (ААВ) and pentaploid (АААВВ) hybrids of S. stoloniferum × S. tuberosum. This set of markers will be used for efficiency assessment of different schemes for S. stoloniferum genetic material introgression into the obtained BC2-BC3 generations after crossing the interspecific hybrids with cultivated potato.

Production of male fertile interspecific hybrides between cultivated potatoes and valuable for breeding allotetraploid species Solanum stoloniferum

The germplasm of valuable for breeding wild allotetraploid potato species Solanum stoloniferumis rarely used because of pre- and postzygotic reproductive barriers with cultivated potatoes. One of the factors that complicate crosses between S. stoloniferum and S. tuberosumis unilateral incompatibility (UI).Here, we present the results of application of original SvSv-lines for overcoming UI in crosses with S. stoloniferum and of generating male fertile hybrids derived from this species. SvSv-lines are F2 S. tuberosum dihaploid× S. verrucosum that are male fertile and have D/γ-type cytoplasm. Since they are hybrids on homozygous for Svgene from S. verrucosum, they do not form SvSv-lines and have the same ability for elimination of prezygotic incompatibility as this species.As a result of pollination seven SvSv-lines were pollinated by 26 accessions of S. stoloniferum and a lot of hybrid seeds have been produced.In spite of low percentage of germination (1.9 %), formed 40 seedlings of interspecific hybrids. The experiment on hybridization between SvSv-lines and S. stoloniferum has been reproduced with the accession PI205522 of the wild species, which had DNA markers of PVY and LB resistance genes and “sterile” type cytoplasm W/γ: 950 hybrid seeds and 12 viable seedlings were produced. The genome of the seedlings was doubled by colchicine treatment, which generated hexaploids (F1) that formed highly fertile pollen and set seeds from self-pollination. We were able to cross them as females with the variety Katahdin. Produced pentaploid hybrids (BC1) were readily backcrossed by potato variety Quarta. Seedlings of BC2 were then backcrossed by potato varieties as female and, some of them, as male parents. The substantial part of F1, BC1 and BC2 plants of interspecific hybrids were male fertile (produced a lot functionally fertile pollen).