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We genotyped strawberry cultivars by two newly selected and two previously reported SSR markers. All four markers produced interpretable electropherograms from 75 accessions consisting of 72 Fragaria × ananassa cultivars or lines and three octoploid Fragaria species accessions. These SSR markers were highly polymorphic; in particular, one of the newly developed markers, FxaHGA02P13, was capable of distinguishing all of the accessions except for a mutant strain that was derived from another accession in the set. When two markers were combined, all 48 full-sib individuals could be distinguished. Fingerprinting patterns were reproducible between multiple samples, including the leaves, sepals, and fruit flesh of the same accession. Principal-coordinate analysis of the 75 accessions detected several groups, which reflect taxon and breeding site. Together with other available markers, these SSR markers will contribute to the management of strawberry genetic resources and the protection of breeders’ rights.

Male sterility is defined as the loss of pollen fertility, and it represents a plant reproductive isolation symptom, along with self-incompatibility. It plays an important role in the efficient production of F 1-hybrid seeds, which results in affordable seed prices for farmers. Male sterile cultivated strawberry Fragaria × ananassa Duch. plants were found in an F 1 population and reciprocal backcrossed populations derived from a cross between 'Fukuoka S6' and 'Kaorino'. Male sterile plants were clearly distinguished from male fertile plants in those populations based on the anther color. The pollen of the male sterile plants was a lighter yellow color and not maturely shaped compared with pollen of male fertile plants. Genotyping was performed using EST-SSR markers in the three populations. Quantitative trait locus analyses for pollen fertility were conducted independently using three kinds of populations, and this revealed that male sterility was controlled by three independent chromosomal regions in these populations, which corresponded to chromosome 4 in the wild strawberry (Fragaria vesca) genome. One region was derived from 'Fukuoka S6' and the other two regions from 'Kaorino'. The segregation patterns of fertile and sterile plants in each population clearly supported the three gene theory of male sterility in cultivated strawberries. The accumulation of recessive alleles at the three regions led to male sterility, and the existence of a dominant allele in at least one region resulted in fertile pollen. Male sterile plants were also found in two self-pollenated populations derived from 'Fukuoka S6' and 'Kaorino', and the effects of the three regions were validated. The adaptability levels of the three genes with different genetic backgrounds were also evaluated using core collection cultivars and selected lines derived using recurrent selection. We also detected flanking DNA markers for the three regions associated with male sterility. The use of these markers, which are in the vicinity of quantitative trait loci and responsible for malesterility, could increase the efficiency of producing seed-propagated strawberry F 1-hybrids.

Cultivated strawberry is the most widely consumed fruit crop in the world, and therefore, many breeding programs are underway to improve its agronomic traits such as fruit quality. Strawberry cultivars were vegetatively propagated through runners and carried a high risk of infection with viruses and insects. To solve this problem, the development of F1 hybrid seeds has been proposed as an alternative breeding strategy in strawberry. In this study, we conducted a potential assessment of genomic selection (GS) in strawberry F1 hybrid breeding. A total of 105 inbred lines were developed as candidate parents of strawberry F1 hybrids. In addition, 275 parental combinations were randomly selected from the 105 inbred lines and crossed to develop test F1 hybrids for GS model training. These populations were phenotyped for petiole length, leaf area, Brix, fruit hardness, and pericarp color. Whole-genome shotgun sequencing of the 105 inbred lines detected 20,811 single nucleotide polymorphism sites that were provided for subsequent GS analyses. In a GS model construction, inclusion of dominant effects showed a slight advantage in GS accuracy. In the across population prediction analysis, GS models using the inbred lines showed predictability for the test F1 hybrids and vice versa, except for Brix. Finally, the GS models were used for phenotype prediction of 5,460 possible F1 hybrids from 105 inbred lines to select F1 hybrids with high fruit hardness or high pericarp color. These F1 hybrids were developed and phenotyped to evaluate the efficacy of the GS. As expected, F1 hybrids that were predicted to have high fruit hardness or high pericarp color expressed higher observed phenotypic values than the F1 hybrids that were selected for other objectives. Through the analyses in this study, we demonstrated that GS can be applied for strawberry F1 hybrid breeding.