BackgroundJapanese cedar (Cryptomeria japonica) is an important tree for Japanese forestry. Male-sterile marker development in Japanese cedar would facilitate selection of male-sterile plus trees, addressing the widespread social problem of pollinosis and facilitating the identification of heterozygotes, which are useful for breeding.ResultsThis study used next-generation sequencing for single-nucleotide polymorphism discovery in libraries constructed from several organs, including male-sterile and male-fertile strobili. The single-nucleotide polymorphisms obtained were used to construct a high-density linkage map, which enabled identification of a locus on linkage group 9 strongly correlated with male-sterile trait. Expressed sequence tags corresponding to 11 marker loci from 5 isotigs were associated with this locus within 33.4-34.5 cM. These marker loci explained 100% of the phenotypic variation. Several homologs of these sequences are associated with male sterility in rice or Arabidopsis, including a pre-mRNA splicing factor, a DEAD-box protein, a glycosyl hydrolase, and a galactosyltransferase. These proteins are thus candidates for the causal male-sterile gene at the ms-1 locus. After we used a SNaPshot assay to develop markers for marker-assisted selection (MAS), we tested F2 progeny between male-sterile and wild-type plus trees to validate the markers and extrapolated the testing to a larger plus-tree population. We found that two developed from one of the candidates for the causal gene were suitable for MAS.ConclusionsMore than half of the ESTs and SNPs we collected were new, enlarging the genomic basis for genetic research on Japanese cedar. We developed two SNP markers aimed at MAS that distinguished individuals carrying the male-sterile trait with 100% accuracy, as well as individuals heterozygous at the male-sterile locus, even outside the mapping population. These markers should enable practical MAS for conifer breeding.Electronic supplementary materialThe online version of this article (10.1186/s12864-018-4581-5) contains supplementary material, which is available to authorized users.

Pollen-mediated gene flow was analyzed in a small, fragmented, natural population of Fagus crenata Blume by surveying five microsatellite markers in seedlings derived from open-pollinated crosses. Paternity of 162 seedlings derived from two maternal trees was assigned by the maximum-likelihood method using CERVUS 2.0, and pollen dispersal patterns within the study site were determined. Most of the trees within the site sired seeds, although we found evidence of limited pollen-mediated gene flow from outside the stand; 92% of the matings that generated the seedlings occurred between trees within the population, and 8% of the matings were mediated by pollen derived from trees located outside the study site. Although the pollen-mediated gene flow within the site was not strongly limited, mating frequencies of paternal trees were found to be weakly negatively correlated with their distance from the mother trees, positively correlated with their stem diameter at breast height, and uncorrelated with their relatedness to the mother trees.