Isesaki, Japan
Isesaki, Japan

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Oikawa E.,Tohoku University | Takuno S.,Tohoku University | Takuno S.,University of California at Irvine | Izumita A.,Sakata Seed Corporation | And 4 more authors.
Molecular Breeding | Year: 2011

In F1 hybrid breeding of Brassica vegetables utilizing the self-incompatibility system, identification of S genotypes in breeding lines is required. In the present study, we developed S-tester lines of 87 S haplotypes, i. e., 42 S haplotypes in B. rapa and 45 S haplotypes in B. oleracea. With these materials, we established a simple, efficient, and reliable dot-blot technique for S genotyping for 40 S haplotypes of B. rapa and and 33 of B. oleracea using allele-specific oligonucleotide probes and allele-specific primer pairs designed from sequences of each SP11 allele. In this method, DNA fragments amplified using multiplex primer pairs with digoxigenin-dUTP were hybridized with dot-blotted allele-specific oligonucleotide probes with distinct signals. In addition, we developed a screening method for identification of plants harboring a particular S haplotype using a labeled allele-specific oligonucleotide probe. This method is considered to be useful for purity testing of F1 hybrid seeds. © 2010 Springer Science+Business Media B.V.


An F2 population was developed from a cross between a mur-cytoplasmic male sterile broccoli line and a restorer Chinese kale line. Phenotypic analysis of F2 plants indicated that the pollen fertility is controlled by two genes and segregated in a duplicate gene interaction mode with a ratio of 15:1. A total of 236 single nucleotide polymorphism (SNP) markers were developed utilizing 1,448 primers designed for production of expressed sequence tag (EST)-SNP markers of Raphanus sativus and analyzed by the dot-blot technique in 205 F2 individuals. A linkage map was constructed with a total of 142 markers and these markers were assigned to nine linkage groups together with simple sequence repeat markers mapped previously on the published linkage maps of Brassica oleracea. The linkage map spanned 909 cM with an average marker distance of 6.4 cM. A fertility restorer locus (Rfm1) was mapped on LG1, corresponding to chromosome 3, along with a flower color locus at a distance of 25 cM. SNP markers flanking the Rfm1 locus were BoCL2642s at a distance of 2.5 cM on one side and BoCL2901s at a distance of 7.5 cM on the other side. All the SNP markers showed homology with Arabidopsis thaliana and Brassica rapa genome sequences. Three pentatricopeptide repeat genes of the P-subfamily, particularly expressed in buds of the restorer line, were identified and these genes could be potential candidate fertility restorer genes. © 2012 Springer Science+Business Media B.V.


Kifuji Y.,Tohoku University | Kifuji Y.,Kaneko Seeds Co. | Hanzawa H.,Kaneko Seeds Co. | Terasawa Y.,Kaneko Seeds Co. | And 2 more authors.
Euphytica | Year: 2013

One hundred sixty-one EST-SNP markers were newly developed for analysis of QTLs for resistance to black rot caused by Xanthomonas campestris pv. campestris by determining EST sequences of a resistant line obtained from cabbage 'Early Fuji' and a susceptible broccoli line. A linkage map consisting of nine linkage groups was constructed with a total of 209 markers, including these new SNP markers and previously reported DNA markers. F2 plants grown in a field for 1 month were inoculated by spraying bacteria of race 1, and disease severity of each plant was recorded. Three QTLs, i. e., QTL-1, QTL-2, and QTL-3, were detected on linkage group C2, C4 and C5, respectively. QTL-1, which showed the highest LOD score and additive effect, was again detected in another F2 population used the next year, suggesting QTL-1 to be a major QTL. QTL-2 and QTL-3 could be minor QTLs influenced by environmental factors. The genomic region harboring QTL-1 showed synteny with a region from 5.3 to 7.4 Mb from the short arm end of chromosome 5 of Arabidopsis thaliana, which is rich in TIR-NBS-LRR family genes. The identified SNP markers in QTL-1 are considered to be useful in marker-assisted selection for black rot resistance in Brassica oleracea lines. © 2012 Springer Science+Business Media Dordrecht.


Shirasawa S.,Tohoku University | Kifuji Y.,Tohoku University | Kifuji Y.,Kaneko Seeds Co. | Komiya R.,Tohoku University | And 4 more authors.
Molecular Breeding | Year: 2013

Stable genic male sterility (GMS), which is not influenced by environmental factors, has not been used for F1 hybrid seed production because male-sterile inbred lines cannot be developed and male-sterile plants must be selected from segregating populations every time. However, the stability of male sterility may provide a reliable system for F1 seed production without contamination of selfed seeds. A genic male-sterile mutant in rice (Oryza sativa L.), C204, which was selected from progeny of the cultivar 'Koshihikari' irradiated by gamma rays, has shorter and whiter anthers than those of 'Koshihikari' and has no pollen grains. Segregation analysis of C204 suggested the male sterility of this mutant to be controlled by a recessive allele of a single gene. Linkage analysis of a mutated gene responsible for the male sterility revealed the gene to be in a region of ca. 75 kb on the long arm of chromosome 9. The nine genes predicted in the 75-kb region were sequenced, and compared with the published Nipponbare genome sequences. A single-base deletion was found in the first exon of a C204 allele of Os09g0493500, which encodes an NAD-dependent epimerase/dehydratase family protein, resulting in a frameshift causing a premature stop codon. A dot-blot single nucleotide polymorphism marker for detection of the single-base deletion in Os09g0493500 was developed. We herein propose an F1 hybrid seed production system using stable GMS with a simple selection method of GMS plants. © 2013 Springer Science+Business Media Dordrecht.


Nukui S.,Niigata University | Kitamura S.,Niigata University | Kitamura S.,Kaneko Seeds Co. | Hioki T.,Niigata University | And 6 more authors.
Breeding Science | Year: 2011

Fertile plants undergoing male gametogenesis can be treated with nitrous oxide (N 2O) gas to obtain 2n male gametes. N 2O treatment is also expected to restore the fertility of interspecific hybrids through meiotic restitution or mitotic amphidiploidization. However, this technique has few applications to date, and it is unknown how N 2O treatment restores fertility in sterile hybrids. To establish optimal N 2O treatment conditions and determine its cytological mechanism of action, we treated various sized floral buds with N 2O gas at different anther developmental stages from fertile and sterile hybrid lilies. N 2O treatment using the optimal 1- 4 mm floral buds induced mitotic polyploidization of male archesporial cells to produce 2n pollen in fertile hybrid lilies. In sterile hybrid lilies, N 2O treatment doubled the chromosome number in male archesporial cells followed by homologous chromosome pairing and normal meiosis in pollen mother cells (PMC), resulting in restoration of pollen fertility. Backcrossing the resultant fertile pollen to Lilium × formolongi produced many triploid BC1 plants. Thus N 2O treatment at the archesporial cell proliferating stage effectively overcame pollen sterility in hybrid lilies, resulting in fertile, 2n pollen grains that could produce progeny. The procedure presented here will promote interspecific or interploidy hybridization of lilies.


Tomioka K.,Japan National Institute of Agrobiological Science | Sato T.,Japan National Institute of Agrobiological Science | Moriwaki J.,Horticultural Research Institute | Terasawa Y.,Kaneko Seeds Co. | Koganezawa H.,Kaneko Seeds Co.
Journal of General Plant Pathology | Year: 2012

Severe spotting and blighting of leaves were found on bacopa (Sutera cordata), a scrophulariaceous ornamental, in greenhouses in Gunma Prefecture, Japan, from January through February 2007. After we isolated and identified the causal fungus as Colletotrichum destructivum and inoculated host plants with the isolate to confirm pathogenicity, we named this new disease anthracnose of bacopa. © 2012 The Phytopathological Society of Japan and Springer.


Trademark
Kaneko Seeds Co. | Date: 2011-11-22

Fresh vegetables; unprocessed corn; animal foodstuffs; seeds and bulbs; live trees; living grasses plants; natural turf; seedlings; saplings.

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