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Now showing items 17 - 30 of 30

  • A practical method for apple cultivar identification and parent-offspring analysis using simple sequence repeat markers

    Iwanami, Hiroshi   Okada, Kazuma   Yamamoto, Toshiya   Abe, Kazuyuki  

    Differentiation of cultivars with simple sequence repeat (SSR) markers is a very useful technique for the true-to-type characterization of cultivars and clarification of parent-offspring relationships. We developed an SSR marker set for cultivar identification comprising 15 markers that were screened from 46 previously published SSRs. This marker set could be used for apple varieties including Malus x domestica and/or other Malus species. These SSRs successfully characterized 95 apples, including the leading and major founding cultivars used worldwide for modern apple breeding. Therefore, this marker set could be applied to almost all apple cultivars. We also analyzed the parent-offspring relationships of 69 cultivars by considering allele transmissions. This analysis revealed the true parentage of the following seven cultivars: 'Kizashi', 'Chinatsu', 'Honey Queen', 'Haruka', 'Seirin', 'Ozenokurenai', and Morioka #48. This analysis also revealed a parentage discrepancy for 'Hacnine'. From the parent-offspring analysis, two microsatellite mutation events at alleles inherited from pollen parents were observed.
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  • Fine mapping of Co, a gene controlling columnar growth habit located on apple (Malusdomestica Borkh.) linkage group 10

    Moriya, Shigeki   Okada, Kazuma   Haji, Takashi   Yamamoto, Toshiya   Abe, Kazuyuki  

    With 2 figures and 3 tables Abstract Tree architecture and shoot morphology are important factors in apple production systems. Because a columnar growth habit facilitates high-density planting and labour-saving fruit production methods, columnar apple breeding programmes are spreading worldwide. Columnar growth habit is controlled by a single dominant gene, Co, on linkage group 10. Fine mapping of Co was conducted using 1000 F1 progeny plants from 31 populations. Novel simple sequence-repeat (SSR) markers were developed based on the genome sequence of apple Golden Delicious and analysed for linkage to Co. Graphical genotyping of 22 progeny with recombination event in the vicinity of Co identified three SSR markers (Mdo.chr10.12, Mdo.chr10.13 and Mdo.chr10.14) that co-segregated with Co. Mdo.chr10.12 and Mdo.chr10.14 are well suited for marker-assisted selection because the sizes of the alleles linked to Co are different from those found in common ancestors of current apple breeding germplasm. The Co region was delimited to a 196-kb region of the apple genome, between markers Mdo.chr10.11 and Mdo.chr10.15.
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  • Identification of candidate genes responsible for the susceptibility of apple (Malus × domestica Borkh.) to Alternaria blotch

    Moriya, Shigeki   Terakami, Shingo   Okada, Kazuma   Shimizu, Taku   Adachi, Yoshihiko   Katayose, Yuichi   Fujisawa, Hiroko   Wu, Jianzhon   Kanamori, Hiroyuki   Yamamoto, Toshiya   Abe, Kazuyuki  

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  • Identification of candidate genes responsible for the susceptibility of apple (Malus x domestica Borkh.) to Alternaria blotch

    Moriya, Shigeki   Terakami, Shingo   Okada, Kazuma   Shimizu, Taku   Adachi, Yoshihiko   Katayose, Yuichi   Fujisawa, Hiroko   Wu, Jianzhon   Kanamori, Hiroyuki   Yamamoto, Toshiya   Abe, Kazuyuki  

    BackgroundThe mechanism underlying the interaction between host plant and host-selective toxin (HST)-producing Alternaria alternata during infection is of particular interest for sustainable crop production. Alternaria blotch of apple (Malus x domestica Borkh.) caused by A. alternata apple pathotype is a major disease particularly in East Asia, which is the largest producer of apples globally. A single dominant gene, Alt, controls the susceptibility of the apple cultivar Delicious' to Alternaria blotch. In this study, we fine mapped the Alt locus and characterized three potential candidate genes.ResultsWe used 797 F-1 individuals derived from 15 crosses between apple accessions susceptible (Alt/alt) and resistant (alt/alt) to Alternaria blotch to construct physical and genetic maps of the Alt locus located on the top of chromosome 11. Susceptible accessions were derived from Delicious.' To fine map the Alt locus, we constructed a BAC library of Starking Delicious,' a sport of Delicious,' and used graphical genotyping to delimit the Alt locus to a region of 43kb. Three genes predicted within the candidate Alt region were potentially involved in plant defense response, among which the gene encoding a coiled coil-nucleotide binding-leucine rich repeat (CC-NB-LRR) type disease resistance protein was the most promising. Moreover, a 12-bp insertion was uniquely identified in the 5 untranslated region of the Alt-associated allele of this gene, the presence or absence of which co-segregated with the susceptibility or resistance to A. alternata apple pathotype, respectively, among 43 tested cultivars including old ones and founders of modern apple breeding.ConclusionA disease resistance protein has been suggested as a determinant of susceptibility/resistance to HST-producing A. alternata for the first time. Our finding provides new insight into the mechanism of HST-mediated disease control used by A. alternata against host plants.
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  • Genomic dissection of a 'Fuji' apple cultivar:re-sequencing,SNP marker development,definition of haplotypes,and QTL detection

    Kunihisa, Miyuki   Moriya, Shigeki   Abe, Kazuyuki   Okada, Kazuma   Haji, Takashi   Hayashi, Takeshi   Kawahara, Yoshihiro   Itoh, Ryutaro   Itoh, Takeshi   Katayose, Yuichi   Kanamori, Hiroyuki   Matsumoto, Toshimi   Mori, Satomi   Sasaki, Harumi   Matsumoto, Takashi   Nishitani, Chikako   Terakami, Shingo   Yamamoto, Toshiya  

    'Fuji' is one of the most popular and highly-produced apple cultivars worldwide, and has been frequently used in breeding programs. The development of genotypic markers for the preferable phenotypes of 'Fuji' is required. Here, we aimed to define the haplotypes of 'Fuji' and find associations between haplotypes and phenotypes of five traits (harvest day, fruit weight, acidity, degree of watercore, and flesh mealiness) by using 115 accessions related to 'Fuji'. Through the re-sequencing of 'Fuji' genome, total of 2,820,759 variants, including single nucleotide polymorphisms (SNPs) and insertions or deletions (indels) were detected between 'Fuji' and 'Golden Delicious' reference genome. We selected mapping-validated 1,014 SNPs, most of which were heterozygous in 'Fuji' and capable of distinguishing alleles inherited from the parents of 'Fuji' (i.e., 'Rails Janet' and 'Delicious'). We used these SNPs to define the haplotypes of 'Fuji' and trace their inheritance in relatives, which were shown to have an average of 27% of 'Fuji' genome. Analysis of variance (ANOVA) based on 'Fuji' haplotypes identified one quantitative trait loci (QTL) each for harvest time, acidity, degree of water core, and mealiness. A haplotype from 'Delicious' chr14 was considered to dominantly cause watercore, and one from 'Rails Janet' chr1 was related to low-mealiness.
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  • A Self-compatible Pollen-part Mutant of Japanese Pear Produced by Crossing 'Kosui' with Pollen from Gamma-irradiated 'Kosui'

    Sawamura, Yutaka   Mase, Nobuko   Takada, Norio   Sato, Akihiko   Nishitani, Chikako   Abe, Kazuyuki   Masuda, Tetsuo   Yamamoto, Toshiya   Saito, Toshihiro   Kotobuki, Kazuo  

    The self-incompatible (SI) Japanese pear cultivar 'Kosui' was pollinated with pollen collected from a chronically gamma-irradiated 'Kosui' tree, and a progeny was obtained. This progeny resulted in the identification of self-compatible (SC) breeding selection, designated 415-1, which showed 74.4% fruit set in a self-pollination test. PCR-based genetic analysis revealed that 415-1 has S-RNase genotype S4S5, which is the same as that of the parent, 'Kosui'. Pollination trials were used to investigate whether 415-1 harbors a stylar-part mutation or a pollen-part mutation in its SI locus. When 415-1 was pollinated with pollen from cultivars of the same genotype ('Syuugyoku' and 'Oushuu'), no seed-containing fruit were set, indicating that 415-1 contains functional S-4- and S-5-RNase alleles. On the other hand, when 'Syuugyoku' and 'Oushuu' were pollinated with pollen from 415-1, fruit-bearing seeds were produced; therefore, we conclude that 415-1 carries a pollen-part mutation at the SI locus. This new self-compatible breeding selection will be useful for the development of new Japanese pear cultivars with SC.
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  • Aligned genetic linkage maps of apple rootstock cultivar 'JM7' and Malus sieboldii 'Sanashi 63' constructed with novel EST-SSRs

    Moriya, Shigeki   Iwanami, Hiroshi   Kotoda, Nobuhiro   Haji, Takashi   Okada, Kazuma   Terakami, Shingo   Mimida, Naozumi   Yamamoto, Toshiya   Abe, Kazuyuki  

    Identification of markers associated with genes of interest and quantitative trait loci (QTLs), combined with high-density genetic linkage maps, can help reduce labor and costs by enabling marker-assisted selection (MAS). In this study, a dwarfing apple rootstock cultivar 'JM7' (Malus prunifolia x Malus pumila 'Malling 9') and wild apple Malus sieboldii 'Sanashi 63' (section Sorbomalus) were used for constructing genetic linkage maps. Here, a species from section Sorbomalus was used for the first time as a target species in a genome-wide mapping study. We also developed and mapped 137 novel-expressed sequence tag-simple sequence repeat (EST-SSR) markers. The genetic linkage maps of 'JM7' and 'Sanashi 63' consisted of 415 and 310 loci and spanned 998.0 and 981.8 cM, respectively, comparable to the reference map of Malus x domestica 'Discovery'. A BLASTN search revealed that all of the EST-SSR sequences used in this study exhibited very high homology to one or more previously characterized apple genome contigs. Although the most homologous contigs of 89 EST-SSRs were located within the same linkage groups (LGs) identified by mapping analysis, the other 48 EST-SSRs were aligned into contigs positioned in different LGs than those identified by mapping. When search criteria were expanded to include the five most homologous contigs of each EST-SSR, at least one of the top five contigs for 15 of these 48 EST-SSRs corresponded to the LG obtained by mapping. The maps of 'JM7' and 'Sanashi 63' may be useful for analyzing important rootstock characteristics and identifying markers for MAS.
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  • Allelic composition of MdMYB1 drives red skin color intensity in apple (Malus x domestica Borkh.) and its application to breeding

    Moriya, Shigeki   Kunihisa, Miyuki   Okada, Kazuma   Shimizu, Taku   Honda, Chikako   Yamamoto, Toshiya   Muranty, Helene   Denance, Caroline   Katayose, Yuichi   Iwata, Hiroyoshi   Abe, Kazuyuki  

    Since apple fruit skin reddens poorly under warmer climates, new apple cultivars are desired that are adapted to global warming in terms of bearing well-reddened fruit. We developed a simple sequence repeat marker, Mdo.chr9.4, which is suitable for red skin color selection. It amplified four alleles (Mdo.chr9.4-R-0, Mdo.chr9.4-Y-3, Mdo.chr9.4-Y-9, and Mdo.chr9.4-Y-15) distinguished by length. Mdo. chr9.4-R-0 associated with MdMYB1-1 which confers red fruit skin. The presence of Mdo. chr9.4-R-0 was consistent with empirical skin color in all 160 tested accessions. Mdo. chr9.4 was identified as the only significant marker that contributed to red skin color intensity by a genome wide association study (GWAS), and it accounted for 52.0% of phenotypic variation, confirming that MdMYB1 was the major and principal determinant of fruit skin color in apples. Individuals with a homozygous state of Mdo.chr9.4-R-0 (dose 2) were significantly redder than those showing a heterozygote state (dose 1) in both the accession set and full-sib families, indicating a partially dominant effect of MdMYB1-1. Therefore, the selection of dose 2 individuals would target individuals with intensive red skin. We applied Mdo.chr9.4 to several application populations using a time and cost-efficient genotyping system developed in the present study. This system, along with Mdo.chr9.4, provide advanced marker-assisted breeding for intensive red skin color apples adapted to a global warming climate.
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  • Isolation and characterization of multiple F-box genes linked to the S-9- and S-10-RNase in apple (Malus x domestica Borkh.)

    Okada, Kazuma   Moriya, Shigeki   Haji, Takashi   Abe, Kazuyuki  

    Using 11 consensus primer pairs designed from S-linked F-box genes of apple and Japanese pear, 10 new F-box genes (MdFBX21 to 30) were isolated from the apple cultivar 'Spartan' (S9S10). MdFBX21 to 23 and MdFBX24 to 30 were completely linked to the S-9-RNase and S-10-RNase, respectively, and showed pollen-specific expression and S-haplotype-specific polymorphisms. Therefore, these 10 F-box genes are good candidates for the pollen determinant of self-incompatibility in apple. Phylogenetic analysis and comparison of deduced amino acid sequences of MdFBX21 to 30 with those of 25 S-linked F-box genes previously isolated from apple showed that a deduced amino acid identity of greater than 88.0 % can be used as the tentative criterion to classify F-box genes into one type. Using this criterion, 31 of 35 F-box genes of apple were classified into 11 types (SFBB1-11). All types included F-box genes derived from S-3- and S-9-haplotypes, and seven types included F-box genes derived from S-3-, S-9-, and S-10-haplotypes. Moreover, comparison of nucleotide sequences of S-RNases and multiple F-box genes among S-3-, S-9-, and S-10-haplotypes suggested that F-box genes within each type showed high nucleotide identity regardless of the identity of the S-RNase. The large number of F-box genes as candidates for the pollen determinant and the high degree of conservation within each type are consistent with the collaborative non-self-recognition model reported for Petunia. These findings support that the collaborative non-self-recognition system also exists in apple.
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  • Mode of inheritance in fruit acidity in apple analysed with a mixed model of a major gene and polygenes using large complex pedigree

    Moriya, Shigeki   Kotoda, Nobuhiro   Mimida, Naozumi   Takahashi-Sumiyoshi, Sae   Abe, Kazuyuki  

    Fruit acidity is an important characteristic to determine the marketability of apple (Malus x domestica Borkh.). To reveal the mode of inheritance in fruit acidity and to estimate the genetic parameters, we performed segregation analysis using a population from an apple breeding programme. Four models (mixed, Mendelian, polygenic and environmental) were compared to find the most likely mode of the inheritance of acidity. The phenotypic variance of acidity in the population was properly explained using a mixed model of a major gene and polygenes. The genotypic values of the major gene (AA, Aa and aa) were estimated to be 0.45, 0.52 and 0.92 g/100 ml in titratable acidity, respectively. The values of the homozygote (AA) and the heterozygote (Aa) were very close and lower than that of the other homozygote (aa), indicating that an allele of the major gene appeared to have complete dominance with a function of lowering acidity. The estimate of the heritability after accounting for the major gene was moderately high, 0.43, in the mixed model. This means that even with removal of the effect of the major gene, acidity could fluctuate considerably by the effect of polygenes. The proportions of progeny with adequate acidity differed greatly, depending on which genotype was used as parents for the crossing. Therefore, it is very important to know the genotypes of parental cultivars before crossing.
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  • Optimization of Inversion Time for Postmortem Fluid-attenuated Inversion Recovery (FLAIR) MR Imaging at 1.5T: Temperature-based Suppression of Cerebrospinal Fluid.

    Abe, Kazuyuki   Kobayashi, Tomoya   Shiotani, Seiji   Saito, Hajime   Kaga, Kazunori   Tashiro, Kazuya   Someya, Satoka   Hayakawa, Hideyuki   Homma, Kazuhiro  

    PURPOSE: Signal intensity (SI) and image contrast on postmortem magnetic resonance (MR) imaging are different from those of imaging of living bodies. We sought to suppress the SI of cerebrospinal fluid (CSF) sufficiently for fluid-attenuated inversion recovery (FLAIR) sequence in postmortem MR (PMMR) imaging by optimizing inversion time (TI).; MATERIALS AND METHODS: We subject 28 deceased patients to PMMR imaging 3 to 113 hours after confirmation of death (mean, 27.4 hrs.). PMMR imaging was performed at 1.5 tesla, and T1 values of CSF were measured with maps of relaxation time. Rectal temperatures (RT) measured immediately after PMMR imaging ranged from 6 to 32=C2=B0C (mean, 15.4=C2=B0C). We analyzed the relationship between T1 and RT statistically using Pearson's correlation coefficient. We obtained FLAIR images from one cadaver using both a TI routinely used for living bodies and an optimized TI calculated from the RT.; RESULTS: T1 values of CSF ranged from 2159 to 4063 ms (mean 2962.4), and there was a significantly positive correlation between T1 and RT (r =3D 0.96, P < 0.0001). The regression expression for the relationship was T1 =3D 74.4 * RT + 1813 for a magnetic field strength of 1.5T. The SI of CSF was effectively suppressed with the optimized TI (0.693 * T1), namely, TI =3D 0.693 * (77.4 * RT + 1813).; CONCLUSION: Use of the TI calculated from the linear regression of the T1 and RT optimizes the FLAIR sequence of PMMR imaging.=20
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  • Identification of Venturia inaequalis Races in Morioka,Japan and Identification of a Quantitative Trait Locus Associated with Resistance to Apple Scab in 'Akane' Apples

    Goto, Satoshi   Kubota, Takashi   Kunihisa, Mivuki   Tazawa, Junko   Kudo, Tsuyoshi   Kasai, Satoshi   Kudo, Haruka   Okada, Kazuma   Yamamoto, Toshiya   Fukazawa-Akada, Tomoko   Hatsuyama, Yoshimichi   Abe, Kazuyuki  

    Apple scab, which is caused by Venturia inaequalis (Cooke) G. Wint., is a destructive disease that affects apples (Malus x domestica Borkh.). To develop an approach to breeding apples for scab resistance, race distribution of V inaequalis in Morioka. Japan and genetic factors affecting partial scab resistance observed on 'Alcane' were assessed. An inoculation test using several differential hosts suggested that race 1 of V. inaequalis was the predominant race in Morioka, Japan. To characterize the genetic profile of 'Akane' resistance, quantitative trait locus (QTL) analyses that employed two years of natural scab infection scores in a fungicide-free orchard in Kuroishi, Aomori, Japan were performed using an F-1 population derived from the 'Orin' x 'Akane' cross. A QTL allele that explained 20% of the phenotypic variance in scab resistance was detected on chromosome 17 of 'Akane'. In the middle region of chromosome 17 of 'Orin' and in very close proximity to the 'Akane' QTL, a weak QTL allele was also detected, which explained 5% of the phenotypic variance observed. We considered that these effects were caused by alleles of the identical QTL. Haplotyping analysis indicated that this QTL resistance allele from 'Akane' originated from 'Worcester Pearmain'.
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  • Four TFL1CEN-Like Genes on Distinct Linkage Groups Show Different Expression Patterns to Regulate Vegetative and Reproductive Development in Apple (Malusdomestica Borkh.)

    Mimida, Naozumi   Kotoda, Nobuhiro   Ueda, Takanori   Igarashi, Megumi   Hatsuyama, Yoshimichi   Iwanami, Hiroshi   Moriya, Shigeki   Abe, Kazuyuki  

    Recent molecular analyses in several plant species revealed that TERMINAL FLOWER1 (TFL1) and CENTRORADIALIS (CEN) homologs are involved in regulating the flowering time andor maintaining the inflorescence meristem. In apple (Malusdomestica Borkh.), four TFL1CEN-like genes, MdTFL1, MdTFL1a, MdCENa and MdCENb, were found and mapped by a similar position on putatively homoeologous linkage groups. Apple TFL1CEN-like genes functioned equivalently to TFL1 when expressed constitutively in transgenic Arabidopsis plants, suggesting that they have a potential to complement the TFL1 function. Because MdTFL1 and MdTFL1a were expressed in the vegetative tissues in both the adult and juvenile phases, they could function redundantly as a flowering repressor and a regulator of vegetative meristem identity. On the other hand, MdCENa was mainly expressed in fruit receptacles, cultured tissues and roots, suggesting that it is involved in the development of proliferating tissues but not in the control of the transition from the juvenile to the adult phase. In contrast, MdCENb was silenced in most organs probably due to gene duplication by the polyploid origin of apple. The expression patterns of MdTFL1 and MdCENa in apple were also supported by the heterologous expression of -glucuronidase fused with their promoter regions in transgenic Arabidopsis. Our results suggest that functional divergence of the roles in the regulation of vegetative meristem identity may have occurred among four TFL1CEN-like genes during evolution in apple.
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  • Host-guest complexation behavior of resorcinarenes with tetraalkylammonium ions and N-methylpyridinium ions in methanol: The effect of bulky hydrophobic substituents at the extra-annular positions

    Morikawa, Osamu   Yamaguchi, Hiroshi   Katsube, Yoshiko   Abe, Kazuyuki   Kobayashi, Kazuhiro   Konishi, Hisatoshi  

    The host-guest interaction of C-methylresorcin[4]arene and its derivative having four tert-butylsulfanylmethyl groups at the extra-annular positions was studied by H-1 NMR spectroscopy in CD3OD. Based on the association constants (Ka) and the complexation-induced NMR shifts (CIS), it was concluded that the bulky substituents create a deep cavity with a narrow entrance and improve the size and shape selectivity.
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