S-genotype identification based on allele-specific PCR in Japanese pear     

Kenji Nashima  Shingo Terakami  Sogo Nishio  Miyuki Kunihisa  Chikako Nishitani  Toshihiro Saito  Toshiya Yamamoto 《Breeding Science》2015,65(3):208-215

Gametophytic self-incompatibility in Japanese pear (Pyrus pyrifolia Nakai) is controlled by the single, multi-allelic S-locus. Information about the S-genotypes is important for breeding and the selection of pollen donors for fruit production. Rapid and reliable S-genotype identification system is necessary for efficient breeding of new cultivars in Japanese pear. We designed S allele-specific PCR primer pairs for ten previously reported S-RNase alleles (S¹–S⁹ and S^k) as simple and reliable method. Specific nucleotide sequences were chosen to design the primers to amplify fragments of only the corresponding S alleles. The developed primer pairs were evaluated by using homozygous S-genotypes (S¹/S¹–S⁹/S⁹ and S^4sm/S^4sm) and 14 major Japanese pear cultivars, and found that S allele-specific primer pairs can identify S-genotypes effectively. The S allele-specific primer pairs developed in this study will be useful for efficient S-genotyping and for marker-assisted selection in Japanese pear breeding programs.  相似文献   

Purification and characterization of sulfotransferase specific to O-22 of 11-hydroxy saxitoxin from the toxic dinoflagellate Gymnodinium catenatum (dinophyceae)     

Takashi YOSHIDA  Yoshihiko SAKO  Aritsune UCHIDA  Tomonori KAKUTANI  Osamu ARAKAWA  Tamao NOGUCHI  Yuzaburo ISHIDA 《Fisheries Science》2002,68(3):634-642

ABSTRACT: A novel sulfotransferase (O-ST), which transferred the sulfate group of 3'-phosphoadenosine 5'-phosphosulfate (PAPS) to O-22 of 11-α,β-hydroxy saxitoxin (STX) and produced GTX2 + 3, was purified to homogeneity from the cytosolic fraction of clonal-axenic vegetative cells of the toxic dinoflagellate Gymnodinium catenatum GC21V. After four purification steps, including affinity chromatography and anion exchange chromatography, the enzyme was purified 500-fold and the yield was 4%. On affinity chromatography with a PAP-agarose column, O-ST was observed in the bound fraction, and N-ST specific to N-21 of STX and GTX2 + 3 was found in the unbound fraction. The molecular mass of the purified enzyme was determined by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) to be 65 kDa. Gel filtration chromatography showed a native molecular mass of 67 kDa, indicating that O-ST is a monomeric enzyme. The enzyme was optimally active at pH 6.0 and 35°C. O-ST did not require metal cations for its activity. O-ST required PAPS as the sole source of sulfate. O-ST transferred a sulfate group from PAPS to only O-22 of 11-α,β-hydroxy STX and not to N-21 of these toxins. These observations suggested that two ST, N-ST and O-ST, participate in the sulfation of PSP toxins.  相似文献   

Molecular basis of diverse responses to acetolactate synthase-inhibiting herbicides in sulfonylurea-resistant biotypes of Schoenoplectus juncoides     

AKIRA UCHINO  SHIGERU OGATA  HIROSHI KOHARA  SHUICHI YOSHIDA  TOSHIHITO YOSHIOKA  HIROAKI WATANABE 《Weed Biology and Management》2007,7(2):89-96

Sulfonylurea-resistant biotypes of Schoenoplectus juncoides were collected from Nakafurano, Shiwa, Matsuyama, and Yurihonjyo in Japan. All of the four biotypes showed resistance to bensulfuron-methyl and thifensulfuron-methyl in whole-plant experiments. The growth of the Nakafurano, Shiwa, and Matsuyama biotypes was inhibited by imazaquin-ammonium and bispyribac-sodium, whereas the Yurihonjyo biotype grew normally after treatment with these herbicides. The herbicide concentration required to inhibit the acetolactate synthase (ALS) enzyme by 50% (I₅₀), obtained using in vivo ALS assays, indicated that the four biotypes were > 10-fold more resistant to thifensulfuron-methyl than a susceptible biotype. The Nakafurano, Shiwa, and Matsuyama biotypes exhibited no or little resistance to imazaquin-ammonium, whereas the Yurihonjyo biotype exhibited 6700-fold resistance to the herbicide. The Nakafurano and Shiwa biotypes exhibited no resistance to bispyribac-sodium, but the Matsuyama biotype exhibited 21-fold resistance and the Yurihonjyo biotype exhibited 260-fold resistance to the herbicide. Two S. juncoides ALS genes (ALS1 and ALS2) were isolated and each was found to contain one intron and to encode an ALS protein of 645 amino acids. Sequencing of the ALS genes revealed an amino acid substitution at Pro₁₉₇ in either encoded protein (ALS1 or ALS2) in the biotypes from Nakafurano (Pro₁₉₇ → Ser₁₉₇), Shiwa (Pro₁₉₇ → His₁₉₇), and Matsuyama (Pro₁₉₇ → Leu₁₉₇). The ALS2 of the biotype from Yurihonjyo was found to contain a Trp₅₇₄ → Leu₅₇₄ substitution. The relationships between the responses to ALS-inhibiting herbicides and the amino acid substitutions, which are consistent with previous reports in other plants, indicate that the substitutions at Pro₁₉₇ and Trp₅₇₄ are the basis of the resistance to sulfonylureas in these S. juncoides biotypes.  相似文献   

Allelic frequency of PCR-RFLP type of the BoLA-DRB3 in Japanese Holstein herds and the relation to mastitis     

Tatsuyuki YOSHIDA  Harutaka MUKOYAMA  Hiroki FURUTA  Colin W. HOLMES  Motoaki KOSUGIYAMA  Hiroshi TOMOGANE 《Animal Science Journal》2008,79(4):409-416

Holstein Cows ( n  = 702) from 26 dairy herds in the Tama area of Tokyo, Japan were examined for polymorphisms of the BoLA-DRB3 allele using a PCR-RFLP method. Twenty alleles were observed and allelic frequencies ranged from < 1% to 20.3%. Nine alleles ( DRB3.2 * 24, * 16, * 8, * 23, * 22, * 3, * 11 , * 10 and * 7 in order) constituted 90.0% of all alleles. Somatic cell counts (SCC) were used to classify healthy (group 1), mastitis (group 2) and suspected (group 3) cows. Frequencies of DRB3.2 * 11 and DRB3.2 * 23 were slightly higher in group 1 than in group 2, whereas, frequencies of DRB3.2 * 8 and DRB3.2 * 16 were slightly higher in group 2 than in group 1. However, none of the differences in frequencies between the two groups were statistically significant. For combinations of alleles, frequencies of DRB3.2 * 8/ * 23 ( P  < 0.1) and DRB3.2 * 16/ * 24 ( P  < 0.05) were significantly higher in group 2 than in group 1, and their odds ratios were 2.1, 2.5, respectively. However, there were no significant differences between genotypes in their effects for SCC. On the other hand, frequency of DRB3.2 * 23/ * 23 including combinations of DRB3.2 * 23 with minor alleles was significantly higher in group 1 than in group 2 ( P  < 0.01), and the odds ratio was 0.3. Therefore, it was considered that mastitis resistance or susceptibility of cows may vary with the combination of BoLA-DRB3 alleles.  相似文献   

Identification of heat shock protein 70 genes HSPA2, HSPA5 and HSPA8 from the Japanese quail, Coturnix japonica     

Shigehisa IWAMOTO  Shinya SATO  Kazuyoshi HOSOMICHI  Autthaporn TAWEETUNGTRAGOON  Takashi SHIINA  Hisashi MATSUBAYASHI  Hiromi HARA  Yutaka YOSHIDA  Kei HANZAWA 《Animal Science Journal》2008,79(2):171-181

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Rheological Properties of Starch Gels from Wheat Mutants with Reduced Amylose Content     

Tomoko Sasaki  Takeshi Yasui  Chikako Kiribuchi‐Otobe  Takashi Yanagisawa  Masaya Fujita  Kaoru Kohyama 《Cereal Chemistry》2007,84(1):102-107

Wheat lines with reduced amylose content were recently produced by single and double mutation from a low‐amylose line, Kanto 107. They are appropriate for clarifying the influence of amylose content on starch gel properties because of their similar genetic background. When measured using the concanavalin A method (ConA), the total amylose content of isolated starches from Kanto 107 and three mutants (K107Afpp4, Tanikei A6599‐4, K107Wx2) was 24.8, 18.5, 7.1, and 1.7%, respectively. Results of differential scanning calorimetry (DSC) showed that the difference in amylose content strongly affected gelatinization conclusion temperature and enthalpy. We prepared 30 and 40% starch gels and measured their dynamic shear viscoelasticity using a rheometer with parallel plate geometry. Compressive and creep‐recovery tests were conducted under uniaxial compression. The storage shear modulus correlated highly with the amylose content of starch in 30 and 40% starch gels. The creep‐recovery test showed a clear distinction in creep curves among starch samples. When the compressive force required for 50, 80, and 95% strains was compared, starch gels with lower amylose content showed lower compressive force at 50% strain. Waxy starch gel (K107Wx2) showed higher compressive force at strain >80% than other samples due to its sticky property.  相似文献   

Studies on Endophytic Actinomycetes ( I ) <Emphasis Type="Italic">Streptomyces </Emphasis>sp. Isolated from Rhododendron and Its Antifungal Activity     

Masafumi SHIMIZU  Yoshiko NAKAGAWA  Yukio SATO  Tamotsu FURUMAI  Yasuhiro IGARASHI  Hiroyasu ONAKA  Ryuji YOSHIDA  Hitoshi KUNOH 《Journal of General Plant Pathology》2000,66(4):360-366

To survey endophytic actinomycetes as potential biocontrol agents against fungal diseases of rhododendron, young plants of rhododendron were surface-sterilized for use as an isolation source. Nine, six and two isolates, with distinguishing characteristics based on the macroscopic appearance of colonies, were obtained from roots, stems and leaves, respectively, suggesting that various species of actinomycetes grow in the respective organs of this plant as symbionts or parasites. On an agar medium, only isolate R-5 commonly formed a clear growth-inhibition zone against two major fungal pathogens of rhododendron, Phytophthora cinnamomi and Pestalotiopsis sydowiana, indicating that this isolate can produce antifungal material(s). Acetone extracts of a liquid culture of R-5 had a broad antimicrobial spectrum against Gram-positive bacteria, yeast and filamentous fungi. Isolate R-5 was identified as a Streptomyces sp. based on morphological, physiological and chemotaxonomical characteristics. The present results indicate that isolate R-5 is a suitable candidate for the biocontrol of diseases of rhododendron. Received 25 March 2000/ Accepted in revised form 18 May 2000  相似文献   

Development of a Yersinia entomophaga bait for control of larvae of the porina moth (Wiseana spp.), a pest of New Zealand improved grassland systems     

Mark RH Hurst  Jayanthi Swaminathan  David A Wright  Scott Hardwick  Colin M Ferguson  Amy Beattie  Nicola K Richards  Lincoln Harper  Ray A Moss  Vanessa M Cave  Chikako van Koten  Mark R McNeill 《Pest management science》2020,76(1):350-359

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Carotenoid accumulation in Japanese apricot (Prunus mume Siebold & Zucc.): molecular analysis of carotenogenic gene expression and ethylene regulation     

Kita M  Kato M  Ban Y  Honda C  Yaegaki H  Ikoma Y  Moriguchi T 《Journal of agricultural and food chemistry》2007,55(9):3414-3420

To elucidate the regulatory mechanisms of carotenogenesis in Japanese apricot (Prunus mume Siebold & Zucc.), the relationships between carotenoid accumulation and the expression of the carotenogenic genes, phytoene synthase (PmPSY-1), phytoene desaturase (PmPDS), zeta-carotene desaturase (PmZDS), lycopene beta-cyclase (PmLCYb), lycopene epsilon-cyclase (PmLCYe), beta-carotene hydroxylase (PmHYb), and zeaxanthin epoxidase (PmZEP), were analyzed in two cultivars with different ripening traits, 'Orihime' and 'Nanko.' In 'Orihime' fruits, large amounts of carotenoids accumulated on the tree, concomitant with the induction of PmPSY-1 and the downstream carotenogenic genes PmLCYb, PmHYb, and PmZEP. In 'Nanko' fruits, carotenoids accumulated mainly after harvest, correlating with an appreciable induction of PmPSY-1 expression, but the downstream genes were not notably induced, which may explain the lower total carotenoid content in 'Nanko' than in 'Orihime.' In both cultivars, a decrease in PmLCYe expression and increased or constant PmLCYb expression could cause the metabolic shift from beta,epsilon-carotenoid synthesis to beta,beta-carotenoid synthesis that occurs as ripening approaches. Next, the effects of ethylene on the expression of PmPSY-1 and carotenoid accumulation were investigated in 'Nanko' fruits treated with propylene or 1-methylcyclopropene (1-MCP). Propylene treatment induced both ethylene production and carotenoid accumulation. PmPSY-1 was constitutively expressed, but propylene treatment accelerated its induction. 1-MCP treatment caused a slight inhibition of carotenoid accumulation along with the repression, although not complete, of PmPSY-1. Collectively, although PmPSY-1 expression was not exclusively regulated by ethylene, both the notable induction of PmPSY-1 accelerated by ethylene and the subsequent induction of the downstream carotenogenic genes, especially PmLCYb, could be necessary for the massive carotenoid accumulation that occurs during ripening. Furthermore, the switch from PmLCYe expression to PmLCYb expression could cause beta,beta-carotenoid accumulation in both Japanese apricot cultivars.  相似文献   

Association of BoLA‐DRB3 alleles with mastitis resistance and susceptibility in Japanese Holstein cows     

Tatsuyuki YOSHIDA  Hiroki FURUTA  Yasuko KONDO  Harutaka MUKOYAMA 《Animal Science Journal》2012,83(5):359-366

In this study, 714 cows from 26 dairy herds were reclassified as healthy or mastitic cows on the basis of long‐term somatic cell count (SCC) in milk. Cows with more than three consecutive lactation records of SCC from the first or second to fifth lactation, were selected, and their BoLA‐DRB3 (DRB3) alleles were identified using polymerase chain reaction‐restriction fragment length polymorphism (PCR‐RFLP) method. Cows with an SCC of < 200 000 cells/mL in all monthly records were classified as healthy (n = 91). Cows with an SCC of > 300 000 cells/mL in two consecutive tests or four non‐consecutive tests or cows with an SCC of > 500 000 cells/mL in any one test during lactation, regardless of parity, were classified as mastitic (n = 201). Mastitic cows (n = 153) from another 40 herds were considered to be infected if bacteriological testing revealed mastitis pathogens in milk. Their DRB3 alleles were identified using PCR‐sequence‐based typing (PCR‐SBT). The differences in DRB3 allelic frequencies between healthy cows and cows with various degrees of mastitis were re‐investigated. Moreover, the associations of various amino acid motifs in DRB3 alleles with resistance or susceptibility to mastitis pathogens were re‐examined. DRB3.2*8(DRB3*1201) and DRB3.2*16(DRB3*1501) alleles were found to be associated with susceptibility, while DRB3.2*22(DRB3*1101), DRB3.2*23(DRB3*2703), and DRB3.2*24(DRB3*0101) alleles were found to be associated with resistance.  相似文献