Kayo Yoshimatsu

National Institute of Biomedical Innovation, Ibaragi, Ōsaka, Japan

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Publications (50)80 Total impact

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    ABSTRACT: The root of Chinese licorice (Glycyrrhiza uralensis) is used worldwide as a medicinal herb. The goal of this study was to understand changes in the concentrations and compositions of four medicinal compounds—glycyrrhizic acid (GL), liquiritin (LQ), liquiritigenin (LG), and isoliquiritigenin (ISLG)—in the root of Chinese licorice during post-harvest treatment. The effects of post-harvest storage temperatures (−80, −30, −13, 4, and 25°C) and drying temperatures (30, 40, 50, and 60°C) on concentrations of the four medicinal compounds were investigated. GL and LQ concentrations in roots stored at −30 and −13°C for 1–2 weeks tended to be 4% to 13% higher than GL and LQ concentrations in roots dried directly in a vacuum freeze dryer (controls). LG concentrations in roots stored at 4°C for 2 weeks were nearly 60-fold higher and ISLG concentrations at 25°C for 1 week were 10-fold higher than LG and ISLG concentrations in the controls. In addition, low temperature (30 and 40°C) drying compared to vacuum freeze drying (controls) increased LG and ISLG concentrations without decreasing GL and LQ concentrations. This study provided an approach to increase the target compound concentrations in Chinese licorice for different market demands (drugs, cosmetics, and food).
    01/2014; 51(4):149-155. DOI:10.2525/ecb.51.149
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    ABSTRACT: The effective clonal propagation by node culture of perilla (Perilla frutescens L. Britton) was investigated. Nodes were plated on solidified 1/2MS medium with BA. The optimum shoot elongation and propagation was obtained at BA 0.05 and 0.1 mg l-1. Cryopreservation using an aluminum cryo-plate was successfully applied to in vitro grown perilla shoot tips. Excised shoot tips from nodes were precultured at 25 °C for 1 day on 1/2MS medium containing 0.3 M sucrose and embedded on an aluminum cryo-plate with alginate gel. Then the cryo-plate with shoot tips was treated with a mixture of 2 M glycerol and 0.6 M sucrose solution for 20 min at 25 °C for osmo-protection. Then the shoot tips on cryo-plate were dehydrated in PVS2 for 20 min at 25 °C and cooled directly by immersion into liquid nitrogen. The recovery growth after cryopreservation was about 80%. This new method (V-Cryo-plate method) has many advantages and will facilitate the cryo-storage of other medicinal plants.
    Advances in Horticultural Science 01/2014; 28(2):79-83.
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    ABSTRACT: Genetically modified (GM) rice (Oryza sativa) lines, such as insecticidal Kefeng and Kemingdao, have been developed and found unauthorised in processed rice products in many countries. Therefore, qualitative detection methods for the GM rice are required for the GM food regulation. A transgenic construct for expressing cowpea (Vigna unguiculata) trypsin inhibitor (CpTI) was detected in some imported processed rice products contaminated with Kemingdao. The 3' terminal sequence of the identified transgenic construct for expression of CpTI included an endoplasmic reticulum retention signal coding sequence (KDEL) and nopaline synthase terminator (T-nos). The sequence was identical to that in a report on Kefeng. A novel construct-specific real-time polymerase chain reaction (PCR) detection method for detecting the junction region sequence between the CpTI-KDEL and T-nos was developed. The imported processed rice products were evaluated for the contamination of the GM rice using the developed construct-specific real-time PCR methods, and detection frequency was compared with five event-specific detection methods. The construct-specific detection methods detected the GM rice at higher frequency than the event-specific detection methods. Therefore, we propose that the construct-specific detection method is a beneficial tool for screening the contamination of GM rice lines, such as Kefeng, in processed rice products for the GM food regulation.
    Food Chemistry 12/2013; 141(3):2618-24. DOI:10.1016/j.foodchem.2013.05.005 · 3.26 Impact Factor
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    ABSTRACT: The in vitro shoot culture of a T-DNA insertional mutant of Papaver somniferum L. established by the infection of Agrobacterium rhizogenes MAFF03-01724 accumulated thebaine instead of morphine as a major opium alkaloid. To develop a non-narcotic opium poppy and to gain insight into its genetic background, we have transplanted this mutant to soil, and analyzed its alkaloid content along with the manner of inheritance of T-DNA insertion loci among its selfed progenies. In the transplanted T0 primary mutant, the opium (latex) was found to be rich in thebaine (16.3% of dried opium) by HPLC analysis. The analyses on T-DNA insertion loci by inverse PCR, adaptor-ligation PCR, and quantitative real-time PCR revealed that as many as 18 copies of T-DNAs were integrated into a poppy genome in a highly complicated manner. The number of copies of T-DNAs was decreased to seven in the selected T3 progenies, in which the average thebaine content was 2.4-fold that of the wild type plant. This may indicate that the high thebaine phenotype was increasingly stabilized as the number of T-DNA copies was decreased. In addition, by reverse transcription PCR analysis on selected morphine biosynthetic genes, the expression of codeine 6-O-demethylase was clearly shown to be diminished in the T0 in vitro shoot culture, which can be considered as one of the key factors of altered alkaloid composition.
    Pharmaceuticals 12/2012; 5(2):133-154. DOI:10.3390/ph5020133
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    ABSTRACT: Developments in the use of genetically modified plants for human and livestock health and phytoremediation were surveyed using information retrieved from Entrez PubMed, Chemical Abstracts Service, Google, congress abstracts and proceedings of related scientific societies, scientific journals, etc. Information obtained was classified into 8 categories according to the research objective and the usage of the transgenic plants as 1: nutraceuticals (functional foods), 2: oral vaccines, 3: edible curatives, 4: vaccine antigens, 5: therapeutic antibodies, 6: curatives, 7: diagnostic agents and reagents, and 8: phytoremediation. In total, 405 cases were collected from 2006 to 2010. The numbers of cases were 120 for nutraceuticals, 65 for oral vaccines, 25 for edible curatives, 36 for vaccine antigens, 36 for therapeutic antibodies, 76 for curatives, 15 for diagnostic agents and reagents, and 40 for phytoremediation (sum of each cases was 413 because some reports were related to several categories). Nutraceuticals, oral vaccines and curatives were predominant. The most frequently used edible crop was rice (51 cases), and tomato (28 cases), lettuce (22 cases), potato (18 cases), corn (15 cases) followed.
    YAKUGAKU ZASSHI 01/2012; 132(5):629-74. DOI:10.1248/yakushi.132.629 · 0.37 Impact Factor
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    ABSTRACT: Coptis japonica (Cj) rhizomes are used as a crude drug for gastroenteritis, since they accumulate antimicrobial berberine. Berberine also shows various useful bioactivities, including cholesterol-lowering activity. Unfortunately, Cj is a slow-growing plant and more than 5 years are required to obtain a crude drug suitable for the Japanese Pharmacopoeia. To improve alkaloid productivity, we overexpressed the 3'-hydroxy-N-methylcoclaurine 4'-O-methyltransferase (4'OMT) gene in Cj. We established the transgenic plant (named CjHE4') by introducing one copy of Cj4'OMT by Agrobacterium-mediated transformation. The successful overexpression of 4'OMT was confirmed in all tissues of CjHE4' by real-time polymerase chain reaction (PCR) analysis. HPLC analysis revealed that the berberine content of CjHE4' leaves and roots cultivated for 4 months was increased to 2.7- and 2.0-fold, respectively, compared with non-transgenic wild-type (CjWT), and these inductions of alkaloids were stable for at least 20 months. Furthermore, in CjHE4' cultivated for 20 months, the berberine content in medicinal parts, stems and rhizomes was significantly increased (1.6-fold). As a consequence, increased amounts of alkaloids in CjHE4' resulted in the improvement of berberine yields (1.5-fold), whereas CjHE4' showed slower growth than CjWT. These results indicated that 4'OMT is one of the key-step enzymes in berberine biosynthesis and is useful for metabolic engineering in Cj.
    Biological & Pharmaceutical Bulletin 01/2012; 35(5):650-9. DOI:10.1248/bpb.35.650 · 1.85 Impact Factor
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    ABSTRACT: Carapichea ipecacuanha produces various emetine-type alkaloids, known as ipecac alkaloids, which have long been used as expectorants, emetics, and amebicides. In this study, we isolated an O-methyltransferase cDNA from this medicinal plant. The encoded protein (CiOMT1) showed 98% sequence identity to IpeOMT2, which catalyzes the 7'-O-methylation of 7'-O-demethylcephaeline to form cephaeline at the penultimate step of emetine biosynthesis (Nomura and Kutchan, J. Biol. Chem., 285, 7722-7738 (2010)). Recombinant CiOMT1 showed both 7'-O-methylation and 6'-O-methylation activities at the last two steps of emetine biosynthesis. This indicates that small differences in amino acid residues are responsible for distinct regional methylation specificities between IpeOMT2 and CiOMT1, and that CiOMT1 might contribute to two sequential O-methylation steps from 7'-O-demethylcephaeline to emetine.
    Bioscience Biotechnology and Biochemistry 01/2011; 75(1):107-13. DOI:10.1271/bbb.100605 · 1.27 Impact Factor
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    ABSTRACT: Illegal cannabis (Cannabis sativa L.) cultivation is still a social problem worldwide. Fifty inquiries on cannabis that Research Center for Medicinal Plant Resources (Tsukuba Division) received between January 1, 2000 and March 31, 2009 were itemized in to 8 categories; 1: seed identification, 2: plant identification, 3: indoor cultivation, 4: outdoor cultivation, 5: germination and growth characteristics, 6: expected amount of cannabis products derived from illegal cannabis plant, 7: non-narcotic cannabis and 8: usage of medicinal cannabis. Top three inquiries were 1: seed identification (16 cases), 3: indoor cultivation (10 cases) and 4: outdoor cultivation (6 cases). Characteristics of cannabis, namely seed morphology, germination and growth characteristics, and distinction from kenaf (Hibiscus cannabinus L.) that is frequently misjudged as cannabis, were studied to contribute for prevention of illegal cannabis cultivation.
    Yakugaku zasshi journal of the Pharmaceutical Society of Japan 02/2010; 130(2):237-46. DOI:10.1248/yakushi.130.237 · 0.31 Impact Factor
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    ABSTRACT: Cannabis plants show a high Delta9-tetrahydrocannabinol content and are used as a psychoactive drug. Therefore the cultivation of hemp and its possession are prohibited by law in Japan. Meanwhile, Cannabis seeds have been used as a component of shichimi-togarashi (a Japanese spice), bird feed, or a crude drug (mashinin). To exclude the possibility of germination, it is officially noticed that hemp seeds must be killed. However, the number of violators has increased in recent years. To judge the ability of seed germination, a germination test is performed. However, the test requires several days and thus has not been used for on-site inspection. In this study, we developed a rapid detection method to determine the ability of Cannabis seeds to germinate using 2,3,5-triphenyl-2H-tetrazolium chloride (TTC). The principle of the assay is as follows. The endogenous respiratory enzymes in hemp seeds convert added colorless TTC into red 1,3,5-triphenylformazan. Consequently, a living embryo is stained red, while red does not appear in the dead seeds. The reaction was active over a pH range of 8.0-9.0, and the optimum activity was found from 40 to 50 degrees C. Under the optimum conditions, we were able to determine the ability of seeds to germinate based on the presence of color within 20 min. Since this method is rapid and simple, it is applicable to on-site inspections. In addition, it could be used as an alternative technique to the germination test, because erroneous decisions is cannot occur under the assay principle.
    Yakugaku zasshi journal of the Pharmaceutical Society of Japan 12/2008; 128(11):1707-11. DOI:10.1248/yakushi.128.1707 · 0.31 Impact Factor
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    Kaori Touno, Kayo Yoshimatsu, Koichiro Shimomura
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    ABSTRACT: Atropa belladonna hairy roots (clone M8) were successfully cryopreserved by using the vitrification method. A. belladonna hairy root tips were precultured on a half strength of Murashige and Skoog (MS) solid medium with 0.1 mg per L 2,4-D or without phytohormone for 1 day, and then dehydrated with PVS2 solution for 15 minutes prior to immersion into liquid nitrogen for 1 day, 1 week, 1 month and 3 months. Hairy root tips kept in liquid nitrogen were rapidly thawed at 36 degree C in a water bath. The root tips were recultured on half strength MS medium. The hairy root tips, precultured with 2,4-D before cryopreservation, showed a higher survival rate than those precultured without phytohormone. The hairy root tips, precultured with 2,4-D, showed an average survival rate of 83 percent. There was no significant difference in the viability of the hairy roots cryopreserved for different periods. The regrowth of cryopreserved hairy roots was similar to that of untreated hairy roots and tropane alkaloid productivity became stable after 4th subculture. PCR analysis of hairy roots demonstrated the conservation of the T-DNA in cryopreserved hairy roots. These results indicate that cryopreservation by vitrification method is useful to preserve A.belladonna hairy root clone M8.
    Cryo letters 01/2006; 27(2):65-72. · 0.64 Impact Factor
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    ABSTRACT: A rapid and reliable solid-phase extraction method for HPLC analysis of opium alkaloids from Papaver plants was established. Fifty mg of dried and powdered plant sample was extracted with 5 ml of 5% acetic acid for 30 min under sonication. After centrifugation, 3 ml of the supernatant was loaded on a reversed-phase cation-exchange solid-phase extraction cartridge. After seriate washings with 0.1 M hydrochloric acid and methanol, alkaloids were eluted with a mixture of 28% ammonia and methanol (1:19). The eluate was concentrated under nitrogen stream at 40 degrees C and the residue was dissolved in 50% aqueous methanol for high performance liquid chromatographic analysis. With this solid-phase extraction method, the recovery of morphine, codeine, oripavine, thebaine, papaverine, noscapine and sanguinarine was from 99.94 to 112.18% when the standard alkaloids were added to the plant samples. Opium alkaloids of a variety of genus Papaver plants cultivated in a field and phytotron were analyzed by this method.
    CHEMICAL & PHARMACEUTICAL BULLETIN 12/2005; 53(11):1446-50. · 1.38 Impact Factor
  • CHEMICAL & PHARMACEUTICAL BULLETIN 01/2005; 53(11):1446-1450. DOI:10.1248/cpb.53.1446 · 1.38 Impact Factor
  • Plant Biotechnology 01/2005; 22(2):113-118. DOI:10.5511/plantbiotechnology.22.113 · 0.94 Impact Factor
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    ABSTRACT: Co-culture conditions for Duboisia myoporoides-D. leichhardtii hybrid hairy root induction were investigated using leaf explants and Agrobacterium rhizogenes ATCC 15834. The bacteria density and duration of co-culture greatly affected the induction rate; the highest rate of 50% was obtained when the leaf explants were co-cultured for 2 d with 10(6) bacteria. One hairy root clone that showed the fastest root growth was selected and used for comparison study with adventitious roots cultured with 0.5 mg/l indole-3-acetic acid (IAA). The hairy roots cultured in Murashige and Skoog (MS) liquid medium grew well and yielded much more tropane alkaloids (35 mg/l scopolamine and 17 mg/l hyoscyamine) than adventitious roots cultured in 0.5 mg/l IAA after 6 weeks of culture at 25 degrees C in the dark. The hairy and adventitious roots (2.5 cm) grown in liquid media were divided into 5 parts (each 0.5 cm) along the root axis. Distribution of scopolamine and IAA was then determined by enzyme-linked immunosorbent assay (ELISA). Inverse relationship between contents of scopolamine and IAA was observed in the hairy roots; increase of scopolamine and decrease of IAA were proportional to the distance from the root meristem. In contrast, the contents of scopolamine and IAA were relatively constant in the adventitious roots. In shoot regeneration experiments, the hairy and adventitious root segments (1 cm) were placed onto 1/2 MS solid medium containing various concentrations of IAA and BA cultured at 25 degrees C under 16 h light. In adventitious roots, the shoots regenerated on media containing 6-benzyladenine (BA) (0.5 to 5 mg/l), and 100% regeneration was observed in medium with 0.1 mg/l IAA and 2 mg/l BA. On the other hand, shoot regeneration was only observed in 33% of hairy roots cultured on medium containing 5 mg/l BA.
    Biological & Pharmaceutical Bulletin 09/2004; 27(8):1261-5. DOI:10.1248/bpb.27.1261 · 1.78 Impact Factor
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    ABSTRACT: Growth characteristics of Cannabis saliva L. are indispensable factors to verify the statements by the criminals of illegal cannabis cultivation. To investigate growth characteristics of C. sativa, two varieties, cannabidiolic acid (CBDA)-rich (CBDA-type) which being cultivated for fiber production and delta9-tetrahydrocannabinolic acid (THCA)-rich (THCA-type) which is used for drug abuse, were cultivated from seeds under the same growth environment in a phytotron. THCA-type showed high germination rate (100%) whereas only 39% of the CBDA-type seeds germinated 6 days after sowing. Plant height, number of true leaves, number of nodes, number of axillary buds and flowering of these two varieties were periodically observed. THCA-type grew more rapidly (plant height: 125.8 cm for THCA-type, 84.7 cm for CBDA-type, 75 days after cultivation) demonstrating vigorous axillary bud formation and earlier male-flowering (63 days for THCA-type, 106 days for CBDA-type, after sowing). Propagation of THCA-type was tested using the axillary shoot cuttings of female plants either with or without the main stem. All the cuttings with the main stem rooted after 21 days and grew healthily in a phytotron. However, all the newly developed leaves were single instead of palmate. In the field, THCA-type male-flowered after 155 days of cultivation after sowing on March 31. The height of the field-cultivated plants reached 260.9 cm 163 days after sowing. Despite the great differences in final plant heights, the increases of plant height per day during the vegetative growth stage were similar in the field and in the phytotron. Thus estimating the starting time of illegal cannabis cultivation might be possible if the plant is in the vegetative growth stage.
    Kokuritsu Iyakuhin Shokuhin Eisei Kenkyūjo hōkoku = Bulletin of National Institute of Health Sciences 02/2004;
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    ABSTRACT: Transformed root cultures of ipecac (Cephaelis ipecacuanha A. Richard), one of the recalcitrant woody plant species for Agrobacterium-mediated transformation, were established by co-culturing of in vitro petiole segments with Agrobacterium rhizogenes ATCC 15 834. Southern blot analysis of the established roots revealed that only the TL-DNA was integrated into the plant genome without incorporation of the TR-DNA. The transformed roots grew slowly on phytohormone-free solid medium and adventitious shoots were regenerated after over 6 months of culture on HF, half-strength Murashige and Skoog (1/2 MS) medium in the dark. The individually separated transformed shoots developed into plantlets on phytohormone-free solid medium at 25 degrees C under 16 h/day light, and the plants demonstrated wider leaves, shorter internodes and vigorous root growth compared to non-transformed plants. Effects of basal media and auxins on the growth and the ipecac alkaloid production of the transformed roots were investigated either under light or in the dark. The roots cultured in the dark grew well in Gamborg B5 (B5) liquid medium containing 0.5 mg/L IBA and yielded 112 mg/L of cephaeline and 14 mg/L emetine after 8 weeks of culture.
    Planta Medica 12/2003; 69(11):1018-23. DOI:10.1055/s-2003-45149 · 2.34 Impact Factor
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    ABSTRACT: The effect of fertilizer on the yield and yield components of opium from Papaver somniferum L.cv.Ikkanshu were investigated from 1993 to 1995. The weight of opium per one capsule was tendency to increase in the order of peat moss > pulverized charcoal > compound fertilizer, but the differences among the results were not significant. Furthermore, no difference in the capsule weight per 100 m2 and the yields of opium and alkaloids per 100 m2 were detected. The significant difference was only observed when the data were compared among the weights and yields of different years. The difference among the yield of opium was mainly due to the weight of opium per one capsule. The weight of opium per one capsule showed a high correlation with capsule husk weight. The alkaloids contents in opium obtained with a different fertilizer application showed no difference. Morphine content at the first lancing was about 11%, and the value decreased with the order of lancing. On the contrary, the codeine and the thebaine content did not change during lancing and the value were 6-7% and 3%, respectively. The papaverine and the noscapine content decreased in the order of lancing.
    Kokuritsu Iyakuhin Shokuhin Eisei Kenkyūjo hōkoku = Bulletin of National Institute of Health Sciences 02/2003;
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    ABSTRACT: Adventitious root cultures of Physochlaina physaloides were established and their growth and tropane alkaloid production were investigated. Although the roots cultured in Murashige-skoog liquid medium containing 1.0-3.0 mg/l IBA grew well, the addition of lower concentrations of auxins was more suitable for alkaloid production. The best alkaloid yield was obtained with 0.01 mg/l NAA in 1/2 MS liquid medium. In addition, the alkaloid differences between the cultured roots and the regenerated plant cultivated in the soil was studied. Cultured roots in this condition showed higher level of alkaloids when compared to leaf, stem and roots from the regenerated plant (3 months in field conditions).
    Kokuritsu Iyakuhin Shokuhin Eisei Kenkyūjo hōkoku = Bulletin of National Institute of Health Sciences 02/2002;
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    ABSTRACT: Adventitious and hairy root cultures of Physochlaina physaloides were established. These roots grew well and produced high amounts of tropane alkaloids (particularly hyoscyamine and 6 beta-hydroxyhyoscyamine) in auxin-free culture medium. The effects of basal media and temperature on the growth and alkaloid production of these roots were investigated. Both root cultures produced highest amount of tropane alkaloids in B5 medium though the optimum temperature for hairy roots were lower than that for adventitious roots.
    Kokuritsu Iyakuhin Shokuhin Eisei Kenkyūjo hōkoku = Bulletin of National Institute of Health Sciences 02/2002;
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    ABSTRACT: An auxin-independent adventitious root culture of Hyoscyamus niger was established, and the roots were successfully cryopreserved with a high regeneration rate of 93.3 percnt; by vitrification method. The root tips were cultured for 12 to 14 days in phytohormone-free Murashige and Skoog (MS) liquid medium, and were excised and precultured on Woody Plant (WP) solid medium supplemented with 0.3 mol/L sucrose at 25 °C in the dark. After 1 day, they were treated with MS-based loading solution for 10 min, followed by soaking in MS-based PVS2 for 10 min at 0 °C. The treated root tips were immersed in liquid nitrogen (-196 °C). For recovery, the root tips were thawed rapidly at 40 °C and washed with MS medium containing 1 mol/L sucrose prior to plating onto WP solid medium. The regenerated roots were evaluated by their growth and tropane alkaloid production. The growth and alkaloid content of regenerated roots analyzed using HPLC were found to be almost the same as those of non-treated roots.
    Journal of Plant Physiology 06/2001; 158(6-158):801-805. DOI:10.1078/0176-1617-00180 · 2.77 Impact Factor

Publication Stats

409 Citations
80.00 Total Impact Points

Institutions

  • 2005–2013
    • National Institute of Biomedical Innovation
      • • Reserach Center for Medicinal Plant Resources
      • • Tsukuba Division
      Ibaragi, Ōsaka, Japan
  • 2001–2004
    • National Institute of Health Sciences, Japan
      Edo, Tōkyō, Japan
  • 2003
    • Toyo University
      Tōkyō, Japan
  • 1998
    • Nagoya City University
      • Faculty of Pharmaceutical Sciences
      Nagoya-shi, Aichi-ken, Japan
  • 1991
    • The Graduate University for Advanced Studies
      Миура, Kanagawa, Japan
  • 1990
    • Nagoya University
      Nagoya, Aichi, Japan