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61.
Isomaro Yamaguchi Takao Yokota Shigeo Yoshida Nobutaka Takahashi 《Phytochemistry》1979,18(10):1699-1702
The application of high pressure liquid chromatography to the purification and identification of conjugated gibberellins was examined. Two kinds of reversed phase columns, octadesylsilanized and dimethylsilanized silica gel, were useful for the isolation and identification of gibberellin A3 glucoside and gibberellenic acid glucoside from immature seeds of Quamoclit pennata. 相似文献
62.
Fujioka Shozo; Yamaguchi Isomaro; Murofushi Noboru; Takahashi Nobutaka; Kaihara Sumiko; Takimoto Atsushi 《Plant & cell physiology》1983,24(2):235-239
The occurrence of benzoic acid, a flower-inducing factor inLemna species, in L. paucicostata strains 151, 381, 321 andL. gibba G3 was established by several purification steps andfinal use of gas liquid chromatography-selected ion monitoring.Quantitative analyses of benzoic acid were made in non-floweringand flowering Lemna to determine differences in levels. Theendogenous level of benzoic acid was shown to vary dependingon culture conditions, but no positive correlation could befound between the endogenous level of benzoic acid and floweringof Lemna. (Received October 6, 1982; Accepted December 23, 1982) 相似文献
63.
Preparation and Validation of an Antiserum Specific for Non-Derivatized Gibberellins 总被引:1,自引:0,他引:1
Nakajima Masatoshi; Yamaguchi Isomaro; Kizawa Satoru; Murofushi Noboru; Takahashi Nobutaka 《Plant & cell physiology》1991,32(4):505-501
An antiserum recognizing free gibberellins (GAs) was preparedby immunizing rabbits with a GA4-BSA conjugate. A radioimmunoassay(RIA) and an enzyme-linked immunosorbent assay (ELISA) wereset up using this antiserum. This antiserum showed high cross-reactivityto the so-called active GAs, such as GA1, GA3, GA4 and GA7.The range for measurements of these gibberellins extended from30 fmol to 3 pmol in both RIA and ELISA. Extracts from immature seeds of P. vulgaris were subjected todetermination of GA, by RIA and GC/SIM. The two assays providedsimilar results, indicating the high degree of reliability ofthe immunoassay.
2Present address: Department of Agricultural Chemistry, UtsunomiyaUniversity, Mine-machi 350, Utsunomiya, 321 Japan ( Accepted August 8, 1990) 相似文献
64.
Alteration of Hormone Levels in Transgenic Tobacco Plants
Overexpressing the Rice Homeobox Gene OSH1 总被引:8,自引:0,他引:8 下载免费PDF全文
Shinnosuke Kusaba Yuriko Kano-Murakami Makoto Matsuoka Masanori Tamaoki Tomoaki Sakamoto Isomaro Yamaguchi Masashi Fukumoto 《Plant physiology》1998,116(2):471-476
The rice (Oryza sativa L.) homeobox gene OSH1 causes morphological alterations when ectopically expressed in transgenic rice, Arabidopsis thaliana, and tobacco (Nicotiana tabacum L.) and is therefore believed to function as a morphological regulator gene. To determine the relationship between OSH1 expression and morphological alterations, we analyzed the changes in hormone levels in transgenic tobacco plants exhibiting abnormal morphology. Levels of the plant hormones indole-3-acetic acid, abscisic acid, gibberellin (GA), and cytokinin (zeatin and trans-zeatin [Z]) were measured in leaves of OSH1-transformed and wild-type tobacco. Altered plant morphology was found to correlate with changes in hormone levels. The more severe the alteration in phenotype of transgenic tobacco, the greater were the changes in endogenous hormone levels. Overall, GA1 and GA4 levels decreased and abscisic acid levels increased compared with wild-type plants. Moreover, in the transformants, Z (active form of cytokinin) levels were higher and the ratio of Z to Z riboside (inactive form) also increased. When GA3 was supplied to the shoot apex of transformants, internode extension was restored and normal leaf morphology was also partially restored. However, such GA3-treated plants still exhibited some morphological abnormalities compared with wild-type plants. Based on these data, we propose the hypothesis that OSH1 affects plant hormone metabolism either directly or indirectly and thereby causes changes in plant development. 相似文献
65.
Tomonobu Toyomasu Hiroko Tsuji Hisakazu Yamane Masayoshi Nakayama Isomaro Yamaguchi Noboru Murofushi Nobutaka Takahashi Yasunori Inoue 《Journal of Plant Growth Regulation》1993,12(2):85-90
Gibberellin A1 (GA1), 3-epi-GA1 GA17, GA19, GA20, and GA77 were identified by Kovats retention indices and full-scan mass spectra from gas chromatography-mass spectrometry analysis of a purified extract of mature seeds of photoblastic lettuce (Lactuca sativa L. cv. Grand Rapids). Non-13-hydroxylated GAs such as GA4 and GA9 were not detected even by highly sensitive radioimmunoassay. These results show that the major biosynthetic pathway of GAs in lettuce seeds is the early-13-hydroxylation pathway leading to GA1, which is suggested to be physiologically active in lettuce seed germination. Quantification of endogenous GAs in the lettuce seeds by gas chromatography-selected ion monitoring using deuterated GAs as internal standards indicated that the endogenous level of GA1 increased to a level about three times that of dark control 6 h after a brief red light irradiation, and that far-red light given after red light suppressed the effect of red light. The contents of GA20 and GA19 were not affected by the red light irradiation. Evidence is also presented that 3-epi-GA1 is a native GA in the lettuce seeds. 相似文献
66.
Takao Yokota Satoru Watanabe Yoichi Ogino Isomaro Yamaguchi Nobutaka Takahashi 《Journal of Plant Growth Regulation》1990,9(1-4):151-159
Antiserum against the brassinosteroid (BR), castasterone, was produced by immunizing a rabbit with castasterone-carboxymethoxylamine oxime conjugated with bovine serum albumin (BSA). In a radioimmunoassay (RIA), the antiserum recognized a range of naturally occurring BRs with varying specificities. Detection limits of castasterone and brassinolide were approximately 0.3 pmol. This RIA system was successfully used for analyzing endogenous BRs in seeds and stems ofPhaseolus vulgaris L., and showed that stems are quite different from seeds in terms of the species and quantity of the endogenous BRs. 相似文献
67.
Immunomodulation is a means to modulate an organism's function by antibody production to capture either endogenous or exogenous antigens. We have recently succeeded in obtaining gibberellin (GA)-deficient phenotypes in Arabidopsis thaliana by using anti-bioactive GA antibodies. In this study, a single-chain antibody (scFv) against GA(24), a precursor GA, was utilized to repress the biosynthesis of bioactive gibberellins. Stable accumulation of the scFv in endoplasmic reticulum (ER) was achieved by being produced as a fusion with GFP as well as KDEL ER-retention signal. The transgenic plants showed GFP fluorescence in the reticulate cortical ER network in epidermal cells. The GFP-scFv fusion produced in plants maintained its binding activity. The transgenic plants showed GA-deficient phenotypes, including reduced rosette leaf development, delayed flower induction and reduced stem elongation of the main culm, especially in the early stage of inflorescence growth. Contrarily, stem elongation of the main culm at a later stage, or that of lateral shoots was much less affected by scFv production. These phenotypes were different from anti-bioactive GA scFv-producing lines, whose stem elongation was continuously repressed throughout the inflorescence development. The GA-deficient phenotypes were recovered by treatment with GA(24) and bioactive GA(4), the latter being more effective. The transgenic lines contained conspicuously higher endogenous GA(24) and clearly less GA(4) than wild-type plants. The expression of GA 20-oxidase and GA 3-oxidase genes, which are feedback-regulated by GA signaling, were up-regulated in those plants. These results demonstrate that the scFv trapped GA(24) in ER and inhibited metabolism of GA(24) to bioactive GA(4). 相似文献
68.
Preparation of functional single-chain antibodies against bioactive gibberellins by utilizing randomly mutagenized phage-display libraries 总被引:1,自引:0,他引:1
Suzuki Y Ito S Otsuka K Iwasawa E Nakajima M Yamaguchi I 《Bioscience, biotechnology, and biochemistry》2005,69(3):610-619
Screening randomly mutagenized proteins displayed on a phage surface by biopanning is a powerful strategy to obtain evolved clones with improved properties such as higher stability and functionality. We utilized this method to overcome the problem that functional single-chain antibodies against active gibberellins, a class of plant hormones, can not be prepared by some of the conventional methods. Single-chain antibody libraries with random mutations were constructed from two independent anti-bioactive gibberellin monoclonal antibody lines in a phagemid vector, so that the mutagenized scFvs were expressed in a phage-displayed form upon helper phage infection. From both libraries, scFv clones with binding activity to GA(4) were successfully obtained by successive rounds of biopanning against BSA-GA(4), the original immunogen. The results are highly suggestive that this approach might be a general solution when a single-chain antibody does not show binding activity. We found further that a ribosomal frameshift to complement a nonsense mutation frequently occurred in an amber suppressor strain of E. coli TG1, resulting in the display of a functional antibody, while such a nonsense mutant failed to produce a soluble antibody in a non-amber suppressor strain. This result explains at least partly why single-chain antibodies are sometimes functional only in a phage-displayed form, not in a soluble form. 相似文献
69.
70.
An anti-chlorpropham single-chain variable-fragment (scFv) gene was introduced into Arabidopsis in a manner to express the antibody fragment in each of four different subcellular compartments. The accumulation of scFv in transgenic plants was detected by targeting the fragment in the endoplasmic reticulum or apoplastic space, or by expressing the fragment as a glycosylphosphatidylinositol-anchored protein, while no accumulation could be detected by targeting the fragment in the cytosol. Transgenic plants accumulating the scFv gene at a high level in the endoplasmic reticulum had enhanced tolerance to chlorpropham in comparison with the non-transformants. 相似文献