著者

Ryo Nakabayashi Tomoko Nishizawa Tetsuya Mori Hiroshi Sudo Isao Fujii Takashi Asano Kazuki Saito

出版者

Japanese Society for Plant Cell and Molecular Biology

雑誌

Plant Biotechnology (ISSN:13424580)

巻号頁・発行日

pp.19.1002a, (Released:2019-12-18)

参考文献数

6

被引用文献数

8

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Asparaptine, a conjugate of L-arginine and asparagusic acid, was found in green asparagus (Asparagus officinalis) using ultrahigh-resolution metabolomics for sulfur-containing metabolites (S-metabolites), called S-omics. Asparaptine has been shown to inhibit the activity of angiotensin-converting enzyme. Larger amounts of this S-metabolite are therefore required for further analysis; however, there are limitations that asparagus is a perennial plant and its spears, wherein asparaptine accumulates, can be mainly harvested at the spring to summer season. In order to overcome these, we prepared a callus and suspension cell line from green asparagus. Untargeted metabolome analysis using liquid chromatography-tandem mass spectrometry was performed in the materials as well as spears and three calluses derived from wild type Asparagus. The analysis demonstrated that the amount of asparaptine in the callus derived from the green asparagus was more than the others per mg dry weight. The suspension cell line treated with methyljasmonate showed the induction of asparaptine, suggesting that the asparaptine production is modifiable under appropriate culture conditions. The described materials can be utilized for the production of asparaptine and in integrated metabolomics to study the biosynthesis of this S-metabolite, which is currently unknown.

著者

Naoki Yamamoto Hiroyuki Kajiura Shinya Takeno Nobuaki Suzuki Yoshihisa Nakazawa

出版者

Japanese Society for Plant Cell and Molecular Biology

雑誌

Plant Biotechnology (ISSN:13424580)

巻号頁・発行日

pp.14.0609b, (Released:2014-08-23)

参考文献数

33

被引用文献数

1 3

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We established a DNA watermarking system for discriminating transgenic plants. The system contains an encryption algorithm based on a binary system, genetic transformation and a detection algorithm for encrypted DNA watermark sequences using a DNA dot plot. The encryption algorithm converted character strings into nucleic acid sequences through binary digits, and the sequence was designed to be resistant to transition mutations to decipher codes completely. Moreover, the encrypted sequences were capable of taking specific nucleotide sequences in using the algorithmic redundancy of the corresponding DNA. Genetic transformation enables labeling plant genomes with DNA watermarks. The detection algorithm allows finding traces of sequence changes in DNA watermarks, complementing the error protection function of the encryption algorithm. To validate the effectiveness of our DNA watermarking system, we introduced a DNA watermark to the tobacco genome and detected the DNA watermark in PCR products amplified from the genome. This indicates that DNA watermark technology is useful for introducing artificial genetic markers in plant organisms, in particular when several transgenic host plants and transgenes are used. The source codes of the Perl scripts are available in this report.

著者

Yuki Kanesaka Masaaki Okada Shogo Ito Tokitaka Oyama

出版者

Japanese Society for Plant Cell and Molecular Biology

雑誌

Plant Biotechnology (ISSN:13424580)

巻号頁・発行日

pp.19.0531a, (Released:2019-09-07)

参考文献数

27

被引用文献数

7

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The rapid assessment of gene function is crucial in biological research. The CRISPR/Cas9 system is widely used as a tool for targeted gene editing in many organisms including plants. Previously, we established a transient gene expression system for investigating cellular circadian rhythms in duckweed. In this system, circadian reporters and clock gene effectors—such as overexpressors, RNA interference (RNAi), and CRISPR/Cas9—were introduced into duckweed cells using a particle bombardment method. In the present study, we applied the CRISPR/Cas9 system at a single cell level to Arabidopsis thaliana, a model organism in plant biology. To evaluate the mutation induction efficiency of the system, we monitored single-cell bioluminescence after application of the CRISPR/Cas9 system targeting the ELF3 gene, which is essential for robust circadian rhythmicity. We evaluated the mutation induction efficiency by determining the proportion of cells with impaired circadian rhythms. Three single guide RNAs (sgRNAs) were designed, and the proportion of arrhythmic cells following their use ranged from 32 to 91%. A comparison of the mutation induction efficiencies of diploid and tetraploid Arabidopsis suggested that endoreduplication had a slight effect on efficiency. Taken together, our results demonstrate that the transiently introduced CRISPR/Cas9 system is useful for rapidly assessing the physiological function of target genes in Arabidopsis cells.

著者

Sumire Fujiwara Keiko Kigoshi Nobutaka Mitsuda Kaoru Suzuki Masaru Ohme-Takagi

出版者

Japanese Society for Plant Cell and Molecular Biology

雑誌

Plant Biotechnology (ISSN:13424580)

巻号頁・発行日

pp.14.0121a, (Released:2014-03-20)

参考文献数

58

被引用文献数

5 7

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Proper gene expression regulated by transcription factors is essential for plants to achieve proper growth and development. However, the biological functions of many transcription factors remain largely unknown. Furthermore, although there are transcription factors which possess a plant-specific repression domain(s), their biological functions and whether such transcription factors function as transcriptional repressors are unclear. Thus, aiming for searching clues to understand their functions, we generated transgenic plants in which a putative transcriptional repressor fused with a VP16 viral trans-activation domain was expressed constitutively. Several plants with strong morphological phenotypes such as leaf and flower development defects were isolated from those lines expressing potential transcriptional repressors with unknown functions, giving the clue to reveal the yet-to-be analyzed functions of each protein. Reversal of function of the well-known transcriptional and floral repressor SHORT VEGETATIVE PHASE by VP16 fusion was observed, exemplifying successful functional reversion by this system. Plants constitutively expressing VP16 fused WUSCHEL, which is known to function both as a transcriptional activator and repressor, showed both phenotypes reported in its overexpression and loss-of-function lines. Taken together, our data provide examples showing the efficacy of VP16 fusion to provide helpful information to uncover the unknown functions of potential transcriptional repressors. This technique could also be effective to produce “super plants” which obtained strong and useful traits for application by strongly activating genes which are usually silent.

著者

藤本 忠明 京 正晴 宮内 由起夫 真山 滋志

出版者

Japanese Society for Plant Cell and Molecular Biology

雑誌

植物組織培養 (ISSN:02895773)

巻号頁・発行日

vol.7, no.3, pp.177-180, 1990 (Released:2010-04-30)

参考文献数

14

被引用文献数

1 2

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マリーゴールド (Tagetes patula L.) で確立された組織培養系カルスのヘキサン抽出物中に高い殺線虫活性が検出された. 異なるホルモン条件下で誘導, 継代されたカルス系統間で, その殺線虫活性は大きく異なった. 最も高い活性は0.1ppmのNAAを含むMS寒天培地上で誘導, 継代された緑色カルスにおいて見られた. その殺線虫活性およびα-ターチオフェン含量は試験管内栽培された植物体の根においてみられるものに匹敵した. いくつかの異なる系統のカルスのヘキサン抽出物を, HPLC分析した結果, 殺線虫活性はおもにα-ターチオフェン含量に相関するものの, α-ターチオフェン以外の殺線虫物質の存在をも示唆した.

著者

Anung Wahyudi Dinni Ariyani Gang Ma Ryosuke Inaba Chikako Fukasawa Ryohei Nakano Reiko Motohashi

出版者

Japanese Society for Plant Cell and Molecular Biology

雑誌

Plant Biotechnology (ISSN:13424580)

巻号頁・発行日

vol.35, no.4, pp.303-312, 2018-12-25 (Released:2018-12-31)

参考文献数

29

被引用文献数

10

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In this study, two temperature-induced lipocalin genes SlTIL1 and SlTIL2, and a chloroplastic lipocalin gene SlCHL were isolated from ‘Micro-Tom’ tomato. The coding sequences of SlTIL1, SlTIL2 and SlCHL were 558, 558, and 1002 bp, respectively. By TargetP analysis, no characteristic transit peptides were predicted in the proteins of SlTIL1 and SlTIL2, while a chloroplastic transit peptide was predicted in the protein of SlCHL. The subcellular localization results indicated that SlTIL1 and SlTIL2 proteins were major localized in the plasma membrane, while SlCHL was localized in chloroplast. To understand the function of lipocalins, transgenic tomato over-expressed SlTIL1, SlTIL2 and SlCHL and their virus-induced gene silencing (VIGS) plants were generated. The phenotypes were significantly affected when the SlTIL1, SlTIL2 and SlCHL were over-expressed or silenced by VIGS, which suggested that the three lipocalins played important roles in regulating the growth and development of tomato. In addition, the level of ROS (O2− and H2O2) was low in SlTIL1, SlTIL2 and SlCHL over-expressed plants, while it was high in their silenced plants. The changes in the expression of SODs were consistent with the accumulations of ROS, which indicated that lipocalins might have an important role in abiotic oxidative stress tolerance in tomato plants. Especially SlTIL1 and SlTIL2 are localized around their membranes and protect them from ROS. The results will contribute to elucidating the functions of lipocalin in plants, and provide new strategies to improve the tolerance to abiotic stress in tomato plants.

著者

Hiroshi SHINJI Minoru OKADA

出版者

Japanese Society for Plant Cell and Molecular Biology

雑誌

Plant tissue culture letters (ISSN:02895773)

巻号頁・発行日

vol.8, no.2, pp.129-132, 1991 (Released:2010-04-30)

参考文献数

8

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Time course of the growth of cultured Pinellia ternata plants was investigated.Corm-like bodies were cultured in a MS liquid medium containing 0.25mg/l IAA and 0.5mg/l BA by shaking. On the 13th, 27th, 41th, and 54th day, the dry weight of each organ-corm-like body, leaf blade, petiole, root-was measured. From these data, growth parameters and distribution rate of dry matter in each organ were calculated.Relative growth rate was at its maximum at the beginning of culture and then decreased gradually. The growth rate in the dry matter of the whole plant was at its maximum between the 13th-27th day.The time course of the distribution rate of dry matter in each organ indicates that during the first half of the culture period, mainly leaf blades and petioles grow and during the latter half, mainly corm-like bodies grow.Judging from the weight of corm-like bodies obtained for a fixed period and the cost in gaining corm-like bodies, the best culture period was 41 days.

著者

Calvino Martin Kamada Hiroshi Mizoguchi Tsuyoshi

出版者

Japanese Society for Plant Cell and Molecular Biology

雑誌

Plant biotechnology (ISSN:13424580)

巻号頁・発行日

vol.22, no.3, pp.179-183, 2005-09-01

参考文献数

42

被引用文献数

9 4

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Plants in the genus <i>Arabidopsis</i> are facultative LD plants that flower much earlier under LD conditions than SD regimens, with the photoperiod (or LD) pathway contributing to floral acceleration. <i>LHY</i> and <i>CCA1</i> genes, among other factors, have central roles in the circadian clock of <i>Arabidopsis</i>, which plays a key role in measuring day length. <i>GI</i> gene mediates the circadian clock and floral activator genes, <i>CO</i> and <i>FT</i>, to control photoperiodic flowering. <i>GI</i> is required to set the peak phase of <i>CO</i> expression at the end of the light period under LD conditions, so that the <i>CO</i> protein is stabilized and activated by light to increase <i>FT</i> expression. However, recent studies have demonstrated that the role of SDs is not solely to switch off CO activity. For example, GI interacts with SPY, a negative regulator of the GA signal. The flowering times of <i>gi</i> mutants were still significantly later under SD conditions than LD regimes, which suggests that <i>GI</i> has a potential role in accelerating the start of flowering, even under SDs. Over-expression of either <i>FT</i> or <i>TSF</i> genes caused early flowering, and the acceleration of flowering was enhanced under SDs, suggesting that SDs have an additional role to that in the <i>LHY</i>/<i>CCA1</i>-<i>GI</i>-<i>CO</i>-<i>FT</i> pathway. In this short review, we discuss the hidden roles of SDs in controlling flowering based on recent studies of the molecular genetics of flowering time in <i>Arabidopsis</i>.

著者

古野 哲郎 神山 亜紀 明石 智義 臼井 真理子 高橋 武美 綾部 真一

出版者

Japanese Society for Plant Cell and Molecular Biology

雑誌

植物組織培養 (ISSN:02895773)

巻号頁・発行日

vol.10, no.3, pp.275-280, 1993

被引用文献数

10

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西洋タンポポのカルス培養細胞から完全な植物体を再生させた. カルス細胞をNAAとBAを添加した1/2MS培地上明所で培養すると, 一部の培地上で著しいシュートの形成が見られた. ホルモン無添加培地で発根させ, バーミキュライトを経てポット中の土壌に移植したところ, 開花し, 種子を得る事ができた. カルス培養ではトリテルペン酸 (オレアノール酸, ウルソール酸) が顕著に検出されたが, 再分化すると検出されなくなり, 代わりに分化器官ではトリテルペン-3-オール量が増加した. トリテルペン-3-オールの組成をHPLCで解析したところ, カルスではα-およびβ-アミリンが主要な成分であるのに対して, 分化器官ではタラキサステロール, ルペオールなどがさらに見出され, 特に乳液ではタラキサステロールが主成分であった.

著者

大菅 康一 鎌田 博 駒嶺 穆

出版者

Japanese Society for Plant Cell and Molecular Biology

雑誌

植物組織培養 (ISSN:02895773)

巻号頁・発行日

vol.10, no.2, pp.180-183, 1993

被引用文献数

8 19

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ニンジンを対象に不定胚分化に与える細胞密度の影響を定量的に検討した. 2,000個/m<i>l</i>の細胞塊密度で Embryogenic な細胞塊から球状胚及び心臓型胚を誘導し, さらに球状胚を150個/m<i>l</i>以下の密度で培養すると5日後に80~90%の高頻度で魚雷型胚が得られた. 細胞密度は高頻度同調的な不定胚分化に極めて重要なことが明らかになった.