Plant Biotechnology最新文献

{"title":"Selection of a histidine auxotrophic <i>Marchantia polymorpha</i> strain with an auxotrophic selective marker.","authors":"Tatsushi Fukushima,&nbsp;Yutaka Kodama","doi":"10.5511/plantbiotechnology.22.0810a","DOIUrl":"https://doi.org/10.5511/plantbiotechnology.22.0810a","url":null,"abstract":"<p><p><i>Marchantia polymorpha</i> has emerged as a model liverwort species, with molecular tools increasingly available. In the present study, we developed an auxotrophic strain of <i>M. polymorpha</i> and an auxotrophic selective marker gene as new experimental tools for this valuable model system. Using CRISPR (clustered regularly interspaced palindromic repeats)/Cas9-mediated genome editing, we mutated the genomic region for <i>IMIDAZOLEGLYCEROL-PHOSPHATE DEHYDRATASE</i> (<i>IGPD</i>) in <i>M. polymorpha</i> to disrupt the biosynthesis of histidine (<i>igpd</i>). We modified an <i>IGPD</i> gene (<i>IGPDm</i>) with silent mutations, generating a histidine auxotrophic selective marker gene that was not a target of our CRISPR/Cas9-mediated genome editing. The <i>M. polymorpha</i> <i>igpd</i> mutant was a histidine auxotrophic strain, growing only on medium containing histidine. The <i>igpd</i> mutant could be complemented by transformation with the <i>IGPDm</i> gene, indicating that this gene could be used as an auxotrophic selective marker. Using the <i>IGPDm</i> marker in the <i>igpd</i> mutant background, we produced transgenic lines without the need for antibiotic selection. The histidine auxotrophic strain <i>igpd</i> and auxotrophic selective marker <i>IGPDm</i> represent new molecular tools for <i>M. polymorpha</i> research.</p>","PeriodicalId":20411,"journal":{"name":"Plant Biotechnology","volume":null,"pages":null},"PeriodicalIF":1.6,"publicationDate":"2022-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10240916/pdf/plantbiotechnology-39-4-22.0810a.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9946703","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comprehensive identification of terpene synthase genes and organ-dependent accumulation of terpenoid volatiles in a traditional medicinal plant <i>Angelica archangelica</i> L.","authors":"Miki Suenaga-Hiromori,&nbsp;Daisuke Mogi,&nbsp;Yohei Kikuchi,&nbsp;Jiali Tong,&nbsp;Naotsugu Kurisu,&nbsp;Yuichi Aoki,&nbsp;Hiroyuki Amano,&nbsp;Masahiro Furutani,&nbsp;Takefumi Shimoyama,&nbsp;Toshiyuki Waki,&nbsp;Toru Nakayama,&nbsp;Seiji Takahashi","doi":"10.5511/plantbiotechnology.22.1006a","DOIUrl":"https://doi.org/10.5511/plantbiotechnology.22.1006a","url":null,"abstract":"<p><p><i>Angelica archangelica</i> L. is a traditional medicinal plant of Nordic origin that produces an unusual amount and variety of terpenoids. The unique terpenoid composition of <i>A. archangelica</i> likely arises from the involvement of terpene synthases (TPSs) with different specificities, none of which has been identified. As the first step in identifying TPSs responsible for terpenoid chemodiversity in <i>A. archangelica</i>, we produced a transcriptome catalogue using the mRNAs extracted from the leaves, tap roots, and dry seeds of the plant; 11 putative TPS genes were identified (<i>AaTPS1-AaTPS11</i>). Phylogenetic analysis predicted that AaTPS1-AaTPS5, AaTPS6-AaTPS10, and AaTPS11 belong to the monoterpene synthase (monoTPS), sesquiterpene synthase (sesquiTPS), and diterpene synthase clusters, respectively. We then performed in vivo enzyme assays of the AaTPSs using recombinant <i>Escherichia coli</i> systems to examine their enzymatic activities and specificities. Nine recombinant enzymes (AaTPS2-AaTPS10) displayed TPS activities with specificities consistent with their phylogenetics; however, AaTPS5 exhibited a strong sesquiTPS activity along with a weak monoTPS activity. We also analyzed terpenoid volatiles in the flowers, immature and mature seeds, leaves, and tap roots of <i>A. archangelica</i> using gas chromatography-mass spectrometry; 14 monoterpenoids and 13 sesquiterpenoids were identified. The mature seeds accumulated the highest levels of monoterpenoids, with β-phellandrene being the most prominent. α-Pinene and β-myrcene were abundant in all organs examined. The in vivo assay results suggest that the AaTPSs functionally identified in this study are at least partly involved in the chemodiversity of terpenoid volatiles in <i>A. archangelica</i>.</p>","PeriodicalId":20411,"journal":{"name":"Plant Biotechnology","volume":null,"pages":null},"PeriodicalIF":1.6,"publicationDate":"2022-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10240917/pdf/plantbiotechnology-39-4-22.1006a.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9963415","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"RNA-seq study reveals the signaling and carbohydrate metabolism regulators involved in dormancy release by warm stratification in <i>Paris polyphylla</i> var. <i>yunnanensis</i>.","authors":"Bin Yang,&nbsp;Shan Sun,&nbsp;Shengyu Li,&nbsp;Jiali Zeng,&nbsp;Furong Xu","doi":"10.5511/plantbiotechnology.22.0824a","DOIUrl":"https://doi.org/10.5511/plantbiotechnology.22.0824a","url":null,"abstract":"<p><p>Long-term seed dormancy of <i>Paris polyphylla</i> var. <i>yunnanensis</i> limits its large-scale artificial cultivation. It is crucial to understand the regulatory genes involving in dormancy release for artificial cultivation in this species. In this study, seed dormancy of <i>Paris polyphylla</i> var. <i>yunnanensis</i> was effectively released by warm stratification (20°C) for 90 days. The freshly harvested seeds (dormant) and stratified seeds (non-dormant) were used to sequence, and approximately 147 million clean reads and 28,083 annotated unigenes were detected. In which, a total of 10,937 differentially expressed genes (DEGs) were identified between dormant and non-dormant seeds. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) classification revealed that the majority unigenes involved in signaling transduction and carbohydrate metabolism. Of them, the signaling transduction-related DEGs were mainly hormones-, reactive oxygen species (ROS)-, and transcription factor (TF)-related genes. The largest number of signaling transduction-related DEGs were auxin-responsive genes (<i>SAUR</i>, <i>AUX</i>/<i>IAA</i>, and <i>ARF</i>) and AP2-like ethylene-responsive transcription factor (ERF/AP2). Moreover, at least 29 DEGs such as α-amylase (<i>AMY</i>), β-glucosidase (<i>Bglb/Bglu/Bglx</i>), and endoglucanase (<i>Glu</i>) were identified involving in carbohydrate metabolism. These identified genes provide a valuable resource to investigate the molecular basis of dormancy release in <i>Paris polyphylla</i> var. <i>yunnanensis</i>.</p>","PeriodicalId":20411,"journal":{"name":"Plant Biotechnology","volume":null,"pages":null},"PeriodicalIF":1.6,"publicationDate":"2022-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10240920/pdf/plantbiotechnology-39-4-22.0824a.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9963418","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization of γ-glutamyltransferase- and phytochelatin synthase-mediated catabolism of glutathione and glutathione <i>S</i>-conjugates in <i>Arabidopsis thaliana</i>.","authors":"Ryota Inoue,&nbsp;Naoto Nakamura,&nbsp;Chie Matsumoto,&nbsp;Hisabumi Takase,&nbsp;Jiro Sekiya,&nbsp;Rafael Prieto","doi":"10.5511/plantbiotechnology.22.1003a","DOIUrl":"https://doi.org/10.5511/plantbiotechnology.22.1003a","url":null,"abstract":"<p><p>Glutathione (GSH, γ-L-glutamyl-L-cysteinyl-glycine) has been implicated in a multitude of cellular functions, such as protection of cells against oxidative stress, detoxification of xenobiotics via degradation of GSH <i>S</i>-conjugates, and disease resistance. Glutathione also serves as a precursor of phytochelatins, and thereby plays an essential role in heavy metal detoxification. The <i>Arabidopsis</i> genome encodes three functional γ-glutamyltransferase genes (<i>AtGGT1, AtGGT2, AtGGT4</i>) and two phytochelatin synthase genes (<i>AtPCS1</i>, <i>AtPCS2</i>). The function of plant GGT has not yet been clearly defined, although it is thought to be involved in GSH and GSH <i>S</i>-conjugate catabolism. On the other hand, besides its role in heavy metal detoxification, PCS has also been involved in GSH <i>S</i>-conjugate catabolism. Herein we describe the HPLC characterization of GSH and GSH <i>S</i>-conjugate catabolism in <i>Arabidopsis</i> mutants deficient in GSH biosynthesis (<i>pad2-1/gsh1</i>), <i>atggt</i> and <i>atpcs1</i> T-DNA insertion mutants, <i>atggt pad2-1</i>, <i>atggt atpcs1</i> double mutants, and the <i>atggt1 atggt4 atpcs1</i> triple mutant. The results of our HPLC analysis confirm that AtGGT and AtPCS play important roles in two different pathways related with GSH and GSH <i>S</i>-conjugate (GS-bimane) catabolism in <i>Arabidopsis</i>.</p>","PeriodicalId":20411,"journal":{"name":"Plant Biotechnology","volume":null,"pages":null},"PeriodicalIF":1.6,"publicationDate":"2022-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10240914/pdf/plantbiotechnology-39-4-22.1003a.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9589417","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The essential role of the quasi-long terminal repeat sequence for replication and gene expression of an endogenous pararetrovirus, petunia vein clearing virus.","authors":"Kazunori Kuriyama,&nbsp;Midori Tabara,&nbsp;Hiromitsu Moriyama,&nbsp;Hideki Takahashi,&nbsp;Toshiyuki Fukuhara","doi":"10.5511/plantbiotechnology.22.1017a","DOIUrl":"https://doi.org/10.5511/plantbiotechnology.22.1017a","url":null,"abstract":"<p><p>Petunia vein clearing virus (PVCV) is a type member of the genus <i>Petuvirus</i> within the <i>Caulimoviridae</i> family and is defined as one viral unit consisting of a single open reading frame (ORF) encoding a viral polyprotein and one quasi-long terminal repeat (QTR) sequence. Since some full-length PVCV sequences are found in the petunia genome and a vector for horizontal transmission of PVCV has not been identified yet, PVCV is referred to as an endogenous pararetrovirus. Molecular mechanisms of replication, gene expression and horizontal transmission of endogenous pararetroviruses in plants are elusive. In this study, agroinfiltration experiments using various PVCV infectious clones indicated that the replication (episomal DNA synthesis) and gene expression of PVCV were efficient when the QTR sequences are present on both sides of the ORF. Whereas replacement of the QTR with another promoter and/or terminator is possible for gene expression, it is essential for QTR sequences to be on both sides for viral replication. Although horizontal transmission of PVCV by grafting and biolistic inoculation was previously reported, agroinfiltration is a useful and convenient method for studying its replication and gene expression.</p>","PeriodicalId":20411,"journal":{"name":"Plant Biotechnology","volume":null,"pages":null},"PeriodicalIF":1.6,"publicationDate":"2022-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10240922/pdf/plantbiotechnology-39-4-22.1017a.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9582617","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Recombinant MBP-pσ1 expressed in soybean seeds delays onset and reduces developing disease in an animal model of multiple sclerosis.","authors":"Linda M Robles,&nbsp;Laura H Reichenberg,&nbsp;James H Grissom Ⅲ,&nbsp;Richard J Chi,&nbsp;Kenneth J Piller","doi":"10.5511/plantbiotechnology.22.0926a","DOIUrl":"https://doi.org/10.5511/plantbiotechnology.22.0926a","url":null,"abstract":"<p><p>It is estimated that multiple sclerosis (MS) affects over 2.8 million people worldwide, with a prevalence that is expected to continue growing over time. Unfortunately, there is no cure for this autoimmune disease. For several decades, antigen-specific treatments have been used in animal models of experimental autoimmune encephalomyelitis (EAE) to demonstrate their potential for suppressing autoimmune responses. Successes with preventing and limiting ongoing MS disease have been documented using a wide variety of myelin proteins, peptides, autoantigen-conjugates, and mimics when administered by various routes. While those successes were not translatable in the clinic, we have learned a great deal about the roadblocks and hurdles that must be addressed if such therapies are to be useful. Reovirus sigma1 protein (pσ1) is an attachment protein that allows the virus to target M cells with high affinity. Previous studies showed that autoantigens tethered to pσ1 delivered potent tolerogenic signals and diminished autoimmunity following therapeutic intervention. In this proof-of-concept study, we expressed a model multi-epitope autoantigen (human myelin basic protein, MBP) fused to pσ1 in soybean seeds. The expression of chimeric MBP-pσ1 was stable over multiple generations and formed the necessary multimeric structures required for binding to target cells. When administered to SJL mice prophylactically as an oral therapeutic, soymilk formulations containing MBP-pσ1 delayed the onset of clinical EAE and significantly reduced developing disease. These results demonstrate the practicality of soybean as a host for producing and formulating immune-modulating therapies to treat autoimmune diseases.</p>","PeriodicalId":20411,"journal":{"name":"Plant Biotechnology","volume":null,"pages":null},"PeriodicalIF":1.6,"publicationDate":"2022-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10240915/pdf/plantbiotechnology-39-4-22.0926a.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9963419","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The influences of fungal endophytes inoculation on the biochemical status of grape cells of different varieties in vitro.","authors":"Xiaoxia Pan,&nbsp;Tong Li,&nbsp;Changmei Liao,&nbsp;Youyong Zhu,&nbsp;Mingzhi Yang","doi":"10.5511/plantbiotechnology.22.0730a","DOIUrl":"https://doi.org/10.5511/plantbiotechnology.22.0730a","url":null,"abstract":"<p><p>The metabolic patterns of grape cells can be specifically shaped by different strains of dual-cultured fungal endophytes. In this work, a solid co-culture system was furtherly proposed to illustrate the different impacts of endophytic fungi on the biochemical status of grape cells of different varieties. By measuring the metabolic impacts of contact fungal endophytes on grape cells of the varieties 'Rose honey' (RH) and 'Cabernet sauvignon' (CS), we observed that most of the fungal strains used had promoting effects on grape cellular biochemistry parameters. Compared with the control, inoculation with most of the fungal strains increased the superoxide dismutase (SOD) and phenylalanine ammonia-lyase (PAL) activities as well as the total flavonoid (TF) and total phenolics (TPh) contents in both types of grape cells. Among the tested strains, RH34, RH49 and MDR36 had relatively stronger biochemical impacts on grape cells. More interestingly, in addition to the varietal specificity, a certain degree of fungal genus specificity was also observed during the metabolic interactions between fungal endophytes and grape cells, as fungal endophytes from the same genus tended to be clustered into the same group based on the affected biochemical traits. This work revealed the differential biochemical status effects of fungal endophytes on different varietal grape cells and raised the possibility of reshaping grape qualities by applying endophytes.</p>","PeriodicalId":20411,"journal":{"name":"Plant Biotechnology","volume":null,"pages":null},"PeriodicalIF":1.6,"publicationDate":"2022-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10240921/pdf/plantbiotechnology-39-4-22.0730a.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9963412","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Measurement of reactive oxygen species production by luminol-based assay in <i>Nicotiana benthamiana</i>, <i>Arabidopsis thaliana</i> and <i>Brassica rapa</i> ssp. <i>rapa</i>.","authors":"Lalita Jantean,&nbsp;Kentaro Okada,&nbsp;Yaichi Kawakatsu,&nbsp;Ken-Ichi Kurotani,&nbsp;Michitaka Notaguchi","doi":"10.5511/plantbiotechnology.22.0823a","DOIUrl":"https://doi.org/10.5511/plantbiotechnology.22.0823a","url":null,"abstract":"<p><p>Reactive oxygen species (ROS) are critical for plant biological processes. As signaling molecules, ROS regulate plant growth and development through cell expansion, elongation, and programmed cell death. Furthermore, ROS production is induced by microbe-associated molecular patterns (MAMPs) treatment and biotic stresses, and contributes to plant resistance to pathogens. Thus, MAMP-induced ROS production has been an indicator for plant early immune responses or stress responses. One of widely used methods for the measurement is a luminol-based assay to measure extracellular ROS production with a bacterial flagellin epitope (flg22) as a MAMP elicitor. <i>Nicotiana benthamiana</i> is susceptible to a wide variety of plant pathogenic agents and therefore commonly used for ROS measurements. On the other hand, <i>Arabidopsis thaliana</i>, many of genetical lines of which are available, is also conducted to ROS measurements. Tests in an asterid <i>N. benthamiana</i> and a rosid <i>A. thaliana</i> can reveal conserved molecular mechanisms in ROS production. However, the small size of <i>A. thaliana</i> leaves requires many seedlings for experiments. This study examined flg22-induced ROS production in another member of the Brassicaceae family, <i>Brassica rapa</i> ssp. <i>rapa</i> (turnip), which has large and flat leaves. Our experiments indicated that 10 nM and 100 nM flg22 treatments induced high ROS levels in turnip. Turnip tended to have a lower standard deviation in multiple concentrations of flg22 treatment. Therefore, these results suggested that turnip can be a good material from the rosid clade for ROS measurement.</p>","PeriodicalId":20411,"journal":{"name":"Plant Biotechnology","volume":null,"pages":null},"PeriodicalIF":1.6,"publicationDate":"2022-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10240919/pdf/plantbiotechnology-39-4-22.0823a.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9963413","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"High-transcriptional activation ability of bamboo SECONDARY WALL NAC transcription factors is derived from C-terminal domain.","authors":"Shingo Sakamoto,&nbsp;Taiji Nomura,&nbsp;Yasuo Kato,&nbsp;Shinjiro Ogita,&nbsp;Nobutaka Mitsuda","doi":"10.5511/plantbiotechnology.22.0501a","DOIUrl":"https://doi.org/10.5511/plantbiotechnology.22.0501a","url":null,"abstract":"<p><p>The secondary cell wall, which is mainly composed of cellulose, hemicellulose, and lignin, constitutes woody tissues and gives physical strength and hydrophobic properties for resistance against environmental stresses. We cloned and functionally analyzed the homologous transcription factor (TF) genes of SECONDARY WALL NAC (SWN) proteins from Hachiku bamboo (<i>Phyllostachys nigra</i>; <i>PnSWN</i>s). An RT-PCR analysis showed that <i>PnSWN</i>s are expressed in young tissues in bamboo. Their transcriptional activation activities were higher than that of the <i>Arabidopsis</i> NAC SECONDARY WALL THICKENING PROMOTING FACTOR 3 (NST3) TF, which was equivalent to SWN TFs in monocot. PnSWNs preferred to activate the genes related to secondary cell wall formation but not the genes related to programmed cell death. When <i>PnSWN</i>s were expressed in <i>Arabidopsis</i>, they highly induced secondary cell wall formation, like previously-shown rice SWN1. Dissection analysis revealed that this high activity largely depends on C-terminal domain. These results demonstrate that the cloned bamboo SWNs function as regulators of secondary cell wall formation with strong activation ability derived from C-terminal domain, and could be served as new genetic tools for secondary cell wall manipulation.</p>","PeriodicalId":20411,"journal":{"name":"Plant Biotechnology","volume":null,"pages":null},"PeriodicalIF":1.6,"publicationDate":"2022-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9592943/pdf/plantbiotechnology-39-3-22.0501a.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40673513","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Establishment of an efficient transformation method of garden stock (<i>Matthiola incana</i>) using a callus formation chemical inducer.","authors":"Yoshiki Tanahara,&nbsp;Kaho Yamanaka,&nbsp;Kentaro Kawai,&nbsp;Yukiko Ando,&nbsp;Takashi Nakatsuka","doi":"10.5511/plantbiotechnology.22.0602a","DOIUrl":"https://doi.org/10.5511/plantbiotechnology.22.0602a","url":null,"abstract":"<p><p><i>Matthiola incana</i> is an important floricultural plant that blooms from winter to spring, and had been desired to be established a transformation system. This study successfully obtained stable transgenic plants from <i>M. incana</i>. We used <i>Agrobacterium tumefaciens</i> harboring a binary vector containing the β-glucuronidase gene (<i>GUS</i>) under the control of cauliflower mosaic virus 35S promoter to evaluate the transformation frequency of <i>M. incana</i>. We observed that cocultivation with the <i>A. tumefaciens</i> strain GV3101 for 5 days effectively enhanced the infection frequency, assessed through a transient <i>GUS</i> expression area in the seedling. Furthermore, the addition of 100 µM acetosyringone was necessary for <i>Agrobacterium</i> infection. However, we could not obtain transgenic plants on a shoot formation medium supplemented with 1 mg l^-1 6-benzyladenine (BA). For callus formation from the leaf sections, a medium supplemented with 1-50 µM fipexide (FPX), a novel callus induction chemical, was employed. Then, the callus formation was observed after 2 weeks, and an earlier response was detected than that in the BA medium (4-6 weeks). Results also showed that cultivation in a selection medium supplemented with 12.5 µM FPX obtained hygromycin-resistant calli. Thus, this protocol achieved a 0.7% transformation frequency. Similarly, progenies from one transgenic line were observed on the basis of GUS stains on their leaves, revealing that the transgenes were also inherited stably. Hence, FPX is considered a breakthrough for establishing the transformation protocol of <i>M. incana</i>, and its use is proposed in recalcitrant plants.</p>","PeriodicalId":20411,"journal":{"name":"Plant Biotechnology","volume":null,"pages":null},"PeriodicalIF":1.6,"publicationDate":"2022-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9592952/pdf/plantbiotechnology-39-3-22.0602a.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9177145","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}