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1.
Plants (Basel) ; 10(8)2021 Aug 09.
Artigo em Inglês | MEDLINE | ID: mdl-34451680

RESUMO

Weigela subsessilis is used in folk medicine to treat pain and allergic syndromes in Korea. However, the antibacterial and anti-inflammatory activities of W. subsessilis callus extract remain unexplored. In this study, we aimed to evaluate the W. subsessilis callus of pharmacological activity. Therefore, we first established in vitro calluses of W.subsessilis via plant tissue culture methods. We then evaluated the antioxidant and anti-inflammatory effects of W. subsessilis callus extract in lipopolysaccharide (LPS)-treated RAW264.7 macrophage cells. The W. subsessilis callus extract showed antioxidant and anti-inflammatory effects. These effects were regulated via suppression of mitogen-activated protein kinase signaling through LPS-induced translocation of nuclear factor kappa B (NF-κB) p65 from the cytoplasm to the nucleus. W. subsessilis callus extract also showed antibacterial and anti-inflammatory activities in Propionibacterium acnes-treated HaCaT keratinocyte cells. These results indicate that W. subsessilis callus extract has antioxidant, antibacterial and anti-inflammatory activities, suggesting its possible application in the treatment of inflammatory disorders.

2.
Int J Mol Sci ; 22(15)2021 Jul 22.
Artigo em Inglês | MEDLINE | ID: mdl-34360584

RESUMO

Trichostatin A (TSA) is a representative histone deacetylase (HDAC) inhibitor that modulates epigenetic gene expression by regulation of chromatin remodeling in cells. To investigate whether the regulation of chromatin de-condensation by TSA can affect the increase in the efficiency of Cas9 protein-gRNA ribonucleoprotein (RNP) indel formation from plant cells, genome editing efficiency using lettuce and tobacco protoplasts was examined after several concentrations of TSA treatments (0, 0.1, 1 and 10 µM). RNP delivery from protoplasts was conducted by conventional polyethylene glycol (PEG) transfection protocols. Interestingly, the indel frequency of the SOC1 gene from TSA treatments was about 3.3 to 3.8 times higher than DMSO treatment in lettuce protoplasts. The TSA-mediated increase of indel frequency of the SOC1 gene in lettuce protoplasts occurred in a concentration-dependent manner, although there was not much difference. Similar to lettuce, TSA also increased the indel frequency by 1.5 to 1.8 times in a concentration-dependent manner during PDS genome editing using tobacco protoplasts. The MNase test clearly showed that chromatin accessibility with TSA treatments was higher than that of DMSO treatment. Additionally, TSA treatment significantly increased the level of histone H3 and H4 acetylation from lettuce protoplasts. The qRT-PCR analysis showed that expression of cell division-related genes (LsCYCD1-1, LsCYCD3-2, LsCYCD6-1, and LsCYCU4-1) was increased by TSA treatment. These findings could contribute to increasing the efficiency of CRISPR/Cas9-mediated genome editing. Furthermore, this could be applied for the development of useful genome-edited crops using the CRISPR/Cas9 system with plant protoplasts.


Assuntos
Sistemas CRISPR-Cas , Edição de Genes/métodos , Ácidos Hidroxâmicos/farmacologia , Alface/metabolismo , Proteínas de Plantas/metabolismo , Protoplastos/metabolismo , Tabaco/metabolismo , Divisão Celular , Genoma de Planta , Alface/efeitos dos fármacos , Alface/genética , Alface/crescimento & desenvolvimento , Células Vegetais , Proteínas de Plantas/antagonistas & inibidores , Proteínas de Plantas/genética , Inibidores da Síntese de Proteínas/farmacologia , Protoplastos/efeitos dos fármacos , Tabaco/efeitos dos fármacos , Tabaco/genética , Tabaco/crescimento & desenvolvimento
3.
Sci Rep ; 11(1): 15120, 2021 07 23.
Artigo em Inglês | MEDLINE | ID: mdl-34301990

RESUMO

To establish an efficient plant regeneration system from cell suspension cultures of Euonymus alatus, embryogenic callus formation from immature embryos was investigated. The highest frequency of embryogenic callus formation reached 50% when the immature zygotic embryos were incubated on Murashige and Skoog (MS) medium supplemented with 1 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D). At higher concentrations of 2,4-D (over 2 mg/L), the frequency of embryogenic callus formation declined significantly. The total number of somatic embryos development was highest with the 3% (w/v) sucrose treatment, which was found to be the optimal concentration for somatic embryo formation. Activated charcoal (AC) and 6-benzyladenine (BA) significantly increased the frequency of plantlet conversion from somatic embryos, but gibberellic acid (GA3) had a negative effect on plantlet conversion and subsequent development from somatic embryos. Even though the cell suspension cultures were maintained for more than 1 year, cell aggregates from embryogenic cell suspension cultures were successfully converted into normal somatic embryos with two cotyledons. To our knowledge, this is the first successful report of a plant regeneration system of E. alatus via somatic embryogenesis. Thus, the embryogenic cell line and plant regeneration system established in this study can be applied to mass proliferation and production of pharmaceutical metabolite in E. alatus.

4.
Artigo em Inglês | MEDLINE | ID: mdl-34296986

RESUMO

An aerobic, Gram-stain-negative, weak-motile, short-rod-shaped bacterial strain, designated JBR3-12T, was isolated from halophyte Carex pumila plants, and its taxonomic position was investigated by using a polyphasic taxonomic approach. The strain produced a pink pigment on tryptic soy agar and grew optimally at 25 °C, pH 8 and in the presence of 3 % (w/v) NaCl. Results of phylogenetic analysis based on 16S rRNA gene sequences showed that strain JBR3-12T formed a lineage within the genus Pedobacter and was most closely related to Pedobacter sandarakinus DS-27T (98.0 %) and Pedobacter agri PB92T (97.6 %). The DNA G+C content of the genome was 41.3 mol%; the whole genome length was 5 426 070 bp. The major fatty acids of JBR3-12T were iso-C15 : 0, summed feature 3 (comprising C16 : 1 ω6c and/or C16 : 1 ω7c) and iso-C17 : 0 3-OH. The predominant polar lipid was phosphatidylethanolamine. The predominant quinone was menaquinone-7. Based on its phenotypic, phylogenetic and genotypic features, strain JBR3-12T is proposed to represent a novel species of the genus Pedobacter, for which the name is Pedobacter endophyticus sp. nov. The type strain is JBR3-12T (=KCTC 82363T=NBRC 114901T).


Assuntos
Carex (Planta)/microbiologia , Pedobacter/classificação , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Pedobacter/isolamento & purificação , Fosfatidiletanolaminas/química , Pigmentação , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Microbiologia do Solo , Vitamina K 2/análogos & derivados , Vitamina K 2/química
5.
Plant Methods ; 17(1): 21, 2021 Feb 23.
Artigo em Inglês | MEDLINE | ID: mdl-33622383

RESUMO

BACKGROUND: Plants have a remarkable reprogramming potential, which facilitates plant regeneration, especially from a single cell. Protoplasts have the ability to form a cell wall and undergo cell division, allowing whole plant regeneration. With the growing need for protoplast regeneration in genetic engineering and genome editing, fundamental studies that enhance our understanding of cell cycle re-entry, pluripotency acquisition, and de novo tissue regeneration are essential. To conduct these studies, a reproducible and efficient protoplast regeneration method using model plants is necessary. RESULTS: Here, we optimized cell and tissue culture methods for improving protoplast regeneration efficiency in Arabidopsis thaliana. Protoplasts were isolated from whole seedlings of four different Arabidopsis ecotypes including Columbia (Col-0), Wassilewskija (Ws-2), Nossen (No-0), and HR (HR-10). Among these ecotypes, Ws-2 showed the highest potential for protoplast regeneration. A modified thin alginate layer was applied to the protoplast culture at an optimal density of 1 × 106 protoplasts/mL. Following callus formation and de novo shoot regeneration, the regenerated inflorescence stems were used for de novo root organogenesis. The entire protoplast regeneration process was completed within 15 weeks. The in vitro regenerated plants were fertile and produced morphologically normal progenies. CONCLUSION: The cell and tissue culture system optimized in this study for protoplast regeneration is efficient and reproducible. This method of Arabidopsis protoplast regeneration can be used for fundamental studies on pluripotency establishment and de novo tissue regeneration.

6.
Plants (Basel) ; 9(12)2020 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-33287317

RESUMO

Toona sinensis has been traditionally used to treat dysentery, enteritis, flatulence, and itchiness. However, the existence of anti-inflammatory effects of T. sinensis on Propionibacterium acnes-induced skin disease is unknown. In vitro cultures of plant cells and tissues produced under controlled conditions offer a continuous production platform for plant natural products including pigments and anti-inflammatory agents. In this study, we determine the anti-inflammatory activities of an extract of in vitro grown adventitious shoots of T. sinensis on P. acnes, the etiologic agent of skin inflammation. The extract of T. sinensis showed antioxidant and anti-inflammatory activity in LPS-treated RAW264.7 cells. It also had antibacterial activity and anti-inflammatory effects on P. acnes-treated HaCaT cells. In addition, these effects were regulated by suppression of the mitogen-activated protein kinase (MAPK) pathways. These results suggesting the potential application of adventitious shoots of T. sinensis grown with an in vitro proliferation system as a medicine for treating P. acnes-induced inflammatory skin disease.

7.
Plants (Basel) ; 9(12)2020 Dec 07.
Artigo em Inglês | MEDLINE | ID: mdl-33297321

RESUMO

We aimed to develop a novel technology capable of rapidly selecting mutant plant cell lines. Salt resistance was chosen as a rapid selection trait that is easily applicable to protoplast-derived cell colonies. Mesophyll protoplasts were cultured in a medium supplemented with 0, 50, 100, 150, 200, 250, and 300 mM NaCl. At NaCl concentrations ≥ 100 mM, cell colony formation was strongly inhibited after 4 weeks of culture. Tobacco protoplasts irradiated with 0, 50, 100, 200, and 400 Gy were then cultured to investigate the effects of radiation intensity on cell division. The optimal radiation intensity was 50 Gy. To develop salt-resistant tobacco mutant plants, protoplasts irradiated with 50 Gy were cultured in a medium containing 100 mM NaCl. The efficiency of cell colony formation from these protoplasts was approximately 0.002%. A salt-resistant mutant callus was selected and proliferated in the same medium and then transferred to a shoot inducing medium for adventitious shoot formation. The obtained shoots were then cultured in a medium supplemented with 200 mM NaCl and developed into normal plantlets. This rapid selection technology for generating salt-resistant tobacco mutants will be useful for the development of crop varieties resistant to environmental stresses.

8.
Int J Mol Sci ; 21(22)2020 Nov 10.
Artigo em Inglês | MEDLINE | ID: mdl-33182800

RESUMO

Histone acetylation plays an important role in plant growth and development. Here, we investigated the effect of sodium butyrate (NaB), a histone deacetylase inhibitor, on adventitious shoot formation from protoplast-derived calli and cotyledon explants of tobacco (Nicotiana benthamiana) and tomato (Solanum lycopersicum). The frequency of adventitious shoot formation from protoplast-derived calli was higher in shoot induction medium (SIM) containing NaB than in the control. However, the frequency of adventitious shoot formation from cotyledon explants of tobacco under the 0.1 mM NaB treatment was similar to that in the control, but it decreased with increasing NaB concentration. Unlike in tobacco, NaB decreased adventitious shoot formation in tomato explants in a concentration-dependent manner, but it did not have any effect on adventitious shoot formation in calli. NaB inhibited or delayed the expression of D-type cyclin (CYCD3-1) and shoot-regeneration regulatory gene WUSCHEL (WUS) in cotyledon explants of tobacco and tomato. However, compared to that in control SIM, the expression of WUS was promoted more rapidly in tobacco calli cultured in NaB-containing SIM, but the expression of CYCD3-1 was inhibited. In conclusion, the effect of NaB on adventitious shoot formation and expression of CYCD3-1 and WUS genes depended on the plant species and whether the effects were tested on explants or protoplast-derived calli.


Assuntos
Ácido Butírico/farmacologia , Lycopersicon esculentum/efeitos dos fármacos , Lycopersicon esculentum/crescimento & desenvolvimento , Tabaco/efeitos dos fármacos , Tabaco/crescimento & desenvolvimento , Cotilédone/efeitos dos fármacos , Cotilédone/genética , Cotilédone/crescimento & desenvolvimento , Ciclina D/genética , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Genes de Plantas/efeitos dos fármacos , Inibidores de Histona Desacetilases/farmacologia , Lycopersicon esculentum/genética , Proteínas de Plantas/genética , Brotos de Planta/efeitos dos fármacos , Brotos de Planta/genética , Brotos de Planta/crescimento & desenvolvimento , Especificidade da Espécie , Tabaco/genética
9.
Int J Mol Sci ; 21(15)2020 Jul 26.
Artigo em Inglês | MEDLINE | ID: mdl-32722633

RESUMO

Enhancing the competence for plant regeneration in tissue culture studies is an important issue not only for efficient genetic transformation of commercial crops but also for the reproducibility of scientific reports. In this study, we investigated optimization of several tissue culture conditions including plant growth regulators, types and ages of explants, culture densities, and plant position in order to improve the competence of adventitious shoot formation of the tomato (Solanum lycopersicum cv. Micro-Tom). In addition, we examined the differential expression of D-type cyclin (CYCD3-1) and several shoot regeneration regulatory genes from hypocotyl and cotyledon explants of tomato during shoot organogenesis. A treatment of 1 mg L-1 Zeatin and 0.1 mg L-1 Indole-3-acetic acid (IAA) in Murashige and Skoog (MS) medium containing 3% sucrose was optimal for adventitious shoot formation from hypocotyl and cotyledon explants. The younger explants exhibited more shoot formation regardless of explant types. Additionally, those closest to the shoot apical meristem produced more shoots compared to the other regions in the hypocotyl and the cotyledon explants. Gene expression of CYCD3-1, SHOOT MERISTEMLESS (STM), and cytokinin dependent WUSCHEL (WUS) was significantly higher in younger explants than in older ones. Furthermore, an increase in CYCD3-1, STM, and WUS expression was evident at the distal part of hypocotyls and the proximal part of cotyledons compared to other regions. These differential gene expression profiles exhibited good agreement with the results of shoot formation obtained from diverse explants of tomato. These results suggest that temporal and spatial gene expression of shoot regeneration regulatory genes plays an important role in enhancing the competence and the reproducibility of adventitious shoot formation from tomato explants.


Assuntos
Cotilédone/metabolismo , Regulação da Expressão Gênica de Plantas , Hipocótilo/metabolismo , Lycopersicon esculentum/metabolismo , Proteínas de Plantas/biossíntese
10.
Int J Mol Sci ; 21(11)2020 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-32486319

RESUMO

Pterocarpans are derivatives of isoflavonoids, found in many species of the family Fabaceae. Sophora flavescens Aiton is a promising traditional Asian medicinal plant. Plant cell suspension cultures represent an excellent source for the production of valuable secondary metabolites. Herein, we found that methyl jasmonate (MJ) elicited the activation of pterocarpan biosynthetic genes in cell suspension cultures of S. flavescens and enhanced the accumulation of pterocarpans, producing mainly trifolirhizin, trifolirhizin malonate, and maackiain. MJ application stimulated the expression of structural genes (PAL, C4H, 4CL, CHS, CHR, CHI, IFS, I3'H, and IFR) of the pterocarpan biosynthetic pathway. In addition, the co-treatment of MJ and methyl-ß-cyclodextrin (MeßCD) as a solubilizer exhibited a synergistic effect on the activation of the pterocarpan biosynthetic genes. The maximum level of total pterocarpan production (37.2 mg/g dry weight (DW)) was obtained on day 17 after the application of 50 µM MJ on cells. We also found that the combined treatment of cells for seven days with MJ and MeßCD synergistically induced the pterocarpan production (trifolirhizin, trifolirhizin malonate, and maackiain) in the cells (58 mg/g DW) and culture medium (222.7 mg/L). Noteworthy, the co-treatment only stimulated the elevated extracellular production of maackiain in the culture medium, indicating its extracellular secretion; however, its glycosides (trifolirhizin and trifolirhizin malonate) were not detected in any significant amounts in the culture medium. This work provides new strategies for the pterocarpan production in plant cell suspension cultures, and shows MeßCD to be an effective solubilizer for the extracellular production of maackiain in the cell cultures of S. flavescens.


Assuntos
Acetatos/farmacologia , Ciclodextrinas/farmacologia , Ciclopentanos/farmacologia , Oxilipinas/farmacologia , Raízes de Plantas/metabolismo , Pterocarpanos/metabolismo , Sophora/efeitos dos fármacos , Sophora/metabolismo , Biotecnologia , Meios de Cultura , Sinergismo Farmacológico , Flavonoides/análise , Glucosídeos/análise , Compostos Heterocíclicos de 4 ou mais Anéis/análise , Espectroscopia de Ressonância Magnética , Malonatos/análise , Extratos Vegetais/química , Folhas de Planta/metabolismo , Plantas Medicinais , Pterocarpanos/análise
11.
Int J Syst Evol Microbiol ; 70(5): 3541-3546, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32369001

RESUMO

The genus Izhakiella was established and designated as a member of the family Enterobacteriaceae in 2016. Although the taxonomical classification of most members in this family has been relatively resolved after two reclassifications in 2016 and 2017, the classification of the genus Izhakiella remains ambiguous. In this study, a polyphasic approach was used to provide evidence supporting the fact that the genus Izhakiella should no longer be considered a member of Enterobacteriaceae and proposes its reclassification into the family Erwiniaceae. The phylogenetic tree of type species in the families Enterobacteriaceae and Erwiniaceae based on the sequences of the 16S rRNA gene, rpoB housekeeping gene, and the whole-genome comprising the 92 core genes revealed that the genus Izhakiella forms a phylogenetic lineage within the family Erwiniaceae. The average nucleotide identity (ANI) value of the type species with genus Izhakiella was found to be higher for the family Erwiniaceae than that for the family Enterobacteriaceae. Notably, 12 conserved signature indels (CSIs) that are exclusively shared among the Erwiniaceae clade members were found in the type strains of the genus Izhakiella. Based on these analyses, this study suggests the reclassification of I. capsodis and I. australiensis into the family Erwiniaceae.


Assuntos
Enterobacteriaceae/classificação , Gammaproteobacteria/classificação , Filogenia , Técnicas de Tipagem Bacteriana , DNA Bacteriano/genética , Genômica , Mutação INDEL , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
12.
J Microbiol ; 58(5): 357-366, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32221820

RESUMO

Strain KSNA2T, a Gram-negative, moderately halophilic, facultatively anaerobic, motile, rod-shaped bacterium, was isolated from the surface-sterilized stem tissue of a beach morning glory (Cafystegia soldanella) plant in Chuja Island, Jeju-do, Republic of Korea. Phylogenetic analysis based on 16S rRNA gene and whole-genome sequences revealed that strain KSNA2T formed a distinct lineage within the family Enterobacteriaceae, with the highest 16S rRNA gene sequence similarity to Izhakiella australiensis KCTC 72143 (96.2%) and Izhakiella capsodis KCTC 72142T (96.0%), exhibited 95.5-95.9% similarity to other genera in the family Enterobacter-iaceae and Erwiniaceae. Conserved signature indels analysis elucidated that strain KSNA2T was delimited into family Enterobacteriaceae. KSNA2 genome comprises a circular chromosome of 5,182,800 bp with 56.1% G + C content Digital DNA-DNA relatedness levels between strain KSNA2T and 18 closely related species were 19.3 to 21.1%. Average nucleotide identity values were between 72.0 and 76.7%. Growth of strain KSNA2T was observed at 4 to 45°C (optimum, 25°C) and pH 5.0 to 12.0 (optimum, pH 7.0) in the presence of 0 to 11% (w/v) NaCl (optimum, 0-7%). The major cellular fatty acids (> 10%) were C16:o followed by summed feature 8 (C18ω7c and/or C18ω6c), summed feature 3 (C16:1 ω7c and/or C16:1 ω6c), C17:0cyclo, and C14:0. The major isoprenoid quinone was ubiquinone-8 (Q-8). With combined phylogenetic, genomic, phenotypic, and chemotaxo-nomic features, strain KSNA2T represents a novel species of a new genus in the family Enterobacteriaceae, for which the name Jejubacter calystegiae gen. nov., sp. nov. is proposed. The type strain is KSNA2T (= KCTC 72234T = CCTCC AB 2019098T).


Assuntos
Calystegia/microbiologia , Enterobacteriaceae/classificação , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Endófitos/classificação , Endófitos/isolamento & purificação , Enterobacteriaceae/isolamento & purificação , Ácidos Graxos/química , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Ubiquinona/química
13.
J Microbiol ; 58(1): 24-29, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31768939

RESUMO

Strain ATSA2T was isolated from surface-sterilized kimchi cabbage (Brassica rapa subsp. pekinensis) seeds and represents a novel bacterium based on the polyphasic taxonomic approach. A phylogenetic analysis based on 16S rRNA gene sequences showed that strain ATSA2T formed a lineage within genus Saccharibacillus and was most closely to Saccharibacillus deserti WLG055T (98.1%) and Saccharibacillus qing-shengii H6T (97.9%). The whole-genome of ATSA2T comprised a 5,619,468 bp of circular chromosome with 58.4% G + C content. The DNA-DNA relatedness values between strain ATSA2T and its closely related type strains S. deserti WLJ055 and S. qingshengii H6T were 26.0% and 24.0%, respectively. Multiple gene clusters associated with plant growth promotion activities (stress response, nitrogen and phosphorus metabolism, and auxin biosynthesis) were annotated in the genome. Strain ATSA2T was Gram-positive, endospore-forming, facultatively anaerobic, and rod-shaped It grew at 15-37°C (optimum 25°C), pH 6.0-10.0 (optimum pH 8.0), and in the presence of 0-5% (w/v) NaCl (optimum 1%). The major cellular fatty acids (> 10%) of strain ATSA2T were anteiso-C15:0 and C16:0. MK-7 was the major isoprenoid quinone. The major polar lipids present were diphosphatidylglycerol, phosphatidylglycerol, and three unknown glycolipids. Based on its phylogenetic, genomic, phenotypic, and chemotaxo-nomic features, strain ATSA2T is proposed to represent a novel species of genus Saccharibacillus, for which the name is Saccharibacillus brassicae sp. nov. The type strain is ATSA2T (KCTC 43072T = CCTCC AB 2019223T).


Assuntos
Bacillales/classificação , Bacillales/isolamento & purificação , Brassica rapa/microbiologia , Sementes/microbiologia , Técnicas de Tipagem Bacteriana/métodos , Filogenia , RNA Ribossômico 16S/genética
14.
Sci Rep ; 9(1): 16354, 2019 11 08.
Artigo em Inglês | MEDLINE | ID: mdl-31704990

RESUMO

Biocontrol offers a promising alternative to synthetic fungicides for the control of a variety of pre- and post-harvest diseases of crops. Black rot, which is caused by the pathogenic fungus Ceratocytis fimbriata, is the most destructive post-harvest disease of sweet potato, but little is currently known about potential biocontrol agents for this fungus. Here, we isolated several microorganisms from the tuberous roots and shoots of field-grown sweet potato plants, and analyzed their ribosomal RNA gene sequences. The microorganisms belonging to the genus Pantoea made up a major portion of the microbes residing within the sweet potato plants, and fluorescence microscopy showed these microbes colonized the intercellular spaces of the vascular tissue in the sweet potato stems. Four P. dispersa strains strongly inhibited C. fimbriata mycelium growth and spore germination, and altered the morphology of the fungal hyphae. The detection of dead C. fimbriata cells using Evans blue staining suggested that these P. dispersa strains have fungicidal rather than fungistatic activity. Furthermore, P. dispersa strains significantly inhibited C. fimbriata growth on the leaves and tuberous roots of a susceptible sweet potato cultivar ("Yulmi"). These findings suggest that P. dispersa strains could inhibit black rot in sweet potato plants, highlighting their potential as biocontrol agents.


Assuntos
Ascomicetos/crescimento & desenvolvimento , Ipomoea batatas/imunologia , Pantoea/fisiologia , Controle Biológico de Vetores , Doenças das Plantas/prevenção & controle , Folhas de Planta/imunologia , Raízes de Plantas/imunologia , Resistência à Doença , Ipomoea batatas/microbiologia , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Folhas de Planta/microbiologia , Raízes de Plantas/microbiologia
15.
Data Brief ; 26: 104465, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31534997

RESUMO

Saccharibacillus sp. ATSA2 was isolated from Kimchi cabbage seeds grown in Gyeongbuk province in the Republic of Korea. Whole-genome sequencing of Saccharibacillus sp. ATSA2 was performed using the PacBio RSII and Illumina HiSeq platforms, and it features a 5,619,468 bp circular chromosome with 58.4% G + C content. The genome includes 4543 protein-coding genes, 104 RNA genes (70 transfer RNA genes, 30 ribosomal RNA genes, and 4 non-coding RNA), and 73 pseudogenes. Multiple gene clusters associated with stress responses, nitrogen and phosphorus metabolism, and plant hormone biosynthesis were annotated in the genome. The genome information will provide fundamental knowledge of this organism as well as insight for understanding microbial activity in the agricultural application. The whole-genome sequence of Saccharibacillus sp. ATSA2 is available at GenBank/EMBL/DDBJ under accession number CP041217.

16.
J Microbiol ; 57(11): 953-958, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31463785

RESUMO

A strictly aerobic, motile, endospore-forming, rod-shaped bacterium, designated HS21T, was isolated from rhizospheric soil of the Korean fir tree (Abies koreana) from Halla mountain on Jeju island, Korea. Growth of strain HS21T was observed at pH 6.0-8.0 (optimum: pH 7.0), 0-2% (w/v) NaCl and 4-30°C (optimum: 25°C). A comparative analysis of 16S rRNA gene sequences showed that strain HS21T was most closely related to Cohnella luojiensis HY-22RT (97.6%), followed by C. lupini RLAHU4BT (97.4%) and C. collisoli NKM-5T (97.2%). The genome of strain HS21T comprised a circular chromosome of 7,059,027 bp with 44.8% G + C content. The DNA-DNA relatedness values between strain HS21T and C. luojiensis HY-22RT and C. lupini RLAHU4BT were 18.1% and 13.8%, respectively. The major cellular fatty acids (> 5%) of the isolate were anteiso-C15:0, iso-C16:0, C16:0, and iso-C15:0. The polar lipids present were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, lysylphosphatidylglycerol, and three unidentified aminophospholipids. Based on its phenotypic, phylogenetic, genomic, and chemotaxonomic properties, strain HS21T represents a novel species of the genus Cohnella, for which the name Cohnella abietis sp. nov. is proposed. The type strain is HS21T (= KCTC 43028T = CCTCC AB 2019010T).


Assuntos
Abies/microbiologia , Bacillales/classificação , Bacillales/isolamento & purificação , Filogenia , Rizosfera , Microbiologia do Solo , Bacillales/genética , Bacillales/fisiologia , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Lipídeos/química , Lisina/química , Fosfatidilgliceróis/química , Fosfolipídeos/química , RNA Ribossômico 16S/genética , República da Coreia , Nódulos Radiculares de Plantas/microbiologia , Análise de Sequência de DNA , Solo , Sequenciamento Completo do Genoma
17.
Artigo em Inglês | MEDLINE | ID: mdl-33620309

RESUMO

A Gram-stain-positive, facultatively anaerobic, rod-shaped, endospore-forming, oxidase-positive, and catalase-negative strain designated as BRMEA1T was isolated from the surface-sterilized Selaginella involvens roots. Growth of strain BRMEA1T was found to occur at pH 6.0-8.0 (optimum, pH 7.0), 15-50 °C (optimum, 25-30 °C) and in the absence of NaCl. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain BRMEA1T formed a lineage within the genus Neobacillus (family Bacillaceae) and showed the highest sequence similarity to Neobacillus drentensis DSM 15600T (98.3 %) and Neobacillus fumarioli KCTC 13885T (98.2 %), and less than 98.2 % 16S rRNA gene sequence similarity to the other members of the genus Neobacillus. Whole-genome analysis of strain BRMEA1T comprised a circular chromosome (5 632 809 bp in size) with 38.5 mol% G+C content. Digital DNA-DNA hybridization analyses revealed that strain BRMEA1T showed 20.5 and 22.0% genomic DNA relatedness with the closest species, N. drentensis DSM 15600T and N. fumarioli KCTC 13885T, respectively. The whole-genome sequence of strain BRMEA1T showed the presence of 11 specific conserved signature indels for the genus Neobacillus. The major cellular fatty acids (>10 %) of strain BRMEA1T were found to be iso-C15 : 0 and anteiso-C15 : 0, while the major polar lipids were found to be diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol. Polyphasic analysis results revealed that BRMEA1T represents a novel species of the genus Neobacillus, with the proposed name Neobacillus endophyticus sp. nov. The type strain is BRMEA1T (=KCTC 43208T=CCTCC AB 2020071T).

18.
Appl Microbiol Biotechnol ; 102(22): 9563-9575, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30218377

RESUMO

Hairy root cultures (HRCs) are characterized by fast and unlimited root growth, and they have greater genetic stability than other cultivation methods. HRCs are known to accumulate phytochemical levels comparable to those of intact plant. In this study, HRCs of Polygonum multiflorum were established from leaf explants infected with Agrobacterium rhizogenes strain KCCM 11879. Over 60% of the explants showed hairy root induction after 21 days of cultivation on hormone-free MS (Murashige and Skoog Physiol Plant 15:473-479, 1962) medium; induced roots were confirmed by PCR using a rolC-specific primer. Of the six lines of HRCs selected for further analysis, line HR-01 performed best, producing a root biomass (105.2 g L-1 of FW, 9.7 g L-1 of DW), which is 10-fold higher than that of non-transgenic roots. The HR-01 line also showed a significant increase in its total phenolic content (26.64 mg g-1 DW), while non-transgenic roots accumulated 8.36 mg g-1 DW of total phenolic. The levels of phenolic compounds in the HRCs increased more than 2.5-fold following exposure to 50 µM methyl jasmonate for 5 days. Fourier transform infrared (FT-IR) spectroscopic analysis of bioactive accumulation in P. multiflorum enabled discrimination between hairy root and adventitious root cultures. Thus, it is evident from this study that HRCs could be an attractive proposition for large-scale production of root biomass and secondary metabolites of P. multiflorum in bioreactors.


Assuntos
Fallopia multiflora/química , Fenóis/metabolismo , Raízes de Plantas/química , Acetatos/farmacologia , Agrobacterium , Biomassa , Reatores Biológicos , Cromatografia Líquida de Alta Pressão , Meios de Cultura/química , Ciclopentanos/farmacologia , Oxilipinas/farmacologia , Fenóis/análise , Folhas de Planta , Espectroscopia de Infravermelho com Transformada de Fourier , Técnicas de Cultura de Tecidos
19.
J Agric Food Chem ; 66(16): 4099-4105, 2018 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-29630360

RESUMO

Isoflavonoids are a class of biologically active natural products that accumulate in soybean ( Glycine max L.) seeds during development, play vital roles in plant defense, and act as phytoestrogens with important human health benefits. Plant cell suspension cultures represent an excellent source of biologically important secondary metabolites. We found that methyl jasmonate (MJ) treatment increased isoflavone production in soybean suspension cell cultures. To investigate the underlying mechanism, we examined the expression of structural genes ( CHS6, CHS7, CHI1, IFS1, IFS2, IFMaT, and HID) in the isoflavonoid biosynthesis pathways in soybean suspension cells under various abiotic stress conditions. MJ treatment had the most significant effect on gene expression and increased the production of three glycosidic isoflavones (daidzin, malonyldaidzin, and malonylgenistin), with the maximum total isoflavone production (∼10-fold increase) obtained on day 9 after MJ application. MJ treatment significantly increased total phenolic contents and upregulated isoflavonoid biosynthesis genes, shedding light on the underlying mechanism.


Assuntos
Acetatos/farmacologia , Ciclopentanos/farmacologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Isoflavonas/biossíntese , Oxilipinas/farmacologia , Proteínas de Plantas/genética , Soja/efeitos dos fármacos , Células Cultivadas , Proteínas de Plantas/metabolismo , Soja/genética , Soja/metabolismo
20.
Front Plant Sci ; 8: 1965, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29204151

RESUMO

Synechocystis salt-responsive gene 1 (sysr1) was engineered for expression in higher plants, and gene construction was stably incorporated into tobacco plants. We investigated the role of Sysr1 [a member of the alcohol dehydrogenase (ADH) superfamily] by examining the salt tolerance of sysr1-overexpressing (sysr1-OX) tobacco plants using quantitative real-time polymerase chain reactions, gas chromatography-mass spectrometry, and bioassays. The sysr1-OX plants exhibited considerably increased ADH activity and tolerance to salt stress conditions. Additionally, the expression levels of several stress-responsive genes were upregulated. Moreover, airborne signals from salt-stressed sysr1-OX plants triggered salinity tolerance in neighboring wild-type (WT) plants. Therefore, Sysr1 enhanced the interconversion of aldehydes to alcohols, and this occurrence might affect the quality of green leaf volatiles (GLVs) in sysr1-OX plants. Actually, the Z-3-hexenol level was approximately twofold higher in sysr1-OX plants than in WT plants within 1-2 h of wounding. Furthermore, analyses of WT plants treated with vaporized GLVs indicated that Z-3-hexenol was a stronger inducer of stress-related gene expression and salt tolerance than E-2-hexenal. The results of the study suggested that increased C6 alcohol (Z-3-hexenol) induced the expression of resistance genes, thereby enhancing salt tolerance of transgenic plants. Our results revealed a role for ADH in salinity stress responses, and the results provided a genetic engineering strategy that could improve the salt tolerance of crops.

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