Ho Bang Kim · Chang Jae Oh · Nam-Hoon Kim · Cheol Woo Choi · Minju Kim · Sukman Park · Seong Beom Jin · Su-Hyun Yun · Kwan Jeong Song
J Plant Biotechnol 2022; 49(4): 271-291Abstract : Pectin methylesterase (PME) plays an important role in vegetative and reproductive development and biotic/abiotic stress responses by regulating the degree of methyl-esterification of pectic polysaccharides in the plant cell wall. PMEs are encoded by a large multigene family in higher land plant genomes. In general, the expression of plant PME genes shows tissue- or cell-specific patterns and is induced by endogenous and exogenous stimuli. In this study, we identified PME multigene family members (CsPMEs) from the sweet orange genome and report detailed molecular characterization and expression profiling in different citrus tissues and two fruit developmental stages. We also discussed the possible functional roles of some CsPME genes by comparing them with the known functions of PMEs from other plant species. We identified 48 CsPME genes from the citrus genome. A phylogenetic tree analysis revealed that the identified CsPMEs were divided into two groups/types. Some CsPMEs showed very close phylogenetic relationships with the PMEs whose functions were formerly addressed in Arabidopsis, tomato, and maize. Expression profiling showed that some CsPME genes are highly or specifically expressed in the leaf, root, flower, or fruit. Based on the phylogenetic relationships and gene expression profiling results, we suggest that some CsPMEs could play functional roles in pollen development, pollen tube growth, cross incompatibility, root development, embryo/seed development, stomata movement, and biotic/abiotic stress responses. Our results shed light on the biological roles of individual CsPME isoforms and contribute to the search for genetic variations in citrus genetic resources.
Puspa Raj Poudel · Kazuya Koyama · Nami Goto-Yamamoto
J Plant Biotechnol 2022; 49(4): 292-299Abstract : Anthocyanin, an important component in the grape berry skin, strongly affects grape quality. The transcription factors VvMYBA1 and VvMYBA2 (VvMYBA1/2) control anthocyanin biosynthesis. In addition, cultivation and environmental factors, such as light, influence anthocyanin accumulation. The present study aimed to clarify the effect of shading (reduced light condition) on the transcriptomic regulation of anthocyanin biosynthesis using a red-wine grape cultivar, Vitis vinifera ‘Pinot Noir’, and its white mutant, ‘Pinot Blanc’, caused by the deletion of the red allele of VvMYBA1/2. The grape berry skins were analyzed for anthocyanin content and global gene transcription accumulation. The microarray data were later validated by quantitative real-time PCR. A decisive influence of VvMYBA1/2 on the expression of an anthocyanin-specific gene, UDP glucose: flavonoid 3-O-glucosyltransferase, was observed as expected. In contrast, upstream genes of the pathway, which are shared by other flavonoids, were also expressed in ‘Pinot Blanc’, and the mRNA levels of some of these genes decreased in both cultivars on shading. Thus, the involvement of light-sensitive transcription factor(s) other than VvMYBA1/2 was suggested for the expression control of the upstream genes of the anthocyanin biosynthetic pathway. Furthermore, it was suggested that the effects of these factors are different among isogenes.
Hojin Ryu
J Plant Biotechnol 2022; 49(4): 300-306Abstract : The mitogen-activated protein kinase (MAPK) signaling cascade is essential for a wide range of cellular responses in plants, including defense responses, responses to abiotic stress, hormone signaling, and developmental processes. Recent investigations have shown that the stress, ethylene, and MAPK signaling pathways negatively affect the formation of nitrogen-fixing nodules by directly modulating the symbiotic signaling components. However, the molecular mechanisms underlying the defense responses mediated by MAPK signaling in the organogenesis of nitrogen-fixing nodules remain unclear. In the present study, I demonstrate that the Medicago truncatula mitogen-activated protein kinase kinase 5 (MtMKK5)-Medicago truncatula mitogen-activated protein kinase 3/6 (MtMPK3/6) signaling module, expressed specifically in the symbiotic nodules, promotes defense signaling, but not ethylene signaling pathways, thereby inhibiting nodule development in M. truncatula. U0126 treatment resulted in increased cell division in the nodule meristem zone due to the inhibition of MAPK signaling. The phosphorylated TEY motif in the activation domain of MtMPK3/6 was the target domain associated with specific interactions with MtMKK5. I have confirmed the physical interactions between M. truncatula nodule inception (MtNIN) and MtMPK3/6. In the presence of high expression levels of the defense-related genes FRK1 and WRKY29, MtMKK5a overexpression significantly enhanced the defense responses of Arabidopsis against Pseudomonas syringae pv. tomato DC3000 (Pst DC3000). Overall, my data show that the negative regulation of symbiotic nitrogen-fixing nodule organogenesis by defense signaling pathways is mediated by the MtMKK5-MtMPK3/6 module.
Kyung Woon Kim ・ Cheol Ho Hwang
J Plant Biotechnol 2022; 49(4): 307-315Abstract : Artemisinin is a secondary metabolite of Artemisia annua that shows potent anti-malarial, anti-bacterial, anti-viral, and anti-tumor effects. The supply of artemisinin depends on its content in Artemisia annua, in which various environmental factors can affect the plant’s biosynthetic yield. In this study, the effects of different light-emitting diode (LED)-irradiation conditions were tested to optimize the germination and growth of Artemisia annua for the enhanced production of artemisinin. Specifically, the ratio between the red and blue lights in the irradiating LED was varied for investigation as follows: [Red : Blue] = [6 : 4], [7 : 3], and [8 : 2]. Furthermore, additional stress factors like UV-B-irradiation (1,395 μW/cm2), low temperature (4°C), and dehydration were also explored to induce hormetic expressions of ADS, CYP, and ALDH1, which are essential genes for the biosynthesis of artemisinin. Quantitative polymerase chain reaction (qPCR) was used to analyze the expression levels of the respective genes and their correlation with the specified conditions. [8 : 2] LED-irradiation was the most optimal among the tested conditions for the cultivation of Artemisia annua in terms of both fresh and dry weights post-harvest. For the production of artemisinin, however, [7 : 3] LED-irradiation with dehydration for six hours pre-harvest was the most optimal condition by inducing around twofold enhancement in the biosynthetic yield of artemisinin. As expected, a correlation was observed between the expression levels of the genes and the contents of artemisinin accumulated.
Sung-Gyu Lee ・Sang Moon Kang ・Hyun Kang
J Plant Biotechnol 2022; 49(4): 316-324Abstract : Hair loss causes psychological stress due to its effect on appearance. Therefore, the global market for hair loss treatment products is rapidly growing. The present study demonstrated that ginseng berry-derived and sequence-modified peptides promoted the proliferation rate of dermal papilla (DP) cells and keratinocytes, in addition to having antioxidant properties. Moreover, the potential role of these ginseng berry peptides as TGF-β2 antagonists was confirmed through in silico computer docking. In addition to promoting the growth of ,the ginseng berry-derived peptides also promoted the proliferation of keratinocytes experimental Particularly, an unmodified ginseng berry-derived peptide (GB-1) and two peptides with sequence modifications (GB-2 and GB-3) decreased ROS generation and exhibited a protective effect on damaged HaCaT keratinocytes. Computer-aided peptide discovery was conducted to identify the potential interactions of important proteins with transforming growth factor-beta 2 (TGF-β2), a key protein that plays a crucial role in the human hair growth cycle. Our results demonstrated that MAGH, an amino acid sequence present in herbal supplements and plant-based natural compounds, can inhibit TGF-β2.
Hui Yeong Jeong ・Ji Ah Kim
J Plant Biotechnol 2022; 49(4): 325-330Abstract : In this study, a plant tissue culture system was established for Atractylodes spp., an economically valuable medicinal crop in Korea that has low domestic production and is increasingly imported. In particular, Atractylodes ovata was treated with four types of cytokinins, 6-benzylaminopurine (BA), zeatin, kinetin, and thidiazuron (TDZ), in two different concentrations (0.5 and 1.0 mg/L). Among the four types of cytokinins, the BA treatment was effective for the shoot and root growth of A. ovata. Both the 0.5 mg/L and 1.0 mg/L concentrations of BA showed similar results; however, the 1.0 mg/L concentration of BA was more effective in promoting shoot and root growth. The treatments showed that the TDZ treatment was not effective for the shoot and root growth, except for the number of shoots and the fresh weight (FW) of the root; therefore, it was unsuitable for this species. In this study, we established a mass production system of A. ovata. Our results showed that direct in vitro regeneration may make a significant contribution to improving the cultivation of the medicinal plant A. ovata.
Chitpan Kativat ・Witsarut Chueakhunthod ・Piyada Alisha Tantasawat
J Plant Biotechnol 2022; 49(4): 331-338Abstract : Sunflower (Helianthus annuus L.) protoplasts were isolated from seven-day-old etiolated hypocotyls of 10 A line and four-week-old fully expanded young leaves of PI 441983 line in vitro seedlings using an enzymatic method. Purified protoplasts were collected by filtration and floatation in sucrose solution. Semi-solid protoplast culture was performed using the L4 regeneration protocol with various culture media and plating densities to achieve the highest efficiencies for protoplast culture of hypocotyl and mesophyll protoplasts of 10 A and PI 441983 lines, respectively. The concentrations in liquid L’4M medium and different plating densities were evaluated in two types of cytokinins, the adenine-type 6-benzyladenine (BA) and the phenylureatype thidiazuron (TDZ). The highest colony formation was achieved in both sunflower lines when 0.5 mgL-1 BA and 0.5 mgL-1 TDZ were applied with a high plating density (3 × 105 protoplasts mL-1). These conditions led to 38.45% and 39.40% colony formation for hypocotyl protoplasts of the 10 A line and mesophyll protoplasts of the PI 441983 line, respectively. Moreover, many hypocotyl protoplast-derived colonies developed into micro-calli. In addition, superior development of both sunflower protoplasts was observed with all plating densities when BA was used in combination with TDZ. This finding will be applicable to future sunflower hybrid production via somatic hybridization.
Jin-Woo Hwang ・Chan Hwi Park ・Hae-Yeon An ・Ye-Won Jang ・Hyun Kang ・Sung-Gyu Lee
J Plant Biotechnol 2022; 49(4): 339-346Abstract : This study investigates the antibacterial and antioxidant activities of the ethanol extract of Acer tegmentosum Maxim (EATM) against Staphylococcus aureus. The total polyphenol and flavonoid content, 1,1-diphenyl-2-picryl hydrazyl (DPPH) and 2,2’-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging effect, and reducing power of the ferric reducing antioxidant power (FRAP) method were measured to verify the antioxidant activity of the EATM. The antibacterial activity against S. aureus using the EATM was verified by the paper disc method, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) methods. The total polyphenol and flavonoid contents were 266.66 μg GAE/mg and 6.46 μg QE/mg, respectively. The DPPH and ABTS radical scavenging activity showed a concentration-dependent scavenging activity. The RC50 values of the EATM were 21.49 and 12.81 μg/mL, respectively. A FRAP analysis was conducted for evaluating the reducing power of the EATM and an efficacy of 0.73 ± 0.19 mM FeSO4 E/mg was observed. The antibacterial activity of EATM against S. aureus, determined using the paper disc method, showed an inhibitory ring of 3 mm at 2 mg. The MIC was confirmed at a concentration of ≥ 16 mg/mL, while the MBC was confirmed at 32 mg/mL. As the EATM shows antioxidant and antibacterial activities against S. aureus, it can be used as an effective antidote against atopic dermatitis.
Areumi Park ・Yeon-Ji Lee ・Nalae Kang ・Do-Hyung Kang ・Soo-Jin Heo
J Plant Biotechnol 2022; 49(4): 347-355Abstract : Jeju lava seawater’s abundant minerals are known to exert antioxidant effects that remove the free radicals responsible for aging. Therefore, lava seawater reportedly possesses high commercial value as a functional food material. This study compared and analyzed the antioxidant activities of extracts from crops produced in Jeju (carrots, blueberries, and mandarins) using distilled water and lava seawater as solvents. Lava seawater extracts exhibited higher total polyphenol and flavonoid contents of blueberries and mandarins than distilled water extracts. Furthermore, the antioxidant enzymatic and 2, 2-diphenyl-1-picrylhydrazyl radical-scavenging activities of these crops were higher in lava seawater extracts than in distilled water extracts. In particular, using Vero cells, the ROS-scavenging efficacies of blueberries and mandarins were found to be higher in lava seawater extracts. Meanwhile, the antioxidant activities of carrots were higher in lava seawater extracts, despite no difference in total polyphenol and flavonoid contents. These results suggest that lava seawater exhibits favorable potential as a solvent in the functional food industry, and lava seawater-based Jeju crop extracts are potentially useful as functional food ingredients.
Kang-Hyun Choi ・Sun Il Seo ・Haeseong Park ・Ji-hwan Lim・Pyoung Il Kim
J Plant Biotechnol 2022; 49(4): 356-361Abstract : Bacillus sp. is a useful strain for agriculture because it promotes plant growth and controls plant pathogens through a variety of mechanisms. In this study, we obtained a microbial preparation with a high number of viable cells by culturing newly isolated soil bacteria on an optimized medium. Subsequently, we applied this preparation to lettuce to enhance its growth and yield. First, B. amyloliquefaciens ISP-5 was isolated from soil. Next, optimization of culture medium was carried out using 5 L scale fermenters. When culturing B. amyloliquefaciens ISP-5 on this optimized medium, the number of viable cells was approximately 1000 times higher than that obtained from culturing on the commercial medium. Afterwards, the plant growth promotion properties of the ISP-5 strain were evaluated using lettuce as a test plant. Foliar spray treatment of lettuce was carried out by inoculating half the standard concentration suspension (0.5 × 107 cfu/ml). As a result, leaf width increased by 8.6% and leaf length increased by 12.9% compared to the control group. Live weight also increased by 24.2% and dry weight by 23.9%. Considering the results from field test, B. amyloliquefaciens ISP-5 showed potential as a plant growth-promoting bacteria.
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