Microalgae are attracting much attention as promising, eco-friendly producers of bioenergy due to their fast growth, absorption of carbon dioxide from the atmosphere, and production capacity in wastewater and salt water. However, microalgae can only accumulate large quantities of lipid in abiotic stress, which reduces productivity by decreasing cell growth. In this study, the strategy was investigated to increase cell viability and lipid production by overexpressing S-adenosylmethionine (SAM) synthetase (SAMS) in the microalga Chlamydomonas reinhardtii. SAM is a substance that plays an important role in various intracellular biochemical reactions, such as cell proliferation and stress response, and the overexpression of SAMS could allow cells to ithstand the abiotic stress and increase productivity. Compared to wild-type C. reinhardtii, recombinant cells overexpressing SAMS grew 1.56-fold faster and produced 1.51-fold more lipids in a nitrogen-depleted medium. Furthermore, under saline-stress conditions, the survival rate and lipid accumulation were 1.56 and 2.04 times higher in the SAMS-overexpressing strain, respectively. These results suggest that the overexpression of SAMS in recombinant C. reinhardtii has high potential in the industrial-scale production of biofuels and various other high-value-added materials.
Kang, Woo Hyun;Han, Zeesoo;Lee, Seung Jun;Shin, Jong Hwa;Ahn, Tae In;Lee, Joo Young;Kang, Suk Woo;Jung, Sang Hoon;Son, Jung Eek
Journal of Bio-Environment Control
/
v.27
no.1
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pp.94-101
/
2018
Mugwort (Artemisia princeps) is a medicinal plant that has a substance called euphatilin, which is effective for cell damage and gastritis recovery. The objectives of this study were to investigate the annual growth characteristics of Artemisia princeps in greenhouse and to increase the eupatiline content by environmental stresses. Growth and eupatilin content of the plants were compared after 6 weeks of seedling and subsequent 8 weeks of greenhouse cultivation. Photosynthesis of mugwort plants did not saturate even at a relatively high light intensity of $1,200{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$. Growth rate of the plants reached its highest at two weeks after transplanting and began to decrease since 8 weeks after transplanting. The plants showed typical characteristics of a perennial herbaceous plant as they were sensitive to seasonal changes. In particular, the plants showed high growth and eupatilin content in spring and summer as vegetative growth periods, but flowering and wintering caused considerable decreases in growth and eupatilin content in fall and winter. Therefore, application of night interruption is essential for year-round cultivationof the plant. Two stresses and a elicitor were treated: drought stresses by stopping irrigation at 5, 6, 7, and 8 days before harvest; salt stresses with nutrient solution concentrations of 2, 4, 6, 8, and $10dS{\cdot}m^{-1}$ by adding sodium chloride at 3 days before harvest; and foliar applications of methyl jasmonates of 12.5, 25, 50, and $100{\mu}M$ at 3 days before harvest. Significant increase in eupatilin content was observed at drought stresses of 7- and 8-days of irrigation stop and foliar application of $25{\mu}M$ methyl jasmonate, while no significant increase observed at salt stresses. From the results, it was confirmed that the environmental treatments can improve the productivity and quality of Artemisia princeps as a phamaceutical raw material.
A bacterial strain antagonistic to some fungal phytopathogens was isolated from the stem of a Persimmon tree in Yeongam, Korea. This bacterium was identified as Bacillus subtilis by 16S rRNA gene sequencing and designated as B. subtilis GDYA-1. In in vivo experiment, the fermentation broth exhibited antifungal activities against Magnaporthe oryzae on rice plants, Phytophthora infestans on tomato plants, and Puccinia recondita on wheat plants. We isolated one antifungal compound and its chemical structure was determined by mass and $^1H$-NMR spectral data. The antifungal substance was identified as benzoic acid. It inhibited mycelial growth of M. oryzae, Rhizoctonia solani, Sclerotinia sclerotiorum, and P. capsici with minimum inhibition concentration (MIC) values, ranging from 62.5 to 125 ${\mu}g/ml$. Moreover, the substance effectively suppressed Phytophthora blight of red pepper caused by P. capsici in a pot experiment. To the author's knowledge, this is the first report on the antifungal activity of benzoic acid against phytopathogenic fungi. Benzoic acid and B. subtilis GDYA-1 may contribute to environmental-friendly protect crops from phytopathogenic fungi.
This experiment was designed to improve the in vitro production system of apple rootstock M.26 as being influenced by the growth regulators, TDZ, BA, IAA, IBA, zeatin, and GA$_3$. Different levels of potassium humate (KH), known as cytokinin and auxin-like substance, were also supplemented to the MS basal medium along with IBA 0.6 mg/L to find out it effect on root formation in apple rootstock M.26. ID initiate and establish the in vitro multiplication of shoots byway of meristem culture, MS medium added with zeatin 1.0 mg/L was found to be the most suitable, showing the 100% of survival rate of shoot tips. A combination of thidiazuron (TDZ) 0.2 mg/L and NAA 0.5 mg/L promoted the shoot proliferation when shoot tips were used as explants. MS basal medium plus IBA 0.6 mg/L was very effective for root induction, but an addition of potassium humate (250 mg/L) to the medium containing IBA 0.6 mg/L stimulated the induction and proliferation of the rook by far the better.
Barley (Hordeum vulgare cv. Hangmaeg) sprouts are important microgreens that contain high levels of polyphenols and flavonoids, as well as minerals, vitamin and chlorophyll. Barley sprouts were grown for 9 days and growth was checked every 3 days. In this study, the cultivation efficiency according to the nutrient solution treatment was evaluated by analyzing the length of barley sprouts, fresh weight, chlorophyll, and the yield by growth period. In addition, we tried to increase the industrial applicability of germinated barley through analysis of inorganic component, total polyphenol and total flavonoid content of the extract, and functional substance analysis using HPLC. As a result, the growth rate of the nutrient solution treatment group was faster than that of the control group. When harvested on the 9th day of sowing, the nutrient solution treatment group showed a significant increase in yield compared to the control group. And the barley sprout extract of the nutrient solution treatment group had higher total flavonoid content and luteolin content. Also, the efficiency of water was higher than that of ethanol when extracting phenolics from barley sprouts. Therefore, this study suggests that nutrient input is effective for increasing polyphenol content and increasing production in barley sprout hydroponics.
This study was conducted to anticipate nitrate reduction state in tree through measurement of nitrate reductase activity (NRA) and investigate the effect of nitrogen concentrations (100, 200, 400, and 600 $mg\;L^{-1}$) on growth, the nitrogen content of various tissue, and NRA of pear (Pyrus pyrifolia cv. Niitaka) seedlings in sand culture. Nutrient solutions used in this experiment were adjusted to pH 6.5 and fixed the ratio of ammonium and nitrate to 1:3 and trickle-irrigated 3 times a day. Tree height and dry weight of various organs in seedlings were higher in low nitrogen concentration (100 and 200 $mg\;L^{-1}$) than in high nitrogen concentration (400 and 600 $mg\;L^{-1}$). The shoot growth in 600 $mg\;L^{-1}$ was extremely poor by nitrogen over supply. Increasing the nitrogen concentration, the concentration of nitrate-N in leaves and roots were insignificantly changed but that of stems increased. The accumulation of total and reduced nitrogen in all organs with increasing concentrations of nitrogen supply were increased at 30 days after treatment but those of all organs at 60 and 90 days after treatment were highest in 600 $mg\;L^{-1}$, whereas there were no significant changes among other nitrogen concentration. The in vivo (${+NO_3}^-$) NRA of all organs did not relate to nitrogen concentration but the in vivo (${-NO_3}^-$) NRA of leaves except roots increased with increasing the nitrogen concentration. Therefore, the proper nitrogen concentration to promote growth and nitrate reduction of pear tree was 200 $mg\;L^{-1}$.
Insects constitute the largest and most diverse group of animals in the world. They also serve as the hosts or nutrient sources for an immense assemblage of pathogens, parasites, and predators. More than 700 fungal species from 100 genera have adopted an entomopathogenic lifestyle. Although entomopathogenic fungi were studied as only biocontrol agents against a variety of pests in various countries, it has been recently focused their additional roles in nature. They are antagonists to/against plant pathogens, endophytes, and possibly even plant growth promoting agents. The potential antimicrobial effect against fungal plant pathogens by an isolate of entomopathogenic fungi including Beauveria bassiana, Lecanicillium spp., and Isaria fumosorosea have been reported since late 1990s, but wasn't reported pathogenicity of the isolate against pests. Later, a Canadian Lecanicillium sp. isolate and L. longisporium isolated from Vertalec$^{(R)}$ showed simultaneous control effect against both aphid and cucumber powder mildew. Therefore, the antimicrobial activities of 342 fungi isolates collected from various regions and conditions in Korea were evaluated against plant pathogenic fungus Botrytis cinerea using dual culture technique on agar plate. As a result, 186 isolates (54.4%) shown the antifungal activity against B. cinerea. The culture filtrates of selected fungi completely suppressed the growth of the microorganisms, indicating that suppression was due to the presence of antimicrobial substances in the culture filtrate. Mode of action of these fungi against insect involves the attachment of conidia to the insect cuticle, followed by germination, cuticle penetration, and internal dissemination throughout the insect. During infection process, secreted enzymes, proteinous toxins, and/or secondary metabolites secreted by entomopathogenic fungi can be used to overcome the host immune system, modify host behavior, and defend host resources. Recently, secondary metabolites isolated from entomopathogenic fungi have been reported as potential bioactive substances. Generally, most of bioactive substances produced by entomopathogenic fungi have reported low molecular weight (lower than 1,000 g/mol) as peptide and, in contrast the high molecular weight fungal bioactive substances are rare. Most substances based on entomopathogenic fungi were shown antimicrobial activity with narrow control ranges. In our study we analyzed the antimicrobial substances having antagonistic effects to B. cinerea. Antimicrobial substances in our fungal culture filtrates showed high thermostability, high stability to proteolytic enzymes, and hydrophilicity and their molecular weights were differed from substance. In conclusion, entomopathogenic fungi showed pathogenicity against insect pests and culture filtrate of the fungi also shown to antimicrobial activity. In the future, we can use the entomopathogenic fungi and its secondary metabolites to control both insect pest control and plant pathogenic fungi simultaneously.
Lee, Da Young;Min, Jin Woo;Joo, Gwang Sik;Kang, Hee Cheol
Korean Journal of Medicinal Crop Science
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v.25
no.2
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pp.95-101
/
2017
Background: Callus cultivation has the advantage of producing a large amount of tissue of a plant in a laboratory regardless of the environment, for extracting an active substance. In the present study, callus formation was induced in the leaves of the succulent plant Adenium obesum (Forssk.) Roem & Schult. After callus cultivation, anti-inflammatory activity tests were conducted, because leaves and stems of A. obesum have been reported to possess biological activity. Methods and Results: In order to induce callus formation, various concentrations of plant growth factors, such as kinetin, naphtha-leneacetic acid (NAA), 6-benzyladenine (BA), and indole-3-acetic acid (IAA) were added to MS solid medium. The maximum callus proliferation was induced by mixed medium consisting of NAA ($2mg/{\ell}$) and BA ($1mg/{\ell}$). In addition, an elicitor was added to the medium under optimal conditions for initiating suspension culture. After suspension culturing, the activities of the callus extracts were compared and analyzed. The cytotoxicity and anti-inflammatory activity tests revealed that the anti-inflammatory activity of the callus extract and the content of phenolic compounds were elevated after treatment of the callus culture with the elicitior. Conclusions: A. obesum callus might be considered as potential source of biologically active anti-inflammatory material.
This study aims to evaluate the antitumor effect of Adonis multiflora, one of the plants in the Ranunculaceae, on mice to which hepatoma cells were transplanted and to suggest its possibility as a candidate natural substance to replace antitumor drugs. We performed the MTT assay to assess the extract had a decrease in the growth rate of hepatoma cells depending on concentration. In particular, 100 ㎍/㎖ of the extract showed 40% of growth retardation rate. We assessed the autophagy activity to identify the inhibitory autophagy mechanism of tumor cells in the extract. This proved that the activity increases more as the concentration of the extract is higher. We conducted the Western blot test to confirmed the expression of two proteins LC3 and p62. The expression of p62 was in inverse proportion to the concentration of the extract whereas LC3-Ⅱ increased more as the concentration of the extract was higher. This showed that an increase in the autophagy relies on the conentration of the extract. We performed a test to discover the influence of the extracts on hepatoma cells transplanted to mice. The test proved that the extract triggers a significant decrease in the growth rate of tumor cells. Compared to the start of the test, the size of tumor cells with 50, 100 and 200 ㎎/㎏ of the extract respectively increased by 4, 3.7 and 3.5 times whereas in the controlling group by 6.3 times. The size of tumor cells in benign tumor controlling group increased by 3.1 times. This showed a significant decrease in the growth rate of tumor cells compared to the controlling group. We carried out the experiment of influence of the extract on the expression of two proteins LC3 and p62 in the tumor tissue transplanted into mice. The experiment showed that LC3-II increases more as the concentration of the extract is higher. However, there was a rapid decrease in p62 with 200 ㎎/㎏ of the extract compared to the controlling group. In this study, we proved that the autophagy activity of Adonis multiflora extract inhibits the growth of hepatoma cells by in vitro and in vivo experiments. In conclusion, the inhibitory autophagy mechanism of tumor cells in the extract can be used as a new treatment of antitumor.
A phytotoxin was purified by repeated chromatography from liquid cultures of Fusarium oxysporum BG isolated from barnyardgrass. Its chemical structure was determined to be dehydrofusaric acid by mass and NMR spectral analyses. The substance showed a potent phytotoxic activity against growth of duckweed with a $EC_{50}$ value of $1.5{\mu}g/ml$. It also inhibited the root growth of barnyard millet, cress, barnyard grass, and rice cultivar 'Dongjin'. However, it had no inhibitory activity against seed germination of barnyard millet and cress, and the shoot growth of the four plant species.
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