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Confirmation of Drought Tolerance of Ectopically Expressed AtABF3 Gene in Soybean

  • Kim, Hye Jeong;Cho, Hyun Suk;Pak, Jung Hun;Kwon, Tackmin;Lee, Jai-Heon;Kim, Doh-Hoon;Lee, Dong Hee;Kim, Chang-Gi;Chung, Young-Soo
    • Molecules and Cells
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    • v.41 no.5
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    • pp.413-422
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    • 2018
  • Soybean transgenic plants with ectopically expressed AtABF3 were produced by Agrobacterium-mediated transformation and investigated the effects of AtABF3 expression on drought and salt tolerance. Stable Agrobacterium-mediated soybean transformation was carried based on the half-seed method (Paz et al. 2006). The integration of the transgene was confirmed from the genomic DNA of transformed soybean plants using PCR and the copy number of transgene was determined by Southern blotting using leaf samples from $T_2$ seedlings. In addition to genomic integration, the expression of the transgenes was analyzed by RT-PCR and most of the transgenic lines expressed the transgenes introduced. The chosen two transgenic lines (line #2 and #9) for further experiment showed the substantial drought stress tolerance by surviving even at the end of the 20-day of drought treatment. And the positive relationship between the levels of AtABF3 gene expression and drought-tolerance was confirmed by qRT-PCR and drought tolerance test. The stronger drought tolerance of transgenic lines seemed to be resulted from physiological changes. Transgenic lines #2 and #9 showed ion leakage at a significantly lower level (P < 0.01) than ${\underline{n}}on-{\underline{t}}ransgenic$ (NT) control. In addition, the chlorophyll contents of the leaves of transgenic lines were significantly higher (P < 0.01). The results indicated that their enhanced drought tolerance was due to the prevention of cell membrane damage and maintenance of chlorophyll content. Water loss by transpiration also slowly proceeded in transgenic plants. In microscopic observation, higher stomata closure was confirmed in transgenic lines. Especially, line #9 had 56% of completely closed stomata whereas only 16% were completely open. In subsequent salt tolerance test, the apparently enhanced salt tolerance of transgenic lines was measured in ion leakage rate and chlorophyll contents. Finally, the agronomic characteristics of ectopically expressed AtABF3 transgenic plants ($T_2$) compared to NT plants under regular watering (every 4 days) or low rate of watering condition (every 10 days) was investigated. When watered regularly, the plant height of drought-tolerant line (#9) was shorter than NT plants. However, under the drought condition, total seed weight of line #9 was significantly higher than in NT plants (P < 0.01). Moreover, the pods of NT plants showed severe withering, and most of the pods failed to set normal seeds. All the evidences in the study clearly suggested that overexpression of the AtABF3 gene conferred drought and salt tolerance in major crop soybean, especially under the growth condition of low watering.

Effect of SUS316L Bipolar Plate Corrosion on Contact Resistance and PEMFC Performance (SUS316L 분리판 부식에 의한 접촉저항 및 고분자전해질 연료전지 성능에 미치는 영향)

  • Kim, Junseob;Kim, Junbom
    • Applied Chemistry for Engineering
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    • v.32 no.6
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    • pp.664-670
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    • 2021
  • Stainless steel was applied as bipolar plate (BP) of polymer electrolyte membrane fuel cell (PEMFC) due to high mechanical strength, electrical conductivity, and good machinability. However, stainless steel was corroded and increased contact resistance resulting PEMFC performance decrease. Although the corrosion resistance could be improved by surface treatment such as noble metal coating, there is a disadvantage of cost increase. The stainless steel corrosion behavior and passive layer influence on PEMFC performance should be studied to improve durability and economics of metal bipolar plate. In this study, SUS316L bipolar plate of 25 cm2 active area was manufactured, and experiments were conducted for corrosion behavior at an anode and cathode. The influence of SUS316L BP corrosion on fuel cell performance was measured using the polarization curve, impedance, and contact resistance. The metal ion concentration in drained water was analyzed during fuel cell operation with SUS316L BP. It was confirmed that the corrosion occurs more severely at the anode than at the cathode for SUS316L BP. The contact resistance was increased due to the passivation of SUS316L during fuel cell operation, and metal ions continuously dissolved even after the passive layer formation.

Effect of Nardostachyos Rhizoma on Apoptosis, Differentiation and Proliferation in HL-60 cells

  • Ju Sung-Min;Lee Jun;Choi Ho-Seung;Yoon Sang-Hak;Kim Sung-Hoon;Jeon Byung-Hun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.1
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    • pp.163-170
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    • 2006
  • Nardostachyos Rhizoma (N. Rhizoma) belonging to the family Valerianaceae has been anti-arrhythmic effect, and sedation to the central nerve and a smooth muscle. We reported that the water extract of N. Rhizoma induced apoptotic cell death and differentiation in human promyelocytic leukemia (HL-60) cells. Cytotoxicity of N. Rhizoma was detected only in HL-60 cells (IC50 is about 200 ${\mu}g/ml$). The cytotoxic activity of N. Rhizoma in HL-60 cells was increased in a dose-dependent manner. We used several measures of apoptosis to determine whether these processes were involved in N. Rhizoma-induced apoptotic cell death. The high-dose (200 ${\mu}g/ml$) treatment of N. Rhizoma to HL-60 cells showed cell shrinkage, cell membrane blobbing, apoptotic bodies, and the fragmentation of DNA, suggesting that these cells underwent apoptosis. Treatment of HL-60 cells with N. Rhizoma time-dependently induced activation of caspase-3, caspase-8, and caspase-9 and proteolytic cleavage of poly(ADP-ribose) polymerase. Also, we investigated the effect of N. Rhizoma on cellular differentiation and proliferation in HL-60 cells. Differentiation and proliferation of HL-60 cells was determined through expression of CD11b and CD14 surface antigens using flow cytometry and nitroblue tetrazolium (NBT) assay, and through analysis of cell cycle using propidium iodide assay, respectively. N. Rhizoma induced the differentiation of HL-60 at the low-dose (100 ${\mu}g/ml$) treatment, as shown by increased expression of differentiation surface antigen CD11b, but not CDl4 and increased reducing activity of NBT. When HL-60 cells were treated with N. Rhizoma at concentration of $50{\mu}g/ml\;and\;100{\mu}g/ml$, NBT-reducing activities induced approximately 1.5-fold and 20.0-fold as compared with the control. In contrast, HL-60 cells treated with the N. Rhizoma-ATRA combination showed markedly elevated levels of 26.3-fold at $50{\mu}g/ml$ N. Rhizoma-0.1 ${\mu}M$ ATRA combination and 27.5-fold at 50 ${\mu}g/ml$ N. Rhizoma-0.2 ${\mu}M$ ATRA combination than when treated with N. Rhizoma alone or ATRA alone. It may be that N. Rhizoma plays important roles in synergy with ATRA during differentiation of HL-60 cells. DNA flow-cytometry indicated that N. Rhizoma markedly induced a G1 phase arrest of HL-60 cells. N. Rhizoma-treated HL-60 cells increased the cell population in G1 phase from 32.71% to 42.26%, whereas cell population in G2/M and S phases decreased from 23.61% to 10.33% and from 37.78% to 33.98%, respectively. We examined the change in the $p21^{WAF1/Cip1}\;and\;p27^{Kip1}$ proteins, which are the CKIs related with the G1 phase arrest. The expression of the CDK inhibitor $p27^{Kip1},\;but\;not\;p21^{WAF1/Cip1}$ were markedly increased by N. Rhizoma. Taken together, these results demonstrated that N. Rhizoma induces apoptotic cell death through activation of caspase-3, and potently inhibits the proliferation of HL-60 cells via the G1 phase cell cycle arrest in association with $p27^{Kip1}$ and granulocytic differentiation induction .

Antimicrobial, antifungal effect and safety verification using BCOP assay of extracts from Coptis chinensis (황련(Coptis chinensis) 추출물의 항균, 항진균 효과와 BCOP 분석을 이용한 안전성 검증)

  • Kim, Eun-Hee;Jang, Young-Ah;Kim, Sol-Bi;Kim, Han-Hyuk;Lee, Jin-Tae
    • Journal of Applied Biological Chemistry
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    • v.61 no.3
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    • pp.297-304
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    • 2018
  • Coptis chinensis is used in oriental medicine for soothing, anti-inflammation, antimicrobial and antipyretic properties, and its main ingredient berberine is known to have strong antibacterial activity. In this study, we investigated the anti-microbial effect of hot water extract of Coptis chinensis (CW) on skin related microorganism and the airborne microbe, the antifungal effects of fungi, which are frequently detected in residential environments. CW showed antibacterial effect against Propionibacterium acnes, Staphylococcus aureus and Staphylococcus epidermidis, against the airborne microbe, which was collected in four different places. At the concentration of 100 mg/mL, the antimicrobial activity continued for 42 days, showed heat stability without change in the antimicrobial activity even after heat treatment. The MIC and MBC of CW against S. aureus was 0.03, 0.05 mg/mL, against S. epidermidis was 0.50, 0.75 mg/mL and against P. acne was 0.10, 0.15 mg/mL. As a result of measuring the MIC of four kinds of fungi with high detection frequency in the surrounding environment, Gliocladium virens was 65 mg/mL by determined as MIC which can inhibit one hundred percent of mycelial growth. The concentration 90 mg/mL was determined as MIC against Aureobasidium pullulans and 100 mg/mL against Penicilium pinophilum and Chaetomium globosum. CW was considered a safe extract that showed no irritation even in the ocular mucous membrane irritation evaluation test, a patch test. Therefore, these results suggest that Coptis chinensis has antimicrobial, antifungal and safety on human body and can be applied to the development of materials for cosmetic and residential environment industries.

Distribution of Agalmatolite Mines in South Korea and Their Utilization (한국의 납석 광산 분포 현황 및 활용 방안)

  • Seong-Seung Kang;Taeyoo Na;Jeongdu Noh
    • The Journal of Engineering Geology
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    • v.33 no.4
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    • pp.543-553
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    • 2023
  • The current status of domestic a agalmatolite mines in South Korea was investigated with a view to establishing a stable supply of agalmatolite and managing its demand. Most mined agalmatolite deposits were formed through hydrothermal alteration of Mesozoic volcanic rocks. The physical characteristics of pyrophyllite, the main constituent mineral of agalmatolite, are as follows: specific gravity 2.65~2.90, hardness 1~2, density 1.60~1.80 g/cm3, refractoriness ≥29, and color white, gray, grayish white, grayish green, yellow, or yellowish green. Among the chemical components of domestic agalmatolite, SiO2 and Al2O3 contents are respectively 58.2~67.2 and 23.1~28.8 wt.% for pyrophyllite, 49.2~72.6 and 16.5~31.0 wt.% for pyrophyllite + dickite, 45.1 and 23.3 wt.% for pyrophyllite + illite, 43.1~82.3 and 11.4~35.8 wt.% for illite, and 37.6~69.0 and 19.6~35.3 wt.% for dickite. Domestic agalmatolite mines are concentrated mainly in the southwest and southeast of the Korean Peninsula, with some occurring in the northeast. Twenty-one mines currently produce agalmatolite in South Korea, with reserves in the order of Jeonnam (45.6%) > Chungbuk (30.8%) > Gyeongnam (13.0%) > Gangwon (4.8%), and Gyeongbuk (4.8%). The top 10 agalmatolite-producing mines are in the order of the Central Resources Mine (37.9%) > Wando Mine (25.6%) > Naju Ceramic Mine (13.4%) > Cheongseok-Sajiwon Mine (5.4%) > Gyeongju Mine (5.0%) > Baekam Mine (5.0%) > Minkyung-Nohwado Mine (3.3%) > Bugok Mine (2.3%) > Jinhae Pylphin Mine (2.2%) > Bohae Mine. Agalmatolite has low thermal conductivity, thermal expansion, thermal deformation, and expansion coefficients, low bulk density, high heat and corrosion resistance, and high sterilization and insecticidal efficiency. Accordingly, it is used in fields such as refractory, ceramic, cement additive, sterilization, and insecticide manufacturing and in filling materials. Its scope of use is expanding to high-tech industries, such as water treatment ceramic membranes, diesel exhaust gas-reduction ceramic filters, glass fibers, and LCD panels.