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High frequency direct plant regeneration from leaf, internode, and root segments of Eastern Cottonwood (Populus deltoides)

  • Yadav, Rakesh;Arora, Pooja;Kumar, Dharmendar;Katyal, Dinesh;Dilbaghi, Neeraj;Chaudhury, Ashok
    • Plant Biotechnology Reports
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    • v.3 no.3
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    • pp.175-182
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    • 2009
  • Simple, reproducible, high frequency, improved plant regeneration protocol in Eastern Cottonwood (Populus deltoides) clones, WIMCO199 and L34, has been reported. Initially, aseptic cultures established from axillary buds of nodal segments from mature plus trees on MS liquid medium supplemented with $0.25mg\;1^{-1}$ KIN and $0.25mg\;1^{-1}$ IAA. Nodal and internodal segments were found to be extra-prolific over shoot apices during course of aseptic culture establishment, while $0.25mg\;1^{-1}$ KIN concentration played a stimulatory role in high frequency plant regeneration. Diverse explants, such as various leaf segments, internodes, and roots from in vitro raised cultures, were employed. Direct plant regeneration was at high frequency of 92% in internodes, 88% in leaf segments, and 43% in root segments. This led to the formation of multiple shoot clusters on established culture media with rapid proliferation rates. Many-fold enhanced shoot elongation and growth of the clusters could be achieved on liquid MS medium supplemented with borosilicate glass beads, which offer physical support for proliferating shoots leading to faster growth in comparison to semi-solid agar or direct liquid medium. SEM examination of initial cultures confirmed direct plant regeneration events without intervening calli. In vitro regenerated plants induced roots on half-strength MS medium with $0.15mg\;1^{-1}$ IAA. Rooted 5- to 6-week-old in vitro regenerated plants were transferred into a transgenic greenhouse in pots containing 1:1 mixture of vermicompost and soil at $27{\pm}2^{\circ}C$ for hardening and acclimatization. 14- to 15-week-old well-established hardened plants were transplanted to the field and grown to maturity. The mature in vitro raised poplar trees exhibited a high survival rate of 85%; 4-year-old healthy trees attained an average height of 8 m and an average trunk diameter of 25 cm and have performed well under field conditions. The regeneration protocol presented here will be very useful for undertaking genetic manipulation, providing a value addition to Eastern Cottonwood propagation in future.

Effect of Deep Seawater on Expression of μ-Opioid Receptor in Cultured Rat Hippocampal Neurons (배양된 쥐 해마신경세포에서 μ-아편양 수용체의 발현에 대한 해양심층수의 영향)

  • Moon, Il-Soo;Kim, Seong-Ho
    • Journal of Life Science
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    • v.21 no.2
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    • pp.176-182
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    • 2011
  • Deep seawater (DSW) generally refers to seawater at depths equal to or greater than 200 meters. DSW is rich in inorganic materials which have attracted attention for its various applications. In this study we investigated the effects of the DSW upwelled from the East Sea, offshore Yang Yang (KangWon-do, Korea), on the expression of ${\mu}$-opioid receptor (MOR) of cultured rat hippocampal neurons. Neurons were grown in a minimal essential medium containing 10% (v/v) fetal bovine serum and either 25% (v/v) distilled water, or hardness (H) 800, or H 1000 DSW. Cultures grown in the presence of DSW with H 800 and H 1000 exhibited robust MOR immunoreactive signals in both neurons and astrocytes. Interestingly, the increase in MOR immunoreactive signals was more dramatic in astrocytes than in neurons. Statistical analysis revealed that the relative intensities for MOR clusters increased approximately 4-fold in astrocytes cultured in H 800 and H 1000 media. These increases were statistically very significant (p<0.001). In contrast, the increase in intensities for MOR immunoreactive signals was relatively less dramatic in neurons, where only the increase in the H 1000 culture was statistically very significant (p<0.001). These results indicated that DSW promotes expression of MOR in both neurons and astrocytes, and more significantly in the latter.

Effects of various lights, solvents, and zinc protoporphyrin on the chemical behavior of MTT formazan (빛, 용매와 zinc protoporphyrin에 의한 MTT 포마잔의 화학적 동태 변화)

  • Kim, Joo Hyoun;Hong, Jungil
    • Korean Journal of Food Science and Technology
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    • v.50 no.1
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    • pp.1-7
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    • 2018
  • The MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay is commonly used for analyzing the cell viability. In this study, effects of various solvents, different lights, and zinc protoporphyrin (ZnPP) on the chemical behavior of MTT formazan were investigated. The color response of MTT formazan in NaOH was highly pronounced; the absorbance of MTT formazan in 0.1 N NaOH at 550 nm was >2-fold higher than that in water, dimethyl sulfoxide (DMSO), methanol, and ethanol. MTT formazan in DMSO and NaOH (>0.1 N) was relatively stable under fluorescent and UV light at 365 nm; its rapid degradation was induced under UV light at 254 nm in all solvents. ZnPP degraded MTT formazan under light in a time- and concentration-dependent manner; MTT formazan in 0.1 N NaOH was the most sensitive to ZnPP, followed by DMSO. These results suggest that NaOH and DMSO might be suitable media for MTT formazan for monitoring photosensitizing properties.

A New Green-Kerneled Glutinous Rice Mutant Variety, "Nogwonchalbyeo" Developed by Gamma Ray Irradiation (녹색찰현미로 이용 가능한 "녹원찰벼"의 돌연변이 육종)

  • Kang, Si-Yong;Shin, In Chul;Kim, Dong Sub;Lee, Geung-Joo;Kim, Jin-Baek;Lee, Deok Yul;Lee, Sang Young;Lee, Dong Jin
    • Korean Journal of Breeding Science
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    • v.40 no.3
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    • pp.303-307
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    • 2008
  • We bred a new green-kerneled glutinous rice variety that can be cultivated in the whole area of Korea, because only one native green-kerneled glutinous rice cultivar, "Saengdongchalbyeo", has been cultivated in the southern coastal area due to its late heading. The seeds of "Saengdongchalbyeo" were irradiated with 200 Gy of gamma ray in 1995. A promising mutant variety, "Nogwonchalbyeo" ("Wonnong 17") was selected through line selection and regional yield trials. In particular, the new variety revealed at the earlier mid of August compared to that of "Saengdongchalbyeo", the early of September, and it was considerably tolerant to a field lodging due to its shortened culm length. Also, "Nogwonchalbyeo" had a higher ripened grain ratio and 1,000 grain weight compared to the original variety. The brown grain yield of the new variety was about 5.40 MT/ha, which was 11.3% higher than that of the original variety, in the regional yield trials at 3 different fields during 2000~2001. The brown and milled grains of the new rice variety contained 20 to 65% higher amount of total amino acids, respectively than that of the original and two checks. For chlorophyll -a, -b and total chlorophyll, the new variety showed nearly two-fold higher than the checks, and for the carotenoid, it had 5.3 - 7.6 times higher amount. These results showed that the new variety can be cultivated as a special green-kerneled glutinous rice with high functional compounds.

Identification, sequence characterization and expression analysis of the arginine kinase gene in response to laminarin challenge from the Oriental land snail, Nesiohelix samarangae (동양달팽이(Nesiohelix samarangae)의 arginine kinase 유전자 분석 및 발현 패턴에 관한 연구)

  • Jeong, Ji Eun;Lee, Yong Seok
    • The Korean Journal of Malacology
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    • v.29 no.3
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    • pp.171-179
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    • 2013
  • Arginine kinase (ArK) is known to play an important role in most invertebrates the level of ATP by phosphorylation of phosphagens in cell and immuninty in living organisms. ArK has been identified in many kinds of organisms ranging from invertebrate to vertebrate. However, no ArK gene has been cloned and investigated from N. samarangae. This leads us to identify ArK cDNA (NsArK) from the expressed sequence tag (EST) sequencing of N. samarangae. Sequence analysis indicated that the coding region of 1,065 bp contains 355 amino acid residues. Molecular phylogenetic analysis shows that NsArK had very high similarities with mollusca and arthropoda. In an attempt to investigate a potential role of NsArK in the digestive gland of N. samarangae, expression patterns were analyzed. RT-PCR analsysis shows that NsArK mRNA is induced in the rane of 1.2 fold at 6 hr by laminarin when compared with the control. The immunnologial and physiological role of NsArK remains to be further investigated in N. samarangae.

Effect of Korean pine nut oil on hepatic iron, copper, and zinc status and expression of genes and proteins related to iron absorption in diet-induced obese mice

  • Shin, Sunhye;Lim, Yeseo;Chung, Jayong;Park, Soyoung;Han, Sung Nim
    • Journal of Nutrition and Health
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    • v.54 no.5
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    • pp.435-447
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    • 2021
  • Purpose: Body adiposity is negatively correlated with hepatic iron status, and Korean pine nut oil (PNO) has been reported to reduce adiposity. Therefore, we aimed to study the effects of PNO on adiposity, hepatic mineral status, and the expression of genes and proteins involved in iron absorption. Methods: Five-week-old male C57BL/6 mice were fed a control diet containing 10% kcal from PNO (PC) or soybean oil (SBO; SC), or a high-fat diet (HFD) containing 35% kcal from lard and 10% kcal from PNO (PHFD) or SBO (SHFD). Hepatic iron, copper, and zinc content; and expression of genes and proteins related to iron absorption were measured. Results: HFD-fed mice had a higher white fat mass (2-fold; p < 0.001), lower hepatic iron content (25% lower; p < 0.001), and lower hepatic Hamp (p = 0.028) and duodenal Dcytb mRNA levels (p = 0.037) compared to the control diet-fed mice. Hepatic iron status was negatively correlated with body weight (r = -0.607, p < 0.001) and white fat mass (r = -0.745, p < 0.001). Although the PHFD group gained less body weight (18% less; p < 0.05) and white fat mass (18% less; p < 0.05) than the SHFD group, the hepatic iron status impaired by the HFD feeding did not improve. The expression of hepatic and duodenal ferroportin protein was not affected by the fat amount or the oil type. PNO-fed mice had significantly lower Slc11a2 (p = 0.022) and Slc40a1 expression (p = 0.027) compared to SBO-fed mice. However, the PC group had a higher Heph expression than the SC group (p < 0.05). The hepatic copper and zinc content did not differ between the four diet groups, but hepatic copper content adjusted by body weight was significantly lower in the HFD-fed mice compared to the control diet-fed mice. Conclusion: HFD-induced obesity decreased hepatic iron storage by affecting the regulation of genes related to iron absorption; however, the 18% less white fat mass in the PHFD group was not enough to improve the iron status compared to the SHFD group. The hepatic copper and zinc status was not altered by the fat amount or the oil type.

Optimal Ratio of Wnt3a Expression in Human Mesenchymal Stem Cells Promotes Axonal Regeneration in Spinal Cord Injured Rat Model

  • Yoon, Hyung Ho;Lee, Hyang Ju;Min, Joongkee;Kim, Jeong Hoon;Park, Jin Hoon;Kim, Ji Hyun;Kim, Seong Who;Lee, Heuiran;Jeon, Sang Ryong
    • Journal of Korean Neurosurgical Society
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    • v.64 no.5
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    • pp.705-715
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    • 2021
  • Objective : Through our previous clinical trials, the demonstrated therapeutic effects of MSC in chronic spinal cord injury (SCI) were found to be not sufficient. Therefore, the need to develop stem cell agent with enhanced efficacy is increased. We transplanted enhanced Wnt3-asecreting human mesenchymal stem cells (hMSC) into injured spines at 6 weeks after SCI to improve axonal regeneration in a rat model of chronic SCI. We hypothesized that enhanced Wnt3a protein expression could augment neuro-regeneration after SCI. Methods : Thirty-six Sprague-Dawley rats were injured using an Infinite Horizon (IH) impactor at the T9-10 vertebrae and separated into five groups : 1) phosphate-buffered saline injection (injury only group, n=7); 2) hMSC transplantation (MSC, n=7); 3) hMSC transfected with pLenti vector (without Wnt3a gene) transplantation (pLenti-MSC, n=7); 4) hMSC transfected with Wnt3a gene transplantation (Wnt3a-MSC, n=7); and 5) hMSC transfected with enhanced Wnt3a gene (1.7 fold Wnt3a mRNA expression) transplantation (1.7 Wnt3a-MSC, n=8). Six weeks after SCI, each 5×105 cells/15 µL at 2 points were injected using stereotactic and microsyringe pump. To evaluate functional recovery from SCI, rats underwent Basso-Beattie-Bresnahan (BBB) locomotor test on the first, second, and third days post-injury and then weekly for 14 weeks. Axonal regeneration was assessed using growth-associated protein 43 (GAP43), microtubule-associated protein 2 (MAP2), and neurofilament (NF) immunostaining. Results : Fourteen weeks after injury (8 weeks after transplantation), BBB score of the 1.7 Wnt3a-MSC group (15.0±0.28) was significantly higher than that of the injury only (10.0±0.48), MSC (12.57±0.48), pLenti-MSC (12.42±0.48), and Wnt3a-MSC (13.71±0.61) groups (p<0.05). Immunostaining revealed increased expression of axonal regeneration markers GAP43, MAP2, and NF in the Wnt3a-MSC and 1.7 Wnt3a-MSC groups. Conclusion : Our results showed that enhanced gene expression of Wnt3a in hMSC can potentiate axonal regeneration and improve functional recovery in a rat model of chronic SCI.

Molecular physiological inhibitory effects of chloroacetanilide herbicide pretilachlor on marine dinoflagellate Prorocentrum minimum (해양 와편모조류 Prorocentrum minimum에 대한 아세트아닐라이드계 제초제 프레틸라클로르의 분자 생물학적 저해 효과)

  • Hansol Kim;Jang-Seu Ki
    • Korean Journal of Environmental Biology
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    • v.39 no.4
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    • pp.452-462
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    • 2021
  • Pretilachlor (PRE) is a common acetanilide herbicide used worldwide. However, its effects on aquatic organisms, particularly marine photosynthetic life, are not sufficiently known. Herein, we evaluated the toxic effects of PRE by physiological and molecular parameters in the photosynthetic dinoflagellate Prorocentrum minimum. The cell density, pigment content, and photosynthetic parameters (Fv/Fm and PIABS) were considerably decreased with increased PRE exposure time and doses. In addition, photosynthesis-related genes, PmpsbA, PmpsaA, and PmatpB, were significantly upregulated when exposed to 1.0 mg L-1 of PRE for 24 h (p<0.001). In 72 h treatment, the relative gene expression was significantly increased (0.1 and 0.5 mg L-1; p<0.01). In contrast, PmrbcL was decreased or little changed compared to the controls. Reactive oxygen species (ROS) increased after 24 h exposure (p<0.001). However, the transcriptional fold-changes in glutathione S-transferase (GST) were significantly increased (0.5 and 1.0 mg L-1; p<0.001) at 72 h. These findings suggested that the PmGST might be involved in PRE detoxification in P. minimum. In addition, PRE may affect the photosystem function in phytoplankton similar to other acetanilides, causing severe damage or cell death.

Selection of Aerial Spraying Control Agent and Susceptibility of Pinewood Nematode Vectors to Spraying Using Unmanned Aerial Vehicles (소나무재선충 매개충에 대한 항공방제 약제 탐색 및 무인항공기 살포에 의한 매개충의 약제 감수성)

  • Junheon Kim;Sangjune Nam;Jinyoung Song
    • Korean journal of applied entomology
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    • v.62 no.1
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    • pp.43-48
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    • 2023
  • The efficacy of spinetoram sprayed from an unmanned helicopter to control two insect vectors of pine wood nematodes, Monochamus alternatus and M. saltuarius was assessed. By using a ULV (Ultra Low Volume) sprayer, the mortality rates of phenthoate, bifenthrin, etofenprox, and diflubenzuron, which are registered for use against coleopteran insects, indoxacarb and spinetoram, which are registered for use against lepidopteran insects, were evaluated for efficacy as aerial spraying agents. The ULV test, using 33- and 55-fold dilutions of phenthoate, bifenthrin, indoxacarb, and spinetoram demonstrated a 100% insect mortality rate upon contact and feeding mortality on the third day after treatment, while etofenprox and diflubenzuron exhibited a slightly lower mortality rate than the remaining compounds. Consequently, spinetoram was selected as a candidate pesticide for aerial spraying and its efficacy to control the insect vectors was assessed. The x 33 dilution of spinetoram resulted in a 98.6-100% control efficacy against two insect vectors. However, risk assessment for bees following aerial spraying will be necessary before applying the pesticide to control insect vectors.

The Effect of BMI and Physical Ability on Self-efficacy, Quality of Life, and Self-esteem in Overweight and Obese Children (비만도와 체력이 비만 아동의 자기 효능감, 삶의 질, 자아개념에 미치는 영향)

  • Ahn, Hyun-Sun;Chung, Kyong-Mee;Jeon, Justin
    • Korean Journal of Health Psychology
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    • v.16 no.3
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    • pp.537-555
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    • 2011
  • The purpose of this study was two-fold. First, comparisons on the psychological and physical attributes of normal weight, overweight, and obese children were conducted. Second, the influence of BMI and physical fitness interaction on psychological adaptation in children with obesity was explored. Participants were 245 children between the ages of 9 and 13 years (64.5% males). Data on children considered overweight (n = 45) or obese (n = 78) were gathered from the Korean Obese Children's Physical Activity (KOCPA) projects. Normal weight children (n = 122) were recruited from two Seoul elementary schools. Psychological measurements included Weight Efficacy Life-style Questionnaire (WEL), Physical Self-efficacy Scale (PSES), Child Dietary Self-efficacy Scale (CDSS), Self-concept Inventory (SCI), and the Korean version of the Pediatric Quality of Life Intervention TM Version 4.0 Generic Core Scales (PedsQLTM4.0). BMI (kg/m2) and physical fitness (e.g., aerobic endurance, flexibility) were recorded by experts in exercise physiology. Results showed that children in the high BMI group reported poorer psychological adaption and demonstrated lower physical fitness when compared to the remaining groups. Compared to normal weight children, children considered overweight and obese were found to have lower physical self-efficacy, more negative self-concept, and poorer quality of life. Further, these children also had significantly lower physical fitness levels than their normal weight counterparts. Physical fitness was found to have a significant main effect on weight efficacy (WEL), physical self-efficacy (PSES), and quality of life (PedsQL) in children considered overweight or obese. A significant BMI-physical fitness interaction effect was found for self-concept (SCI) only. Children with higher BMI reported poorer self-concept regardless of fitness level whereas children with lower BMI and higher fitness reported more positive self-concept. Implications and limitations are discussed.