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Selection and Genetic Relationship of Salt Tolerant Rice Mutants by in vitro Mutagenesis

  • Song, Jae Young;Kim, Dong Sub;Lee, Myung-Chul;Lee, Kyung Jun;Kim, Jin-Baek;Kim, Sang Hoon;Yun, Song Joong;Kang, Si-Yong
    • Journal of Radiation Industry
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    • v.4 no.4
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    • pp.307-312
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    • 2010
  • Plants have evolved physiological, biochemical and metabolic mechanisms to increase their survival under the adverse conditions. This present study has been performed to select salt-tolerant rice mutant lines through in vivo and in vitro mutagenesis with gamma-rays. For the selection of the salt-tolerant rice mutants, we conducted three times of selection procedure using 1,500 gamma ray mutant lines resulted from an embryo culture of the original rice cv. Dongan (wild-type, WT): first, selection in the a nutrient solution with 171 mM NaCl; second, selection under in vitro condition with 171 mM NaCl; and third, selection in a reclaimed saline land. Based on a growth comparison of the entries, out of the mutant lines, two putative 2 salt tolerant (ST) rice mutant lines, ST-87 and ST-301, were finally selected. The survival rate of the WT, ST-87 and ST-301 were 36.6%, 60% and 66.3% after 7 days in 171 mM NaCl treatment, respectively. The WT and two salt tolerant mutant lines were used to analyze their genetic variations. A total of 21 EcoRI and Msel primer combinations were used to analyze the genetic relationship of among the two salt-tolerant lines and the WT using the ABI3130 capillary electrophoresis system. In the AFLP analysis, a total of 1469 bands were produced by the 21 primer combinations, and 700 (47.6%) of them were identified as having polymorphism. The genetic similarity coefficients were ranged from 0.52 between the ST-87 and WT to 0.24 between the ST-301 and the WT. These rice mutant lines will be used as a control plot for physiological analysis and genetic research on salt tolerance.

Ethanol changes atpB gene expression and proton permeability in Streptococcus mutans (에탄올이 Streptococcus mutans의 atpB 유전자 발현 및 양성자 투과성에 미치는 영향)

  • Cho, Chul Min;Park, Yong Jin;Lee, Sae A;Kim, Jin Bom;Kang, Jung Sook
    • Journal of Korean Academy of Oral Health
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    • v.42 no.4
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    • pp.224-228
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    • 2018
  • Objectives: As a first step to study the anticaries effect of ethanol alone, we investigated the effects of ethanol on the expression levels of the atpB gene and proton permeability of Streptococcus mutans in suspension cultures. Methods: S. mutans UA159 was grown in brain heart infusion medium at either pH 4.8 or 6.8. The total extracted RNA was reverse-transcribed into cDNA using a $Superscript^{TM}$ First-Strand Synthesis System. The resulting cDNA and negative controls were amplified by ABI PRISM 7700 real-time PCR system with SYBR Green PCR Master Mix. For proton flux assay, bacterial suspensions were titrated to pH 4.6 with 0.5 M HCl, and then additional 0.5 M HCl was added to decrease the pH values by approximately 0.4 units. The subsequent increase in pH was monitored using a glass electrode. Ten percent (v/v) butanol was added to the suspensions at 80 min to disrupt the cell membrane. Results: In a concentration-dependent manner, ethanol alone not only decreased the growth rate of S. mutans and the expression of the atpB gene but also increased the proton permeability at both pH 4.8 and 6.8. Conclusions: These findings suggest that ethanol has the potential for an anticaries ingredient. We believe that ethanol may be used together with fluoride and/or other cariostatic agents in order to develop better anticaries toothpastes and/or mouthrinses.

The Effects of Argument-Based Inquiry Activities On Elementary School Students' Claims and Evidence in Science Writing (논의기반 탐구활동이 초등학생의 과학 글쓰기에 나타나는 주장과 증거에 미치는 영향)

  • Park, Jiaeng;Jung, Dojun;Kim, Geonu;Jun, Jaekyoung;Nam, Jeonghee
    • Journal of the Korean Chemical Society
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    • v.64 no.6
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    • pp.389-400
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    • 2020
  • The purpose of this study was to examine the effects of Argument-based Inquiry activities on the claims and evidence in elementary students' science writing. Participants were thirty three fifth grade elementary school students and argument-based inquiry activities on five topics were implemented. We analyzed the Summary Writing samples written by students to investigate the effect of the Argument-based inquiry activities on elementary students' claims and evidence in their science writing, and also analyzed the writing samples of the experimental group to which the Argument-based inquiry activities were implemented, to examine the change of claims and evidence. The results of this study showed that the mean of experimental group was significantly higher than that of the comparison group. As a result of analyzing claims and evidence in Summary Writing of experimental group, the level of claim and evidence has tended to increase gradually as the number of classes progresses.

Two novel mutations in ALDH18A1 and SPG11 genes found by whole-exome sequencing in spastic paraplegia disease patients in Iran

  • Komachali, Sajad Rafiee;Siahpoosh, Zakieh;Salehi, Mansoor
    • Genomics & Informatics
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    • v.20 no.3
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    • pp.30.1-30.9
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    • 2022
  • Hereditary spastic paraplegia is a not common inherited neurological disorder with heterogeneous clinical expressions. ALDH18A1 (located on 10q24.1) gene-related spastic paraplegias (SPG9A and SPG9B) are rare metabolic disorders caused by dominant and recessive mutations that have been found recently. Autosomal recessive hereditary spastic paraplegia is a common and clinical type of familial spastic paraplegia linked to the SPG11 locus (locates on 15q21.1). There are different symptoms of spastic paraplegia, such as muscle atrophy, moderate mental retardation, short stature, balance problem, and lower limb weakness. Our first proband involves a 45 years old man and our second proband involves a 20 years old woman both are affected by spastic paraplegia disease. Genomic DNA was extracted from the peripheral blood of the patients, their parents, and their siblings using a filter-based methodology and quantified and used for molecular analysis and sequencing. Sequencing libraries were generated using Agilent SureSelect Human All ExonV7 kit, and the qualified libraries are fed into NovaSeq 6000 Illumina sequencers. Sanger sequencing was performed by an ABI prism 3730 sequencer. Here, for the first time, we report two cases, the first one which contains likely pathogenic NM_002860: c.475C>T: p.R159X mutation of the ALDH18A1 and the second one has likely pathogenic NM_001160227.2: c.5454dupA: p.Glu1819Argfs Ter11 mutation of the SPG11 gene and also was identified by the whole-exome sequencing and confirmed by Sanger sequencing. Our aim with this study was to confirm that these two novel variants are direct causes of spastic paraplegia.

Aeroelastic modeling to investigate the wind-induced response of a multi-span transmission lines system

  • Azzi, Ziad;Elawady, Amal;Irwin, Peter;Chowdhury, Arindam Gan;Shdid, Caesar Abi
    • Wind and Structures
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    • v.34 no.2
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    • pp.231-257
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    • 2022
  • Transmission lines systems are important components of the electrical power infrastructure. However, these systems are vulnerable to damage from high wind events such as hurricanes. This study presents the results from a 1:50 scale aeroelastic model of a multi-span transmission lines system subjected to simulated hurricane winds. The transmission lines system considered in this study consists of three lattice towers, four spans of conductors and two end-frames. The aeroelastic tests were conducted at the NSF NHERI Wall of Wind Experimental Facility (WOW EF) at the Florida International University (FIU). A horizontal distortion scaling technique was used in order to fit the entire model on the WOW turntable. The system was tested at various wind speeds ranging from 35 m/s to 78 m/s (equivalent full-scale speeds) for varying wind directions. A system identification (SID) technique was used to evaluate experimental-based along-wind aerodynamic damping coefficients and compare with their theoretical counterparts. Comparisons were done for two aeroelastic models: (i) a self-supported lattice tower, and (ii) a multi-span transmission lines system. A buffeting analysis was conducted to estimate the response of the conductors and compare it to measured experimental values. The responses of the single lattice tower and the multi-span transmission lines system were compared. The coupling effects seem to drastically change the aerodynamic damping of the system, compared to the single lattice tower case. The estimation of the drag forces on the conductors are in good agreement with their experimental counterparts. The incorporation of the change in turbulence intensity along the height of the towers appears to better estimate the response of the transmission tower, in comparison with previous methods which assumed constant turbulence intensity. Dynamic amplification factors and gust effect factors were computed, and comparisons were made with code specific values. The resonance contribution is shown to reach a maximum of 18% and 30% of the peak response of the stand-alone tower and entire system, respectively.

Bonding effects of cleaning protocols and time-point of acid etching on dentin impregnated with endodontic sealer

  • Tatiane Miranda Manzoli;Joissi Ferrari Zaniboni;Joao Felipe Besegato;Flavia Angelica Guiotti;Andrea Abi Rached Dantas;Milton Carlos Kuga
    • Restorative Dentistry and Endodontics
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    • v.47 no.2
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    • pp.21.1-21.11
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    • 2022
  • Objectives: This study aimed to investigate the bonding effects of cleaning protocols on dentin impregnated with endodontic sealer residues using ethanol (E) or xylol (X). The effects of dentin acid etching immediately (I) or 7 days (P) after cleaning were also evaluated. For bonding to dentin, universal adhesive (Scotchbond Universal; 3M ESPE) was used. The persistence of sealer residues, hybrid layer formation and microshear bond strength were the performed analysis. Materials and Methods: One hundred and twenty bovine dentin specimens were allocated into 4 groups (n = 10): G1 (E+I); G2 (X+I); G3 (E+P); and G4 (X+P). The persistence of sealer residues was evaluated by SEM. Confocal laser scanning microscopy images were taken to measure the formed hybrid layer using the Image J program. For microshear bond strength, 4 resin composite cylinders were placed over the dentin after the cleaning protocols. ANOVA followed by Tukey test and Kruskal-Wallis followed by Dunn test were used for parametric and non-parametric data, respectively (α = 5%). Results: G2 and G4 groups showed a lower persistence of residues (p < 0.05) and thicker hybrid layer than the other groups (p < 0.05). No bond strength differences among all groups were observed (p > 0.05). Conclusions: Dentin cleaning using xylol, regardless of the time-point of acid etching, provided lower persistence of residues over the surface and thicker hybrid layer. However, the bond strength of the universal adhesive system in etch-and-rinse strategy was not influenced by the cleaning protocols or time-point of acid etching.

DNA Sequence Variation of Candidate Gene for Salt Tolerance in Soybean Mutant

  • Chang Yeok Moon;Byeong Hee Kang;Woon Ji Kim;Sreeparna Chowdhury;Sehee Kang;Seo Young Shin;Wonho Lee;Hyeon-Seok Lee;Bo-Keun Ha
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.259-259
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    • 2022
  • Soil salinity is a major factor that reduces crop yields. The amount of soil affected by salinity is about 83 million hectares (FAO 2000), which is increasing due to the effects of climate change. In soybean [Glycine max (L.) Merr.], nutritional properties such as protein, starch, and sucrose content together with biomass and yield tends to reduce due to excessive salt. As a result of QTL mapping using the 169 F2:3 population from the KA-1285 (salt-tolerant) × Daepung (salt-sensitive) in a previous study, two major QTLs (Gm03_39796778 and Gm03_40600088) related to salt tolerance were found on chromosome 3. In this study, the CDS region of the Gmsalt3 gene was analyzed using the ABI 3730x1 DNA Analyzer (Macrogen, Korea). The sequence of Gmsalt3 gene in KA-1285 was compared with Williams 82.a4.vl and PI483463 (Glycine soja). Two transversions were found at exon6 in KA-1285 and PI483463. Currently, whole genome sequencing and variation analysis using the Illumine Novaseq 6000 machine (Illumina, USA) are in progress. The results of this study can provide useful molecular markers for the selection of salt-tolerant soybeans and can be used as basic data for future salt-tolerant gene research.

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The Genes Expression Patterns Induced by High Temperature in Licorice (Glycyrrhiza uralensis F.) (온도상승에 따른 감초(Glycyrrhiza uralensis Fisch.)의 유전자 발현 양상)

  • Hyeju Seong;Woosuk Jung
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2020.12a
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    • pp.56-56
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    • 2020
  • 감초는 다년생 콩과(Leguminocae) 식물로 국내에서 시중가격이 높은 만주감초가 일부 재배되고 있다. 우리나라에서 감초 재배법이 불완전한 상황에서 한반도의 기후변화에 의한 온도 상승은 약용작물의 생산 및 품질에 많은 영향을 미칠 것으로 예상되므로 본 연구에서는 재배환경 중 온도 조건만 조절할 수 있는 온도구배터널(temperature gradient tunnel system)을 이용하여 4개의 T1(외기온도+0.5~1.3℃), T2(+1.3~2.2℃), T3(+2.2~3.2℃), T4(+3.2~4.0℃) 처리로 온도구배 하여 4년생 만주감초(Glycyrrhiza uralensis F.)를 재배하였다. 지하부가 오래된 모주와 신초1의 경우 저온(T1)과 중간구간(T2, T3)에서 초장과 총화수가 우세하였고, 번식이 가장 늦은 신초2의 경우 중간구간(T2, T3)에서의 생육 및 개화반응이 뚜렷했다. 각 온도처리구마다 3개의 감초 개체를 선발하여 모주의 정단으로부터 5개의 성엽을 채취하였다. Reverse transcription quantitative PCR (RT-qPCR)은 AccuPower® GreenStarTM RT-qPCR Master Mix (Bioneer, Korea)를 이용하여 진행되었다. Primer 디자인은 NCBI Primer-blast 프로그램을 사용해 제작하였고 ABI StepOne real time system (Applied Biosystem)의 melting curve analysis에서 one-peak test를 통해 gene specific primer임을 확인하였다. 각 온도처리구의 감초 잎에서 RNA를 추출하였고, RT-qPCR을 통해 감초의 유전자 발현양상을 비교, 분석하였다. Phytochrome interacting factor 4 (PIF4)는 식물 호르몬을 유발하는 전사조절을 조정함으로써 고온 신호전달에 핵심적인 역할을 수행한다. 활성화된 Phytochrome B(PhyB)는 PIF4의 활성을 억제한다고 알려졌다. Eukaryotic initiation factors(eIFs)는 mRNA 번역 개시인자로 유전자 발현과 특정 단백질 생산을 조절하는 역할을 한다. 본 결과는 온도조건에서 반응하는 생리적 변화를 전사체 수준에서 조사 분석하여 생리해석의 기초자료로 활용, 국내 감초 재배를 위한 환경조건 구명 및 적지 선정 기초자료로서 활용을 기대한다.

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Impact of vitamin D supplementation as COVID-19 vaccine adjuvant on sperm parameters and sex hormones in men with idiopathic infertility: Two separate pre-post studies

  • Mahtab Zarepoor;Alireza Nazari;Soheila Pourmasumi
    • Clinical and Experimental Reproductive Medicine
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    • v.51 no.2
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    • pp.125-134
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    • 2024
  • Objective: Vitamin D deficiency is a major problem for human health worldwide. The mechanisms of vitamin D in the male reproductive system are unknown. After coronavirus disease 2019 (COVID-19) vaccines were developed, doubts were raised about their possible effects on male fertility. Based on vitamin D's function in the immune system, its potential role as an adjuvant for COVID-19 vaccines is intriguing. The aims of this study were to assess the effects of vitamin D first on sperm parameters and sex hormones, and then as an immune adjuvant on sperm parameters and sex hormones after study participants had received their second doses of COVID-19 vaccines. Methods: Phase 1 (before the COVID-19 pandemic) included 72 men with idiopathic infertility, and phase 2 had 64 participants who received two doses of COVID-19 vaccines. Both groups were instructed to take 50,000 IU of vitamin D twice monthly for 3 months. Sperm parameters and sex hormones were assessed pre-and post-supplementation. Results: Regular vitamin D intake for 3 months significantly increased the participants' vitamin D levels (p=0.0001). Both phases showed a positive correlation between vitamin D intake and sperm parameters. Vaccination had no negative effects on sperm parameters and sex hormones. Vitamin D was associated with follicle-stimulating hormone (p=0.02) and testosterone (p=0.0001) in phase 2 after treatment. Conclusion: Our results support vitamin D supplementation as an immune adjunct to COVID-19 vaccination for improving sperm parameters and hormone levels. COVID-19 vaccination is not harmful for male fertility potential, and vitamin D is an effective factor for male fertility.

No association between endothelin-1 gene polymorphisms and preeclampsia in Korean population

  • Kim, Shin-Young;Park, So-Yeon;Lim, Ji-Hyae;Yang, Jae-Hyug;Kim, Moon-Young;Park, Hyun-Young;Lee, Kwang-Soo;Ryu, Hyun-Mee
    • Journal of Genetic Medicine
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    • v.5 no.1
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    • pp.34-40
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    • 2008
  • Purpose : Preeclampsia is a major cause of maternal and perinatal mortality and morbidity and is considered to be a multifactorial disorder involving a genetic predisposition and environmental factors. Endothelin-1 (ET-1) is a potent vasoconstrictor peptide, and alterations in the ET-1 system are thought to play a role in triggering the vasoconstriction seen with preeclampsia. The aim of this study was to examine the frequency of the 4 common single-nucleotide polymorphisms (SNPs) (c.1370T>G, c.137_139delinsA, c.3539+2T>C, and c.5665G>T) of the ET-1 gene in normotensive and preeclamptic pregnancies and to investigate whether these SNPs are associated with preeclampsia in pregnant Korean women. Methods : We analyzed blood samples from 206 preeclamptic and 216 normotensive pregnancies using a commercially available SNapShot kit and an ABI Prism 3100 Genetic analyzer. Results : There were no significant differences in genotype or allele frequencies of the 4 SNPs in the ET-1 gene between preeclamptic and normotensive pregnancies. The respective frequencies of the 3 haplotypes (TDTG, GDCT, and TICT; >10% haplotype frequency) were 61%, 13% and 13%, respectively, in preeclampsic pregnancies and 62%, 14% and 12%, respectively, in normotensive pregnancies. The frequencies of these haplotypes were similar for both groups. Using multiple logistic regression analysis, we did not observe an increase in the risk of preeclampsia for the 4 SNPs of the ET-1 gene under either a recessive or dominant model. Conclusion : This study suggests that the 4 SNPs of the ET-1 gene are not associated with an increased risk for preeclampsia in pregnant Korean women.

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