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Crossbreeding and parental lineage influences the diversity and community structure of rice seed endophytes

  • Walitang, Denver I.;Halim, MD Abdul;Kang, Yeongyeong;Kim, Yongheon;Sa, Tongmin
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.161-161
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    • 2017
  • Seed endophytes are very remarkable groups of bacteria for their unique abilities of being vertically transmitted and conserved. As plants attain hybrid vigor and heterosis in the process of crossbreeding, this might also lead to the changes in the community structure and diversity of plant endophytes in the hybrid plants ultimately affecting the endophytes of the seeds. It would be interesting to characterize how seed endophyte composition change over time. The objective of this study is to gain insights into the influence of natural crossbreeding and parental lineage in the seed bacterial endophytic communities of two pure inbred lines exploring contributions of the two most important sources of plant endophytes - colonization from external sources and vertical transmission via seeds. Total genomic DNA was isolated from rice seeds and bacterial DNA was selectively amplified by PCR. The diversity of endophytic bacteria was studied through Terminal-Restriction Fragment Length Polymorphism (T-RFLP) analysis. Diversity between the original parents and the pure inbred line may show significant differences in terms of richness, evenness and diversity indices. Heat maps reveal astonishing contributions of both or either parents (IR29 ${\times}$ Pokkali and AT401 ${\times}$ IR31868) in the shaping of the bacterial seed endophytes of the hybrid, FL478 and IC32, respectively. Most of the T-RFs of the subsequent pure inbred line could be traced to any or both of the parents. Comparison of common and genotype-specific T-RFs of parents and their offspring reveals that majority of the T-RFs are shared suggesting higher transmission of bacterial communities common to both parents. The parents influence the bacterial community of their offspring. Unique T-RFs of the offspring also suggest external sources of colonization particularly as the seeds are cultivated in different ecogeographical locations. This study showed that host parental lines contributed greatly in the shaping of bacterial seed endophytes of their offspring. It also revealed transmission and potential conservation of core seed bacterial endophytes that generally become the dominant microbiota in the succeeding generations of plant hosts.

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Rapid detection of shiga-toxin producing E. coli by bacteriophage amplification assay (박테리오파지 증폭 기법을 활용한 시가 독소 생성 병원성 대장균의 신속 검출)

  • Baek, Da-Yun;Park, Jong-Hyun;Cho, Seok-Cheol;Lee, Young-Duck
    • Korean Journal of Food Science and Technology
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    • v.52 no.1
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    • pp.103-108
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    • 2020
  • Shiga toxin-producing Escherichia coli (STEC) is an important pathogenic bacteria and can cause severe foodborne disease. For STEC detection, conventional culture methods have disadvantages in the fact that conventional culture takes a long time to detect and PCR can also detect dead bacteria. To overcome these problems, we suggest a bacteriophage amplification assay, which utilizes the ability of bacteriophages to infect living cells and their high specificity. We used a combination of six bacteriophages infecting E. coli to make the bacteriophage cocktail and added ferrous ammonium sulfate as a virucidal agent to remove free-bacteriophages. When cherry tomato and paprika were artificially inoculated with the cocktail at a final concentration of around 3 log CFU/mL and were enriched for at least 5 h in mTSB broth with Novobiocin, approximately 2-3 log PFU/mL were detected through the bacteriophage amplification assay. Therefore, bacteriophage amplification assay might be convenient and a useful method to detect STEC in a short period of time.

The Effects of Armeniacae Arnarum Semen and Platycodi Radix on IL-4, IL-5, IL-6 in Asthma Model (행인(杏仁)과 길경(桔梗)이 Asthma model 내(內)의 Cytokine IL-4, IL-5, IL-6에 미치는 영향(影響))

  • Jung, Hee-Jae;Jung, Sung-Ki;Rhee, Hyung-Koo;Chung, Wook
    • The Journal of Internal Korean Medicine
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    • v.21 no.1
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    • pp.31-38
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    • 2000
  • Background : Nowadays asthma is considered to be the inflammatory disease characterized by airway hyperresponsiveness and pulmonary eosinophilia, and mediated by Th lymphocytes expressing the Th2 cytokine pattern. In many recent studies, molecular biological methods have been used to investigate the role of cytokines in pathogenesis and new therapeutic targets of asthma. Objective : We aimed to identify the effect of Armeniacae Arnarum Semen and Platycodi Radix on the transcriptional activities of cytokine IL-4, IL-5 and IL-6 involved in asthma model. Materials and Methods : RBL-2H3 cell lines were used. Cells were stimulated with calcium inophore for maximal gene expression. After 24 hours of Armeniacae Arnarum Semen and Platycodi Radix-treatment, total cellular RNAs were collected using Trizol solution method. Then transcriptional activities of IL-4, IL-S and IL-6 were measured by RT-PCR with electrophoresis. Results : In IL-4 study, Armeniacae Arnarum Semen treated group showed 48.4% of transcriptional activities compared to the control group and Platycodi Radix treated group showed 45.4% of transcriptional activities compared to the control group. In IL-5 study, Armeniacae Arnarum Semen treated group showed 52.7%of transcriptional activities compared to the control group and Platycodi Radix treated group showed 60.2% of transcriptional activities compared to the control group. In IL-6 study, Armeniacae Amarum Semen treated group showed 42.3% of transcriptional activities compared to the control group and Platycodi Radix treated group showed 69.1% of transcriptional activities compared to the control group. Conclusion : This study shows that Armeniacae Arnarum Semen and Platycodi Radix have the inhibitory effect on the transcription of IL-4, IL-5 and IL-6 gene expression in RBL-2H3 cell lines. Advanced studies are required to investigate the mechanisms of inhibition by herbal medicine in asthma model.

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Changes in mRNA Expression of Obesity-related Genes by GyeongshinhaeGihwan 1 (GGT1) in hGHTg (human growth hormone transgenic) obese Female Rats (암컷 hGHTg 비만 쥐에서 경신해지환(輕身解脂丸) (GGT1)에 의한 비만관련 유전자 mRNA 발현의 변화)

  • Yoon, Ki-Hyeon;Yoon, Mi-Chung;Kim, Hoon;Shin, Soon-Shik
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.2
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    • pp.383-387
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    • 2006
  • To investigate the effect of GyeongshinhaeGihwan 1(GGT1) frequently used as an anti-obesity herbal medicine in oriental medicine on the expression of obesity-related genes, we measured the changes in mRNA levels of these genes by GGT1 in human growth hormone transgenic (hGHTg) obese female rats, and these effects by GGT1 were compared with those of reductil (RD), an anti-obesity drug approved by FDA. Rats received once daily oral administrations of autoclaved water, RD, or GGT1 for 8 weeks. At the end of study, rats were sacrificed and tissues were harvested. Total RNA from adipose tissue, liver and kidney was prepared and the mRNA levels for LPL (lipoprotein lipase), $PPAR{\gamma}$ (peroxisome proliferator activated receptor-gamma), $PPAR{\delta}$ (peroxisome proliferator activated receptor-delta), leptin, $TNF{\alpha}$ (tumor necrosis factor-alpha), and internal standard G3PDH (glyceraldehyde-3-phosphate dehydrogenase) were analyzed by RT-PCR. Compared with control group, $PPAR{\gamma}$ mRNA levels of liver and kidney were decreased in both RD and GGT1 groups, and the effects were more prominent in GGT1 group than in RD group, suggesting that GGT1 is effective in the inhibition of lipid storage by decreasing the $PPAR{\gamma}$ expression. $PPAR{\delta}$ mRNA levels of adipose tissue were increased by RD and GGT1 compared with DW, and the magnitude of increase were higher in GGT1 group than in RD group, indicating that GGT1 stimulates fatty acid oxidation and energy metabolism by activating $PPAR{\delta}$ expression. GGT1 group had higher concentrations of serum leptin, a well-known inhibitor of appetite, than control and RD groups. However, The mRNA levels of leptin, LPL, and $TNF{\alpha}$ were not changed by GGT1. These results indicate that GGT1 can prevent obesity in hGHTg obese female rats by down-regulating and up-regulating the mRNA expression of $PPAR{\gamma}$ and $PPAR{\delta}$, respectively, and that this anti-obesity effects were more pronounced in GGT1 group compared with RD group. In addition, GGT1 seems to inhibit obesity by increasing the circulating leptin levels.

Association of the G134A and G184C Polymorphisms in the CYP1A1 Gene with Lung Cancer Incidence

  • Ryu, Doug-Young;Huang, Ming-Ai;Park, Chang-Bo;Chang, Soo-Im;Im, Ruth;Choi, Seong-Jin;Kim, Na-Young;Park, In-Won;Choi, Byoung-Whui;Kim, Jae-Yeol;Shin, Jong-Wook;Choi, Jae-Chul;Choi, Byung-Sun;Park, Jung-Dock
    • Toxicological Research
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    • v.24 no.2
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    • pp.109-112
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    • 2008
  • The G184C and G134A single nucleotide polymorphisms(SNPs) of the CYP1A1 gene result in Ala62Pro and Gly45Asp substitutions, respectively. Here, we tested whether these SNPs are associated with an alteration in lung cancer incidence. We examined 80 Korean subjects with lung cancer and 240 age- and sex-matched controls. For each subject, the CYP1A1 gene was PCR amplified and sequenced. We observed that the odds ratio(OR) for lung cancer was 3.37 higher in subjects with the G184C polymorphism than in controls(95% confidence interval(CI), $0.89{\sim}12.73$, P=0.07). In contrast, the OR for lung cancer was 1.23 in subjects with the G134A polymorphism compared to controls(95% CI, $0.68{\sim}2.20$, P=0.49). The G184C polymorphism exacerbated the effects of smoking on lung cancer development. Gene-smoking interaction analyses revealed that past or present smokers with the G184C polymorphism had a higher incidence of lung cancer(OR, 24.72; 95% CI, $4.48{\sim}136.31$; P<0.01) than control smokers(OR, 6.65; 95% CI, $2.72{\sim}16.28$; P<0.01). However, there was only a slight difference in the ORs for lung cancer between control smokers and smokers with the G134A polymorphism. These findings suggest that the G184C polymorphism, but not the G134A polymorphism, is associated with an increased risk of lung cancer.

Genetic variation of halophyte New Zealand spinach (Tetragonia tetragonioides) accessions collected in Korea using an AFLP marker (AFLP 마커를 이용한 국내수집 염생식물 번행초 유전다양성 평가)

  • Jeon, Yongsam;Jin, Yong-Tae;Choi, Seo-Hee;Park, Nuri;Kim, In-Kyung;Lee, Ka Youn;Choi, Jong-Jin;Lee, Geung-Joo
    • Journal of Plant Biotechnology
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    • v.43 no.2
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    • pp.157-163
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    • 2016
  • This study was conducted to investigate the potential use of New Zealand spinach (Tetragonia tetragonioides) as a new vegetable crop which will be cultivated in salt-affected soils such as reclaimed areas. New Zealand spinach ecotypes native to Korea were collected across the Southern, Western and Eastern seashore regions of the Korean peninsula, among which fifty-five accessions were later further propagated and evaluated genetically by using an AFLP (amplified fragment length polymorphism) marker. Based on the AFLP analysis performed to uncover the genetic diversity of the collected ecotypes, enzymatic cleavage of the extracted DNA was implemented based on 12 EcoRI and MseI combinations. A total of 1,279 alleles (107 alleles per EcoRI and MseI enzyme combination) were successfully amplified, among which 62 alleles per enzyme combination were polymorphic (58%). The AFLP analysis indicated that the rate of genetic dissimilarity was 29% among the New Zealand spinach collections, which were clustered into the 7 genetic diversity group. This is the first report on the genetic variation in the genus Tetragonia, and the basic information can be applied to select parental lines for enhancing the segregation spectrum of the new halophytic vegetable plant grown in salt-affected areas.

Gene expression and SNP identification related to leaf angle traits using a genome-wide association study in rice (Oryza sativa L.) (GWAS 분석을 이용한 벼 지엽각 관련 SNP 동정 및 발현 분석)

  • Kim, Me-Sun;Yu, Yeisoo;Kang, Kwon-Kyoo;Cho, Yong-Gu
    • Journal of Plant Biotechnology
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    • v.45 no.1
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    • pp.17-29
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    • 2018
  • This study was conducted to investigate a morphological trait in 294 rice accessions including Korean breeding lines. We also carried out a genome-wide association study (GWAS) to detect significant single nucleotide polymorphism markers and candidate genes affecting major agronomic traits. A Manhattan plot analysis of GWAS using morphological traits showed that phenotypic and statistical significance was associated with a chromosome in each group. The significance of SNPs that were detected in this study was investigated by comparing them with those found previously studied QTL regions related to agronomic traits. As a result, SNP (S8-19815442), which is significant with regard to leaf angle, was located in the known QTL regions. To observe gene mutations related to leaf angle in a candidate gene, Os08g31950, its sequences were compared with sequences in previously selected rice varieties. In Os08g31950, a single nucleotide mutation occurred in one region. To compare relative RNA expression levels of candidate gene Os08g31950, obtained from GWAS analysis of 294 rice accessions and related to lateral leaf angle, we investigated relative levels by selecting 10 erect leaf angle varieties and 10 horizontal leaf angle varieties and examining real-time PCR. In Os08g31950, a high level of expression and various expression patterns were observed in all tissues. Also, Os08g31950 showed higher expression levels in the erect leaf angle variety group and higher expression rates in the leaf than in the root. The candidate gene detected through GWAS would be useful in developing new rice varieties with improved yield potential through future molecular breeding.

Prevalence of Feline Panleukopenia Virus in Stray and Household Cats in Seoul, Korea (한국의 서울에 사는 길 고양이와 집 고양이에서 고양이 범백혈구감소증 바이러스의 유병률)

  • Kim, Seung-Gon;Lee, Kyo-Im;Kim, Ha-Jung;Park, Hee-Myung
    • Journal of Veterinary Clinics
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    • v.30 no.5
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    • pp.333-338
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    • 2013
  • We investigated the prevalence of feline panleukopenia virus (FPV) in stray and household cats in different regions of Seoul, Republic of Korea. Blood samples were collected from a total of 200 cats (100 stray cats and 100 household cats) and examined by polymerase chain reaction (PCR). The overall prevalence of FPV was 2%. Among test-positive cats, 3% (3/100) were stray cats and 1% (1/100) was a household cat. The incidence of FPV was higher in juvenile cats (< 1 year, 1.5%) than in adult cats (> 1-year-old, 0.5%). The FPV-positive rates of healthy infected cats and sick cats were 1.9% (3/156) and 2.2% (1/44), respectively. We found the positive rate of vaccinated and unvaccinated cats to be 1.3% (1/77) and 2.4% (3/123), respectively. Unlike antibody tests, FPV antigen tests detected current infections in stray and household cats. Therefore, these tests can help in disease diagnosis and treatment. To our knowledge, our study is the first to survey the prevalence of FPV in different cat populations across Seoul. We found a high prevalence of FPV infection in stray and juvenile cats. Therefore, proper vaccination and surveillance are important to prevent FPV outbreaks.

Anti-Inflammatory Effects of Vegetable Soup in Murine Macrophage RAW 264.7 Cells (야채스프의 RAW 264.7 세포에서 항염증 효과)

  • Sim, Jae-Geun;Lee, Jae-Hyeok;Shin, Tae-Yong;Shin, Hye-Young;Jeong, Sook-Hee;Kim, Mi-Hye;Ku, Ho-Jun;Park, Jeong-Suk
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.8
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    • pp.1097-1101
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    • 2010
  • The aim of the present study is to investigate the anti-inflammatory effect of a vegetable soup (VS). The present study was designed to determine the effect of the vegetable soup on pro-inflammatory factors such as NO, iNOS and TNF-$\alpha$ in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. The cell toxicity was determined by MTS assay. To evaluate the anti-inflammatory effect of vegetable soup, amount of NO was measured using the NO detection kit and the iNOS expression was measured by reverse transcriptase polymerase chain reaction (RT-PCR). Also, proinflammatory cytokines were measured by ELISA kit. The results showed that the vegetable soup reduced NO, iNOS and TNF-$\alpha$ production without cytotoxicity. Our results suggest that the vegetable soup may have an anti-inflammatory property through suppressing inflammatory mediator productions and appears to be useful to develop the functional food realted to anti-inflammation.

Effects of Petasites japonicus and Momordica charantia L. Extracts on MC3T3-E1 Osteoblastic Cells (머위(Petasites japonicus)와 여주(Momordica charantia L.) 추출물의 MC3T3-E1 조골세포 증식 및 분화에 미치는 효과)

  • Ji, Suk-Hee;Ahn, Do-Hwan;Jun, Mi-Ra
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.2
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    • pp.203-209
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    • 2010
  • In this study, the effects of Petasites japonicus and Momordica charantia L. extracts on MC3T3-ET1 osteoblastic cells were investigated. Since the activity of osteoblastic cell is one of the important factors for bone formation, the cellular proliferation of osteoblast was evaluated by MTT and alkaline phosphatase (ALP) activity. Compared to control, the cell proliferation was elevated to 114% and 112% by the treatment of Petasites japonicus and Momordica charantia L. extracts, respectively at the concentration of $10\;{\mu}g/mL$. The cell differentiation was also measured by alkaline phosphatase (ALP) activity at 3, 7, 14, and 27 days treatments with one of the extracts, respectively. As results, the ALP activity was significantly increased at 3 days, compared to control (p<0.05). To evaluate the effect of Petasites japonicus and Momordica charantia L. extracts on bone nodule formation, MC3T3-E1 cells were cultured in $\alpha$-MEM for 3, 14, and 21 days and then stained by alizarin red. To determine the expression patterns of bone-related proteins during the MC3T3-E1 osteoblast-like cell differentiation, osteoblast cells were cultured in $\alpha$-MEM for 24 hr. RNA was extracted and RT-PCR analysis was performed to examine the expression of OPG, RANKL and osteocalcin. Petasites japonicus extract exhibited the significant increment of osteocalcin compared with the positive control, which suggests that Petasites japonicus may have beneficial effects on bone health through the proliferation of osteoblast cells.