• Title/Summary/Keyword: phenotypic stability

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Phenotypic characterization of pre-harvest sprouting resistance mutants generated by the CRISPR/Cas9-geminiviral replicon system in rice

  • Jong Hee Kim;Jihyeon Yu;Jin Young Kim;Yong Jin Park;Sangsu Bae;Kwon Kyoo Kang;Yu Jin Jung
    • BMB Reports
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    • v.57 no.2
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    • pp.79-85
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    • 2024
  • Pre-harvest sprouting is a critical phenomenon involving germination of seeds in the mother plant before harvest under relative humid conditions and reduced dormancy. In this paper, we generated HDR mutant lines with one region SNP (C/T) and an insertion of 6 bp (GGT/GGTGGCGGC) in OsERF1 genes for pre-harvest sprouting (PHS) resistance using CRISPR/Cas9 and a geminiviral replicon system. The incidence of HDR was 2.6% in transformed calli. T1 seeds were harvested from 12 HDR-induced calli and named ERF1-hdr line. Molecular stability, key agronomic properties, physiological properties, and biochemical properties of target genes in the ERF1-hdr line were investigated for three years. The ERF1-hdr line showed significantly enhanced seed dormancy and pre-harvest sprouting resistance. qRT-PCR analysis suggested that enhanced ABA signaling resulted in a stronger phenotype of PHS resistance. These results indicate that efficient HDR can be achieved through SNP/InDel replacement using a single and modular configuration applicable to different rice targets and other crops. This work demonstrates the potential to replace all genes with elite alleles within one generation and greatly expands our ability to improve agriculturally important traits.

A Study on Proliferation and Phenotypical Stability of Schwann Cell on Keratin/PLGA Film (케라틴이 첨가된 PLGA 필름에서 케라틴 함량별 SC세포의 증식 및 형태유지에 관한 연구)

  • Oh, A-Young;Kim, Soon-Hee;Kim, Yun-Tae;Jeon, Na-Ri;Yang, Jae-Chan;Lee, Sang-Jin;Yoo, James-J.;Van Dyke, Mark;Shin, Hyung-Sik;Rhee, John-M.;Khang, Gil-Son
    • Polymer(Korea)
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    • v.33 no.2
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    • pp.118-123
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    • 2009
  • Keratin contains regulatory molecules that can enhance neuronal cell activity. We fabricated keratin/ PLGA films using 0, 10, 20, and 50 wt% of keratin using solvent casting method. We measured the contact angle of each film and cell proliferation was assayed by counting the cells attached on the film. Adhered cell morphology was confirmed by scanning electron microscope. RT-PCR was conducted to evaluate the gene expression of NF, NSE, and S-100, the Schwann cell markers. The keratin content of 20 and 50 wt% provided higher wettability than PLGA. The 20 wt% keratin was better in cell adhesion and proliferation of SCs than other keratin/PLGA films. The phenotypic stability of SC was maintained with the keratin content of 10 and 20 wt%.

Epigenetic Responses Programmed by Prenatal Stress : $F_1$ Male Rat Model (출생 전 스트레스에 의해 프로그램된 후생학적 반응 : $F_1$ 수컷 흰쥐 모델)

  • Lee, Sung-Ho
    • Development and Reproduction
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    • v.12 no.2
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    • pp.117-124
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    • 2008
  • The efficient strategies to cope with unpredictable and/or harmful environmental changes have been developed by every organism in order to ensure its survival and continuity of it's own species. As a results, all living things on earth maintain dynamically internal stability via a process termed 'homeostasis' among physiological parameters despite of external environment changes. Stress is an emotional and physical response to threat homeostasis. Stress may have not only transient but rather permanent effect on the organism; recent evidence clearly show that prenatal stress could organize or imprint permanently physiological systems without any change in genetic codes, a process known as 'epigenetic programming'. In this review, a series of reproduction-associated events occurred in prenatally stressed male rats such as alteration in the structure of sexually dimorphic brain regions, modification of neurotransmitter metabolism, changes in reproductive endocrine status, and finally, disorders of sexual behavior will be introduced. The fetal brain is highly sensitive to prenatal programming and glucocorticoids in particular have powerful brain-programming properties. The chronic hyperactivation of fetal brain by maternal stress-induced glucocorticoid input will provide new program via increasing the neuroplasticities. This 'increased neuroplasticities' will be the basis for the 'increased phenotypic plasticities' rendering the organism's better adaptation to environmental challenges. In conclusion, organism who experienced 'harsh' environment in his fetal life seems to give up a certain portion of reproductive competence to make good chance of survival in his future life by epigenetic (re)programming.

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Effect of Genetic-Environmental Interaction on Quality of Wheat (소맥(小麥) 품질특성(品質特性)의 유전(遺傳) 및 환경적(環境的) 변이(變異))

  • Chang, Hak-Gil;Kim, Chang-Sik;Hah, Duk-Mo;Shin, Hyo-Sun
    • Korean Journal of Food Science and Technology
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    • v.18 no.1
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    • pp.31-37
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    • 1986
  • Seven cultivars of hard and soft wheat were evaluated by regression analysis for five bread quality characteristics to determine varietal response to environments. The regression coefficients were used as the measure of adaptability, and determination coefficients were used as the measure of stability by models of Eberhart and Russell. Phenotypic, genotypic and environmental correlation coefficient estimated for 6 characters tested in this experiments. Statistical analyses confirmed the strong influence of environment on five bread quality. A significant positive correlation exists between protein content, sedimentation value, pelshenke value and specific loaf volume. High heritability was found for sedimentation value ($h^2=0.747$), protein content ($h^2=0.557$) and specific loaf volume ($h^2=0.551$).

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Study of Viral Effects of the Mycovirus (LeV) and Virus-Free Commercial Line in the Edible Mushroom Lentinula edodes

  • Kim, Jung-Mi;Song, Ha-Yeon;Yun, Suk-Hyun;Lee, Hyun-Suk;Ko, Han-Kyu;Kim, Dae-Hyuk
    • 한국균학회소식:학술대회논문집
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    • 2015.11a
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    • pp.37-37
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    • 2015
  • dsRNA was found in malformed cultures of Lentinula edodes strain FMRI0339, one of the three most popular sawdust cultivated commercial strains of shiitake, and was also found in healthy-looking fruiting bodies and actively growing mycelia. Cloning of the partial genome of the dsRNA revealed the presence of the RdRp sequence of a novel L. edodes mycovirus (LeV), and sequence comparison of the cloned amplicon showed an identical sequence to known RdRp genes of LeV found in strain HKA. The meiotic stability of dsRNA was examined by measuring the ratio of the presence of dsRNA among sexual monokaryotic progeny. More than 40% of the monokaryotic progeny still contained the dsRNA, indicating the persistence of dsRNA during sexual reproduction. Comparing the mycelia growth of monokaryotic progeny suggested that, although variations in the growth rate existed among progeny and virus infection was observed in highly actively growing progeny, there appeared to be a tendency toward a lower frequency of virus incidence in actively growing progeny. This study attempted to cure the edible mushroom L. edodes strain FMRI0339 of the L. edodes mycovirus (LeV) in order to obtain an isogenic virus-free fungal strain as well as a virus-infected strain for comparison. Mycelial fragmentation, followed by being spread on a plate with serial dilutions resulted in a virus-free colony. Viral absence was confirmed with gel electrophoresis after dsRNA-specific virus purification, Northern blot analysis, and PCR using reverse transcriptase (RT-PCR). Once cured, all of fungal cultures remained virus-free over the next two years. Interestingly, the viral titer of LeV varied depending on the culture condition. The titer from the plate culture showed at least a 20-fold higher concentration than that grown in the liquid culture. However, the reduced virus titer in the liquid culture was recovered by transferring the mycelia to a plate containing the same medium. In addition, oxygen-depleted culture conditions resulted in a significant decrease of viral concentration, but not to the extent seen in the submerged liquid culture. Although no $discernable phenotypic changes in colony morphology were observed, virus-cured strains showed significantly higher growth rates and mycelial mass than virus-infected strains. We were also explored effects of LeV on fruiting body formation and mushroom yield. The fruiting body formation yield of virus-free L. edodes was larger than virus-infected L. edodes. These results indicate that LeV infection has a deleterious effect on mycelial growth and fruiting body formation. In addition, we have been investigated host-parasite interaction between L. edodes and its mycovirus interaction to study viral mechanism by establishment of proteomics.

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Isolation and Characterization of Bacillus Strain as a Potential Biocontrol Agent (환경친화적 미생물농약으로서의 잠재성을 가진 세균의 분리 및 특성)

  • Lee, Ye-Ram;Lee, Sang-Mee;Jang, Eun-Young;Hong, Chang-Oh;Kim, Keun-Ki;Park, Hyean-Cheal;g Lee, Sang-Mon;Kim, Young-Gyun;Son, Hong-Joo
    • Journal of Life Science
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    • v.25 no.12
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    • pp.1408-1414
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    • 2015
  • In this study, to retain a stable bacterial inoculant, Bacillus strains showing antifungal activity were screened. The improved production, antifungal mechanism, and stability of the antifungal metabolite by a selected strain, AF4, a potent antagonist against phytopathogenic Botrytis cinerea, were also investigated. The AF4 strain was isolated from rhizospheric soil of hot pepper and identified as Bacillus subtilis by phenotypic characters and 16S rRNA gene analysis. Strain AF4 did not produce antifungal activity in the absence of a nitrogen source and produced antifungal activity at a broad range of temperatures (25-40℃) and pH (7-10). Optimal carbon and nitrogen sources for the production of antifungal activity were glycerol and casein, respectively. Under improved conditions, the maximum antifungal activity was 140±3 AU/ml, which was higher than in the basal medium. Photomicrographs of strain AF4-treated B. cinerea showed morphological abnormalities of fungal mycelia, demonstrating the role of the antifungal metabolite. The B. subtilis AF4 culture exhibited broad antifungal activity against several phytopathogenic fungi. The antifungal activity was heat-, pH-, solvent-, and protease-stable, indicating its nonproteinous nature. These results suggest that B. subtilis AF4 is a potential candidate for the control of phytopathogenic fungi-derived plant diseases.