• Title/Summary/Keyword: Seed proteins

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Development of Gene-based Markers for the Allelic Selection of the Restorer-of-fertility Gene, Rfo, in Radish (Raphanus sativus)

  • Kim, Sunggil;Lim, Heerae;Cho, Kang-Hee;Park, Pue Hee;Park, Suhyung;Sung, Soon-Kee;Oh, Daegeun;Kim, Ki-Taek
    • Korean Journal of Breeding Science
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    • v.41 no.3
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    • pp.194-204
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    • 2009
  • Cytoplasmic male sterility (CMS) and fertility restoration have been utilized as valuable tools for $F_1$-hybrid seed production in many crops despite laborious breeding processes. Molecular markers for the selection of CMS-related genes help reduce the expenses and breeding times. A previously reported genomic region containing the Ppr-B gene, which is responsible for restoration of fertility and corresponds to the Rfo locus, was used to develop gene-based or so-called "functional" markers for allelic selection of the restorer-of-fertility gene (Rfo) in $F_1$-hybrid breeding of radish (Raphanus sativus L.) Polymorphic sequences among Rfo alleles of diverse breeding lines of radish were examined by sequencing the Ppr-B alleles. However, presence of Ppr-B homolog, designated as Ppr-D, interferes on specific PCR amplification of Ppr-B in certain breeding lines. The organization of Ppr-D, resolved by genome walking, revealed extended homology with Ppr-B even in the promoter region. Interestingly, PCR amplification of Ppr-D was repeatedly unsuccessful in certain breeding lines implying the lack of Ppr-D in these radishes. Ppr-B could only be successfully amplified for analysis through designing primers based on the sequences unique to Ppr-B that exclude interference from Ppr-D gene. Four variants of Rfo alleles were identified from 20 breeding lines. A combination of three molecular markers was developed in order to genotype the Rfo locus based on polymorphisms among four different variants. These markers will be useful in facilitating $F_1$-hybrid cultivar development in radish.

Overexpression of a Chromatin Architecture-Controlling ATPG7 has Positive Effect on Yield Components in Transgenic Soybean

  • Kim, Hye Jeong;Cho, Hyun Suk;Pak, Jun Hun;Kim, Kook Jin;Lee, Dong Hee;Chung, Young-Soo
    • Plant Breeding and Biotechnology
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    • v.5 no.3
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    • pp.237-242
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    • 2017
  • AT-hook proteins of plant have shown to be involved in growth and development through the modification of chromatin architecture to co-regulate transcription of genes. Recently, many genes encoding AT-hook protein have been identified and their involvement in senescence delay is investigated. In this study, soybean transgenic plants overexpressing chromatin architecture-controlling ATPG7 gene was produced by Agrobacterium-mediated transformation and investigated for the positive effect on the important agronomic traits mainly focusing on yield-related components. A total of 27 transgenic soybean plants were produced from about 400 explants. $T_1$ seeds were harvested from all transgenic plants. In the analysis of genomic DNAs from soybean transformants, ATPG7 and Bar fragments were amplified as expected, 975 bp and 408 bp in size, respectively. And also exact gene expression was confirmed by reverse transcriptase-PCR (RT-PCR) from transgenic line #6, #7 and #8. In a field evaluation of yield components of ATPG7 transgenic plants ($T_3$), higher plant height, more of pod number and greater average total seed weight were observed with statistical significance. The results of this study indicate that the introduction of ATPG7 gene in soybean may have the positive effect on yield components.

Identification of ABSCISIC ACID (ABA) signaling related genes in Panax ginseng

  • Hong, Jeongeui;Kim, Hogyum;Ryu, Hojin
    • Journal of Plant Biotechnology
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    • v.45 no.4
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    • pp.306-314
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    • 2018
  • Korean ginseng (Panax ginseng) has long been cultivated as an important economic medicinal plant. Owing to the seasonal and long-term agricultural cultivation methods of Korean ginseng, they are always vulnerable to various environmental stress conditions. ABSCISIC ACID (ABA) is an essential plant hormone associated with seed development and diverse abiotic stress responses including drought, cold and salinity stress. By modulating ABA responses, plants can regulate their immune responses and growth patterns to increase their ability to tolerate stress. With recent advances in genome sequencing technology, we first reported the functional features of genes related to canonical ABA signaling pathway in P. ginseng genome. Based on the protein sequences and functional genomic analysis of Arabidopsis thaliana, the ABA related genes were successfully identified. Our functional genomic characterizations clearly showed that the ABA signaling related genes consisting the ABA receptor proteins (PgPYLs), kinase family (PgSnRKs) and transcription factors (PgABFs, PgABI3s and PgABI5s) were evolutionary conserved in the P. ginseng genome. We confirmed that overexpressing ABA related genes of P. ginseng completely restored the ABA responses and stress tolerance in ABA defective Arabidopsis mutants. Finally, tissue and age specific spatio-temporal expression patterns of the identified ABA-related genes in P. ginseng tissues were also classified using various available RNA sequencing data. This study provides ABA signal transduction related genes and their functional genomic information related to the growth and development of Korean ginseng. Additionally, the results of this study could be useful in the breeding or artificial selection of ginseng which is resistant to various stresses.

Molecular discrimination of Panax ginseng cultivar K-1 using pathogenesis-related protein 5 gene

  • Wang, Hongtao;Xu, Fengjiao;Wang, Xinqi;Kwon, Woo-Saeng;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.43 no.3
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    • pp.482-487
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    • 2019
  • Background: The mixed-cultivation of different Panax ginseng cultivars can cause adverse effects on stability of yield and quality. K-1 is a superior cultivar with good root shape and stronger disease resistance. DNA markers mined from functional genes are clearly desirable for K-1, as they may associate with major traits and can be used for marker-assisted selection to maintain the high quality of Korean ginseng. Methods: Five genes encoding pathogenesis-related (PR) proteins of P. ginseng were amplified and compared for polymorphism mining. Primary, secondary, and tertiary structures of PR5 protein were analyzed by ExPASy-ProtParam, PSSpred, and I-TASSER methods, respectively. A coding single nucleotide polymorphism (SNP)-based specific primer was designed for K-1 by introducing a destabilizing mismatch within the 3' end. Allele-specific polymerase chain reaction (PCR) and real-time allele-specific PCR assays were conducted for molecular discrimination of K-1 from other cultivars and landraces. Results: A coding SNP leading to the modification of amino acid residue from aspartic acid to asparagine was exploited in PR5 gene of K-1 cultivar. Bioinformatics analysis showed that the modification of amino acid residue changed the secondary and tertiary structures of the PR5 protein. Primer KSR was designed for specific discrimination of K-1 from other ginseng cultivars and landraces. The developed real-time allele-specific PCR assay enabled easier automation and accurate genotyping of K-1 from a large number of ginseng samples. Conclusion: The SNP marker and the developed real-time allele-specific PCR assay will be useful not only for marker-assisted selection of K-1 cultivar but also for quality control in breeding and seed programs of P. ginseng.

Anti-Obesity Effect of Parthenocissus tricuspidata (Siebold & Zucc.) Planch Stem Extracts (소나무 담쟁이덩굴(Parthenocissus tricuspidata (Siebold & Zucc.) Planch) 줄기 추출물의 항비만 효과)

  • Yeo, Joo Ho;Yu, Ju Hyeong;Choi, Min Yeong;Lee, Jae Won;Geum, Na Gyeong;An, Mi-Yun;Jung, Chuleui;Jeong, Jin Boo
    • Korean Journal of Plant Resources
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    • v.35 no.2
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    • pp.380-384
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    • 2022
  • In this study, we investigated in vitro anti-obesity activity of Parthenocissus tricuspidata (Siebold & Zucc.) Planch stem (PTS) in mouse adipocytes 3T3-L1 cells. PTS inhibited lipid accumulation and reduced the expression level of the proteins such as C/EBPα, PPARγ and FABP4 associated with adipogenesis in 3T3-L1 cells. In addition, PTS increased lipolysis-related protein expression such as p-AMPK, ATGL and p-HSL, and reduced perilipin-1 in 3T3-L1 cells. Taken together, PTS may have anti-obesity activity through inhibiting adipogenesis and inducing lipid lipolysis in adipocytes. Based on these results, PTS is expected to be used as a potential functional agent for anti-obesity.

Prevention of UV-induced Skin Damage by Activation of Tumor Suppressor Genes p53 and $p14^{ARF}$

  • Petersen, R.;John, S.;Lueder, M.;Borchert, S.
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.338-351
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    • 2003
  • UV radiation is the most dangerous stress factor among permanent environmental impacts on human skin. Consequences of UV exposure are aberrant tissue architecture, alterations in skin cells including functional changes. Nowadays new kinds of outdoor leisure-time activities and changing environmental conditions make the question of sun protection more important than ever. It is necessary to recognize that self-confident consumers do not consider to change their way of life, they demand modern solutions on the basis of new scientific developments. In the past one fundamental principle of cosmetics was the use of physical and organic filter systems against damaging UV-rays. Today new research results demonstrate that natural protecting cell mechanisms can be activated. Suitable biological actives strongly support the protection function not from the surface but from the inside of the cell. A soy seed preparation (SSP) was proven to stimulate natural skin protective functions. The major functions are an increased energy level and the prevention of DNA damage. These functions can I be defined as biological UV protection. The tumor suppressor protein p53 plays a key role in the regulation of DNA repair. p53 must be transferred into the phosphorylated form to work as transcription factor for genes which are regulating the cell cycle or organizing DNA repair. A pretreatment with SSP increases the phosphorylation rate of p53 of chronically UV-irradiated human keratinocytes significantly. According to the same test procedure SSP induces a dramatic increase in the expression of the tumor suppressor protein p14$^{ARF}$ that is supporting the p53 activity by blocking the antagonist of p53, the oncoprotein Mdm2. Mdm2, a ubiquitin E3-ligase, downregulates p53 and at the same time it prevents phosphorylation of p53. The positive influence of the tumor suppressor proteins explains the stimulation of DNA repair and prevention of sunburn cell formation by SSP, which was proven in cell culture experiments. In vivo the increased skin tolerance against UV irradiation by SSP could be confirmed too. We have assumed, that an increased repair potential provides full cell functionality.y.

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Selection of Resistant Varieties to Aspergillus flavus by Determination of Aflatoxin B1 Content in Korean Peanut (Arachis hypogaea L.) Accessions

  • Seungah Han;Byeong-Cheol Kim;Jungmin Ha;Tae-Hwan Jun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.68 no.3
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    • pp.175-187
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    • 2023
  • Peanuts, also known as groundnuts (Arachis hypogaea L.), are globally recognized as a vital oilseed crop. Peanuts are rich in proteins (e.g., arginine), oils (e.g., oleic acid and linoleic acid), fiber, vitamins (e.g., niacin and tocopherol), and carbohydrates and are consumed worldwide. However, the presence of aflatoxin (AF) has garnered substantial attention since its initial discovery as the causative agent of Tukey's X disease in the United Kingdom in 1960. Among the 18 aflatoxins identified, aflatoxin B1 (AFB1) has the highest toxic activity and causes hepatocellular carcinoma. It is classified as Group I by the International Agency for Research on Cancer (IARC) of the World Health Organization (WHO). The present study was conducted to evaluate aflatoxin B1 resistance of 102 peanut accessions and select putative aflatoxin B1-resistant peanut accessions to aflatoxin B1. One hundred and one Korean germplasms harvested in 2020 were inoculated with A. flavus to identify aflatoxin-resistant cultivars, and the aflatoxin B1 concentration was measured using an ultra-performance liquid chromatography-photodiode array detector. Twenty-six accessions with aflatoxin B1 concentrations lower than those of the check plant 55-437 were chosen for the development of aflatoxin-resistant varieties in Korea. As Korean aflatoxin-resistant varieties have not yet been developed, the findings of the present study are expected to provide useful information for the development of aflatoxin-resistant cultivars.

Difference in Electrophoretic Phenotypes of Rice Cultivars Selected to Oxyfluorfen (Oxyfluorfen에 대한 내성(耐性) 및 감수성(感受性) 수도품종(水稻品種)의 전기영동(電氣泳動) 표현형(表現型) 차이(差異))

  • Kuk, Y.I;Guh, J.O.;Lee, D.J.;Kim, Y.J.
    • Korean Journal of Weed Science
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    • v.8 no.2
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    • pp.199-207
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    • 1988
  • The study was intended to know any relations between the rice tolerance to oxyfluorfen and varietal speciation in seed protein composition or any enzymatical allelies with or without chemical treatment. Rice varieties used were Chokoto, Aichiasahi, Agabyeo, IR 3941 and Tablei as the tolerant group, and Mushakdanti, Weld Pally, HP 1033, HP 857, and HP 907 as the susceptible, respectively. Electrophoretic methods used were SDS-PAGE for seed protein, 7% PAGE for isozymes (acid phosphatase and peroxidase from rice seedling) and changes in isoenzyme activity (malate dehydrogenase, peroxidase and esterase) as affected by oxyfluorfen treatment ($10^{-4}M$) was also studied. The results are summarized as follows. -Among 19 bands separated in seed proteins, two different rice groups selected in terms of tolerance were clustered in dissimilarity. This was based on 2 facts in that G band was not present in susceptible varieties and that less activity of H, N, O, P, Q, Rand S band was shown. -Among 4 bands separated in acid phosphatase, the presence of (band and lower activity of B band was specific for tolerant varieties. For 4 minor bands separated in peroxidase, the tolerant varieties had no activity in B band and higher activity in A, C, D bands. -Time-course study of isozymes as affected by $10^{-4}M$ oxyfluorfen showed that Chokoto, the tolerant varieties, had little activity in A band and consistently higher activities in Band C bands for malate dehydrogenase. For 5 bands separated in peroxidase, B band was not found in Chokoto while A, C, D, and E bands were consistently present. Esterase was separated into about 4 bands in which Chokoto had maintained higher activities in A, C and D bands.

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Iron fortification of grains by introducing a recombinant gene of ferritin with seed promoters in rice (종자 특이 프로모터와 대두 Ferritin 유전자에 의한 벼 종실의 철분강화)

  • Cho, Yong-Gu;Kim, Hyung-Keun;Choi, Jang-Sun;Jung, Yu-Jin;Kang, Kwon-Kyoo
    • Journal of Plant Biotechnology
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    • v.36 no.1
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    • pp.87-95
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    • 2009
  • The recombinant DNAs, pGBF, pGTF, and pZ4F, using soybean ferritin gene have constructed with the promoters derived from seed proteins, glutelin, globulin, and zein. The recombinant ferritin genes were transformed into rice plant by Agrobacterium-mediated transformation. Iron contents and agronomic traits have been evaluated in the transgenic progenies. The embryogenic calli survived from second selection medium were regenerated at the rates of 19.2% with pGBF, 15.0% with pGTF, and 18.4% with pZ4F in Donganbyeo and 6.7% with pGBF, 11.7% with pGTF, and 3.4% with pZ4F in Hwashinbyeo. The introduction of ferritin gene in putative transgenic rice plants was confirmed by PCR and Southern blot analysis and also the expression of ferritin gene was identified by Northern blot and Western blot analysis. The iron accumulation in transgenic rice grains of the transgenic rice plant, T1-2, with zein promoter and ferritin gene contained 171.4 ppm showing 6.4 times higher than 26.7 ppm of Hwashinbyeo seed as wild type rice, but the transgenic plants with globulin and glutelin showed a bit higher iron contents with a range from 2.1 to 3.0 times compare to wild type grain. The growth responses of transgenic plants showed the large variances in plant height and number of tillers. However, there were some transgenic plants having similar phenotype to wild type plants. In the T1 generation of transgenic plants, plant height, culm length, panicle length, and number of tillers were similar to those of wild type plants, but ripened grain ratio ranged from 53.3% to 82.2% with relatively high variation. The transgenic rice plants would be useful for developing rice varieties with high iron content in rice grains.

Difference in Electrophoretic Phenotypes of rice Cultivars Selected to Bensulfuron (Bensulfuron에 대(對)한 내성(耐性) 및 감수성(感受性) 수도품종(水稻品種)의 전기영동(電氣泳動) 표현형(表現型) 차이(差異))

  • Kuk, Y.I.;Guh, J.O.;Kim, Y.J.;Lee, D.J.
    • Korean Journal of Weed Science
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    • v.8 no.3
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    • pp.250-257
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    • 1988
  • The study was intended to know any relations between the rice tolerance to bensulfuron and varietal speciation in seed protein composition or any enzymatical allelies with or without chemical treatment. Rice varieties used were UCP-28, Chinsurah Boro II, Fukunohama, Fadehpur-2, IR 14252-13-2-2-5 as the tolerant group, and HP 93(3) FA, HP94(9) FA, Padilabou Alumbis, KH-17854, and IR 1846-2841-1 as the susceptible, respectively. Electrophoretic methods used were SDS-PAGE for seed protein, 7% PAGE for isozymes (acid phosphatase, peroxidase, malate dehydrogenase, and esterase from rice seedling) and variation in isoenzyme profiles (malate dehydrogenase, peroxidase, and esterase) as affected by different concentrations of bensulfuron(0, $10^{-6}$, $10^{-5}$ and $3{\times}10^{-5}M$) was also studied. The results are summarized as follows. -Among 16 bands separated in seed proteins, two different rice groups selected in terms of tolerance to bensulfuron were clustered in dissimilarity, which was based on relatively larger area in whole peaks and higher activities in N, O, P bands for the tolerant group. -Among isozymes obtained from rice seedlings without chemical treatments, the following specificities were obtained. The tolerant varieties had the relatively higher activity in D band out of 4 peroxidase bands. Malate dehydrogenase was separated into 3 bands and only tolerant varieties had A band and higher activities in Band C bands. Esterase was separated into 3-4 bands with higher activities in A and B bands for tolerant varieties. There were one major band accompanied by 2-3 minor bands for acid phosphatase in which only tolerant varieties had the B band. -The effect of Bensulfuron concentration on the isozyme activities showed that the activity of C band in peroxidase was not present in tolerant varieties which was contrary to the increased activities in susceptible varieties. However, D band was gradually disappeared only in susceptible varieties as the concentration of bensulfuron was increased. For malate dehydrogenase in the susceptible varieties, major bands D, E and F kept consistantly higher activities while minor bands A, B and C disappeared sensitively. Among 5 bands of esterase separated, D band was present only in the tolerant varieties while E band only in the susceptible. The activities in A, C, E bands were sharply decreased in the susceptible varieties as the concentration of bensulfuron was increased.

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