• Title/Summary/Keyword: chloroplast

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Development of HRM Markers Based on Identification of SNPs from Next-Generation Sequencing of Sanguisorba officinalis, Sanguisorba tenuifolia f. alba (Trautv. & Mey.) Kitam and Sanguisorba tenuifolia Fisch. ex Link (오이풀, 흰오이풀, 긴오이풀의 NGS 기반 유전체 서열의 완전 해독 및 차세대 염기서열 재분석으로 탐색된 SNP 기반 HRM 분자표지 개발)

  • Sim, Mi-Ok;Jang, Ji Hun;Jung, Ho-Kyung;Hwang, Taeyeon;Kim, Sunyoung;Cho, Hyun-Woo
    • The Korea Journal of Herbology
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    • v.34 no.6
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    • pp.91-97
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    • 2019
  • Objective : To establish a reliable tool between for the distinction of original plants of Sanguisorbae Radix, we analyzed the complete chloroplast genome sequence of Sanguisorbae Radix and identified single nucleotide polymorphisms (SNPs). Materials and methods : The chloroplast genome sequence of Sanguisorba officinalis, Sanguisorba tenuifolia f. alba (Trautv. & Mey.) Kitam and Sanguisorba tenuifolia Fisch. ex Link obtained using next-generation sequencing technology were described and compared with those of other species to develop specific markers. Candidate genetic markers were identified to distinguish species from the chloroplast sequences of each species using Modified Phred Phrap Consed and CLC Genomics Workbench programs. Results : The structure of the chloroplast genome of each sample that had been assembled and verified was circular, and the length was about 155 kbp. Through comparative analysis of the chloroplast sequences, we found 220 nucleotides, 158 SNPs, and 62 Indel (insertion and/or deletion), to distinguish Sanguisorba officinalis, Sanguisorba tenuifolia f. alba (Trautv. & Mey.) Kitam and Sanguisorba tenuifolia Fisch. ex Link. Finally, 15 specific SNP genetic markers were selected for the verification at positions. Avaliable primers for the dried herb, which is used as medicine, were used to develop the PCR amplification product of Sanguisorbae Radix to assess the applicability of PCR analysis. Conclusion : In this study, we found that Fendel-qPCR analysis based on the chloroplast DNA sequences can be an efficient tool for discrimination of Sanguisorba officinalis, Sanguisorba tenuifolia f. alba (Trautv. & Mey.) Kitam and Sanguisorba tenuifolia Fisch. ex Link.

Analysis of Chloroplast Mutants of Arabidopsis Induced by Diepoxybutane (Diepoxybutane에 의해 유도된 애기장대 엽록체 돌연변이체의 분석)

  • 윤용휘;이정훈;박해진;강용원;이경민;신동현;이인중;김학윤;김달웅
    • Journal of Life Science
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    • v.12 no.4
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    • pp.399-406
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    • 2002
  • The Arabidopsis mutants involved in chloroplast development were induced by seed treatment of diepoxybutane which was rarely known mutagenic compound in plant mutagenesis. Three kinds of mutants designated as iml, gev, and yev were represented by the characteristics of variegated leaves, green vein with yellow leaves, and yellow green vein with green leaves respectively. We investigated the ultrastructure of chloroplast in mutated regions using transmission electron microscopy. The ultrastructure of chroloplast in wildtype showed regularly stacked grana thylakoid and stroma thylakoid while iml, gev and yev mutants displayed different shapes of grana stacking and stroma stacking of chloroplasts. Genetic analysis of three chloroplast mutants exhibit that divergent traits were ruled by a single recessive nuclear gene.

Development of PCR-based markers for discriminating Solanum berthaultii using its complete chloroplast genome sequence

  • Kim, Soojung;Cho, Kwang-Soo;Park, Tae-Ho
    • Journal of Plant Biotechnology
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    • v.45 no.3
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    • pp.207-216
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    • 2018
  • Solanum berthaultii is one of the wild diploid Solanum species, which is an excellent resource in potato breeding owing to its resistance to several important pathogens. On the other hand, sexual hybridization between S. berthaultii and S. tuberosum (potato) is limited because of their sexual incompatibility. Therefore, cell fusion can be used to introgress various novel traits from this wild species into the cultivated potatoes. After cell fusion, it is crucial to identify fusion products with the aid of molecular markers. In this study, the chloroplast genome sequence of S. berthaultii obtained by next-generation sequencing technology was described and compared with those of five other Solanum species to develop S. berthaultii specific markers. A total sequence length of the chloroplast genome is 155,533 bp. The structural organization of the chloroplast genome is similar to those of the five other Solanum species. Phylogenic analysis with 25 other Solanaceae species revealed that S. berthaultii is most closely located with S. tuberosum. Additional comparison of the chloroplast genome sequence with those of the five Solanum species revealed 25 SNPs specific to S. berthaultii. Based on these SNPs, six PCR-based markers for differentiating S. berthaultii from other Solanum species were developed. These markers will facilitate the selection of fusion products and accelerate potato breeding using S. berthaultii.

Production of Human Serum Albumin in Chloroplast-Transformed Tobacco Plants

  • Ko, Suk-Min;Kim, Hyun-Chul;Yoo, Byung-Ho;Woo, Je-Wook;Chung, Hwa-Jee;Choi, Dong-Woog;Liu, Jang-R.
    • Journal of Plant Biotechnology
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    • v.33 no.4
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    • pp.233-236
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    • 2006
  • Human serum albumin (HSA) is the most abundant protein in plasma and is the most often used intravenous protein in many human therapies. However, HSA is currently extracted only from plasma because commercially feasible recombinant expression systems are not available. This study attempted to develop an efficient system for recombinant HSA production by chloroplast transformation of tobacco. A HSA cDNA was isolated from a cDNA library constructed with human liver tissue. Chloroplast transformation vectors were constructed by introducing various regulatory elements to HSA regulatory sequences. Vectors were delivered by particle bombardment into leaf explants and chloroplast-transformed plants were subsequently regenerated into whole plants. Southern blot analysis confirmed that the HSA cDNA was incorporated between rps12 and orf70B of the chloroplast genome as designed. Western blot analysis revealed that hyper-expression and increasing the stability of HSA were achieved by modification of the regulatory sequences using the psbA5'UTRs in combination with elements of the 14 N-terminal amino acids of the GFP and the FLAG tag. However, only plants transformed with the vector containing all of these elements were able to accumulate HSA.

Complete Chloroplast Genome Sequence of Korean Endermic Species, Pseudostellaria longipedicellata

  • Kim, Yongsung;Heo, Kyeong-In;Lee, Sangtae;Park, Jongsun
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.40-40
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    • 2018
  • Pseudostellaria Pax (Caryophyllaceae) is a small genus distributed in temperate region. It consists of 25 species presenting high diversity in Asia. Pseudostellaria longipedicellata S. Lee, K. Heo & S. C. Kim was first announced as new species in 2012. Morphological characters of P. longipedicellata are closely related to those of Psedusotellaria palibiniana and Psedusotellaria okmotoi. These are distinguished from P. longipedicellata by shorter pedicel and puberulent pedicels, respectively and by being distributed allopatically between P. longipedicellata and rest of species. The complete chloroplast genome of P. longipedicellata was successfully rescued from raw reads generated by HiSeq2000. Its total length is 149,626 bp consisting of four regions: large single copy (LSC) region (81,292 bp), small single copy (SSC) region (16,984bp), and inverted repeats (IRs; 25,765 bp per each). It contained 126 genes (81 coding DNA sequence (CDS), eight rRNAs, and 37 tRNAs); 18 genes (seven CDS, four rRNAs, and seven tRNAs) are duplicated in inverted repeat regions. The overall GC content of P. longipedicellata is 36.5% and in the LSC, SSC, and IR regions were 34.3%, 29.3%, and 42.4%, respectively. Based on phylogenetic analysis of chloroplast genomes of P. longipedicellata and relatives species presents clear phylogenetic positions of Pseudostellaria genus. This chloroplast genome will be an important sequence resources for further researches of Pseudostellaria genus.

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Effects of Light on the Pigment Production and Chloroplast Development of Ginseng Hairy Roots (인삼 모상근의 색소 생성 및 엽록체 발달에 미치는 광의 효과)

  • 양덕조;최혜연
    • Journal of Ginseng Research
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    • v.21 no.1
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    • pp.28-34
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    • 1997
  • The effects of light on the pigment production and chloroplast development were examined on ginseng hairy roots cultured in 1/2MS liquid medium. The chlorophyll and carotenoid production were increased from 1,000 to 3,500 lux condition, but decreased drastically in 7,000 lux condition. The anthocyanin production was significantly increased with increment light intensity(1,000∼7,000 lux). The thylakoid membrane of chloroplast was proplastid in dark condition and it began to develop into thylakoid membrane in 1,000 lux condition and then intact thylakoid membrane was developed in 3,500 lux condition. However, the development of thylakoid membrane in 7,000 lux condition was inhibited comparing to 3,500 lux condition. The total chlorophyll production in blue light condition were high comparing to other wavelength and same as 40% of total chlorophyll on white light(3,500 lux) condition. The chlorophyll and carotenoid production by sucrose concentration were high in 3% sucrose condition and anthocyanin production was high in 4% condition. The production of chlorophyll and carotenoid by light periods was high when explants were cultured in dark condition for 1 week and then transferred to light condition for 4 weeks. Our results suggest that pigment production and chloroplast development could be accelerated by light Intensity of specific wavelength in cultures of ginseng hairy root.

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The complete chloroplast genome of Diarthron linifolium (Thymelaeaceae), a species found on a limestone outcrop in eastern Asia

  • KIM, Sang-Tae;OH, Sang-Hun;PARK, Jongsun
    • Korean Journal of Plant Taxonomy
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    • v.51 no.4
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    • pp.345-352
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    • 2021
  • Diarthron linifolium Turcz. is an annual herb usually found in sandy soil or limestone areas. Plants in the genus Diarthron are known to have toxic chemicals that may, however, be potentially useful as an anticancer treatment. Diarthron linifolium is a unique species among the species of the genus distributed in Korea. Here, we determine the genetic variation of D. linifolium collected in Korea with a full chloroplast genome and investigate its evolutionary status by means of a phylogenetic analysis. The chloroplast genome of Korean D. linifolium has a total length of 172,644 bp with four subregions; 86,158 bp of large single copy and 2,858 bp of small single copy (SSC) regions are separated by 41,814 bp of inverted repeat (IR) regions. We found that the SSC region of D. linifolium is considerably short but that IRs are relatively long in comparison with other chloroplast genomes. Various simple sequence repeats were identified, and our nucleotide diversity analysis suggested potential marker regions near ndhF. The phylogenetic analysis indicated that D. linifolium from Korea is a sister to the group of Daphne species.

Effect of Carbon Sources on the Synthesis of Phospholipid and Fatty Acid Composition in Chloroplast of Chlorella ellipsoidea (Chlorella ellipsoidea 엽록체의 인지질 생합성 및 지방산 조성에 미치는 탄소원의 효과)

  • 정효선
    • Journal of Plant Biology
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    • v.33 no.1
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    • pp.49-54
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    • 1990
  • Chlorella cells were cultured with M4N media treated with glucose (5 mM) sucrose (10 mM) and raffinose (30 mM). Phospholipids and their fatty acid compositions were analyzed in the chloroplast isolated from cultured Chlorella cells. Growth rate was prominently raised in the treatment with raffinose. Glucose was the most excellent carbon source in the biosynthesis of total lipid, phosphatidylcholine(PC), phosphatidylethanolamine(PE), phosphatidylinositol(PI) of the chloroplast. Also, the major fatty acids were palmitic, linoleic and linolenic acid during the biosynthesis of phospholipid in the control and in the treatment with carbon sources.

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Photoproduction of Hydrogen in Polyvinylalcohol-Iimmobilized Spinach Chloroplsats with Platinum Catalysts (Polyvinylalcohol에 고정한 시금치 엽록체와 백금 촉매를 이용한 광수소 발생)

  • 박인호
    • Journal of Plant Biology
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    • v.36 no.4
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    • pp.313-319
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    • 1993
  • Photoproduction of hydrogen by free and polyvinylalcohol (PVA)-immobilized spinach chloroplasts was investigated. Immobilization of chloroplast with PVA increased the functional stability of the chloroplast during storage. PVA-immobilized chloroplasts preserved photosynthetic electron transport activity much better than free chloroplasts. The hydrogen production of free chloroplast decreased to 17% of initial activity after storage of six days. The hydrogen production of the PVA-immobilized chloroplast, however, showed 44% of initial activity after storage of 15 days. The maximal rate of hydrogen production was accomplished at 2$^{\circ}C$ under the light intensity above 116 $\mu$E.m-2.s-1. The amount of hydrogen produced was proportional to the chlorophyll concentration. The hydrogen production was inhibited by DCMU treatment, indicating hydrogen production is dependent on photosynthetic electron transport. These results suggest that PVA is a good candidate for the immobilization matrix of chloroplasts for the photoproduction of hydrogen.

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Detection of DNA Rearrangement in Rice Using a Cosmid Library

  • Mun, Eun-Pyo;Nahm, Baek-Hie
    • Animal cells and systems
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    • v.1 no.4
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    • pp.629-635
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    • 1997
  • Previously we reported the migration and rearrangement of a chloroplast gene cluster into mitochondria. The exact genomic locations of the clusters, modes of the gene rearrangement and mechanisms of the interorganellar migration of the clusters have yet to be understood. The detailed analysis needs to include a larger region of DNA surrounding each cluster. To study DNA rearrangement and migration in more detail a cosmid library was constructed using the total rice genomic DNA including nuclear, chloroplast and mitochondrial DNA. From this cosmid library, a sub-library was obtained by selecting the clones hybridized to various regions of chloroplast DNA. According to the hybridization pattern 136 clones from the sub-library were classified into 29 groups. Detailed analysis of these clones revealed that in addition to authentic chloroplast DNA, the clones contain its homologs resulted from rearrangement and mutation. We analyzed two clones in detail, which contain different rp12 homologs resulted from rearrangement and/or migration, respectively.

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