• Title/Summary/Keyword: Plastid genome

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Plastid Transformation of Soybean Suspension Cultures

  • Zhang, Xing-Hai;Archie R.Portis. Jr.;Jack M.Widholm
    • Journal of Plant Biotechnology
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    • v.3 no.1
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    • pp.39-44
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    • 2001
  • Plastid transformation was attempted with soybean [Glycine max (L.) Merr.] leaves and photoautotrophic and embryogenic cultures by particle bombardment using the transforming vector pZVII that carries the coding sequences for both subunits of Chlamydomonas reinhardtii Rubisco and a spectinomycin resistance gene (aadA). Spectinomycin resistant calli were selected from the bombarded leaves but the transgene was not present, indicating that the resistance was due to mutations. The Chlamydomonas rbcL and rbcS genes were shown to be site-specifically integrated into the plastid genome of the embryogenic cells with a very low transformation efficiency. None of the transformed embryogenic lines survived the plant regeneration process so no whole plants were recovered. This result does indicate that it should be possible to insert genes into the plastid genome of the important crop soybean if the overall methods are improved.

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The complete plastid genome of Scopolia parviflora (Dunn.) Nakai (Solanaceae)

  • Park, Jin Hee;Lee, Jungho
    • Korean Journal of Plant Taxonomy
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    • v.46 no.1
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    • pp.60-64
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    • 2016
  • Scopolia parviflora of the family Solanaceae is an endemic species of Korea and a traditional Korean medicinal plant. The plastid genome was sequenced by next-generation sequencing (NGS) method. The characterized cp genome is 156,193 bp in size; the large single-copy (LSC) region is 86,364 bp, the inverted repeat (IR) is 25,905 bp, and the small single copy (SSC) region is 18,019 bp. The overall GC content of the plastid genome amounts to 37.61%. The cp genome contains 113 genes and 21 introns, including 80 proteincoding genes, four RNA genes, 30 tRNA genes, 20 group II introns, and one group I intron. A phylogenetic analysis showed that Scopolia parviflora was closely related to Hyoscyamus niger.

Improved plastid transformation efficiency in Scoparia dulcis L.

  • Kota, Srinivas;Hao, Qiang;Narra, Muralikrishna;Anumula, Vaishnavi;Rao, A.V;Hu, Zanmin;Abbagani, Sadanandam
    • Journal of Plant Biotechnology
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    • v.46 no.4
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    • pp.323-330
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    • 2019
  • The high expression level of industrial and metabolically important proteins in plants can be achieved by plastid transformation. The CaIA vector, a Capsicum-specific vector harboring aadA (spectinomycin resistance), is a selectable marker controlled by the PsbA promoter, and the terminator is flanked by the trnA and trnI regions of the inverted repeat (IR) region of the plastid. The CaIA vector can introduce foreign genes into the IR region of the plastid genome. The biolistic method was used for chloroplast transformation in Scoparia dulcis with leaf explants followed by antibiotic selection on regeneration medium. Transplastomes were successfully screened, and the transformation efficiency of 3 transgenic lines from 25 bombarded leaf explants was determined. Transplastomic lines were evaluated by PCR and Southern blotting for the confirmation of aadA insertion and its integration into the chloroplast genome. Seeds collected from transplastomes were analyzed on spectinomycin medium with wild types to determine genetic stability. The increased chloroplast transformation efficiency (3 transplastomic lines from 25 bombarded explants) would be useful for expressing therapeutically and industrially important genes in Scoparia dulcis L.

Plastid Transformation in the Monocotyledonous Cereal Crop, Rice (Oryza sativa) and Transmission of Transgenes to Their Progeny

  • Lee, Sa Mi;Kang, Kyungsu;Chung, Hyunsup;Yoo, Soon Hee;Ming Xu, Xiang;Lee, Seung-Bum;Cheong, Jong-Joo;Daniell, Henry;Kim, Minkyun
    • Molecules and Cells
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    • v.21 no.3
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    • pp.401-410
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    • 2006
  • The plastid transformation approach offers a number of unique advantages, including high-level transgene expression, multi-gene engineering, transgene containment, and a lack of gene silencing and position effects. The extension of plastid transformation technology to monocotyledonous cereal crops, including rice, bears great promise for the improvement of agronomic traits, and the efficient production of pharmaceutical or nutritional enhancement. Here, we report a promising step towards stable plastid transformation in rice. We produced fertile transplastomic rice plants and demonstrated transmission of the plastidexpressed green fluorescent protein (GFP) and aminoglycoside 3′-adenylyltransferase genes to the progeny of these plants. Transgenic chloroplasts were determined to have stably expressed the GFP, which was confirmed by both confocal microscopy and Western blot analyses. Although the produced rice plastid transformants were found to be heteroplastomic, and the transformation efficiency requires further improvement, this study has established a variety of parameters for the use of plastid transformation technology in cereal crops.

Expression of the Glyphosate Resistant Gene, cp4-epsps, through Plastid Transformation in Rice (Oryza sativa L.) (벼 색소체 형질전환을 이용한 글리포세이트 저항성 유전자 cp4-epsps의 발현)

  • Kang, Kyung-Su;Kim, Min-Kyun
    • Journal of Plant Biotechnology
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    • v.33 no.2
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    • pp.75-84
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    • 2006
  • Heteroplasmic rice plastid transformant was generated using suspension cells as bombardment materials. PCR analyses confirmed incorporation of aadA and cp4-epsps genes into the rice plastid genome by homologous recombination events via the flanking sequences of the trnI and trnA. Transplastomic calli were actively proliferated when cultured on AAM2 medium supplemented with various concentrations (500-3000 mg/L) of streptomycin in dark condition, and transplastomic suspension cells showed resistance to nonselective herbicide, glyphosate. Through 'agarose pie selection' method, heteroplastomic calli, containing considerably high level of transplastome and expressing the CP4 EPSPS protein, were obtained. They were further regenerated to green shoots with healthy roots.

국립공원 북한산의 환경평가에 관하여 - 도봉산지역 일대를 중심으로-

  • 박봉규
    • Proceedings of the Botanical Society of Korea Conference
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    • 1985.08b
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    • pp.35-48
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    • 1985
  • Plastids, which are organelles unique to plant cells, bear their own genome that is organized into DNA-protein complexes (nucleoids). Regulation of gene expression in the plastid has been extensively investigated because this organelle plays an important role in photosynthesis. Few attempts, however, have been made to characterize the regulation of plastid gene expression at the chromosomal structure, using plastid nucleoids. In this report, we summarize the recent progress in the characterization of DNA-binding proteins in plastids, with special emphasis on CND41, a DNA binding protein, which we recently identified in the choloroplast nucleoids from photomixotrophically cultured tobacco cells. CND41 is a protein of 502 amino acids which consisted of a transit peptide of 120 amino acids and a mature protein of 382 amino acids. The N-terminal of the 'mature' protein has lysine-rich region which is essential for DNA-binding. CNA41 also showed significant identities to some aspartyl proteases. Protease activity of purified CND41 has been recently confirmed and characterized. On the other hand, characterization of accumulation of CND41 both in wild type and transgenic tobacco with reduced amount of CND41 suggests that CND41 is a negative regulator in chloroplast gene expression. Further investigation indicated that gene expression of CND41 is cell-specifically and developmentally regulated as well as sugar-induced expression. The reduction of CND41 expression in transgenic tobacco also brought the stunted plant growth due to the reduced cell length in stem. GA3 treatment on apical meristem reversed the dwarf phenotype in the transformants. Effects of CND41 expression on GA biosynthesis will be discussed

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Development of a Plastid DNA-Based Maker for the Identification of Five Medicago Plants in South Korea

  • Kim, Il Ryong;Yoon, A-Mi;Lim, Hye Song;Lee, Sunghyeon;Lee, Jung Ro;Choi, Wonkyun
    • Proceedings of the National Institute of Ecology of the Republic of Korea
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    • v.3 no.4
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    • pp.212-220
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    • 2022
  • DNA markers have been studied and used intensively to identify plant species based on molecular approaches. The genus Medicago belongs to the family Fabaceae and contains 87 species distributed from the Mediterranean to central Asia. Five species of Medicago are known to be distributed in South Korea; however, their morphological characteristics alone cannot distinguish the species. In this study, we analyzed the phylogenetic relationships using collected five species of Medicago from South Korea and 44 taxa nucleotide information from NCBI. The constructed phylogenetic tree using gibberellin 3-oxidase 1 and tRNALys (UUU) to maturase K gene sequences showed the monophyly of the genus Medicago, with five species each forming a single clade. These results suggest that there are five species of Medicago distributed in South Korea. In addition, we designed polymerase chain reaction primers for species-specific detection of Medicago by comparing the plastid sequences. The accuracy of the designed primer pairs was confirmed for each Medicago species. The findings of this study provide efficient and novel species identification methods for Medicago, which will assist in the identification of wild plants for the management of alien species and living modified organisms.

Plastid genome of Aster altaicus var. uchiyamae Kitam., an endanger species of Korean asterids

  • Park, Jihye;Shim, Jaekyung;Won, Hyosig;Lee, Jungho
    • Journal of Species Research
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    • v.6 no.1
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    • pp.76-90
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    • 2017
  • Aster altaicus var. uchiyamae Kitam. is an endemic taxon of Korea and is protected by law as an endanger taxon. The genetic information of A. altaicus var. uchiyamae is unavailable in Genbank. Here we sequenced chloroplast genome of A. altaicus var. uchiyamae. The cp-genome of Aster altaicus var. uchiyamae was 152,446 bps in size: LSC was 84,240 bps, IR 25,005 bps, SSC 18,196 bps. The cp-genome contains 112 genes and 21 introns consisted of 79 protein coding genes(PCGs), 4 RNA genes, and 29 tRNA genes, with 20 group II introns and one group I intron. There were three pseudo-genes including ${\psi}$-ycf1, ${\psi}$-rps19, and ${\psi}$-trnT_GGU. Eighteen genes, five introns, and parts of two genes and an intron are found within the IR, which has two copies. The cp-DNA of Aster altaicus var. uchiyamae is distinguished from A. spathulifolius, only known cp-genome of the genus Aster, by 172 SNP in genic regions of 43 PCGs and 21 indels in 11 PCGs and SSU. The chloroplast genome sequence was deposited at GenBank (KX35265).

Phylogenomics and its Growing Impact on Algal Phylogeny and Evolution

  • Adrian , Reyes-Prieto;Yoon, Hwan-Su;Bhattacharya, Debashish
    • ALGAE
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    • v.21 no.1
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    • pp.1-10
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    • 2006
  • Genomic data is accumulating in public database at an unprecedented rate. Although presently dominated by the sequences of metazoan, plant, parasitic, and picoeukaryotic taxa, both expressed sequence tag (EST) and complete genomes of free-living algae are also slowly appearing. This wealth of information offers the opportunity to clarify many long-standing issues in algal and plant evolution such as the contribution of the plastid endosymbiont to nuclear genome evolution using the tools of comparative genomics and multi-gene phylogenetics. A particularly powerful approach for the automated analysis of genome data from multiple taxa is termed phylogenomics. Phylogenomics is the convergence of genomics science (the study of the function and structure of genes and genomes) and molecular phylogenetics (the study of the hierarchical evolutionary relationships among organisms, their genes and genomes). The use of phylogenetics to drive comparative genome analyses has facilitated the reconstruction of the evolutionary history of genes, gene families, and organisms. Here we survey the available genome data, introduce phylogenomic pipelines, and review some initial results of phylogenomic analyses of algal genome data.