• Title/Summary/Keyword: Molecular Breeding

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Agroinfiltration-based Potato Virus X Replicons to Dissect the Requirements of Viral Infection

  • Park, Sang-Ho;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • v.22 no.4
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    • pp.386-390
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    • 2006
  • Extensive research of the Potato virus X(PVX) has been performed in in vitro transcription system using the bacteriophage T7 promoter. We constructed an efficient T-DNA based binary vector, pSNU1, and modified vectors carrying PVX replicons. The suitability of the construct to transiently express PVX RNA using Agrobacterium tumefaciens was tested by analysis of infectivity in plants. The expressed PVX RNA was infectous and systemically spread in three plant species including Nicotiana benthamiana, N. tabacum cv. Xanthi-nc, and Capsicum annuum cv. Chilsungcho. The PVX full length construct, pSPVXp31, was caused severe mosaic symptoms on N. benthamiana, severe necrotic lesions on C. annuum while milder symptoms and delayed mosaic symptoms were appeared on the systemic leaves on N. tabaccum. RT-PCR analysis confirmed the presence of PVX RNAs on both inoculated and systemic leaves in all three plant species tested. Our results indicated that PVX replicons were efficiently expressed PVX RNA in at least three tested species. Further investigation win be needed to elucidate the mechanism of PVX replication, translation, movement and assembly/disassembly processes.

Identification of New Microsatellite Markers in Panax ginseng

  • Kim, Joonki;Jo, Beom Ho;Lee, Kyoung Lyong;Yoon, Eui-Soo;Ryu, Gi Hyung;Chung, Ki Wha
    • Molecules and Cells
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    • v.24 no.1
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    • pp.60-68
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    • 2007
  • Microsatellites, also called simple sequence repeats (SSR), are very useful molecular genetic markers commonly used in crop breeding, species identification and linkage analysis. In the present study, we constructed a microsatellite-enriched genomic library of Panax ginseng, and identified 251 novel microsatellite sequences. Tri-nt repeat units were the most abundant (46.6%), followed by di-nt repeats (35.5%). The $(AG)_n$ motif was most common (23.1%), followed by the $(AAC)_n$ motif (22.3%). From the genotyping of 94 microsatellites using marker-specific primer sets, we identified 11 intraspecific polymorphic markers as well as 14 possible interspecific polymorphic markers differing between P. ginseng and P. quinquefolius. The exact allele structures of the polymorphic markers were determined and the alleles were named. This study represents the first report of the bulk isolation of microsatellites by screening a microsatellite-enriched genomic library in P. ginseng. The microsatellite markers could be useful for linkage analysis, genetic breeding and authentication of Panax species.

Prospects for Plant Biotechnology and Bioindustry in the 21st Century: Paradigm Shift Driven by Genomics (21세기 식물생명공학과 생물산업의 전망: 유전체 연구에 의한 Paradigm Shift)

  • Liu, Jang-Ryol;Choi, Dong-Woog;Chung, Hwa-Jee
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2002.04b
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    • pp.19-25
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    • 2002
  • Biotechnology in the 21st century will be driven by three emerging technologies: genomics, high-throughput biology, and bioinformatics. These technologies are complementary to one another. A large number of economically important crops are currently subjected to whole genome sequencing. Functional genomics for determining the functions of the genes comprising the given plant genome is under progress by using various means including phenotyping data from transgenic mutants, gene expression profiling data from DNA microarrays, and metabolic profiling data from LC/mass analysis. The aim of plant molecular breeding is shifting from introducing agronomic traits such as herbicide and insect resistance to introducing quality traits such as healthful oils and proteins, which will lead to improved and nutritional food and feed products. Plant molecular breeding is also expected to aim to develop crops for producing human therapeutic and industrial proteins.

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Molecular and Biological Characterization of an Isolate of Cucumber mosaic virus from Glycine soja by Generating its Infectious Full-genome cDNA Clones

  • Phan, Mi Sa Vo;Seo, Jang-Kyun;Choi, Hong-Soo;Lee, Su-Heon;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • v.30 no.2
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    • pp.159-167
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    • 2014
  • Molecular and biological characteristics of an isolate of Cucumber mosaic virus (CMV) from Glycine soja (wild soybean), named as CMV-209, was examined in this study. Comparison of nucleotide sequences and phylogenetic analyses of CMV-209 with the other CMV strains revealed that CMV-209 belonged to CMV subgroup I. However, CMV-209 showed some genetic distance from the CMV strains assigned to subgroup IA or subgroup IB. Infectious full-genome cDNA clones of CMV-209 were generated under the control of the Cauliflower mosaic virus 35S promoter. Infectivity of the CMV-209 clones was evaluated in Nicotiana benthamiana and various legume species. Our assays revealed that CMV-209 could systemically infect Glycine soja (wild soybean) and Pisum sativum (pea) as well as N. benthamiana, but not the other legume species.

Breeding of New Strains of Mushroom by Basidiospore Chemical Mutagenesis

  • Lee, Ji-A;Kang, Hyeon-Woo;Kim, Sang-Woo;Lee, Chang-Yun;Ro, Hyeon-Su
    • Mycobiology
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    • v.39 no.4
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    • pp.272-277
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    • 2011
  • Chemical mutagenesis of basidiospores of Hypsizygus marmoreus generated new mushroom strains. The basidospores were treated with methanesulfonate methylester, an alkylating agent, to yield 400 mutant monokaryotic mycelia. Twenty fast-growing mycelia were selected and mated each other by hyphal fusion. Fifty out of the 190 matings were successful (mating rate of 26.3%), judged by the formation of clamp connections. The mutant dikaryons were cultivated to investigate their morphological and cultivation characteristics. Mutant strains No. 3 and No. 5 showed 10% and 6% increase in fruiting body production, respectively. Eight mutant strains showed delayed and reduced primordia formation, resulting in the reduced production yield with prolonged cultivation period. The number of the fruiting bodies of mutant No. 31, which displayed reduced primordial formation, was only 15, compared to the parental number of 65. Another interesting phenotype was a fruiting body with a flattened stipe and pileus. Dikaryons generated by mating with the mutant spore No. 14 produced flat fruiting bodies. Further molecular biological studies will provide details of the mechanism. This work shows that the chemical mutagenesis approach is highly utilizable in the development of mushroom strains as well as in the generation of resources for molecular genetic studies.

Prospects for Plant Biotechnology and Bioindustry in the 21s1 Century: Paradigm Shift Driven by Genomics (21세기 식물생명공학과 생물산업의 전망 : 유전체 연구에 의한 Paradigm Shift)

  • Liu, Jang-Ryol;Choi, Dong-Woog;Chung, Hwa-Jee
    • Journal of Plant Biotechnology
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    • v.29 no.3
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    • pp.145-150
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    • 2002
  • Biotechnology in the 21st century will be driven by three emerging technologies: genomics, high-throughput biology, and bioinformatics. These technologies are complementary to one another. A large number of economically important crops are currently subjected to whole genome sequencing. Functional genomics for determining the functions of the genes comprising the given plant genome is under progress by using various means including phenotyping data from transgenic mutants, gene expression profiling data from DNA microarrays, and metabolic profiling data from LC/mass analysis. The aim of plant molecular breeding is shifting from introducing agronomic traits such as herbicide and insect resistance to introducing quality traits such as healthful oils and proteins, which will lead to improved and nutritional food and feed products. Plant molecular breeding is also expected to aim to develop crops for producing human therapeutic and industrial proteins.

Molecular Identification and Fine Mapping of a Major Quantitative Trait Locus, OsGPq3 for Seed Low-Temperature Germinability in Rice

  • Nari Kim;Rahmatullah Jan;Jae-Ryoung Park;Saleem Asif;Kyung-Min Kim
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.283-283
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    • 2022
  • Abiotic stresses such as high/low temperature, drought, salinity, and submergence directly or indirectly influence the physiological status and molecular mechanisms of rice which badly affect yield. Especially, the low temperature causes harmful influences in the overall process of rice growth such as uneven germination and the establishment of seedlings, which has become one of the main limiting factors affecting rice production in the world. It is of great significance to find the candidate genes controlling low-temperature tolerance during seed germination and study their functions for breeding new rice cultivars with immense low-temperature tolerance during seed germination. In this study, 120 lines of Cheongcheong/Nagdong double haploid population were used for quantitative trait locus analysis of low-temperature germinability. The results showed significant difference in germination under low different temperature conditions. In total, 4 QTLs were detected on chromosome 3, 6, and 8. A total of 41 genes were identified from all the 4 QTLs, among them, 25 genes were selected by gene function annotation and further screened through quantitative real time polymerase chain reaction. Based on gene function annotation and level of expression under low-temperature, our study suggested OsGPq3 gene as a candidate gene controlling viviparous germination, ABA and GA signaling under low-temperature. This study will provide a theoretical basis for marker-assisted breeding.

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Genomic Differentiation Among Oyster Mushroom Cultivars Released in Korea by URP-PCR Fingerprinting

  • Kang, Hee-Wan;Park, Dong-Suk;Park, Young-Jin;You, Chang-Hyun;Lee, Byoung-Moo;Eun, Moo-Yong;Go, Seong-Joo
    • Mycobiology
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    • v.29 no.2
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    • pp.85-89
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    • 2001
  • URP primers of 20 mer derived from repetitive sequence of rice were used to assess genetic variation of oyster mushroom consisting of 10 cultivars of Pleurotus ostreatus, two cultivars of P. florida and two cultivars of P. sajor-caju which were registered in Korea. URP2F and URP38F primers produced cultivar-specific PCR polymorphic bands in the Pleurotus species. UPGMA cluster analysis using the URP-PCR data showed that 14 Pleurotus cultivars are genetically clustered into large three groups. The URP-PCR data provided important information for more efficient breeding strategies of Pleurotus cultivars.

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Transcriptome-based identification of water-deficit stress responsive genes in the tea plant, Camellia sinensis

  • Tony, Maritim;Samson, Kamunya;Charles, Mwendia;Paul, Mireji;Richard, Muoki;Mark, Wamalwa;Stomeo, Francesca;Sarah, Schaack;Martina, Kyalo;Francis, Wachira
    • Journal of Plant Biotechnology
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    • v.43 no.3
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    • pp.302-310
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    • 2016
  • A study aimed at identifying putative drought responsive genes that confer tolerance to water stress deficit in tea plants was conducted in a 'rain-out shelter' using potted plants. Eighteen months old drought tolerant and susceptible tea cultivars were each separately exposed to water stress or control conditions of 18 or 34% soil moisture content, respectively, for three months. After the treatment period, leaves were harvested from each treatment for isolation of RNA and cDNA synthesis. The cDNA libraries were sequenced on Roche 454 high-throughput pyrosequencing platform to produce 232,853 reads. After quality control, the reads were assembled into 460 long transcripts (contigs). The annotated contigs showed similarity with proteins in the Arabidopsis thaliana proteome. Heat shock proteins (HSP70), superoxide dismutase (SOD), catalase (cat), peroxidase (PoX), calmodulinelike protein (Cam7) and galactinol synthase (Gols4) droughtrelated genes were shown to be regulated differently in tea plants exposed to water stress. HSP70 and SOD were highly expressed in the drought tolerant cultivar relative to the susceptible cultivar under drought conditions. The genes and pathways identified suggest efficient regulation leading to active adaptation as a basal defense response against water stress deficit by tea. The knowledge generated can be further utilized to better understand molecular mechanisms underlying stress tolerance in tea.

Genetic Diversity Analysis of Proso millet (Panicum miliaceum) Germplasm Using EST-SSR Markers

  • Lee, Myung-Chul;Choi, Yu-Mi;Yun, Hyemyeong;Shin, Myoung-Jae;Lee, Sukyeung;Oh, Sejong
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.10a
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    • pp.43-43
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    • 2019
  • The collection, evaluation and conservation of crop germplasm have been treated as one of the basics to breeding program. An understanding of genetic relationships among germplasm resources is vital for future breeding process like yield, quality, and resistance. In the present study, EST-SSR markers were employed to assess the polymorphism and genetic diversity of 192 accessions of Proso millet preserved in the National Agrobiodiversity Center of RDA. We evaluated the efficiency of EST-SSR markers developed for proso millet species. A total of 98 alleles were detected with an average allele number of 4.5 per locus among 192 proso millet millet accessions using 22 EST-SSR markers. The averaged values of gene diversity ($H_E$) and polymorphism information content (PIC) for each EST-SSR marker were 0.362 and 0.404 within populations, respectively. Our results showed the moderate level of the molecular diversity among the proso millet accessions from diverse countries. A phylogenetic tree revealed three major groups of accessions that did not correspond with geographical distribution patterns with a few exceptions. The less correlation between the clusters and their geographic location might be considered due to their type difference. Our study provided a better understanding of genetic relationships among various germplasm collections, and it could contribute to more efficient utilization of valuable genetic resources. The EST-SSR markers developed here will serve as a valuable resource for genetic studies, like linkage mapping, diversity analysis, quantitative trait locus/association mapping, and molecular breeding.

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