• Title/Summary/Keyword: Environmental clone

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Symptom Determinant as RNA3 of Lily Isolates of Cucumber mosaic virus on Zucchini Squash

  • Cho, Seung-Kook;Ahn, Hong-Il;Kim, Min-Jea;Choi, Jang-Kyung;Ryu, Ki-Hyun
    • The Plant Pathology Journal
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    • v.20 no.3
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    • pp.212-219
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    • 2004
  • Three isolates of Cucumber mosaic virus (CMV) from lily plants showing mosaic and distortion symptoms were detected by reverse-transcriptase polymerase chain reaction (RT-PCR) using primers specific to Cucumovirus genus namely, LK-CMV, LK4-CMV, and LKS-CMV. Restriction enzymes patterns of the RT-PCR products revealed that the lily isolates belonged to subgroup IA of CMV. In terms of biological properties, the lily isolates have highly similar but distinct pathogenicity as reported in other lily strains and ordinary strains of CMV. To characterize the molecular properties, cDNAs containing coat protein (CP) gene and 3' non-coding region (NCR) of RNA3 for the isolates were cloned and their nucleotide sequences were determined. The CP similarity (218 amino acids) was highly homologous (>97%) with that of subgroup I CMV strains. However, an additional 20-nulcleotide long segment was only present in 3' NCR of lily isolates, which form an additional stem-loop RNA structure. By using chimeric construct exchange cDNA containing 3'NCR of LK-CMV into the full-length cDNA clone of RNA3 of Fny-CMV, this additional segment may prove to be significant in the identification and fitness of the virus in lily plants. The pathology of zucchini squash infected by F1F2L3-CMV, a pseudorecombinant virus was showed to change drastically the severe mosaic and stunting symptom into a mild chlorotic spot on systemic leave, compared with Fny-CMV. To delimit the sequence of RNA3 affected the pathology, various RNA3 chimeras were constructed between two strains of CMV. The symptom determinants of F1F2L3-CMV were mapped to the positions amino acid 234, 239, and 250 in 3a movement protein (MP). RNA3 chimeras changed the sequences encoding three amino acids were resulted in alteration of systemic symptom.

Genome Mapping Technology And Its Application In Plant Breeding (작물 육종에서 분자유전자 지도의 이용)

  • 은무영
    • Proceedings of the Botanical Society of Korea Conference
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    • 1995.07a
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    • pp.57-86
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    • 1995
  • Molecular mapping of plant genomes has progressed rapidly since Bostein et al.(1980) introduced the idea of constructing linkage maps of human genome based on restriction fragment length polymorphism (RFLP) markers. In recent years, the development of protein and DNA markers has stimulated interest for the new approaches to plant improvement. While classical maps based on morphological mutant markers have provided important insights into the plant genetics and cytology, the molecular maps based on molecular markers have a number of inherent advatages over classical genetic maps for the applications in genetic studies and/or breeding schemes. Isozymes and DNA markers are numerous, discrete, non-deleterious, codominant, and almost entirely free of environmental and epistatic interactions. For these reasons, they are widely used in constructing detailed linkage maps in a number of plant species. Plant breeders improve crops by selecting plants with desirable phenotypes. However a plant's phenotyes is often under genetic control, positioning at different "quantitative trait loci" (QTLs) together with environmental effects. Molecular maps provide a possible way to determine the effect of the individual gene that combines to produce a quantitative trait because the segregation of a large number of markers can be followed in a single genetic cross. Using market-assisted selection, plants that contain several favorable genes for the trait and do not contain unfavourable segments can be obtained during early breeding processes. Providing molecular maps are available, valuable data relevant to the taxonomic relationships and chromosome evolution can be accumulated by comparative mapping and also the structural relationships between linkage map and physical map can be identified by cDNA sequencing. After constructing high density maps, it will be possible to clone genes, whose products are unknown, such as semidwarf and disease resistance genes. However, much attention has to be paid to level-up the basic knowledge of genetics, physiology, biochemistry, plant pathology, entomology, microbiology, and so on. It must also be kept in mind that scientists in various fields will have to make another take off by intensive cooperation together for early integration and utilization of these newly emerging high-techs in practical breeding. breeding.

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Microbial Community in the TPH-Contaminated Aquifer for Hot Air Sparging using Terminal-Restriction Fragment Length Polymorphism (유류오염대수층 고온공기분사공정시 제한효소다형성 미생물 군집)

  • Lee, Junho;Park, Kapsong
    • Journal of Korean Society on Water Environment
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    • v.24 no.1
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    • pp.19-29
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    • 2008
  • Hot air sparging is a groundwater remediation technique, in which organic contaminants volatilized into hot air from the saturated to vadose zone. In the laboratory diesel (10,000 mg TPH/kg) was spiked in contaminated saturated aquifer soil. The hot air ($34.9{\pm}2.7^{\circ}C$) was injected in intermittent (Q=1,500 mL/min, 10 minute injection and 10 minute idle) modes. We performed microcosm tests using the groundwater samples to assess TPH reductive remediation activity. For Terminal-Restriction Fragment Length Polymorphism (T-RFLP) analysis of eubacterial communities in sludge of wastewater treatment plants and soil of experiment site, the 16S rDNA was amplified by Polymerase Chain Reaction (PCR) from the sludge and the soil. The obtained 16S rDNA fragments were digested with Msp I and separated by electrophoresis gel. We found various sequence types for hot air sparging experiment with sludge soil samples that were closely related to Bacillus (149 bp, Firmicutes), Methlobacterium (149 bp, Euryarchaeotes), Pseudomonas (492 bp, ${\gamma}$-Proteobacteria), etc., in the clone library. In this study we find that TPH-water was reduced to 78.9% of the initial value in this experiment aquifer. The results of the present study suggests that T-RFLP method may be applied as a useful tool for the monitoring in the TPH contaminated soil fate of microorganisms in natural microbial community.

Effects of Nitrogen Fertilization and Thinning on Growth and Nutrition of Current Needle and Twigs in Pinus koraiensis (질소(窒素) 시비(施肥)와 간벌(間伐)이 잣나무 당년생(當年生) 침엽(針葉)과 소지(小枝)의 생장(生長) 및 양분(養分)에 미치는 영향(影響))

  • Son, Yowhan;Hwang, Jae Woo;Lee, Do Hyeung;Kim, Jong Sung;Yang, Soo Young
    • Journal of Korean Society of Forest Science
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    • v.90 no.1
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    • pp.28-35
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    • 2001
  • To investigate the effects of nitrogen fertilization and thinning on growth and nutrition of current-year needle and twigs in Pinus koraiensis we treated two clone banks with three levels of nitrogen fertilizer in Yongin and Chuncheon, and also thinned two plantations with three intensities in Yangpyeong. The fertilization and thinning effects greatly varied depending on study sites and stand ages. However, dry weights of needle and twig increased following low nitrogen fertilization in older stands. Moderate thinning increased dry weight and nitrogen concentration of twigs in the 12-year-old stand.

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Characteristics and Germination of Xanthoceras sorbifolia Bunge Seeds Originated from Inner Mongolia and Liaoning, China

  • An, Chan-Hoon;Lee, Hyun-Seok;Yin, Zhi-Yang;Yi, Jae-Seon
    • Journal of Forest and Environmental Science
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    • v.27 no.3
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    • pp.151-156
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    • 2011
  • Seeds of Xanthoceras sorbifolia Bunge were collected from two plantations and two superior trees in Inner Mongolia: and one plantation and one superior tree in Liaoning, China in late August, 2011. Yellowhorn or goldenhorn is an important tree species, from the aspects of source of edible oil and biodiesel and pioneering capacity of degraded and desert land. Characteristics investigated were seed length, width, and weight: weight and volume of 1,000 seeds: and weight and volume of one-liter seeds. The seeds of Qingsonglingxiang No. 1, growing alone in an open space, showed the highest values in seed length (16.08 mm), width (14.48 mm) and weight (1.40 g), while those of Tree No. 160 in Ar Khorqin Banner were the lowest ones: that is, 11.48mm for length, 11.81 mm for width, and 0.73 g for weight, respectively. Traits of seeds varied quite much between trees and among areas; for example, Tree No. 38 and No. 160 produced quite different seeds in several traits, although they are adjacent to each other in the same farm. Weight of 1,000 seeds varied from 718.0 g to 1,010.1 g and volume from 0.76 L to 1.52 L. Weight of one-liter seeds were 522.3 g to 688.2 g, while the number of seeds were 603 to 935. Seeds which were soaked in the water at $4^{\circ}C$ for 2 days showed the highest germination rate (89%) in a 30-day test, which was about 10% to 40% higher than those of non-treatment and dipping treatment at $36^{\circ}C$ followed by keeping under room temperature for 2 days. 81% of seeds in the wet sand at room temperature germinated, while 23% of seeds deprived of seed coat germinated. It is necessary to understand seed traits to select superior clones or provenances for the increased, unfluctuating production of seed.

Screening and Isolation of a Gene Encoding 4-Hydroxyphenylpyruvate Dioxygenase from a Metagenomic Library of Soil DNA (토양의 DNA로부터 4-Hydroxyphenylpyruvate Dioxygenase 유전자 탐색 및 분리)

  • Yun, Sang-Soon;Lee, Jung-Han;Kim, Soo-Jin;Kim, Sam-Sun;Park, In-Cheol;Lee, Mi-Hye;Koo, Bon-Sung;Yoon, Sang-Hong;Yeo, Yun-Soo
    • Applied Biological Chemistry
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    • v.48 no.4
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    • pp.345-351
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    • 2005
  • To access the natural products of uncultured microorganisms, we constructed and screened the metagenomic DNA libraries by using a cosmid vector and DNA inserts isolated directly from soil. Initial screening of the libraries in Escherichia coli resulted in the isolation of several clones that produce a dark brown color when grown in LB medium. One of the positive clones, designed pYS85C, was transposon mutagenized and the DNA surrounding the transposon insertions in cosmids that no longer conferred the production of brown pigment to E. coli was sequenced. Annotation of the pYS85C sequence obtained from the transposon mutagenesis experiment indicated a single 393 amino acid open reading frame (ORF) with a molecular mass of about 44.5 kDa, predicted to be a 4-hydroxyphenylpyruvate dioxygenases (HPPDs), was responsible for the observed brown pigment. In a BLAST search against deposited sequence, the translated protein from this ORF showed moderate-level identity (>60%) to the other known HPPDs and was most conserved in the C-terminal region of the protein. These results show that genes involved in natural product synthesis can be cloned directly from soil DNA and expressed in a heterologous host, supporting the idea that this technology has the potential to provide novel natural products from the wealth of environmental microbial diversity and is a potentially important new tool for drug discovery.

Phenotypic and molecular characteristics of second clone (T0V2) plants of the LeLs-antisense gene-transgenic chrysanthemum line exhibiting non-branching (무측지성 국화 형질전환 계통 영양번식 제2세대의 형태적 및 분자생물학적 특성)

  • Lee, Su Young;Kim, Jeong-Ho;Cheon, Kyeong-Seong;Lee, Eun Kyung;Kim, Won Hee;Kwon, O Hyeon;Lee, Hye Jin
    • Journal of Plant Biotechnology
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    • v.40 no.4
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    • pp.192-197
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    • 2013
  • This study examined the phenotypic and molecular characteristics of the $2^{nd}$ clone ($T_0V_2$) plants of LeLs-antisense gene-transgenic chrysanthemum line (LeLs80) that exhibited non-branching, proving the relevance of these characteristics as a factor for use in environmental risk assessment. Results of the Southern blot analysis showed that three copies of the LeLs-antisense gene were introduced into the transgenic line, and northern analysis showed that the transcripted gene was normally expressed in the transgenic line. A flanking T-DNA sequencing method was used to determine that sequences of 184 and 464 bps flanked the LeLs-antisense gene in the transgenic line. These sequences, respectively, matched the 35S promoter for expression of the npt II gene and the NOS terminator for expression of the LeLs-antisense gene within the pCAMBIA 2300 vector.

Hypersensitivity of Somatic Mutations and Mitotic Recombinations Induced by Heterocyclic amines and Aflatoxin $B_1$ in Transgenic Drosophila (형질전환 초파리에서 Heterocyclic Amines와 Aflatoxin $B_1$에 의한 체세포 돌연변이 유발의 고감수성에 관한 연구)

  • 최영현;유미애;이원호
    • Korean journal of applied entomology
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    • v.35 no.4
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    • pp.315-320
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    • 1996
  • The effects of 2-arnino-3-methyIimidazo[4,5-fq]u inoline (IQ), 2-amino-6dimethyl-dipyrido[l,2-a;3',2'-d] imidazole (Glu-P-1) and aflatoxin B1 (AFBI) on the mitotic recombinations and somatic chromosome mutations were investigated using the transgenic Drosophila bearing a chimeric gene consisting of a promoter region of Drosophila actin 5C gene and rat DNA polymerase $. For investigating mitotic recombinations and the somatic chromosome mutations, the heterozygous (mwhl+) strain possessing or lacking transgene pol P was used. The spontaneous frequency of small mwh spots, due to deletion or nondisjunction etc., in the non-transgenic w strain and the transgenic plpol $1-130 strain was 0.351 and 0.606, respectively. The spontaneous frequency (0.063) of large mwh spots, arising mostly from somatic recombination between the centromere and the locus mwh, in the transgenic plpol $1-130 strain, was about three times higher than that (0.021) of the non-transgenic w strain. The mutant clone frequencies of two types induced by two heterocyclic mines (IQ and Glu-P-1) and AFBl in the transformant pbol PI-130 were two or three times higher than those in the host strain w. These mean that rat DNA polymerase P participates at least in the somatic chromosome mutations and mitotic recombination processes. And the present results suggest that the transgenic Drosophl!~ used in this study can be used as a hypersensitive, in vivo short-term assaying system for various environmental mutagens.

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Molecular Analysis of Bacterial Communities Distributed in Sea Water of Whitening Areas of Jeju Coast (제주연안 갯녹음(백화) 지역의 해수에 분포하는 세균군의 분자생물학적 분석)

  • 강형일;강봉조;김미란;윤병준;이동헌;오덕철
    • Korean Journal of Microbiology
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    • v.38 no.2
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    • pp.127-132
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    • 2002
  • In this study, the bacterial communities distributed in sea water of the whitening areas of Gangjeong and Seongsan, Jeju-do have been analysed using the PCR amplification of 16S rRNA to obtain fundamental data and information on relationship of the whitening phenomenon and microbial ecosystem. In Gangjeong, diverse bacteria such as Alcanivorax, Paracoccus, Damselae, Pseudomonas, Rhodowlum, Silicibacter, Suiftobacter, and Roseobacter have been found, and Alcanivorax was the most abundant clone. The most abundant clone from Seongsan was Pseudomonas, of which Pseudomonas tolaasii and Pseudomonas mandeli were most abundantly occurred in the frequency of approx44% and 24%, respectively. Approx4% of the bacterial clones closest to Verrucomicrobiales and other unidentified clones were also fecund in Seongsan, suggesting there is a great discrepancy between bacterial communities from the whitening areas of Seongsan and Gangjeong. The mean temperature, chlorine concentration, pH, and dissolved oxygen (DO) of the sea water of Gangieong and Seongsan in August of 2001 (sampling period) was $27^{\circ}C$~$27.5^{\circ}C$, 30.24~30.60%, pH 8.23~8.36,7 .20~7.28 mg/ι, suggesting other environmental factors except far the factors mentioned above might result in difference of bacterial communities distributed in both areas.

Oxidative Stress and Antioxidant Responses in Poplar Clones Irrigated with Livestock Waste Leachate (양돈폐수 처리에 의한 포플러클론의 산화스트레스와 항산화반응)

  • Je, Sun-Mi;Yeo, Jin-Gi;Woo, Su-Young
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.13 no.3
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    • pp.140-147
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    • 2011
  • We studied the influence of livestock waste leachate on oxidative damage and antioxidative responses in poplar clones in August which increase the demand of antioxidants because of high temperature and high light during this period. We measured ion leakage, antioxidant enzyme activities (APX, GR), and carotenoid contents. Oxidative damage and antioxidative responses by treated livestock waste leachate in poplar clones showed various results. We divided poplar clones into three groups using the criteria based on ion leakage which represent cell damage induced oxidative stress. Eco 28, 62-10, Bonghwa1 and Dorskamp belonged to the first group in which the cell damaged level was lower than that of the control. The results suggest that this group augmented for demand of antioxidative in summer because high concentration of nitrogen induced by treatment of live stock wastes acted as environmental stress. Consequently, they failed to keep up the homeostasis of reactive oxygen species. The second group in which the cell damaged level was similar to that of the control was Suwon, 72-30 and 72-31 clones. Finally, 97-18 clone belonged to the third group in which the cell damaged level was lower than that of the control group. In this case, nitrogen treated by livestock waste leakage decreased oxidative stress. 97-18 clone was the clones with the least damage in summer oxidative stresses treated by livestock waste leakage. These results suggest that the high concentration nitrogen due to the livestock waste leakage can act differently upon the clones. We speculate that the added oxidation damage in the summer (growing season) may have an effect on the total fresh weight and also influence the purification ability for livestock waste leakage. However, further studies are needed for the confirmation.