Lee, Seung Cho;Parent, Jean-Sebastien;Ernst, Evan;Berger, Frederic;Grimanelli, Daniel;Martienssen, Robert A.
Proceedings of the Korean Society of Crop Science Conference
/
2017.06a
/
pp.20-20
/
2017
Plant genomes include heterochromatic loci that consist of repetitive sequences and transposable elements. LTR retrotransposon is the major class of transposons in advanced plants in terms of proportion in plant genome. The elements contribute not only to genome size but also to genome stability and gene expression. A number of cases have been reported transposon insertions near genic regions affect crop traits such as fruit pigments, stress tolerance, and yields. Functional LTR retrotransposons produce extrachromosomal DNA from genomic RNA by reverse transcription that takes place within virus-like-particles (VLPs). DECREASED DNA METHYLATION 1 (DDM1) plays important roles in maintaining DNA methylation of heterochromatin affecting all sequence contexts, CG, CHG, and CHH. Previous studies showed that ddm1 mutant exhibits massive transcription of retrotransposons in Arabidopsis, but only few of them were able to create new insertions into the genome. RNA-dependent RNA POLYMERASE 6 (RDR6) is known to function in restricting accumulation of transposon RNA by processing the transcripts into 21-22 nt epigenetically activated small interfering RNA (easiRNA). We purified VLPs and sequence cDNA to identify functional LTR retrotransposons in Arabidopsis ddm1 and ddm1rdr6 plants. Over 20 LTR copia and gypsy families were detected in ddm1 and ddm1rdr6 sequencing libraries and most of them were not reported for mobility. In ddm1rdr6, short fragments of ATHILA gypsy elements were detected. It suggests easiRNAs might regulate reverse transcription steps. The highest enriched element among transposon loci was previously characterized EVADE element. It has been reported that active EVADE element is more efficiently silenced through female germline than male germline. By genetic analyses, we found ddm1 and rdr6 mutation affect maternal silencing of active EVADE elements. DDM1-GFP protein accumulated in megaspore mother cell but was not found in mature egg cell. The fusion protein was also found in early embryo and maternal DDM1-GFP allele was more dominantly expressed in the embryo. We observed localization of DDM1-GFP in Arabidopsis and DDM1-YFP in maize and found the proteins accumulated in dividing zone of root tips. Currently we are looking at cell cycle dependency of DDM1 expression using maize system. Among 10 AGO proteins in Arabidopsis, AGO9 is specifically expressed in egg cell and shoot meristematic cells. In addition, mutation of AGO9 and RDR6 caused failure in maternal silencing, implying 21-22 nt easiRNA pathway is important for retrotransposon silencing in female gametophyte or/and early embryo. On the other hand, canonical 24 nt sRNA-directed DNA methylation (RdDM) pathways did not contribute to maternal silencing as confirmed by this study. Heat-activated LTR retrotransposon, ONSEN, was not silenced by DDM1 but the silencing mechanisms require RdDM pathways in somatic cells. We will propose distinct mechanisms of LTR retrotransposons in germline and somatic stages.
Lee, Seung Cho;Parent, Jean-Sebastien;Ernst, Evan;Berger, Frederic;Grimanelli, Daniel;Martienssen, Robert A.
Proceedings of the Korean Society of Crop Science Conference
/
2017.06a
/
pp.97-97
/
2017
Plant genomes include heterochromatic loci that consist of repetitive sequences and transposable elements. LTR retrotransposon is the major class of transposons in advanced plants in terms of proportion in plant genome. The elements contribute not only to genome size but also to genome stability and gene expression. A number of cases have been reported transposon insertions near genic regions affect crop traits such as fruit pigments, stress tolerance, and yields. Functional LTR retrotransposons produce extrachromosomal DNA from genomic RNA by reverse transcription that takes place within virus-like-particles (VLPs). DECREASED DNA METHYLATION 1 (DDM1) plays important roles in maintaining DNA methylation of heterochromatin affecting all sequence contexts, CG, CHG, and CHH. Previous studies showed that ddm1 mutant exhibits massive transcription of retrotransposons in Arabidopsis, but only few of them were able to create new insertions into the genome. RNA-dependent RNA POLYMERASE 6 (RDR6) is known to function in restricting accumulation of transposon RNA by processing the transcripts into 21-22 nt epigenetically activated small interfering RNA (easiRNA). We purified VLPs and sequence cDNA to identify functional LTR retrotransposons in Arabidopsis ddm1 and ddm1rdr6 plants. Over 20 LTR copia and gypsy families were detected in ddm1 and ddm1rdr6 sequencing libraries and most of them were not reported for mobility. In ddm1rdr6, short fragments of ATHILA gypsy elements were detected. It suggests easiRNAs might regulate reverse transcription steps. The highest enriched element among transposon loci was previously characterized EVADE element. It has been reported that active EVADE element is more efficiently silenced through female germline than male germline. By genetic analyses, we found ddm1 and rdr6 mutation affect maternal silencing of active EVADE elements. DDM1-GFP protein accumulated in megaspore mother cell but was not found in mature egg cell. The fusion protein was also found in early embryo and maternal DDM1-GFP allele was more dominantly expressed in the embryo. We observed localization of DDM1-GFP in Arabidopsis and DDM1-YFP in maize and found the proteins accumulated in dividing zone of root tips. Currently we are looking at cell cycle dependency of DDM1 expression using maize system. Among 10 AGO proteins in Arabidopsis, AGO9 is specifically expressed in egg cell and shoot meristematic cells. In addition, mutation of AGO9 and RDR6 caused failure in maternal silencing, implying 21-22 nt easiRNA pathway is important for retrotransposon silencing in female gametophyte or/and early embryo. On the other hand, canonical 24 nt sRNA-directed DNA methylation (RdDM) pathways did not contribute to maternal silencing as confirmed by this study. Heat-activated LTR retrotransposon, ONSEN, was not silenced by DDM1 but the silencing mechanisms require RdDM pathways in somatic cells. We will propose distinct mechanisms of LTR retrotransposons in germline and somatic stages.
When plants are exposed to subfreezing temperatures ice crystals are forming within extracelluar space in leaves. The growth of ice crystal is closely related to the degree of freezing injury. It was shown that an antifreeze protein binds to an ice nucleator through hydrogen bonds to prevent growth of ice crystal and also reduces freezing damage. The antifreeze proteins in plants are similar to PR proteins but only the PR proteins induced upon cold acclimation were shown to have dual functions in antifreezing as well as antifungal activities. Three of the genes encoded for CLP, GLP, and TLP were isolated from barley and Kentucky bluegrass based on amino acid sequence revealed after purification and low temperature-inducibility as shown in analysis of the protein. The deduced amino acid of the genes cloned showed a signal for secretion into extracellular space where the antifreezing activity sup-posed to work. The western analysis using the antisera raised against the antifreeze proteins showed a positive correlation between the amount of the protein and the level of freeze tolerance among different cultivars of barely. Besides it was revealed that TLP is responsible for a freeze tolerance induced by a treatment of trinexapac ethyl in Kentucky bluegrass. Analysis of an overwintering wild rice, Oryza rufipogon also showed that an acquisition of freeze tolerance relied on accumulation of the protein similar to CLP. The more direct evidence for the role of CLP in freeze tolerance was made with the analysis of the transgenic tobacco showing extracellular accumulation of CLP and enhanced freeze tolerance measured by amount of ion leakage and rate of photosynthetic electron transport upon freezing. These antifreeze proteins genes will be good candidates for transformation into crops such as lettuce and strawberry to develop into the new crops capable of freeze-storage and such as rose and grape to enhance a freeze tolerance for a safe survival during winter.
This study was conducted to examine the effect of several postharvest treatments on the storable quality of three apple cultivars, 'Tsugaru', 'jonagold', and 'Hokudo' which are difference in harvest season. Ethylene evolution level showed cultivar-dependent pattern during storage and there was no correlation between the amount of ethylene evolution and storage period among cultivars. Polyethylene film(PE) wrapping was effective in the inhibition of water loss and functional ceramic film was also effective in the prevention of ethylene accumulation inside the wrap during 1 to 3 months of cold storage compared to those of simple PE wrapping. Ethylene scrubber and wax treatment positively contributed to keep the fruit firmness 10% higher than that of control fruit during storage. Waxing enhanced the appearance of 'jonagold' even after simulated marketing period through inhibiting wax accumulation on the fruit surface.
Journal of Advanced Marine Engineering and Technology
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v.34
no.4
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pp.508-514
/
2010
The objective of this work presented here was experimental study of steadystate and cold start exhaust nano-sized particle characteristics from common rail diesel engine. The effect of the diesel oxidation catalyst (DOC) on the particle number reduction was insignificant, however, particle number concentration levels were reduced by 3 orders of magnitude into the downstream of diesel particulate filter (DPF). In high speed and load conditions, natural regeneration of trapped particle occurred inside DPF and it was referable to increase particle number concentration. As fuel injection timing was shifted BTDC $6^{\circ}CA$ to ATDC $4^{\circ}CA$, particle number concentration level was slightly reduced, however particle number and size was increased at ATDC $9^{\circ}CA$. Nucleation type particle reduced and accumulation type particle was increased on EGR condition.
Kim, Byoung-Soo;Kim, Eun-Jung;Lee, Hae-June;Han, Sang-Jin;Choi, Tae-Hyun;Lee, Yun-Sil;Cheon, Gi-Jeong
Bulletin of the Korean Chemical Society
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v.31
no.9
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pp.2649-2655
/
2010
$PKC{\delta}$-catalytic V5 Heptapeptide (FEQFLDI, FP7) interacts with heat shock protein 27 (HSP27) and inhibits HSP27-mediated resistance to cell death against various stimuli including radiation therapy. Here, we prepared radio-iodinated heptapeptide and further investigated its uptake properties in HSP27 expression cells. Peptide sequence of FP7 and a negative control peptide (WSLLEKR, QP7) was modified by substituting their C-terminus residue to tyrosine (FP6Y and QP6Y) to label radio-iodine. Iodinated peptides were confirmed by LC mass analysis with cold iodine reaction mixture. Accumulation of [$^{125}I$]iodo-FP6Y and [$^{125}I$]iodo-QP6Y in NCI-H1299 cell line, with higher level of HSP27, and NCI-H460 cell line, with lower level of HSP27, was measured by NaI(Tl) scintillation counter. The modification of substituting C-terminus residue of FP7 to tyrosine (FP6Y) did not affect its interaction with HSP27. Accumulation of [$^{125}I$]iodo-FP6Y in NCI-H1299 cells was 3 fold higher than in NCI-H460 cells. The novel radio-iodinated FP6Y would be used as a tracer for targeting HSP27 protein.
For the production of Nucleic acid derivatives by microorganisms, adenineless mutants were induced from Bacillus subtilis with the mutagens, ultraviolet rays and diethylsulfate. And total strains of 62 adenineless mutants were isolated. These mutants were found to have an ability of accumulating u. v. absorbing substances in the culture broth. They were analyzed to be hypoxanthine, uracil, and other unknown compound by means of two dimensional thin layer chromatography, u. v. absorption spectra, and I. R. absorption spectrum. Mutant BS-137 was screened out to accumulate hypoxanthine, which was isolated from the culture broth through activated carbon column and recrystallize from cold water. The effect of the medium compositions on hypoxanthine accumulation were examined for the mutant BS-137. Glucose appeared to be the best carbon source, but molasses could be substitute for it, NaNO$_3$ and yeast extract were especially good sources of nitrogen.
Ephedra distachya cultures have been known to accumulate two major $\rho-coumaroylamino$ acids (p-coumaroylgicine and $\rho-coumaroylamino$ -D-alanine) by D-Ala treament. When D-Ala was added together with serial concentrations of yeast-derived elicitor, the accumulation of $\rho-coumaroylamino$ -D-Ala $(\rho-CDA)$ was geatly increased in an additive manner. In feeding experiments, $[1-^14C]$-D-Ala was incorporated into $\rho-CDA$at a rate of 2.2% or 2.3% of added radioactivity, indicating that exogenous D-Ala served as a precursor of the conjugate. $[1-^14C]$-L-Ala wasalso incorporated into p-CDA (0.23%) in the elicitor treated cultures. This fact suggested that at least a part of $(\rho-CDA)$ was produced from active conversion of L-Ala by the elecitation. In order to investigate a possible role of D-Ala as an elicitor of $\rpo-coumaroylamino$ acids $(\rpo-CAA)$, cold D-Ala was added together with labeled L-Ala. Although L-Ala seemed to be incorporated into $\rho-CDA$ by this treatment, the incorporation ratio was too small (0.054%) to draw a clear conclusion. However, the amount of $\rpo-coumaroylglycine$, which did not use D-Ala as a substrate, was also sightly increased by D-Ala treatment irrespective of the presence of elicitor, suggesting that exogenous D-Ala might act as an elicitor of$\rpo-CAA$ as well as a precusor substrate of $\rho-CDA$.
In Korean peninsula as well as in Western European Countries, we can find, reddish bands in sandy deposits of various origin. These bands (called "$B_t$ Band"), composed of fine material, are regularly intervalled and parallel to topography. This study is tring to elucidate their origin and associated milieux, with a group of methods and techniques (field survey, micromorphological analysis, granulometry, X-ray diffractometry of clay minerals...). $B_t$ Bands are not sedimentological origin. They were formed by superposition of two type accumulations in the sandy profile, triggered by different mechanisms: 1) accumulation in banded form, concerning silty heterometric material and micas, oweing to cryophoretic forces operating in course of deep freezing of sandy deposits: 2) illuviation of ferrugineous clays in the precedants bands, migrated in chelate state by soil water. The first processes are associated with very cold and humid climate in which freeze soil more than 3m in depth. The second, cold and humid climate in which podzolizattion is generalized. In case of South Korea and Western Europe, these two types of climates are not present circumstance, but Recent Quaternary (probably since the Wurm).the Wurm).
Korean Journal of Agricultural and Forest Meteorology
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v.6
no.1
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pp.49-60
/
2004
Northern Gyeonggi Province(NGP), consisting of 3 counties, is the northernmost region in South Korea adjacent to the de-militarized zone with North Korea. To supplement insufficient spatial coverage of official climate data and climate atlases based on those data, high-resolution digital climate models(DCM) were prepared to support weather- related activities of residents in NGP Monthly climate data from 51 synoptic stations across both North and South Korea were collected for 1981-2000. A digital elevation model(DEM) for this region with 30m cell spacing was used with the climate data for spatially interpolating daily maximum and minimum temperatures, solar irradiance, and precipitation based on relevant topoclimatological models. For daily minimum temperature, a spatial interpolation scheme accommodating the potential influences of cold air accumulation and the temperature inversion was used. For daily maximum temperature estimation, a spatial interpolation model loaded with the overheating index was used. Daily solar irradiances over sloping surfaces were estimated from nearby synoptic station data weighted by potential relative radiation, which is the hourly sum of relative solar intensity. Precipitation was assumed to increase with the difference between virtual terrain elevation and the DEM multiplied by an observed rate. Validations were carried out by installing an observation network specifically for making comparisons with the spatially estimated temperature pattern. Freezing risk in January was estimated for major fruit tree species based on the DCMs under the recurrence intervals of 10, 30, and 100 years, respectively. Frost risks at bud-burst and blossom of tree flowers were also estimated for the same resolution as the DCMs.
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