Development of transgenic plant increasing crop yield or disease resistance is good way to solve the world food shortage. However, the persistence of marker genes in crops leads to serious public concerns about the safety of transgenic crops. In the present paper, we developed marker-free transgenic rice inserted high molecular-weight glutenin subunit (HMW-GS) gene ($D{\times}5$) from the Korean wheat cultivar 'Jokyeong' using Agrobacterium-mediated co-transformation method. Two expression cassettes comprised of separate DNA fragments containing only the $D{\times}5$ and hygromycin resistance (HPTII) genes were introduced separately into Agrobacterium tumefaciens EHA105 strain for co-infection. Each EHA105 strain harboring $D{\times}5$ or HPTII was infected into rice calli at a 3: 1 ratio of EHA105 with $D{\times}5$ gene and EHA105 with HPTII gene expressing cassette. Then, among 66 hygromycin-resistant transformants, we obtained two transgenic lines inserted with both the $D{\times}5$ and HPTII genes into the rice genome. We reconfirmed integration of the $D{\times}5$ and HPTII genes into the rice genome by Southern blot analysis. Wheat $D{\times}5$ transcripts in $T_1$ rice seeds were examined with semi-quantitative RT-PCR. Finally, the marker-free plants containing only the $D{\times}5$ gene were successfully screened at the $T_1$ generation. These results show that a co-infection system with two expression cassettes could be an efficient strategy to generate marker-free transgenic rice plants.
Rice flour is used in many food products. However, dough made from rice lacks extensibility and elasticity, making it less suitable than wheat for many food products such as bread and noodles. The high-molecular weight glutenin subunits (HMW-GS) of wheat play a crucial role in determining the processing properties of the wheat grain. This paper describes the development of marker-free transgenic rice plants expressing a wheat Glu-Dy10 gene encoding the HMG-GS from the Korean wheat cultivar 'Jokyeong' using Agrobacterium-mediated co-transformation. Two expression cassettes, consisting of separate DNA fragments containing Glu-1Dy10 and hygromycin phosphotransferase II (HPTII) resistance genes, were introduced separately into Agrobacterium tumefaciens EHA105 for co-infection. Each EHA105 strain harboring Glu-1Dy10 or HPTII was infected into rice calli at a 3: 1 ratio of Glu-1Bx7 and HPTII. Among 290 hygromycin-resistant $T_0$ plants, we obtained 29 transgenic lines with both the Glu-1Dy10 and HPTII genes inserted into the rice genome. We reconfirmed the integration of the Glu-1Dy10 gene into the rice genome by Southern blot analysis. Transcripts and proteins of the Glu-1Dy10 in transgenic rice seeds were examined by semi-quantitative RT-PCR and Western blot analysis. The marker-free plants containing only the Glu-1Dy10 gene were successfully screened in the $T_1$ generation.
Korean Journal of Agricultural and Forest Meteorology
/
v.16
no.3
/
pp.219-226
/
2014
Evaluation of tree responses to temperature elevation is critical for a development of forest management techniques coping with climate change. We conducted a study on the growth responses of Betula costata, Fraxinus rhynchophylla, and Quercus variabilis seedlings to open-field artificial warming. Artificial warming set-up using infra-red heater was built in 2012 and the temperature in warmed plots was regulated to be consistently $3^{\circ}C$ higher than that of control plots. The seeds of three species were sown, and the responses of growth, biomass allocation, and net photosynthetic rate of newly-germinated seedlings on the open-field artificial warming were determined. As a result, the growth responses of the seedlings differed with the species. B. costata showed decreases in the height to diameter ratio (H/D ratio), biomass, root weight to shoot weight ratio, and net photosynthetic rate. However, root collar diameter (RCD), height, biomass, and net photosynthetic rate of Q. variabilis were increased, while the response of F. rhynchophylla was rather obscure. There was no significant difference between warmed and control plots in seedling growth for 3 species in July, whereas, RCD, height, and H/D ratio of Q. variabilis were increased and H/D ratio of B. costata was decreased in November under warming. Species-specific growth responses to warming were similar to the species-specific responses of net photosynthetic rate and biomass allocation; therefore, net photosynthetic rate and biomass allocation might attribute to growth responses to warming. Besides, a relatively obvious response in autumn compared to summer might be affected by the phenological change following artificial warming. Species-specific responses of three deciduous species to warming in this study could be applied to the development of adaptive forest management policies to climate change.
Park, Cheol-Ho;Yu, Chang-Yeon;Kim, Dong-Wook;Cho, Hye-Kyeong;Park, Kyeong-Suk;Seo, Jeong-Sik;Ahn, Sang-Deuk;Jang, Byeong-Ho
Korean Journal of Medicinal Crop Science
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v.2
no.1
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pp.60-66
/
1994
This study was conducted to determine the optimum conditions of inducing callus, proliferating callus, forming somatic embryos, and regenerating plantlets via somatic embryogenesis, for the purpose of producing artificial seeds and substantially developing plant factory technologies that can be employed to all seasons production of Bupleurum plants. Callus was efficiently induced from leaf tissues at three leaf stage in the MS medium supplemented with 2, 4-D 2mg /1 and thidiazuron(TDZ) 0.lmg /1. Callus induction from leaf tissues at maturity was mostly effective in the mixture of 2,4- D 2mg /1 and TDZ 1.0mg /1 while that from flower bud tissues was fairly good in the MS medium containing 2,4-D 1 or 2mg /1.Callus was formed in 15 to 20 days after culture initiation in the MS media supplemented with 2, 4- D 1-2mg /1 and TDZ 0.l-1.0mg /1. Such hormones as kinetin 3mg /1, GA 1mg /1, and the mixture of GA 1mg /1 and TDZ 1mg /1 effected markedly to proliferate the callus cells.The optimum temperature and light intensity for callus culture were found to be $25^{\circ}C$ and 3000 Lux, respectively. Direct plant regeneration from cultured callus was fairly made on hormone-free MS or half-strength MS medium. Somatic embryogenesis was most frequently observed in hormone-free media:60 somatic embryos per 20ml in MS medium and 28 somatic embryos per 20ml in half -strength MS medium. There were three stages-globular, heart, and torpedo-in development of somatic embryos, among which globular stage was more frequently observed in MS medium rather than in half-strength MS medium. Somatic embryos induced from suspension culture fairly differentiated a number of shoots and roots on hormone-free and half-strength MS solid medium.
Differing in water conditions, the dry matter weight per plant was highest at 0 cm flooding depth, and was decreased at above 2 cm flooding depths. The shoot and spikes per pot developed best at flooding depths of 0 and 2 cm, but worst at -5cm or above 4cm flooding depths. The dry matter weight of shoots was linearly increased, but the weight of roots was sharply decreased according to high temperature after flower initation. The shoots and spikes per pot developed more effectively at 25$^{\circ}C$ than at 35 or 15$^{\circ}C$. The number of shoot and spike per pot were decreased according to higher shading. The effects of shading of 25-45% were not significantly small. The natural white and yellow spectra were the most effective to increase dry matter weight, shoots and spikes per pot. The dry matter weights of shoot and root per plant were not significantly different among at: 50 and 75% clay. The number of shoots were best and continuously increased at 75% clay, and the increments at 0 and 100% clays showed the lag period at early stage. The pattern of spikes was similar to shoots with less difference in various clay composition. The dry matter weight per plant in paddy field and upland field was basically similar. The numbers of shoot and spike were increased in response to increased fertilizer levels up to 20Kg per 10a of each component.
A water culture experiment was conducted to investigate the effect of As content in a culture solution on the water status and growth of rice plants. Rice (Oryza sativa L. Line. Iri 316) seeds were germinated in bentonite and cultivated there for 30 days. Rice seedlings were transplanted into 3.5l pots containing the culture solution on May 1, 1985 and allowed to grow without As treatment for one month. Afterwards, they were grown in a culture solution maintaining the final concentration of As, 0, 1, 5, 10 and 15mg/1 renewing in the solution dissolved sodium arsenate at intervals of 3 to 7 days. Plants were cultivated in the green house during the growing period and harvested 60 days after As treatment. The results obtained were as follows: Transpiration of rice plants was decreased with the increase of the As level in the culture solution. Stomatal diffusive resistance and leaf temperature increased with increase of As levels though the humidity and the air flow rate in leaf decreased. Air flow rate, transpiration and stomatal diffusive resistance showed a highly significant correlation with As contents in shoots and roots of rice plants: Espally The air flow rate and transpiration revealed a significantly higher correlation with As contents in the root than that in the shoot, but diffusive resistance showed adverse tendency. High levels of As in the culture solution depressed plant height, no. of tillers, leaf width and dry weight of plant remarkably. Typical symptoms of As toxicity were root discoloration, and necrosis of leaf tips and margins, and leaf rolling during the sunny daytime were also another symptoms.
Park, Won-Zei;Lee, Kyong-Jae;Han, Bong-Ho;Jang, Jae-Hoon
Journal of the Korean Institute of Landscape Architecture
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v.40
no.2
/
pp.112-129
/
2012
The aim of this study was to provide basic data in managing the project that was carried out on Bulgwangcheon in a nature-friendly way to improve the conditions around the areas, which was brought to completion in 2002, based on changes in ecological characteristics. For this propose, this study examined documents related to the project, compared physical and enviromnental changes before and after the project was conducted and analyzed changes in the stream ecosystem. The result showed that in areas that effluent water was often observed, especially when it rained, the river wall was washed away and vegetation was found damaged. As for actual vegetation, this study compared planting coverage of each section of the research area and actual vegetation charts. The results indicated that Lespedeza spp., Aster koraiensis among mixed seeds that were planted in the reservoir path were almost swept away while Festuca arundinacea dominated the areas. Phragmites communis, Miscanthus sacchariflorns and Salix gracilistyla which had been planted in a small number were also almost washed out though a small number of them were left to form a colony. After examining the topography and structure of the plant community, this study found that areas where mixed seed were planted had changed into two types of vegetation: First type of area is dominated by P and R which are usually raised in apron with abundant floating particles. The second type of area is dominated by dry gramineous plant such as F and A. Areas around low flow channel where Phragmites communis, Miscanthus sacchariflorus and Salix gracilistyla planting construction method is applied was washed away with the width of low flow channel reduced. Though P, M and S formed a small community in some areas around the low flow area, they were in small number and in composition of simple plant species. Two ways were suggested in this study to manage the stream in an ecological way. First, adequate revetment construction methods should be applied by monitoring the flow of the stream as well as considering the flood control of urban streams. Second, target vegetation communities that are suitable for the environment of the stream should be chosen and be plantedconstantly with high density. At the same time, ornamental native plants shouldn't be planted as they have been and disturbing vegetation should be removed.
Kim, Sun;Ryu, Jin-Hee;Kim, Young-Joo;Jeong, Jae-Hyeok;Lee, Su-Hwan;Oh, Yang-Yeol;Kim, Young-Doo;Kim, Jae-Hyen
KOREAN JOURNAL OF CROP SCIENCE
/
v.61
no.2
/
pp.113-118
/
2016
This experiment was conducted to identify the variations in inorganic nutrients and plant growth in millet (Panicum miliaceum L.) due to soil salinity. The soil series was Munpo and soil texture was silt loam. The experimental soil was amended so that the soil had salinities of $0.8dS\;m^{-1}$, $1.6dS\;m^{-1}$, $3.2dS\;m^{-1}$ and $4.8dS\;m^{-1}$. Millet was transplanted 15 days after sowing. As soil salinity increased, the degree of reduced growth was in the order of seed production > root dry matter > plant dry matter > culm length > tiller number > stem thickness > Panicle length. Seed production was decreased to 18.9% in soil salinity of $1.6dS\;m^{-1}$, 36.9% in of $3.2dS\;m^{-1}$, and 50.7% in EC of $4.8dS\;m^{-1}$. Root dry matter decreased to 35.8% in EC of $3.2dS\;m^{-1}$, and to 40.5% in EC of $4.8dS\;m^{-1}$. As soil salinity increased, Total nitrogen content increased in all aboveground parts, roots and seeds. However, There was no difference in CaO, $P_2O_5$, $K_2O$ and, MgO in soils of different salinity. On the other hand, $Na_2O$ content was higher in the order roots> shoots> seed, and in the case of roots, $Na_2O$ content increased to 1.02% in soil salinity of $4.8dS\;m^{-1}$. However, up to soil salinity of $1.6dS\;m^{-1}$, the $Na_2O$ content of the seed was similar to that in plant grown in the Control conditions($0.8dS\;m^{-1}$). In conclusion, taking into consideration economic factors, millet could be cultivated in soil with salinities of up to approximately $1.6dS\;m^{-1}$, and seed produced from reclaimedland would be suitable for human consumption.
Kang, Eun Seon;Ha, Sun Mi;Cheong, Seoung Ryong;Seo, Myeong Whoon;Park, Su hyoung;Kwack, Yong-Bum;Choi, Keun Jin;Chae, Won Byoung
Korean Journal of Environmental Agriculture
/
v.34
no.3
/
pp.204-209
/
2015
BACKGROUND: Radish (Raphanus sativus L.) is cultivated worldwide and one of important vegetables in Korea where year-round production of radish is possible. Most of radishes in autumn are cultivated with double-row spacing except for Gangwon-do where sing-row spacing is predominant. However, no research has been conducted on double-row spacing in radish cultivation so far. This study was conducted to reveal the optimum double-row spacing in autumn cultivation of radish. METHODS AND RESULTS: Using top two popular autumn radish cultivars 'S' and 'C', seeds were sown in spacing between rows of $55{\times}25$, $45{\times}25$, $35{\times}25$ and $25{\times}25cm$, and that within rows of $35{\times}28$, $35{\times}25$, $35{\times}22$ and $35{\times}19cm$. Plants were harvested 58 days after sowing and leaf weight, length and number, and root weight, length sugar content and pithiness were investigated. In the spacing between rows, no significant difference was observed in leaf weight, length and number in both cultivars; however, 25 cm of spacing between rows significantly reduced the root length and weight in 'S' and 'C' cultivars, respectively. In spacing within rows, 28, 25 and 22 cm did not affect fresh root weight in both cultivars, producing appropriate radish roots of 1,500g on average. However, 19 cm of spacing within rows did not reduced fresh root weight in 'S' cultivar but did significantly in 'C' cultivar (1148.3 g). Sugar contents and pithiness of roots were also affected by spacing but its effect was very small and different between cultivars. CONCLUSION: It is suggested that optimum double-row spacing in autumn radish cultivation is 35 cm and 22 cm of spacing between and within rows, respectively, the spacing that did not reduce the yield and quality of radish roots in two popular autumn radish cultivars.
In order to analyze the lipid composition ana chemical characteristics of rapeseed oil in relation to content of erucic acid, three recommendable cultivars(MOK-PO 70, MOK-PO DANGYO 12 and 14) seeds were used. The composition of lipid was confirmed by column chromatography and TLC, and the composition of fatty acid in total and neutral lipid was determined by GLC, and the results were as follows. 1. The crude fat was 44.0% to 45.3% in all cultivars. 2. The iodine value, saponification value, acid value and nonsaponifiable content were 100 to 101, 176 to 184, 0.7 to 0.9 and 1.0 to 1.2, respectively, with no remarkable difference among three cultivars. 3. The content of neutral lipid was 95.3% to 96.3% of total lipid, and others were compound lipids. 4. The content to triglyceride was 92. 1% to 92.5% of total lipid. Diglyceride and sterol ester were contained 1.1% and 1.2%, respectively. There was a small quantity of free fatty acid and free sterol and monoglyceride was little. 5. The erucic acid was the highest (26.4%), in MOK-PO DANGYO 12 among the composition of fatty acid, while those of MOK-PO 70 and MOK-PO DANGYO 14 were so small as 1.0% and 3.9%. The content of erucic acid was decreased with the increase of oleic acid, the content of oleic acid in MOK-PO DANGYO 12 was 30%, 63% in MOK-PO 70 and, 60% in MOK-PO DANGYO 14. The content of linoleic acid and linolenic acid showed no remarkable difference but that of eicosenoic acid was 11.7% in MOK-PO DANGYO 12 as the highest. The ratio of the fatty acid content in eluted neutral lipid and the total oil showed no remarkable differences.
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