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Development the Gene Based Genomic DNA library and STS primer sets in winter cereals
Poo-Reum Oh,Hwa-Young Heo,Byung-Moo Lee,Jong-Jin Hwang,Si-Ju Kim 한국작물학회 2008 한국작물학회 학술발표대회 논문집 Vol.2008 No.10
The purpose of this study is to develop the EEG (Euchromatin Enriched Genomic) DNA library of wheat, barley, rye and oat. Mcr A and Mcr BC system in DH5 alpha bacteria cell line and Kuemkangmil, Olbori, Olhomil and Olgwiri were used for materials in our experiments. EEG colonies have been constructed by using junk DNA exclusion. We analyzed the genetic information of the colonies using blast searches of NCBI and GRAMENE web sites. One hundred eighty-four, 65, 79 and 119 STS primer pairs were developed using sequencing data of selected colonies in Kuemkangmil, Olbori, Olhomil and Olgwiri respectively. Twenty-eight and forty-two percent of designed primer pair showed polymorphism using six endoucleases in Kuemkangmil, Olbori, Olhomil and Olgwiri germplasm respectively. These primers could be useful for specific allele tagging in mapping populations and germplasm and for the study of functional genomics of wheat, barley, rye and oat.
Han, Si-Nae,Oh, Poo-Reum,Kim, Hong-Sig,Heo, Hwa-Young,Moon, Jun-Cheol,Lee, Sang-Kyu,Kim, Kyung-Hee,Seo, Yong-Weon,Lee, Byung-Moo The Korean Society of Crop Science 2007 Journal of crop science and biotechnology Vol.10 No.2
Antibiotics used for suppressing Agrobacterium in plant transformation procedure might have negligible effects on plant tissues and regeneration. The effects of antibiotics on growth suppression of Agrobacterium and plant regeneration were investigated for enhancing Agrobacterium-mediated transformation using wheat mature embryos. Antibiotics tested, except carbenicillin, were able to suppress that embryos were coated with a layer of Agrobacterium cells in callus induction medium. Agrobacterium growth was suppressed minimally at 50 mg/l of timentin, while cefotaxime and clavamox were completely suppressed at relative high concentration of 250 mg/l. In the treatment of carbenicillin, initiation of growth suppression of Agrobacterium occurred at 750 mg/l of concentration because Agrobacterium KYRT1 contains the carbenicillin resistant gene. In Agrobacterium inoculation, effects of antibiotics were significantly different on the rate of callus induction and shoot formation. Almost embryos were induced calli at 50 mg/l of timentin whereas callus induction rate was achieved above 90% at 100 mg/l and 250 mg/l of cefotaxime and clavamox, respectively. Shoot formation rate was higher in the treatment of timentin than that of cefotaxime and clavamox at 500 mg/l of concentration, respectively. Timentin can be used as a good antibiotics in Agrobacterium-mediated wheat transformation.
Jun-Cheol Moon,Hyung-Ho Park,Poo-Reum Oh,Jong-Jin Hwang,Dea-Wook Kim,Jang Hoon Sung,Kwang-Geun Park,Ji-Ung Jeong,Wook-Han Kim 한국작물학회 2010 한국작물학회 학술발표대회 논문집 Vol.2010 No.04
To develop the STS (Sequence Tagged Site) marker for discrimination between oat cultivars used the EEG (Euchromatin Enriched Genomic) DNA library in oat. The EEG DNA library was constructed the Mcr A and Mcr BC system in DH5 alpha bacterial cell line. About 3,500 EEG colonies had been constructed by using junk DNA exclusion. About 800 colonies were selected that included insert DNA more than 500 bp. It was analyzed the genetic information by using blast searches of NCBI web site. More than three hundred STS primer sets were developed using sequencing data of selected colonies and about 90 primer sets which showed single band were selected in Olgwiri. It was applied to twelve oat cultivars including Olgwiri and has been shown polymorphism at 15%. PCR product which amplified with selected STS primer was treated with six endonucleases and was showed polymorphic bands. These primers could be useful for specific allele tagging in mapping populations and germplasm and for the study of functional genomics of oat.
Plant Regeneration from Sliced Mature Embryo Fragments of Wheat Cultivars
Kyung-Hee Kim,Moon-Seok Kang,Young-Up Kwon,Sang-Kyu Lee,Jung-Hun Moon,Sinae Han,Poo-Reum Oh,Byung-Moo Lee 韓國作物學會 2005 Korean journal of crop science Vol.50 No.5
Mature embryos were aseptically excised with a scalpel and sliced in fragments measuring 0.5 mm in diameter (sliced mature embryo fragment; 4 ~~ 5 fragments/one embryo). Sliced mature embryo fragments of six wheat cultivars were cultured to develop an efficient method of callus induction and plant regeneration. Callus derived from sliced mature embryo fragments showed a good capacity to embryogenesis and regeneration. Furthermore sliced mature embryo fragments decreased contamination from fungi and bacteria. The high efficiency of callus induction were obtained Keumkangmil and Bob-white. For plant regeneration, selected embryogenic calli were transferred to two types regeneration media. An average number of green spots per callus was 4 to 5 in regeneration media after about one week. Percentage of plant regeneration showed high in regeneration medium containing 0.1 mg/l 2,4-D and 5 mg/l zeatin. Especially, Keumkangmil (27.5~% ) and Bobwhite (33.3~% ) showed high regeneration efficiency. This regeneration system from sliced mature embryo fragments may provide an effective and convenient explant for plant transformation studies
Plant Regeneration from Sliced Mature Embryo Fragments of Wheat Cultivars
Kim Kyung-Hee,Kang Moon-Seok,Kwon Young-Up,Lee Sang-Kyu,Moon Jung-Hun,Han Sinae,Oh Poo-Reum,Lee Byung-Moo The Korean Society of Crop Science 2005 Korean journal of crop science Vol.50 No.5
Mature embryos were aseptically excised with a scalpel and sliced in fragments measuring 0.5 mm in diameter (sliced mature embryo fragment; 4 ${\~}$ 5 fragments/one embryo). Sliced mature embryo fragments of six wheat cultivars were cultured to develop an efficient method of callus induction and plant regeneration. Callus derived from sliced mature embryo fragments showed a good capacity to embryogenesis and regeneration. Furthermore sliced mature embryo fragments decreased contamination from fungi and bacteria. The high efficiency of callus induction were obtained Keumkangmil and Bobwhite. For plant regeneration, selected embryogenic calli were transferred to two types regeneration media. An average number of green spots per callus was 4 to 5 in regeneration media after about one week. Percentage of plant regeneration showed high in regeneration medium containing 0.1 mg/l 2,4-D and 5 mg/l zeatin. Especially, Keumkangmil ($27.5\%$) and Bobwhite ($33.3\%$) showed high regeneration efficiency. This regeneration system from sliced mature embryo fragments may provide an effective and convenient explant for plant transformation studies.