논문 : 유세포 분석을 통한 현사시나무 3배체 선발 및 계통별 기내생장 특성

배은경 ( Eun Kyung Bae ),이효신 ( Hyo Shin Lee ),이재순 ( Jae Soon Lee ),노은운 ( Eun Woon Noh ) 한국임학회 2012 한국산림과학회지 Vol.101 No.2

세대기간이 긴 임목에 있어 3배체는 바이오매스 생산과 분자생물학적 연구 분야에서 매우 유용한 연구재료이다. 1970년대 육성된 현사시나무 4배체와 일반 2배체 간의 인공교배를 통해 3배체가 육성되었다. 유세포 분석법을 이용해 이들 F1의 배수성을 확인한 결과 14개체 중 10개체가 3배체로 선발되었다. 그 가운데 3계통의 3배체(Line-1, Line-17, Line-18)는 기내배양시 2배체에 비해 잎이 크고, 엽형이 다소 변형된 형태적 특징이 있었다. 특히 ``Line-18`` 은 줄기가 2배체에 비해 굵었으며, 뿌리 생육에 있어서 발근이 더뎠고 뿌리수도 다른 3배체나 2배체에 비해 적은 특징을 보였다. 그러나 순화율은 모두 100%에 달하였다. 본 연구결과는 3배체 현사시나무가 바이오매스 생산 및 형질전환 연구를 위한 좋은 재료가 될 수 있음을 보여주는 결과이며, 또한 유세포 분석법이 배수체 육종시 배수체를 확인하는데 효율적인 방법임을 시사한다. Triploids are a useful tool for biomass production and molecular breeding of trees with a long life span, Triploids of the poplar ``Hyunsasi`` (Populus alba × P. glandulosa) have been developed by crossing between female diploids and a male tetraploid. The tetraploid was developed around the 1970s at Korea Forest Research Institute by colchicine-induced chromosome doubling. Seedlings of the F1 generation were analyzed using flow cytometry to verify their ploidy status. The mean relative fluorescence index of 3 F1 poplars, labeled as Line-1, Line-17, Line-18, were approximately 1.5 times higher than those of diploid poplars, and the results clearly indicated that they were triploids, The phenotype of the F1 poplars included larger leaves and thicker stem than diploids, and abnormal leaf morphology, especially in the triploid ``Line-18``, Three triploid lines developed roots more slowly and had less roots than diploid. However, 3 poplar cytotypes (2x, Line-1, Line-17) rooted within 10 days on MS medium, In contrast, compared with the 3cytotypes, the Line-18 showed about 80% and 70% in the rooting rate and the number of roots, The triploid poplars could be directly utilized for biomass production and with their sterility, they could serve as basic material for genetic transformation. In addition, flow cytometric analysis proved to be an effective and reliable method for screening forest trees for their ploidy level.

현사시나무 3배체의 핵 DNA 함량 및 조직학적 특성

배은경 ( Eun Kyung Bae ),이효신 ( Hyo Shin Lee ),이재순 ( Jae Soon Lee ),최영임 ( Young Im Choi ),박소영 ( So Young Park ) 한국산림과학회 2013 한국산림과학회지 Vol.102 No.2

Herein we analyzed the nuclear DNA content and the histological characteristics of the triploid of the ‘Hyunsasi’ (Populus alba × P. glandulosa F1) which were developed for biomass production and molecular breeding research. The flow cytometric analysis showed that the nuclear DNA content of the 3 triploids were 1.6 times greater than those of the diploid. In terms of histological characteristics, the cross-section area of the stem of ‘Line-18’ was 1.6 times larger than that of the diploid. The area of pith, and cortex and phloem of the stem of ‘Line-18’ was also 1.6 and 2.0 times larger than that of the diploid, respectively. Moreover, the length and area of guard cell of ‘Line-18’ was 1.2 times larger than that of the diploid. These results helps to understand the cytological characteristics of the triploid poplar clones, and further investigations in the growth rate and wood properties of the triploids growing in the field will determine whether the triploid poplars are good candidates for molecular breeding programs and for the improvement of industrial biomass productivity.

유전자조작, 균주분리 Ochrobactrum anthropi JW-2 유래의 Paraquat 내성유전자 PqrA의 주변 유전자군 분석

배은경 ( Eun Kyung Bae ),이효신 ( Hyo Shin Lee ),원성혜 ( Sung Hye Won ),이병현 ( Byung Hyun Lee ) 한국미생물생명공학회 2006 한국미생물·생명공학회지 Vol.34 No.1

현사시나무에서 Formate Dehydrogenase cDNA의 분리와 특성 구명

이효신 ( Eun Kyung Bae ),배은경 ( Hyo Shin Lee ),이재순 ( Jae Soon Lee ),최영임 ( Young Im Choi ),윤서경 ( Seo Kyung Yoon ),어수형 ( Soo Hyung Eo ) 한국산림과학회 2013 한국산림과학회지 Vol.102 No.3

Formate dehydrogenase (FDH), catalyzing the oxidation of the formate ion to carbon dioxide, is known as the stress protein in response to drought, low temperature and pathogen infection. To study the functions of FDH in poplar (Populus alba × P. glandulosa), we isolated a FDH cDNA (PagFDH1) and examined its expressional characteristics. The PagFDH1 is 1,499 base pairs long and encodes a putative 388 amino acid protein with an expected molecular mass of 42.5 kDa. The PagFDH1 protein has N-terminal mitochondria signal peptide and NAD+ binding domain. Southern blot analysis indicated that a single copy of the PagFDH1 is present in the poplar genome. PagFDH1 is expressed highly in the suspension cells (especially in the lag and early exponential phases) and moderately in roots, flowers and leaves. ABA-mediated enhanced expression of PagFDH1 in response to drought and salt stress treatments indicates that the gene product could play an important role in the development of stress resistant trees.

오차드그래스(Dactylis glomerata L.)의 품종에 따른 종자유래의 캘러스 형성률과, 캘러스 크기 및 식물체 재분화 효율의 비교

배은경,이인애,김기용,이병현,손대영,이효신,정민섭,조진기 Plant molecular biology and biotechnology research 2002 Plant molecular biology and biotechnology research Vol.2002 No.-

Comparison results of callus formation ratio from seed explants, callus sizes, regeneration ratios from callus and regeneration efficiency [calculated by following formular; callus formation ratio(%) × regeneration ratio(%)/100]for 27 ochardgrass (Dactylis glomerata L.)varieties imported and Hapsung 2 developed in Korea are as follows; 1. Among ochardgrass varieties showing more than 50% callus formation ratios, the descending order of callus formation ratio after bedding the seed explants for 4 weeks was 93>Sparta>Pizza>Condor>Lidaglo>Glorus>Hapsung2>Frode. 2. The callus sizes after bedding for 4 weeks were in the range of φ 0.43cm~4.2cm in which there was 10 times size difference between the largest one and the smallest one but most of them were between φ2.5cm~4cm. 3. The regeneration ratio from callus among varieties were in the range of 0~36% and descending order of the upper 6 varieties was Plano>Akimidori>Justus>Lidacta>Currie>Hall mark. 4. The regeneration efficiency which is calculated by the ratios of regeneration from seed explant numbers was between 0 to 17.4% among which Justus showed the highest value in the 4-week treatment. 5. The correlation between callus formation ratios and the callus sizes, callus formation ratios and regeneration efficiency, and callus sizes and regeneration efficiency were r=0.5765, r=0.6365 and r=0.6246, respectively in 4-week callus and all the correlations were significant on the 1% level. 6. In 6-week callus, the descending order callus formation ratios from seed sxplants for the best 6 varieties was Condor>Sparta>93M>Justus>Potomac>Lidaglo>Frode. 7. The callus sizes formed were between φ1.5~5.7cm in which Sparta, the largest one of φ5.7cm was five times larger than the smallest one. The callus size of the control variety. Hapsung 2 was φ3.8cm, which belonged to larger size. 8. Regeneration ratio showed a great deviation among varieties from 6-week old calli by showing from 0% to 100% in which all the calli were regenerated in Plano while no callus was regenerated in Juno, 9. The range of regeneration effciency was between 0~28% among varieties in which the values from 6-week callus treatment were larger than those from 4-week callus treatment. Especially, the value of Potomac in 6-week was 3 times larger than that in 4-week. 10. The correlation between callus formation ratios and the callus sizes, callus formation ratios and regeneration efficiency, and callus sizes and regeneration effciency were r=0.8369, r=0.6683 and r=0.5937, respectively in 6-week callus, and all the correlations were significant on the 1% level.

Ascorbate Peroxidase 유전자의 도입에 의한 식물의 형질전환

이인애,이효신,배은경,김기용,이병헌,손대영,조진기 Plant molecular biology and biotechnology research 2002 Plant molecular biology and biotechnology research Vol.2002 No.-

This study was conducted to obtain the transformed tobacco (Nicotiana tubacum) plants with cytosolic ascorbate peroxidase gene(ApxSC7) using Agrobacterium tumefaciens LBA4404. A cDNA encoding the cytosolic ascorbate peroxidase of strawberry, ApxSC7, was introduced into tobacco plants via Agrobacterium-mediated gene transfer system. The expression vector, pIG-AP8, harboring ApxSC7 gene was used for production of transgenic tobacco plants. A large number of transgenic plants were regenerated on a medium containing hygromycin. Integration of ApxSC7 gene was confirmed by PCR and Southern blot analyses with genomic DNA. Northern blot analyses revealed that the pIGap8 gene was constitutively expressed.