• 한국자원식물학회지
• /
• 제17권3호
• /
• pp.257-264
• /
• 2004

Selection of stress-tolerant ginseng lines in fields is very difficult because it is almost impossible to control properly the environmental conditions of soil. On the contrary, it can be studied with ease to search for stress-tolerant ginseng lines through in vitro culture because of easy manipulation of stress conditions. This study was conducted for the selection of ginseng pure lines tolerant to salt stress. Murashige ＆amp; Skoog(MS) media with 2.5 folds of KNO$_3$, NH$_4$NO$_3$, MgSO$_4$.7$H_2O$, KH$_2$PO$_4$, and CaC1$_2$.2$H_2O$ was established for the selection of ginseng pure lines tolerant to salt stress in vitro. Among 88 ginseng pure lines bred by Korea Ginseng and Tobacco Research Institute, Punggi Hwangsuk, 78093, 82886, 78135, 86024 and KG104 lines was tolerant to salt stress. For the stable production of quality Korean ginseng, genetic tolerance to salt stress is one of important factors since relatively high salt concentrations in the ginseng nursery soil environment of Korea. Ginseng inbred pure lines were tested for their tolerance to salt stress through in vitro culture technique.

• Journal of Plant Biotechnology
• /
• 제29권4호
• /
• pp.247-252
• /
• 2002

1.5% NaCl이 함유된 배지에 캘러스를 치상하여 방사선을 처리한 결과 무처리구보다 처리구 (30, 50 Gy)에서 캘러스 생존율 및 재분화율이 증가하여 내염성 캘러스를 선발하는데 방사선의 적용이 효과적임을 알 수 있었고, 내염성 캘러스에서 재분화된 M$_3$세대 종자에서 내염성 계통들은 모품종보다 초장, 근장 및 근수의 생육이 우수하여 이러한 계통은 내염성 연구를 위한 유용한 유전자원으로 이용할 수 있을 것으로 생각된다. 또한 RAPD 기술은 대조구와 내염성 캘러스에서 재분화된 계통을 구분하는데 유용한 기술임을 알 수 있었다.

• 한국약용작물학회지
• /
• 제10권4호
• /
• pp.263-268
• /
• 2002

Acifluorfen-tolerant callus lines of Solanum ptycanthum were isolated by stepwise selection. Growth of the unselected line was completely inhibited at 0.5 uM. while some selected lines grew at 8 uM acifluorfen. Twenty-two of twenty-five acifluorfen-tolerant callus lines regenerated shoots. Many of the regenerated somaclones were variants, differing in leaf shape, leaf color, number of flower parts, flower color, and fertility. The acifluorfen tolerant S. ptycanthum callus lines differed.

• Journal of Plant Biotechnology
• /
• 제5권1호
• /
• pp.51-57
• /
• 2003

Doubled haploids have long been recognized as a valuable tool in plant breeding since it not only offers the quickest method of advancing heterozygous breeding lines to homozygosity, but also increased the selection efficiency over conventional procedures due to better discrimination between genotypes within any one generation. Salt tolerant mutants were obtained in rice the variety, 'Hawsungbyeo', through in vitro mutagenesis of in vitro cultured anther-derived calli. Various doses (30, 50, 70 and 90 Gy) of gamma ray were applied to investigate the effect of radiation on callus formation on medium containing 1% NaCl, green plant regeneration, frequency of selected doubled haploid mutants and of the salt tolerant screen. It was demonstrated that the dose of 30 and 50 Gy gamma rays had significant effects on callus formation, regeneration and selection of salt tolerance. No tolerant lines were obtained from non-mutagenized cultures. From gamma ray irradiated cultures, five tolerant lines ($M_2$generation) at germination stage and 13 tolerant lines ($M_3$genoration) at seedling stage were obtained. The frequency of salt tolerant mutants indicates that anther culture applied in connection with gamma rays is an effective way to improve salt tolerance.

• 방사선산업학회지
• /
• 제4권4호
• /
• pp.307-312
• /
• 2010

Plants have evolved physiological, biochemical and metabolic mechanisms to increase their survival under the adverse conditions. This present study has been performed to select salt-tolerant rice mutant lines through in vivo and in vitro mutagenesis with gamma-rays. For the selection of the salt-tolerant rice mutants, we conducted three times of selection procedure using 1,500 gamma ray mutant lines resulted from an embryo culture of the original rice cv. Dongan (wild-type, WT): first, selection in the a nutrient solution with 171 mM NaCl; second, selection under in vitro condition with 171 mM NaCl; and third, selection in a reclaimed saline land. Based on a growth comparison of the entries, out of the mutant lines, two putative 2 salt tolerant (ST) rice mutant lines, ST-87 and ST-301, were finally selected. The survival rate of the WT, ST-87 and ST-301 were 36.6%, 60% and 66.3% after 7 days in 171 mM NaCl treatment, respectively. The WT and two salt tolerant mutant lines were used to analyze their genetic variations. A total of 21 EcoRI and Msel primer combinations were used to analyze the genetic relationship of among the two salt-tolerant lines and the WT using the ABI3130 capillary electrophoresis system. In the AFLP analysis, a total of 1469 bands were produced by the 21 primer combinations, and 700 (47.6%) of them were identified as having polymorphism. The genetic similarity coefficients were ranged from 0.52 between the ST-87 and WT to 0.24 between the ST-301 and the WT. These rice mutant lines will be used as a control plot for physiological analysis and genetic research on salt tolerance.

• Journal of Plant Biotechnology
• /
• 제4권3호
• /
• pp.123-127
• /
• 2002

Herbicide tolerant rice (Oryza sativa L. cv. Ilpumbyeo) cell lines were selected from $\gamma$-ray-irradiated anther-derived cell cultures. The anther-derived cell clusters were small (300 to 400 ${\mu}{\textrm}{m}$ in diameter) and uniform ones that were screened by miracloth filtering. The cell suspensions were very efficient to plate one layer onto agar medium and to screen target cell lines. Herbicide tolerant cell lines were selected by 5 mg/L cyhalofop butyl (CHB) treatment by using the small cell suspensions on agar N6 medium containing 1 mg/L 2,4-D and 0.2 mg/L kinetin. Of the cell lines, one line (CHB-1) showed stable tolerance at 10 mg/L concentration after 6-month culture without herbicide suspension. Growth stability of CHB-1 was similar to that of control cell line on 10 mg/L CHB containing medium. In this experiment we established herbicide tolerant cell line selection system by using anther-derived uniform-cell suspensions with $\gamma$-ray-irradiation.

• Journal of Plant Biotechnology
• /
• 제38권1호
• /
• pp.1-8
• /
• 2011

최근 시설재배의 증가나 새만금 간척지의 개발 등 염류 농도가 높은 토양에서의 작물 재배에 대한 수요가 점차적으로 증가하고 있는 상황에서 고염환경 하에서 세포내로 주입된 염 분자를 제한된 구역으로 격리하여 염류내성을 주도록 하는 anti-porter 유전자 TANHX, HVNHX를 우리나라 주요 화훼작물이며 수출유망 작물인 국화로 도입하는 분자육종이 백마 등 5품종에 대하여 이루어졌다. 선발 마커를 이용한 배지선발 과정을 통하여 선발된 기내 식물체 390개체를 대상으로 토양 순화를 거쳐 284계통의 국화 형질전환체가 획득되었으며, 얻어진 식물체 중 임의로 40계통을 선발하고 PCR을 거쳐 진성 여부를 확인한 결과 30계통에서 target band가 검출되어 75%의 배지 발 성공률을 나타내었다. PCR 분석 선발 계통을 포함하여 토양 순화된 284계통을 대상으로 직접적으로 NaCl 0.2 ~ 1.2% (300 mM) 범위로 내염성 생물검정을 실시한 결과 NaCl 0.8% (200 mM) 농도에서도 생존 및 생장이 가능한 15계통이 선발되었으며 이중 7계통은 NaCl 1.2%(300 mM) 내에서도 생존이 가능하였다. 내염성 특성형질 도입을 위하여 anti-porter 유전자 HVNHX가 도입되어 선발된 형질전환 계통의 스트레스 저항성 정도 및 세포 형태적 특성변화가 관찰되었다. 선발된 계통은 NaCl 1.2% (300 mM) 처리 생존가능 7계통을 포함하여 NaCl 0.8% (200 mM) 관수 처리 하에서 생존 및 생장이 가능한 15계통이었다. 세포형태 특성은 전자현미경 (SEM)을 이용하여 형질전환체 및 비 형질전환체의 공변세포를 염 처리 후 관찰함으로서 이루어졌는데 형질전환체, 비형질전환체 모두 무처리에 비해 NaCl 처리한 식물체의 공변세포의 두께가 두꺼워지고 조직 치밀도가 증가하였으며 형질전환체의 경우 비 형질전환체에 비해 두꺼워지는 정도나 조직의 치밀도 증가 정도가 높아 염에 대한 내성이 강화되었음을 알 수 있었고 염 처리 후 세포의 생존정도 비교를 통한 내염 스트레스에 대한 저항성 정도를 측정하고자 TTC 검정을 실시한 결과 강 내염성 계통의 TTC 수치가 높게 나왔으며 NaCl 처리 농도가 높아질수록 TTC 수치가 낮아지는 경향이었으나 강 내염성 7계통은 1.2% NaCl 처리에서도 0.206 ~ 0.331로 비형질전환체의 0.046 중내염성 계통의 0.114 ~ 0.193에 비해 높은 세포생존 비율을 나타내었다. 또한 식물이 스트레스에 대항하기 위하여 분비하는 아미노산인 Proline의 함량을 계통별로 측정한 결과 강 내염성 형질전환 계통이 높게 나왔으며 NaCl 처리 농도가 높아질수록 증가하는 경향을 나타내어 강 내염성 7계통은 1.2% NaCl 처리에서 2.255 ~ 2.638 mg/kg로 중 내염성 형질전환 계통의 1.496 ~ 2.125에 비해 높게 형성되었다.

• Journal of Plant Biotechnology
• /
• 제6권4호
• /
• pp.205-212
• /
• 2004

The objectives were to compare the salt tolerance levels in the parental rice cultivar, Dongjinbyeo, and induced mutagenesis derived its lines for plant height, MDA, ATPase, POD, and 2-dimensional protein electrophoresis pattern in NaCl-containing hydroponic nutrient solutions. Rice plants isolated from a population of rice (Oryza sativa L. cv. Dongjinbyeo) mutation lines, which were generated in combination with in vitro selection and gamma-ray, exhibited salt tolerance. Line No. 18 had the longest plant, whereas NaCl-sensitive line (No. 25) had the shortest plant. The parent, and the sensitive line showed severe damage from salt stress. Tolerant lines (No. 18, 50) had a lower malonaldehyde (MDA) content than the sensitive one (Dongjinbyeo, No. 25) during salt stress. Several proteins showed significant quantitative variation through 2DE; phosphoribulokinase, peroxidase, oxygen evolving enhancer 1 and the $H^+-ATPase$, which are known to be involved in salt tolerance. The effect of salt on peroxidase and $H^+-ATPase$ activity in the seedlings of two groups with contrasting genotypes of rice was studied. A greater activity was recorded in the tolerant lines as compared to the sensitive ones (P<0.05, Duncan's test). The results indicate that salt tolerant lines expressed more salt stress-inducible proteins associated with salt tolerance than the sensitive lines during salt stress.

• 한국육종학회지
• /
• 제41권1호
• /
• pp.25-31
• /
• 2009

벼 내염성 유묘검정에서 실용성, 경제성, 정밀성이 우수한 검정체계를 확립하고벼 자포니카 내염성 중간모본 및 품종개발을 위한 내염성 유전자원 및 품종을 선발 이용하고자 본 시험을 수행하였다. 1. 증류수 이용 Yoshida 양액에 NaCl를 첨가한 염처리는 0.3%, 0.5%, 0.7%로 희석한 바닷물로 Yoshida 양액을 제조한 염처리 보다 유묘 내염성 계통을 선발하는데 효율적이었다. 2. 수돗물로 양액 제조 처리는 증류수로 양액 제조 처리와 유묘 내염성 검정 달관평가에서 차이가 없었다. 3. 년차 및 지역간 내염성 재연성을 확인하기 위하여 '02년, '03년 계화도출장소 하계 및 '04년 국제미작연구소에서 건기에 유묘 내염성을 검정한 결과 IR73571-3B-11-3-K2 등 6계통의 3개년('02, '03 및 '04) 평균 유묘 내염성은 3~4.2로 Pokkali 내염성 3과 비슷하였고 년차 및 지역간 안정적 유묘내염성을 보였다. 4. Pokkali 및 FL478과 내염성이 비슷한 IR73571-3B-11-3-K2 등 6계통의 출수기는 8.16~8.21일로 동진벼와 같거나 2~6일 늦었으며 간장은 대부분의 계통이 동진벼 보다 7~17 cm작았으나 수장은 6~9 cm 길었다. 등숙율 및 수량은 동진벼 보다 각각 52.2~14.7%, 50~5% 낮았다.

• Journal of Microbiology and Biotechnology
• /
• 제18권5호
• /
• pp.805-814
• /
• 2008

Liming of acidic soils can prevent aluminum toxicity and improve crop production. Some maize lines show aluminum (Al) tolerance, and exudation of organic acids by roots has been considered to represent an important mechanism involved in the tolerance. However, there is no information about the impact of liming on the structures of bacterial and fungal communities in Cerrado soil, nor if there are differences between the microbial communities from the rhizospheres of Al-tolerant and Al-sensitive maize lines. This study evaluated the effects of liming on the structure of bacterial and fungal communities in bulk soil and rhizospheres of Al-sensitive and Al-tolerant maize (Zea mays L.) lines cultivated in Cerrado soil by PCR-DGGE, 30 and 90 days after sowing. Bacterial fingerprints revealed that the bacterial communities from rhizospheres were more affected by aluminum stress in soil than by the maize line (Al-sensitive or Al-tolerant). Differences in bacterial communities were also observed over time (30 and 90 days after sowing), and these occurred mainly in the Actinobacteria. Conversely, fungal communities from the rhizosphere were weakly affected either by liming or by the rhizosphere, as observed from the DGGE profiles. Furthermore, only a few differences were observed in the DGGE profiles of the fungal populations during plant development when compared with bacterial communities. Cloning and sequencing of 16S rRNA gene fragments obtained from dominant DGGE bands detected in the bacterial profiles of the Cerrado bulk soil revealed that Actinomycetales and Rhizobiales were among the dominant ribotypes.