• Horticultural Science & Technology
• /
• v.28 no.5
• /
• pp.810-817
• /
• 2010

We observed abnormal morphology of cotyledons occurring in seedlings derived from open-pollinated and cross-pollinated tetraploid grapes and selected aneuploids, especially hypo- and hyper-tetraploid in seedlings with abnormal morphology of cotyledons. Five types of morphologically abnormal cotyledons were observed. In open-pollination of four tetraploid grapes, the frequency of abnormal cotyledons was 1.6% (49 of 3029 seeds). Percentage of aneuploids in the seedlings of abnormal cotyledons was 20.4% (10 of 49 seedlings). Aneuploids in open-pollination consisted of three (4n = 4x-2), four (4n = 4x-1), and three (4n = 4x+1) seedlings. In cross-pollination of tetraploids, the frequency of abnormal cotyledons was 3.4% (59 of 1729 seeds). Percentage of aneuploids in the seedlings with abnormal cotyledons was 22.0% (13 of 59 seedlings). Aneuploids from cross-pollination of tetraploids consisted of two (4n = 4x-2), nine (4n = 4x-1), one (4n = 4x+1), and one (4n = 4x+3) seedlings. According to the results, although the abnormal cotyledon morphology of seedlings obtained from crossing between tetraploid grapes appeared at low rate (2.3%), aneuploid seedlings occurred at high rate (22.0%); therefore, it indicated that this selection strategy might be very efficient in the initial seedling stage.

• The Korean Journal of Mycology
• /
• v.35 no.1
• /
• pp.43-46
• /
• 2007

It was investigated that systemic acquired resistance (SAR) was induced in plant treated with a elicitor, which was derived from a non-virulent fungus. The elicitor, a hyphal cell wall component derived from fungus, induced a production of phytoalexin and a generation of reactive oxygen species (ROS) in potato treated with its low level concentrations. The effect of the fungus-derived elicitor was better than that of virulent pathogen-derived elicitor, which was well known in potato. These results, therefore, suggested potentcial use of fungus-derived elicitor as a new plant protector for commercial development.

• Proceedings of the Plant Resources Society of Korea Conference
• /
• 2019.04a
• /
• pp.56-56
• /
• 2019

본 연구는 식용과 약용으로 이용되는 국내 자생식물인 야산고비(Onoclea sensibilis var. interrupta Maxim.)를 기내에서 대량증식하기 위한 조건을 구명하기 위하여 수행되었다. 무가온 온실에서 수집한 포자를 기내에서 발아시켜 전엽체를 획득하였으며, 계대배양하며 실험의 재료로 사용하였다. 전엽체의 대량증식을 위해 전엽체 0.3g을 메스로 균일하게 다진 후 증류수 1ml와 함께 배지에 고루 펼쳐서 배양하는 방법을 사용하였으며, 증식에 미치는 배지의 영향을 확인하기 위하여 1/4, 1/2, 1, 2배로 조절한 MS배지를 조성하여 8주간 배양하였다. 이후, 증식이 우수한 배지를 선정하여 sucrose와 질소급원의 농도를 조절하였으며, 활성탄을 첨가하여 증식에 미치는 영향을 확인하였다. 배지종류 실험의 결과, 생체중이 1MS에서 10.2g으로 초기 접종량에 비해 가장 많이 증가하였으며, 현미경을 이용하여 관찰한 결과에서도 정상적인 전엽체의 형태인 심장형으로 발달하였다. 증식이 우수하였던 1MS배지에 sucrose의 농도를 조절하여 배양한 결과에서는 1%의 처리구에서 가장 증식률이 좋았으며, 질소급원의 경우 30mM의 농도로 조절한 처리구가 가장 좋은 결과를 보였다. 배지 내 활성탄의 첨가는 처리구당 증가된 전엽체의 생체중이 유의적인 차이를 보이지 않았다. 포자체 대량 형성을 위한 적정 배양토의 혼합조건을 확인하기 위하여 원예상토, 피트모스, 펄라이트 및 마사토의 혼합비율을 5종류로 달리하여 조성하여 사각분($7.5{\times}7.5{\times}7.5cm$)에 충진하였다. 조성한 배양토에 기내에서 배양한 전엽체 1g을 증류수와 함께 블렌더를 이용하여 10초간 분쇄하여 토양표면에 고루 분주하였다. 이후 12주간 재배한 결과, 모든 토양조합에서 포자체가 형성되었다. 그중 원예상토와 마사토를 2:1(v:v)로 혼합한 토양에서 포트 당 405.0개로 가장 많은 포자체가 형성되었으며, 전체적인 생육 또한 비교적 양호한 결과를 보였다. 따라서 야산 고비의 전엽체 대량증식에 적합한 배지는 경제성과 생육수준을 고려하여 1%의 sucrose와 질소급원의 농도를 30mM로 조절한 1MS 배지가 적합하며, 포자체 대량생산을 위해서는 원예상토, 마사토를 2:1(v:v)로 혼합한 토양이 적합하다고 판단되었다.

• Korean Journal of Plant Tissue Culture
• /
• v.22 no.5
• /
• pp.283-290
• /
• 1995

This experiment was canted out to obtain embryogenic callus and to understand developmental mechanism of somatic embryogenesis in Oenanthe javanica ($B_{L.}$) DC. experiments included the examination of explant source and media for embryogenic callus production and the observation of developmental pattern of embryogenic cells and non-embryogenic cells. Embryogenic calli were formed on zygotic pro-embryos together with their endosperms when they were cultured on Ms media containing 1.0mg/L 2,4-D. Embryogenic calli were also formed on the intact surface in vitro grown stem or petiole segmentsafrer 6-8 weeks of culture, whereas non-embryogenic calli were formed on cut surfaces of the stem and petiole after 2 weeks of culture. Non-embryogenic calli were rhizogenic in suspension and solid media culture.

• Korean Journal of Plant Tissue Culture
• /
• v.22 no.6
• /
• pp.351-355
• /
• 1995

A truncated Polygalacturonase (PG) cDNA was fused in reverse orientation to the CaMV 35S promoter of the binary vector pCA643, and introduced into tomato cells by Agrobaderium - mediated transformation. Transformed cells were selected using kanamycin as select agent then regenerated into plants. After selfed, one transgenic line (T9), was germinated and grown on MS medium containing 1 mg/mL of kanamycin Genomic Southern analysis of a T9 progeny with labelled PG2 cDNA probe showed a single antisense PC fragment as well as the endogenous PG2 gene, suggesting that PC antisense gene was integrated into tomato genome. Northern blot analysis demonstrated that the antisense RNA was produced from the transgene at much tiger level than the endogenous PG2 gene. Polygalacturonase activity analysis of the fruit from transgenic plants demonstrated that the antisense transgene expression caused 4 to 60% reduction of endogenous PG activity.

• Journal of Plant Biotechnology
• /
• v.38 no.1
• /
• pp.1-8
• /
• 2011

Recently the increasing of vinyl and green houses and development of reclaimed land including Saemangeum induced the need for breeding salt-tolerant crops which can survive and grow in high salinity soil. So we try to develop salt-tolerant transgenic chrysanthemum (Dendranthema grandiflorum.) lines by using anti-porter gene TANHX and HVNHX. Through marker selection and plant regeneration step, we could get 284 putative transgenic chrysanthemum lines. On selected putative transgenic plants, 40 candidates were used for genetic analysis and 30 lines could be made up of target size band on PCR, so about 75% of marker selected lines were decided as real transgenic lines. Selected 284 transgenic lines were also used for salt-tolerance test as a range of NaCl 0.2 ~ 1.2% (300 mM). As a result of salt-tolerance test, 15 selected transgenic lines could live and grow on the continuous supply of 0.8% (200 mM) NaCl solution and another 7 lines were could survive under 1.2% (300 mM) NaCl solution. This salt-tolerant transgenic lines under salt stress also lead a cell alternation especially a guard cell. A stressed guard cell be swelled and grow larger in proportion to NaCl concentration. TTC test for cell viability on transgenic chrysanthemum lines pointed out that more strong salt-tolerant lines can be live more than another under same salt stress. The numerical value of strong salt-tolerant 7 transgenic lines were 0.206 ~ 0.331 under 1.2% NaCl stress, and then it's value is more larger than middle salinity lines' 0.114 ~ 0.193 and non-transgenic's 0.046. And the proline contents as indicated stress compound also pointed out that HVNHX introduced salt-tolerant transgenic lines were less stressed than other under same salt stress. The contents of strong salt-tolerant transgenic lines were 2.255 ~ 2.638 mg/kg and it is much higher than that of middle salinity lines' 1.496 ~ 2.125.

• Journal of Plant Biotechnology
• /
• v.37 no.4
• /
• pp.511-516
• /
• 2010

The process to acquire intron-GUS gene-expressed transformants from somatic embryos (including embryogenic calli) of Rosa hybrida cv. 'Sweet Yellow' using Agrobacterium-meditated transformation method was reported in this study. Somatic embryos including embryogenic calluses were infected with Agrobacterium tumefaciens AGL1 strain (O.D = 0.7~1.6) including intron-GUS gene for 30 min, and were co-cultured for 3 days. After co-cultivation, they were cultured on embryo germination medium (EGM) supplemented with $250\;mg{\cdot}L^{-1}$ cefotaxim at $4^{\circ}C$ for 7 days. Then, transient GUS gene expression was observed. Shoots were regenerated from the shoot primodia induced from the intron-GUS gene-transferred either somatic embryos or embryogenic calli cultured on EGM supplemented with both cefotaxim $250\;mg{\cdot}L^{-1}$ and ppt $2\;mg{\cdot}L^{-1}$. Before induction of rooting from shoots cultured on shoot growing medium supplemented with both cefotaxim $250\;mg{\cdot}L^{-1}$ and ppt $2\;mg{\cdot}L^{-1}$, the shoots were cultured on multi-shoot induction medium supplemented with both cefotaxim $250\;mg{\cdot}L^{-1}$ and ppt $2\;mg{\cdot}L^{-1}$ to induce multi-shoots. When expression of the gene from a part of the multi-shoots was identified by GUS transient assay, the putative transgenic multishoots were transferred to rooting medium supplemented with cefotaxim $250\;mg{\cdot}L^{-1}$. After the formation of healthy roots, transgenic plantlets were transferred to the greenhouse after acclimatization. The expression rate of the intron-GUS gene in the multi-shoots was 100%.

• Korean Journal of Plant Tissue Culture
• /
• v.26 no.3
• /
• pp.163-169
• /
• 1999

This study was conducted to produce herbicide resistant plants by transferring phosphinothricin acetyltransferase (PAT) gene into Populus alba $\times$ Populus glandulosa No .3 using Agrobacterium tumefaciens MP 90/PAT. Leaf segments from in vitro grown shoots of hybrid poplar No. 3 were soaked in a AB medium containing Agrobacterium tumefaciens MP 90/PAT for 10 min and cocultivated for 2 days on MS medium containing 1.0 mg/L 2,4-D and 0.2mg/L kinetin (CIM). Putative transformed calli could be selected after cocultivation of leaf segments on CIM supplemented with 50mg/L kanamycin and 500mg/L cefotaxime for 3 weeks. The selected calli were cultured on CIM supplemented with 50 mg/L kanamycin and 500 mg/L cefotaxime for 5~8 weeks before transfer to WPM containing 1.0mg/L zeatin, 0.1mg/L BAP, 50 mg/L kanamycin and 500mg/L cefotaxime for shoot regeneration. Shoots were regenerated from the callus after 4 week cultivation, and the regenerants were grown on the same medium for 7~l0 weeks. The plants rooted on 1/2 WPM containing 0.2 mg/L IBA and 50 mg/L kanamycin. To confirm the gene insertion into plants, GUS activity was detected by histochemical assay in the transformed plants. Finally, the presence of both NPT II and PAT genes from the transgenic plants were confirmed by PCR amplification with the gene specific primers and subsequent PCR-Southern blot with DIG-labeled PAT gene probe. After acclimatization in pots for 4 weeks, the plants were sprayed by 3 mL/L of Basta to test resistance to the herbicide. The transgenic plants remained green, whereas all the control plants died after one week.

• Horticultural Science & Technology
• /
• v.29 no.6
• /
• pp.617-622
• /
• 2011

The study was conducted to determine the efficiency in producing spontaneous polyploids in some mandarin hybrids with different seed embryony. Seed formation by open pollination, frequency of spontaneous polyploids, and plant growth characteristics were evaluated in four mandarin hybrids with polyembryony such as 'Amakusa', 'Haruka', 'Hayaka', and 'Seminole' and two with monoembryony such as 'Benibae' and 'Harehime'. The mean number of the developed seeds per fruit was 10.0 and frequency of small seeds was 25.1%. Polyploids were selected from plants germinated in vitro by a flow cytometry and confirmed by chromosome analysis. One triploid was produced from 'Harehime', one tetraploid, 'Amakusa', and one tetrapoid, 'Benibae'. There were little differences in leaf shape, thickness, petiole length, and internode length between diploids and polyploids such as tri- or tetraploid. However, polyploids had larger stomata and lower density of stomata in abaxial epidermis than diploids. SPAS indicating chlorophyll content and photosynthetic rate were significantly affected by ploidy level. The results indicated that spontaneous polyploids might be produced by open pollination in some mandarin hybrids and monoembryony had higher frequency in polyploid occurrence than polyembryony.

• Proceedings of the Plant Resources Society of Korea Conference
• /
• 2018.04a
• /
• pp.82-82
• /
• 2018

본 연구는 국내에 자생하는 자원식물들의 살초활성을 조사하여 식물생장억제물질을 활용한 환경친화형 제초제 개발에 요구되는 기초 자료를 확보하기 위해 수행되었다. 국내 자생 식물 101종을 식물체 부위별로 나누어 MeOH로 추출하여 획득된 시료 129점에 대해 돌피(Echinocholoa crus-galli P.B. var. formosensis Ohwi)를 이용한 살초활성 검정을 실시하였다. 시료를 소량의 methanol로 녹여 직경 5.5cm petri dish 상의 Whatman No. 2 여지에 균일하게 흡착시킨 후, fume hood 내에서 용매를 제거하고 1mL의 증류수를 첨가하였다. 돌피 종자를 15립씩 3반복으로 파종하여 $28^{\circ}C$, 5,000lux의 growth chamber에 치상하고, 7일 후 시험구 당 균일하게 자란 10개의 유묘를 선발해 초장 및 근장을 조사하였으며, 살초활성 평가를 위해 대조구 개체의 초장과 근장에 대한 생장억제율을 산출하였다. $1,000{\mu}g/mL$ 농도에서 101종 129점 MeOH 추출물들의 돌피 유묘 뿌리 생장에 대한 억제율을 조사한 결과, 80% 이상을 나타낸 식물 시료는 가죽나무 줄기, 누린내풀 지상부, 단풍취 뿌리, 두릅나무 지상부, 백양꽃 전초, 백양꽃 지하부, 병조희풀 경엽부, 산사나무 잎 가지, 삽주 뿌리, 상사화 전초, 상사화 지하부, 석산 뿌리, 애기송이풀 전초 등 17점이었다. 이들 중 90% 이상 고활성을 나타낸 단풍취 뿌리, 두릅나무 지상부, 백양꽃 전초, 상사화 지하부, 석산 뿌리 유래의 추출물에는 새로운 환경친화형 제초제 개발소재로써 활용할 수 있는 살초활성물질들이 함유되어 있는 것으로 생각된다.