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Haeyoung Na(나해영),Changhoo Chun(전창후) 한국원예학회 2009 원예과학기술지 Vol.27 No.2
한국의 토종 참나물은 종자번식의 어려움 때문에 원예작물로 재배되기 어려워 아직도 자연채취에 의존하고 있다. 토종 참나물의 재배화를 위해서는 번식 방법의 한계를 극복하고 우량 묘의 안정된 공급 체계의 확립 하여야 하는데 이를 위하여 배발생을 이용한 대량 급속 증식 방법의 개발이 필요하다. 참나물의 엽병을 재료로 하여 형성된 배발생 캘러스를 이용하여 체세포배 형성을 시키기 위한 최적 조건은 배지강도 X0.5 MS, sucrose농도 30g L⁻¹의 배지에 kinetin 0.01에서 0.1㎎ L⁻¹를 첨가한 액체배지에 광도은 PPF 20과 50μ㏖ m⁻²s⁻¹에서 현탁배양하는 것이 가장 효율적이었으며 체세포배의 발아에는 배지강도 X0.25 MS, sucrose 30g L⁻¹, IAA 0.05 혹은 BA0.01㎎ L⁻¹를 첨가하여 광조건은 PPF 20과 50μ㏖ m⁻²s⁻¹에서 배양하는 것이 가장 효과적이었다. Formation and germination of somatic embryo (SE) are affected by the concentration of plant growth regulators; MS medium strength, sucrose concentration, and the photosynthetic photon flux (PPF) level. The MS medium strength and sucrose concentration for an optimal condition of the SE formation and germination of Pimpinella brachycarpa were x0.5, sucrose 30 g L⁻¹ and x0.25, sucrose 30 g L⁻¹, respectively. When 0.05 ㎎ L⁻¹ of kinetin was treated, the percentage of normal somatic embryo (NSE) formation was the greatest and the percentage of hyperhydric somatic embryo (HSE) formation was the lowest. SE germination rate was greater in treatment of IAA 0.05 ㎎ L⁻¹ or BA 0.01 ㎎ L⁻¹ than those of other treatments. Moreover, better SE formation and germination were found at dimmer lighting (20 μ㏖ m⁻² s⁻¹ of PPF) than that of higher PPFs. SE formation and germination were also retarded at higher PPFs. Optimum PPF for SE formation and germination was 20 μ㏖ m⁻² s⁻¹. After forming SE from EC 0.1 g for 4 weeks, EC 0.1 g resulted in 700 total SE formation. Culturing four SEs for four weeks resulted in a secondary SE formation, germination and eventually 52 plantlets. These results indicate that a huge proliferation rate was achieved at up to 8 weeks of EC at liquid medium suspension culture and SE at solid medium culture. Through the experiment, after inducing EC from EC, the mass of EC was increased by 25 times (from 0.05 to 1.30 g) when each of the nutritional and chemical factors was controlled at the optimized condition.
배추의 소포자 배양에 영향을 미치는 배지, AgNO₃, Activated Charcoal 및 NAA
나해영(Haeyoung Na),박수형(Suhyoung Park),황귀영(Guiyoung Hwang),윤무경(Moo-Kyoung Yoon),전창후(Changhoo Chun) 한국원예학회 2009 원예과학기술지 Vol.27 No.4
육종기간을 단축시키기 위한 효율적인 방법의 하나인 소포자배양을 개발하기 위하여 배추 소포자배양 최적조건을 구명하고자 하였다. 소포자배 유기를 위한 최적 조건은 유전자형에 따라 다르지만 뿌리혹병의 race 4에 저항성을 나타내는 ‘ER 6-1-4’의 소포자 배양은 배지농도, 식물생장조절물질, 미량 요소 그리고 활성탄의 농도에 영향을 받았다. 소포자배의 발생은 배지농도 NLN 배지 표준농도에서 다른 배지 농도에 비교하여 효과적이었다. 그리고 0.5배 NLN 배지에 미량요소 표준 농도의 0.25 배 농로 농도로 첨가하였을 때 배 발생률을 2배 이상 증가시킬 수 있었다. AgNO₃를 0.1㎎ℓ?¹를 첨가한 처리구에서 소포자배의 발생률이 가장 높았으며 그 이상의 농도에서는 형성이 억제되었다. 또한 activated charcoal을 petri dish 당 15, 20mg를 첨가한 처리구에서 activated charcoal를 첨가하지 않은 처리구에 비해 20배 이상의 소포자 배를 얻을 수 있었으며 기형배의 형성을 억제할 수 있었다. NAA를 0.05㎎ℓ?¹를 첨가한 처리구에서는 배의 발아율이 높을 뿐 아니라 묘의 생육도 가장 좋았으며 기형식물체의 발생률 또한 가장 낮은 것을 확인할 수 있었다. Optimum conditions for the microspore embryo induction of Brassica rapa were determined in order to develop a microspore culture protocol to efficiently shorten breeding time. Each optimal condition for microspore embryo induction was dependent on the genotypes used. The microspore embryo induction was affected by the concentration of plant growth regulators, NLN medium strength, micro element, and activated charcoal level. The NLN medium strength for an optimal condition of the microspore embryo formation of Brassica rapa was a single NLN medium strength (1×). The addition of a half strength of micro elements to a half strength (0.5 ×) of NLN medium was mostly efficient for the formation of microspore embryos. When 0.1 ㎎ ℓ?¹ of AgNO₃ was added to the half strength NLN medium, the number of normal microspore embryo formation increased and the number of hyperhydric microspore embryo (HSE) decreased while the uniformity of microspore embryos was also enhanced. The addition of activated charcoal not only increased the induction rate of microspore embryos by 200%, but also decreased the induction rate of hyperhydric embryos. These results indicate that the microspore embryo formation rate of Brassica rapa can be significantly increased with significantly reduced chances of HSE.
Haeyoung Na(나해영),Changhoo Chun(전창후) 한국원예학회 2009 원예과학기술지 Vol.27 No.3
체세포배를 이용하여 소식물체를 얻는 각 단계의 최적환경 구명시 수많은 소식물체를 얻을 수 있다. 그러나 배발생과정을 통해 얻어진 소식물체는 기외환경에 이식되기 위해서는 많은 노동력과 경비의 소요를 필요로하는 복잡한 순화과정이 필요하며 만약 순화과정이 생략된다면 생존율이 극히 저조할 것이다. 그래서 체세포배와 소식물체를 이용하여 photoautotrophic시스템과 photomixotrophic 시스템을 이용하여 묘의 생산을 시도하였다, Photomixotrophic 시스템에서는 torpedo와 cotyledonary torpedo 단계의 체세포배 그리고 소식물체의 발아율은 모두 75?¹00%로 발아율은 매우 높았으나 기외로 이식한 후 3일 이내에 모두 고사하였으며 기공도 이식 24시간 후부터는 완전히 괴사하여 파괴괴는 것을 확인할 수 있었다. 그러나 CO2 7,500㏖ moL?¹ 으로 조절된 photoautotrophic 시스템에서는 torpedo 단계의 체세포배의 발아는 30%로 발아율은 낮았지만 순화과정을 생략하고 기외로 바로 이식하여도 생존율은 100%였으며 기공의 상태도 정상인 것을 확인할 수 있었다. Pimpinella brachycarpa somatic embryos were cultured photoautotrophically at different CO₂ concentrations, and conversion of the somatic embryos was compared to those cultured photomixotrphically. The growth rate of plantlet in photoautotrophic system did not show significant difference with that of plantlet in photomixotrophic system. While somatic embryo at torpedo stage didn’t germinate at all at CO₂ concentration 400, 1,500 and 4,500 μ㏖ ㏖?¹, the germination rate at CO₂ concentration 7,500 μ㏖ ㏖?¹ was 31.3%. The germination rate of somatic embryos in cotyledonary torpedo stage increased as CO₂ concentration rate increased, showing that high concentration of CO₂ increases the photoautotrophic ability. When transplanting plants germinated at photomixotrophic system to ex vitro, the survival rate at three days after ex vitro was 0%. All explants withered in 24 hours. However, the survival rate of plants germinated at photoautotrophic system after transplanting to ex vitro was 100%, and it was possible to transplant without acclimatization process. Photoautotrophic system was applicable to P. brachycarpa for development of micropropagation system using somatic embryos being provided with the CO₂ concentration properly.
미나리 종자번식 체계 확립을 위한 최적 채종단계, 층적처리 및 종자 세척방법 구명
배은지(Eunji Bae),나해영(Haeyoung Na) 한국육종학회 2015 한국육종학회지 Vol.47 No.2
The influence of seed maturity, stratification, and seed washing method on seed germination were investigated for establishment of seed propagation system in dropwort (Oenanthe stolonifera DC). Seed germination rate was increased as period of the seed ripening increased, especially the seed over 40 days after flowering showed the highest germination rate. The optimum period of cold stratification treatment to promote seed germination was about 8 weeks. The seed treated by a cold wet stratification with sand exhibited higher germination rate compared to the seed treated by a cold wet stratification without sand. The stratified seeds that were stored in the ground having fluctuating temperature were improved up to 19%, while stratification stored under constant temperature(4℃) did not improve germination rate. In addition, 8 weeks of stratification period under fluctuating temperature in the ground was most effective. Sodium hypochlorite, vital oxide, and distilled water were used as chemicals to remove the germination inhibitor of seed coat. Water washing of seed coat after soaking in 0.5% sodium hypochlorite and 0.0005% vital oxide for three minutes and 60 minutes respectively could effectively remove the germination inhibitors in the seed coat.