• 식물조직배양학회지
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• 제24권3호
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• pp.125-128
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• 1997

'Gala' 잎조직의 신초 재분화에 효과적인 cytokinin 종류와 농도는 BA 4.0 ㎎/L였고, auxin 종류와 농도는 IAA, IBA, 및 NAA 각각 0.1 ㎎/L였다. BA와 NAA를 농도별로 조합 처리하여 'Gala'의 잎, 절간 및 엽병의 신초 재분화 정도를 비교한 결과, 잎조직은 0.5 ㎎/L NAA + 6.0 ㎎/L BA와 0.5㎎/L NAA + 0.8 ㎎/L BA 처리구에서, 절간조직은 0.1 ㎎/L NAA + 0.8 ㎎/L BA와 1.0 ㎎/L NAA + 0.8 ㎎/L BA 처리구에서 신초 재분화율이 가장 높게 나타났으며, 엽병조직은 0.1 ㎎/L NAA + 4.0 ㎎/L BA 처리구에서 가장 높았다.

• Journal of Plant Biotechnology
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• 제48권2호
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• pp.93-99
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• 2021

This study was conducted to develop an Agrobacterium-mediated genetic transformation protocol for the carnation cv. "Jinju" to counteract its ethylene sensitivity. The new protocol involves the use of an improved shoot regeneration medium, optimized minimal concentrations of the selective agent, a pre-culture period, and co-cultivation periods. Silver nanoparticles (NAg) added at a concentration of 2.0 μM to the Murashige and Skoog (MS) basal shoot regeneration medium supplemented with 0.1 mg/L indole-3-butyric-acid (IBA) and 0.2 mg/L thidiazuron (TDZ) improved the shoot regeneration efficiency, number of shoots per explant, and plant growth compared to the control without the addition of NAg. The phosphinothricin (PPT) concentration of 1.0 mg/L was determined to be the minimal and optimal concentration for the selection of putative transgenic plants. When the explants were infected with Agrobacterium cells harboring the acdS gene, the explants that were pre-cultured for three days induced more putative transgenic plants than those that were co-cultivated for four days. Therefore, we expect that the results of this study will benefit researchers who are developing genetic transformations of carnations.

• Journal of Plant Biotechnology
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• 제4권3호
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• pp.129-134
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• 2002

We report the development of a new selection system for the transformation of pepper plants by mannose. In order to achieve this, we first tested several factors related to regeneration conditions. Among the 30 inbred lines examined, line P9l5 was able to generate shoots at the highest rate from both cotyledons and hyporotyls in MS media. A dosage curve for optimizing the selection conditions was established by mixing mannose (range 0-50 g/L) and sucrose (range 0-30 g/L). The least selection pressure on shoot formation was created by a mixture of sucrose and mannose at 20 g/L and 15 g/L, respectively, and the threshold for ultimate tissue death was 50 g/L of mannose irrespective of the sucrose concentration. However, we found that mannose itself was not the sole inhibitor of pepper shoot development. High concentrations of sucrose (30 g/L) contributed additively to the inhibition of shoot formation at higher mannose concentrations. Genotype preference is a major factor that enhances regeneration ability in mannose media, as was observed in MS media. P9l5 and P410 line had high regeneration rates under mannose selection conditions in the presence of Agrobacterium infection. Different virulence levels of Agrobacterium strains did change the regeneration rates, probably due to interaction with the specificities of the inbred lines. Taken together, P9l5 offers the best pepper inbred line for transformation and we recommend a selection condition of 20 g/L of sucrose and 15 g/L or more of mannose up to 50 g/L in media.

• Journal of Plant Biotechnology
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• 제46권4호
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• pp.297-302
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• 2019

Hibiscus syriacus L., the national flower of Korea, is a commonly grown ornamental species found in parks, gardens and along roadsides in Korea. This study sought to establish a plant regeneration system of H. syriacus 'Nanpa' using mature leaves as an explant. Sterilized mature leaf explants were cultured on modified MS medium with combinations of NAA and BAP at various concentrations for 6 weeks. Among the various media evaluated, modified MS media containing 0.46 mg/L NAA and 0.5 mg/L BAP (CI-1) or 0.92 mg/L NAA and 1 mg/L BAP (CI-4) were the most effective for callus formation. Whitish-yellow calluses were observed on CI-1 medium, while green calluses formed on CI-4 medium. The whitish-yellow callus derived from CI-1 medium showed a higher frequency of shoot induction than the green callus derived from CI-4 on modified MS medium containing 0.5 mg/L BAP. Among the various media evaluated in this study, modified MS medium containing 0.46 mg/L NAA and 0.5 mg/L BAP was the most optimal for shoot-forming callus production. Our findings show that mature leaves of H. syriacus 'Nanpa' can be used for in vitro plant regeneration, and the regeneration system described here may be a powerful tool for molecular breeding of H. syriacus 'Nanpa' in the future.

• 한국약용작물학회지
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• 제15권1호
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• pp.62-66
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• 2007

한국의 전통적인 민간 약재로 사용되어져온 흰민들레의 잎절편체로 부터 기관형성을 통한 재분화를 시도하였다. 잎 절편체를 BA (0-4mg/L)와 2,4-D (0-1mg/L)가 혼합 첨가된 MS 기본 배지에 치상하여 배양한 결과, 배양 3주후부터 신초가 발생되었다. 신초 분화율은 2mg/L BA 첨가된 조건에서 가장 높았으며, 배양 4주째 4mg/L BA 첨가된 조건에서는 캘러스가 유기되었다. 잎령에 따른 부정아 형성율, 발생빈도, 건량, 생중량을 조사한 결과, 발아 후 7주된 잎 절편체에서 평균 11.5개의 신초를 형성하였다. 유도된 신초는 0.5mg/L NAA가 첨가된 배지에서 발근되었으며, 화분에 옮겨 완전한 식물체로 재분화 되었다.

• 한국약용작물학회지
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• 제14권2호
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• pp.92-95
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• 2006

A method for plant regeneration via organogenesis from Pelagonium inquinans leaf disc has been developed. Mature leaf explants were collected from field-grown plants and used for the induction of adventitious shoot regeneration on Murashige and Skoog (MS) medium supplemented with 3% (w/v) sucrose plus plant growth regulators. Maximum shoot organogenesis, with $11.8{\pm}1.5$ shoots (98.6%) per leaf disc, was obtained with $2\;mg/l$ $N^6-benzyladenine$ (BA) and $0.5\;mg/l$ ${\alpha}-naphthyleneacetic$ acid (NAA) in 30 days. For rooting, the in vitro proliferated and elongated shoots were excised into 1.5-2 cm in length microcutting, which were plated individually on an half-strength MS (1/2MS) medium supplemented with 2% (w/v) sucrose plus various concentrations of indole-3-butyric acid (IBA). Shoots rooted with a frequency of 100% following culture on 1/2MS medium containing $0.5\;mg/l$ IBA.

• Journal of Plant Biology
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• 제32권4호
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• pp.313-322
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• 1989

A system was established for induction of multi-shoots and plant regeneration from mesophyll protoplasts of alfalfa, Medicago sativa L. cv. Vernal. Different hormonal effects were tested at each step of protoplast culture, i.e. cell division in modified Kao's liquid medium (K566-7). calli formation on SH semi solid medium, and multi-shoot regeneration from calli on SHa and SHb solid media. Frequency of multi-shoots and plant regeneration was affected by various combinations of phytohormones in final step. The evaluation of multi-shoots induction systems via protoplast culture was discused.

• 한국초지조사료학회지
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• 제28권3호
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• pp.171-176
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• 2008

본 연구에서는 버즈풋 트레포일의 Empire 품종을 이용하여 조직 절편체의 종류 및 식물생장조절물질 첨가가 캘러스로부터 식물체 재분화에 미치는 영향을 조사하였다. 각각의 잎과 줄기 절편체를 배지에 치상한 후 배양한 결과, 모든 처리구에서 5-7일 후부터 캘러스 형성이 시작되었다. 캘러스를 유도함에 있어 첨가되는 auxin류로 NAA를 첨가 우수한 결과가 나타났다. 1 mg/L NAA와 1 mg/L BA가 첨가된 CM4 배지에서 잎 절편체 92%, 줄기 절편체 88%로 가장 높게 나타났으며 잎 절편체 배양하는 것이 더 줄기 절편체를 배양하는 것 보다 더 우수한 결과가 나타났다. 캘러스 배양 6주 후 캘러스로부터 shoot이 유도되기 시작하였으며 잎절편체를 치상하여 배양한 것은 대부분 multiple shoot으로 유도 되었다. 줄기신장 후 뿌리 유도를 위해 2$\sim$3 cm 이상 재분화 식물체의 shoot을 절단하여 1 mg/L IBA 첨가된 처리구에서 배양한 결과, 잎 절편체에서 유도된 shoot에서 98.3%, 줄기 절편체에서 유도된 shoot에서 93.3%로 조사되었다. 또한, 캘러스의 형성 없이 직접 뿌리를 내리고 뿌리의 발달도 잘 되었다. 본 연구를 통하여 확립된 재분화 시스템은 분자육종을 통한 신품종 버즈풋 트레포일의 개발에 유용하게 이용될 수 있을 것이다.

• 한국잡초학회지
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• 제8권2호
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• pp.182-186
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• 1988

제초제(除草劑) bialaphos 처리(處理)에서 살아남은 개체(個體)로부터 다시 bialaphos가 처리(處理)된 배지(培地)에서 callus의 유도(誘導), 생장(生長) 및 direct shooting, ammonia 함량(含量) 등(等)으로 내성(耐性)을 재검정(再檢定)하여 얻어진 약간의 결과(結果)를 요약(要約)하면 다음과 같다. 1. Bialaphos가 0.5~0.75 ppm의 저농도(低濃度)로 처리(處理)된 배지(培地)에서 생장(生長)한 callus로부터 분화(分化)된 식물체(植物體) 수(數)는 품종(品種)에 관계없이 분화(分化)가 잘 되었으나 1.0 ppm 처리구(處理區)에서 계대배양(繼代培養)된 것 가운데서는 KA 101만이 분화(分化)되었다. 2. 분화식물체(分化植物體) 가운데 bialaphos에 내성(耐性)인 개체(個體)를 재선발(再選拔)하기 위해 bialaphos 100.0 ppm 처리구(處理區)에 치상시(置床時) 모두 고사(枯死)하였으며 bialaphos 10.0 ppm 처리구(處理區)에서는 NC82 품종(品種) 2.43%. KA 101 2.76%. BY4 0.78%가 살아남아 품종간(品種間)에 다소 차이(差異)가 있었다. 3. Bialaphos 10.0 및 100.0 ppm의 처리구(處理區)에서 고사(枯死)한 shoot에 함유(含有)된 ammonia 함량(含量)은 무처리구(無處理區)보다 약 15배(倍) 정도(程度) 높아 shoot 의 고사원인(枯死原因)이 ammonia 축적(蓄積)에 의한 것임을 입증(立證)할 수 있었다. 4. NC 82 품종(品種)의 TP 1 (내성식물체(耐性植物體) 개체(個體)가 비선발개체(非選拔個體)보다 bialaphos가 2.5 ppm 첨가(添加)된 callus 증식배지(增殖培地)에서 callus 증식률(增殖率)이 2.14배(倍)나 높았고, bialaphos가 10.0 ppm 첨가(添加)된 shoot 분화배지(分化培地)에서는 샤레당(當) 11 개체(個體)의 shoot를 분화(分化)시켜 다른 개체(個體)보다 내성(耐性)이 컸다.

• Plant Biotechnology Reports
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• 제4권1호
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• pp.85-94
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• 2010

In vitro shoot regeneration of gladiolus in three different culture systems, viz., semi-solid agar (AS), membrane raft (MR), and duroplast foam liquid (DF) cultures was evaluated following the kinetics of shoot multiplication and hyperhydricity at optimized growth regulator combinations. Compared to the AS system, matrixsupported liquid cultures enhanced shoot multiplication. The peak of shoot multiplication rate was attained at 18 days of incubation in the MR and DF systems, whereas the maximum rate in the AS system was attained at 21 days. An early decline in acceleration trend was observed in liquid cultures than the AS culture. The hyperhydric status of the regenerated shoots in the different culture systems was assessed in terms of stomatal attributes and antioxidative status. Stomatal behavior appeared to be normal in the AS and MR systems. However, structural anomaly of stomata such as large, round shaped guard cells with damage in bordering regions of stomatal pores was pronounced in the DF system along with a relatively higher $K^+$ ion concentration than in the AS and MR systems. Antioxidative status of regenerated shoots was comparable in the AS and MR systems, while a higher incidence of oxidative damages of lipid membrane as evidenced from malondialdehyde and ascorbate content was observed in the DF system. Higher oxidative stress in the DF system was also apparent by elevated activities of superoxide dismutase, ascorbate peroxidase, and catalase. Among the three culture systems, liquid culture with MR resulted in maximum shoot multiplication with little or no symptoms of hyperhydricity. Shoots in the DF system were more prone to hyperhydricity than those in the AS and MR systems. The use of matrix support such as membrane raft as an interface between liquid medium and propagating tissue could be an effective means for rapid and efficient mass propagation with little or no symptoms of hyperhydricity.