• Plant Biotechnology Reports
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• 제4권1호
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• pp.53-59
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• 2010

A specific deleted version of ARABIDOPSIS RESPONSE REGULATOR1 (ARR1) lacking the signal receiver domain (1.152 amino acids)-coding sequence, referred to as $ARR1{\Delta}DDK$, was amplified using Arabidopsis thaliana cDNA prepared from adult leaves and transferred into the genome of Nicotiana tabacum cv. Samsun under the transcriptional control of a ${\beta}$-estradiol-inducible expression system. The ectopic expression of $ARR1{\Delta}DDK$ affected the morphology of transgenic seedlings and their segments in vitro. In the presence of an inducer, ${\beta}$-estradiol, ectopic expression of $ARR1{\Delta}DDK$ induced only the formation of soft, pseudo-bulbous tissue in the root tip region of intact seedlings, which appeared similar to callus generated on a hypocotyl segment in the presence of 2,4-D and 6-benzyladenine (BA), both at $1\;{\mu}M$. Those callus tissues on the root tip region could not generate shoots unless $1\;{\mu}M$ BA was supplied. In segment culture, ectopic expression of $ARR1{\Delta}DDK$ induced calluslike tissue around the cut-end of cotyledon and hypocotyl segments with occasional shoot formation, suggesting that the expression of $ARR1{\Delta}DDK$ could substitute for the effects of cytokinin on these segments. Additionally, treatment with only ${\beta}$-estradiol induced NtWUS, a WUS ortholog in tobacco, which was detected during the process of callus tissue formation in the root tip region and also in cotyledon or hypocotyl segments. These findings suggest that the NtWUS might be associated in the transdifferentiation process caused by the functional regulation of $ARR1{\Delta}DDK$ in transgenic tobacco seedlings.

• 식물조직배양학회지
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• 제21권4호
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• pp.221-225
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• 1994

주요 약용작물로 사용하고 있는 천궁의 경정을 기내배양 하여 건전한 종경을 대량으로 증식하는데 적함한 배양조건을 구명하기 위하여 실험한 결과를 요약하면 7월에 채취한 재료를 1%의 NaOCI에 20분간 표면살균한 후, carbenicillin 500 mg/L, BA 1.0 mg/L 와 $GA_3$ 1.0 mg/L가 첨 가된 1/2 MS배지가 오염율 감소와 신초 재분화율을 높이는데 효과적이었고, 재분화한 신초의 기내증식은 BA 0.5 mg/L와 당 60 g/L가 함유된 1/2 MS가 양호하였으며 이들 유묘는 1/2 MS에 NAA 3.0 mg/L 단용배지에서 배양하였을때 발근하였다.

• 한국잡초학회지
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• 제6권1호
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• pp.25-32
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• 1986

생장조절물질(生長調節物質) Callus 유도(誘導)에 미치는 영향(影響)을 구명(究明)키 위하여 식물체(植物體)의 종자(種子) 또는 Shoot-tip으로부터 Callus 유도(誘導)를 검정(檢定), 유도(誘導)된 Callus로부터 식물체분화(植物體分化), 제초제(除草劑)가 Callus 생체중량증가(生體重量增加)에 미치는 영향(影響) 및 Succinate dehydrogenase의 TTC 반응검정(反應檢定)을 하여 얻어진 결과(結果)는 다음과 같다. 1. MS 기본배지(基本培地)에 2,4-D 함량(含量)은 올방개와 너도방동산이의 종자(種子)는 1.0ppm, 너도방동산이의 shoot-tip은 4.0pm, 삼강벼와 물피에서는 2ppm이 Callus 유도(誘導)를 위한 과정농도(過定濃度)였다. 2. MS기본배지(基本培地)에다 2,4-D와 BA와 TIBA를 첨가시(添加時) BA는 2,4-D 단독처리(單獨處理)보다 억제적(抑制的)이나 TIBA 처리(處理)는 0.3 또는 0.5ppm 첨가구(添加區)가 2,4-D 단독처리구(單獨處理區)보다 효과적(效果的)이었다. 삼강벼, 물피, 너도방동산이 종자(種子) 및 올방개 shoot-tip에서는 2,4-D 1.0ppm+TIBA 0.5ppm, 너도방동산이 shoot-tip에서는 2,4-D 1.0ppm+TIBA 0.3ppm 일 때가 가장 높았다. 3. Callus로부터 식물체분화율(植物體分化率)은 2,4-D에 Kinetin 첨가구(添加區)가 NAA에 Kinetin 첨가구(添加區)보다 효과적(效果的)이었고 물피는 2,4-D 0.8ppm+Kinetin 8.0 ppm에서 50%, 올방개는 2,4-D 1.6ppm+Kinetin 16.0ppm에서 33.3%가 분화(分化)가 되었으나 너도방동산이는 분화(分化)가 되지 않았다. 4. 제초제(除草劑) Londax 및 Basta는 농도(濃度)가 증가(增加)될수록 삼강벼, 물피, 올방개 및 너도방동산이의 Callus 유도(誘導) 또는 증식(增殖)을 억제(抑制)시켰고 $10^{-3}$M에서는 거의 100%가 억제(抑制)되었으나 저농도(低濃度)에서는 식물체간(植物體間)에 상이(相異)한 반응(反應)을 보였으며 양제초제(兩除草劑])에서 삼강벼가 물피나 너도방동산이보다 내성(耐性)을 보였다. 5. TTC에 대(對)한 Succinate dehydrogenase의 반응(反應)은 공시식물(供試植物)의 전(全) Callus가 모든 처리구(處理區)에서 정반응(正反應)(+)을 보여서 Callus가 생존(生存)해 있음을 알 수 있었다.

• 생물환경조절학회지
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• 제29권1호
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• pp.1-8
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• 2020

고구마 바이러스 무병묘의 기내급속증식을 위한 적정 생장 조절물질 및 sucrose 농도, 최소생장 기내보존(15℃)에 미치는 광의 영향과 생존율 및 기내생장 특성 등을 조사하였다. 고구마 무병묘의 마디배양은 0.2 mg·L^-1 BA 첨가배지에서 줄기신장, 줄기직경, 잎수, 뿌리수, 생체중 및 건물중 등이 가장 양호하였다. 배양부위 및 배지물리성에 따른 적정 sucrose 농도는 마디배양은 5% sucrose를 첨가한 고체배지에서, 정단배양은 3% sucrose를 첨가한 액체배지에서 줄기두께, 잎수, 뿌리수, 뿌리길이, 생체중 및 건물중 등의 생육에 가장 효과적이었다. 15℃ 저온항온기에서 고구마 무병묘의 최소생장 기내보존은 암상태에서는 3개월 내에 모두 고사하였으나, 적색:청색(7:3) 혼합 LED (150±5 μmol·m^-2·s^-1 PPFD)에서는 5개월까지 100% 생존하였다. 따라서 고구마 무병묘의 최소생장 기내보존에는 광이 필요하며, 샬레에 밀식(10 개체/샬레)할 경우, 좁은 공간에서 대량보존이 가능하였다.

• 식물조직배양학회지
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• 제25권3호
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• pp.159-163
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• 1998

배 '신고'의 경정배양을 통한 급속대량증식법을 개발하기 위하여 모수의 수세와 채취시기 및 배지에 첨가되는 생장 조절제 NAA와 BA, sucrose가 경정배양의 각 단계의 생육에 미치는 영향을 구명하고자 하였다. 신초를 채취하는 모수의 수세와 채취시기에 있어서, 중 정도의 수세를 가진 모수로부터 6월에 채취한 경정이 배양확립단계에 가장 적합하였다. 신초생장은 BA 1.0, 2.0㎎/L의 단독처리구에서 가장 좋았으며, NAA의 첨가는 저조한 신초생장과 과도한 callus 발생을 야기하였다. 신초증식단계에서는 BA 단독처리로는 신초의 대량증식이 불가능하였으며, BA 2.0㎎/L와 NAA 0.01㎎/L의 혼합처리구에서 발근단계에 필요한 크기의 신초를 다량으로 얻을 수 있었고, 30g/L 의 sucrose 첨가가 효과적이었다. 발근단계에는 NAA 0.1㎎/L 를 첨가한 1/4MS배지에서 높은 발근율(96%), 1, 2차 근수와 근장을 나타내었으며, 이상의 과정을 통하여 정상적인 소식물체를 얻을 수 있었다.

• 식물조직배양학회지
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• 제27권1호
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• pp.51-55
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• 2000

감나무 (Diospyros kaki Thunb.)의 기내번식에 효과적인 배지 및 생장조정제를 구명하고자 일목계차랑 (Ichikikeijiro), 도근조생 (Tonawase), 평핵무 (Hiratenenasi) 3품종을 공시재료로 하여 시험한 결과, 신초의 생존에 가장 효과적인 배지는 MS 배지였으며 MS 배지내 질소원의 함량을 1/2∼1배로 조정한 경우가 신초의 증식 및 신장에 효과적이었다. 1/2N-MS배지에 2 mg/L zeatin 처리시 0.9 cm로 가장 많은 신초 신장을 보였고, 5 mg/L 처리시 증식된 신초수가 6.2개로 가장 많았다. 증식된 신초의 발근은 동일 배지에 1 mg/L IBA 처리시 가장 효과적이었다.

• 한국자원식물학회지
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• 제30권6호
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• pp.612-617
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• 2017

The preservation of pear germplasm, like that of other clonal germplasms, is difficult because it requires conservation of whole plants or their tissues. Among the currently available methods for long-term conservation of clonal germplasm, cryopreservation of shoot tips is the most reliable and cost- and space-effective option. Alginate-coated axillary shoot tips from in vitro-grown pear were conserved successfully in liquid nitrogen (LN) following dehydration. Shoot recovery from cryopreserved shoot tips was improved greatly after 8 weeks of cold acclimation, but recovery decreased slightly after then. The highest regeneration rate was observed when in vitro shoot tips were preincubated in MS (Murashige and Skoog) medium with 0.3 M sucrose for 48 h, and when alginate-coated shoot tips were precultured in MS medium with increasing sucrose concentrations (0.5 M and 0.7 M) for 8 and 16 h, respectively. When the encapsulated beads were dehydrated for up to 7 h [25% water content (fresh weight basis)] under laminar flow, the highest regeneration rate was observed in "BaeYun No. 3" (55.7%) and "Whanggeum" (43.3%) after warming from LN. This technique is useful as a practical procedure to cryopreserve plant material that is sensitive to freezing of the surrounding cryoprotectant medium. Therefore, this technique appears to be promising for the cryopreservation of shoot tips from in vitro-grown plantlets of pear germplasm.

• 한국자원식물학회지
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• 제31권6호
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• pp.684-694
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• 2018

This study describes the successful establishment of a cryopreservation protocol for Citrus limon cultivars: 'Frost Eureka limon' and 'Cook Eureka limon', using a droplet-vitrification method. The shoot tips that were excised from in vitro grown seedlings of the two cultivars were preserved in liquid nitrogen (LN) and successfully regenerated into whole plants. Excised shoot tips were pre-cultured for 1 or 2 days in 0.3 M and 0.5 M sucrose solutions at $25^{\circ}C$ and incubated in a loading solution (LS) composed of 17.5% glycerol + 17.5% sucrose in Murashige and Skoog (MS) medium for 40 min at $25^{\circ}C$. Prior to direct immersion in LN for 1 h, the shoot tips were dehydrated with plant vitrification solution 2 (PVS2) at $0^{\circ}C$ or PVS3 at $25^{\circ}C$. The frozen shoot tips were re-warmed and unloaded with 1.2 M sucrose in $\text\tiny{^1/_2}$ MS for 30 min at $25^{\circ}C$. Shoot tips were post-cultured overnight on survival medium and then micrografted onto 'trifoliate orange' (Poncirus trifoliate (L.) Raf. seedling rootstocks for recovery and to produce whole plants. The highest regrowth rates were 53.5% and 50.3% for cryopreserved shoot tips of 'Frost Eureka limon' and 'Cook Eureka limon', respectively, when pre-cultured in 0.3 M and 0.5 M sucrose concentrations in a sequencing manner, with LS and treated with PVS2 for 60 min at $0^{\circ}C$. We also investigated whether the ammonium ion concentration on post-culture medium affected the viability of the cryopreserved Citrus shoot tips. The viability of cooled samples, following culturing on woody plant media (WPM) containing $\text\tiny{^1/_4}$ ammonium nitrate overnight before micrografting, was the highest (70.3%) in 'Frost Eureka limon'. The study described here is a cost-effective and safe method to conserve Citrus fruit cultivars, for the improvement and large-scale multiplication of fruit plants and for breeding disease resistance.

• Biotechnology and Bioprocess Engineering:BBE
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• 제7권4호
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• pp.206-211
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• 2002

Calli obtained from a shoot-tip of garlic, Allium sntivum L., were encapsulated using a calcium alginate gel. Some of the encapsulated calli were cultured on a 1/2 MS medium supplemented with 3% sucrose, 10$\^$-5/ kinetin, and 5 ${\times}$ 10$\^$-6/ M NAA whereas the remainder was stored for 40 days at 4$^{\circ}C$. All the naked calli regenerated on the solid medium, while 95% of the encapsulated calli regenerated, and 88% of the encapsulated calli regenerated after 40 days of storage at 4$^{\circ}C$. The capsule matrix delayed the germination time of the encapsulated calli, yet activated the shoot formation of the artificial garlic seeds. The shoot length of the encapsulated garlic calli was much longer than that of the naked garlic calli. The encapsulated garlic calli were dried in a laminar airflow cabinet and the conversion frequency of the dried artificial garlic seeds on a 1/2 MS medium remained at 93% with a water Loss of Less than 50%.

• 한국식물병리학회지
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• 제14권5호
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• pp.507-512
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• 1998

Molecular analysis has been done for characterization of the interactions between three beet curly top virus (BCTV) strains and two Arabidopsis ecotypes in terms of virus inducible disease symptoms and infectivities. The total DNA was isolated from three tissues (shoot tips, infection origins and roots) of virus infected plants and this DNA was analyzed by quantitatively and qualitatively to elucidate virus movement and symptom development. CTV-Worland infected Col-O and Sei-O showed only symptom shown in hypersusceptible ecotype Sei-O by BCTV-worland was shoot tip stunting. Kinetics of virus DNA accumulation of three different viruses indicated that roots contained more virus DNA than shoot tips or infection origins, and that disease symptom severity was strongly correlated with virus DNA accumulation. These results suggest that the mild and Worland-specific symptoms shown in Sei-O by BCTV-worland are caused by the interactions of host factors provided by hypersusceptible ecotype and viral factors of mild strain.