• Journal of Plant Biotechnology
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• 제31권1호
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• pp.31-35
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• 2004

멸종위기에 직면한 피뿌리풀 (Stellera roseaN.)의 기내증식법을 개발하고자 액아 마디를 MS 배지에 BAP와 zeatin을 처리하여 다경 (multiple shoot)를 유도하고 기내발근에 미치는IBA및 NAA처리 효과를 조사하였다. 액아 마디로부터 다경유도는 BA가 현저히 양호한 반면 줄기의 생장은 zeatin이 BA보다 효과적이었다. 증식된 줄기로부터 기내 발근은 오옥신 처리로 가능하였으나 발근율은 대체로 저조하였고 오옥신의 전처리 기간에 따라 차이를 보였다. IBA가 NAA보다 다소 좋은 발근효과를 보였고 1.0mg/L 농도로 15일간 배양시 30%까지 발근되었다. NAA역시 처리농도 및 처리기간에 따라 발근율에 차이를 보였다. 발근묘는 인공토양에서 51%가 활착되어 정상생장이 가능하였다. 이상의 결과는 발근 및 순화조건을 좀더 개선하면 피뿌리풀의 효율적인 기내번식이 가능함을 시사해준다.

• Journal of Plant Biotechnology
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• 제50권
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• pp.56-62
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• 2023

35S cauliflower mosaic virus 프로모터의 조절을 받고 인트론이 포함된 β-Glucuronidase (gus) gene과 35S cauliflower mosaic virus (enhanced) 프로모터의 조절을 받는 blpR유전자가 있는 pCAMBIA3301 벡터가 포함된 AGL-1 균주를 사용하였다. 아그로박테리움을 이용한 형질전환 체계와 PPT (D-L-phosphinothricin) 선발을 통하여 나리 인편조직으로부터 형질전환 식물체가 획득되었다. 본 연구에서 나리 레드플레임'품종의 인편조직에 선발 및 목적유전자로 바스타 제초제저항성 유전자인 blpR 유전자를 도입하였다. 상기 실험 결과, 20분의 접종시간과 5일간의 아그로박테리움과의 공동배양이 100개의 접종된 인편개체에서 각각 24, 27개의 높은 PPT 저항성 개체가 관찰되었고 신초까지 형성된 인편을 19.6 및 22.7개를 생산하는 우수한 형질전환 결과를 보여주었다. 이렇게 제초제를 이용하여 선발되었을 뿐만 아니라 도입된 reporter 유전자인 gus도 발현되었음을 확인하였고 선발유전자이자 목적유전자인 blpR 유전자도 PCR 검정을 통해 도입되었음을 확인하였다. 12주 이상의 선발과정을 거치고 gus 및 PCR 검정을 거친 형질전환 개체들은 발근 배지를 거쳐 순화 후 화분으로 이식하여 높은 활착율을 보여주었다. 결론적으로 본 연구에서 확립한 프로토콜을 이용하면 평균 20% 이상의 형질전환 효율을 나타내고 본 연구에 기술된 아그로박테리움 매개 형질전환 체계에 향후 보완이 필요하지만, 우수 품종개발을 위한 나리 육종 프로그램에 기여할 수 있을 것으로 판단된다.

• Plant Biotechnology Reports
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• 제4권2호
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• pp.173-178
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• 2010

An efficient protocol for shoot regeneration was developed for sesame (Sesamum indicum L.) internodes using the transverse thin cell layer (tTCL) culture method. The frequency of shoot regeneration and the number of adventitious buds produced from regenerated shoots depend significantly on explant age, thickness of the tTCL sections, and the phytohormones supplemented to the culture medium. A combination of 6-benzyladenine (2.0 $mg\;l^{-1}$) and a-naphthaleneacetic acid (0.5 $mg\;l^{-1}$) was found to be the best phytohormone combination for shoot bud induction, with the maximum number of shoots obtained when the tTCL sections were 0.5-1.0 mm thick and derived from 4- to 6-week-old seedlings of sesame. Well-developed shoots were rooted on MS medium without phytohormones, and 80% of the regenerated plantlets were successfully established in soil.

• Plant Biotechnology Reports
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• 제4권2호
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• pp.101-107
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• 2010

An efficient plant regeneration system has been developed for figleaf gourd (Cucurbita ficifolia Bouch$\'{e}$), which is exclusively used as a rootstock for cucumber. The protocol is based on results obtained from a series of culture experiments involving different parts of the cotyledons and various media. The culture of cotyledon explants was critical for the enhancement of shoot regeneration frequency. The lower parts of the cotyledon excised at the plumule base were found to display a markedly enhanced production of adventitious shoots compared to other cotyledon regions. Culture in silver nitrate-supplemented Murashige and Skoog (MS) medium was not beneficial for shoot regeneration and suppressed root regeneration. Efficient shoot regeneration was obtained on MS medium containing 1.0 $mg\;l^{-1}$ zeatin and 0.1 $mg\;l^{-1}$ indole-3-acetic acid. Regenerated shoots successfully elongated and rooted in medium containing 0.1 $mg\;l^{-1}$ 1-naphthalene-acetic acid after 10-15 days of subculturing. The plantlets were satisfactorily acclimatized in a greenhouse and grew into normal plants without any morphological alterations.

• 한국약용작물학회지
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• 제14권5호
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• pp.278-281
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• 2006

The effect of plant growth regulators was investigated on in vitro shoot proliferation from axillary bud explants of Hypericum erectum. To determine the optimal cytokinin for proliferation of axillay buds, we carried out screening four cytokinins (BA, kinetin, 2iP, TDZ). When nodal segments were cultured on MS medium supplemented with $4.5\;{\mu}M$ TDZ (thidiazuron), a number of shoots were induced. Our results indicated that the addition of TDZ to culture medium resulted in the induction of significantly more axillary buds than in the addition of other cytokinins. The optimal concentration of TDZ for proliferation of axillary buds was $10\;{\mu}M$. 92% of shoots spontaneously rooted without any plant growth regulator (PGR) and formed whole plantlets within one month. More than 95% of these regenerants survived and they did not show any detectable variation in morphology or growth characteristics compared to their donor plants.

• Journal of Plant Biology
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• 제39권2호
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• pp.93-98
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• 1996

In order to enhance the system of potato transformation and further regeneration, potato was transformed using the Agrobacterium tumefaciens harboring $\beta$-glucuronidase (GUS) gene. We found that a series fo modified medium ttained 100% shoot regeneration within 5 weeks after the preincubated explants on stage I medium were infected with Agrobacterium. Callus appeared at the cut edges of stem segments on stage II medium, mainly at the basal parts. Some explants started to form shoots after two to three weeks on stage III medium containing kanamycin (50 mg/L). When transferred to MS medium containing 200 mg/L kanamycin, 81% of the transformed shoots formed roots at the cut edge of the plantlets. In contrast, untrasformed shoots never rooted and became yellowish after few weeks under the same conditions. Southern and northern analysis indicated in vitro shoot regeneration on the callus derived from the potato explants, which were incubated with Agrobacteria. The regeneration cycle was shortened after the transformatin and finally the transformation efficiency was highly enhanced.

• Journal of Plant Biotechnology
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• 제8권1호
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• pp.15-19
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• 2006

An efficient protocol for in vitro multiple shoot bud induction and plant regeneration from mature green cotyledon derived callus tissues of Acacia sinuata has been developed. Callus formation occurs at all the concentrations of thidiazuron (TDZ) in Murashige and Skoog's (MS) medium, but 0.6 ${\mu}M$ proved to be the best with maximum callus formation frequency. Supplementation of TDZ in combination with indole-acetic acid (IAA) in MS media accelerates shoot bud organogenesis in differentiating callus tissues with 60-70% conversion of shoot buds into shoot Most efficient shoot organogenesis was recorded when TDZ induced calli were subcultured at different concentrations of 6-benzyla-denine (BA). Optimum shoot bud induction and plant regeneration from callus was achieved when 0.6 ${\mu}M$ (TDZ) induced calli were subcultured at 3.0 ${\mu}M$ (BA) where $16.6{\pm}0.74$ shoots/unit callus on obtained. Rooting in in vitro differentiated shoots was achieved when transferred to medium containing different concentration of indole-3-butyric acid (IBA) in full & half strength MS medium. The well rooted plantlets were hardened and transferred to net house with 90% survival rate.

• Plant Biotechnology Reports
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• 제4권3호
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• pp.229-235
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• 2010

In this study, we established an in vitro regeneration system to maximize the recovery of leafy perilla (Perilla frutescens L. Britton) plantlets as part of developing a molecular biotechnology-based metabolic engineering program for this crop plant. Hypocotyl segments including the apical buds were used as explants for the direct production of shoots without an interim callus phase. The number of shoots produced from the apical buds peaked within 3-4 weeks, and the shoots were subsequently cultured on Murashige and Skoog (MS) media supplemented with 2 mg $1^{-1}$ benzylaminopurine (BA). Spontaneous rhizogenesis was observed after 7-10 days of culture on MS media without hormonal additives. The rooted shoots developed into normal plants in soil after hardening on distilled water for 3-4 days. The average plantlet regeneration frequency was higher for the apical buds (64.33%) than for the top (15.66%), middle (4%), and basal (1.33%) segments of the hypocotyls. This regeneration system demonstrates a capacity for high-frequency plantlet recovery and thus should be considered for use in the genetic manipulation of leafy perilla.

• 한국산림과학회지
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• 제95권2호
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• pp.155-159
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• 2006

An efficient method for in vitro propagation of the medicinal plant Hovenia duleis, was established. Plantlets for micropropagation of H. dulcis were obtained from in vitro germinated seeds. The effectiveness of various levels of cytokinins (BAP, Kinetin and TDZ) on multiple shoot formation from stem nodes was tested. BAP (1.0 mg/L) treatment induced highest number of multiple shoots. The growth pattern of plantlet on various culture media was undertaken. The shoot elongation was optimal on 2MS basal medium without growth regulators. The in vitro rooting ability of H. dulcis shoots was examined with two-auxins IAA and IBA. The IAA (1.0 mg/L) treatments induced earliest rooting with maximum number of roots and root growth. Rooted shoots were transferred directly to small pots with artificial soil and such established plant exhibited a normal growth pattern similar to wild plantlet.

• 한국산림과학회지
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• 제80권4호
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• pp.416-419
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• 1991

15년생 자작나무의 근주 맹아지 액아를 이식절편체로하여 기내배양시켜 효과적으로 증식시킬 수 있었다. 줄기증식에는 WPM에 BAP 0.5와 1.0mg/l 첨가한 배지가 효과적이었다. 기내발근에는 GD배치를 이용하였고 0.2mg/l IBA 첨가시 100% 발근되었다. 이렇게 증식된 식물체는 토양에 이식하여 95% 이상 활착되었으며 활착후 정상적인 생장을 보였다. 이상의 결과는 자작나무 성숙목의 기내 대량 증식 가능성을 시사한다.