일본잎갈나무 체세포배 유래 식물체의 초기 신초와 뿌리 길이, 순화용 기질이 생존율에 미치는 영향

이나념 ( Na Nyum Lee ),윤아영 ( A Young Yun ),김지아 ( Ji Ah Kim ),김태동 ( Tae Dong Kim ),김용욱 ( Yong Wook Kim ),한심희 ( Sim Hee Han ) 한국산림과학회(구 한국임학회) 2020 한국산림과학회지 Vol.109 No.4

본 연구는 체세포배 발아 식물체의 세포 라인별, 순화용 기질별 생육 특성을 분석하여, 일본잎갈나무 조직 배양묘의 순화 중 생존율을 높이려는 방안을 마련하고자 실시하였다. 체세포배 발아 식물체는 라인 L14-66, L16-18, L17-B4를 사용하였으며, 순화용 기질은 낙엽송 전용 상토, 원예용 상토, 엘리포트, 피트플러그를 사용하였다. 평균 초기 신초와 뿌리 길이는 L14-66 라인이 가장 짧았고, L17-B4 라인이 가장 길었다. 세포 라인별 평균 생존율은 L17-B4 라인이 87.0%로 가장 높았고, L14-66 라인이 64.3%로 가장 낮았다. 순화용 기질별 생존율은 엘리포트와 피트플러그에서 각각 88.5%, 88.9%로 가장 높았다. L14-66 라인의 초기 신초 길이는 낙엽송 전용 상토(r = 0.852), 원예용 상토(r = 0.692), 엘리포트(r = 0.867)에서 생존율과 상관이 높았으나, 전 질소 함량이 높은 피트플러그에서는 상관이 없었다. L17-B4 라인의 초기 신초 이는 모든 순화용 기질에서 생존율과 상관이 없었다. L14-66 라인의 초기 뿌리 길이는 낙엽송 전용 상토(r = 0.986), 엘리포트(r = 0.846), 피트플러그(r = 0.802)에서 생존율과 상관이 높았으며, 유식물체의 생존율은 초기 뿌리 길이가 길수록 높았다. L17-B4 라인의 초기 뿌리 길이는 낙엽송 전용 상토(r = 0.896)와 엘리포트(r = 0.696)에서만 생존율과 상관을 보였다. 결론적으로, 유식물체의 생존율을 높이기 위해서 신초 길이보다 뿌리 길이를 우선 고려해야 하며, 피트플러그와 같이 기질 내 질소 함량이 높은 재료를 사용하거나 순화 과정 중에 질소 시비를 높여 주는 것이 좋다. 또한, 생존율을 높이기 위해 초기 생장이 빠른 라인 개발도 함께 이루어져야 한다. We analyzed the growth characteristics of each cell line and acclimating substrate of Larix kaempferi plantlets regenerated from somatic embryos, with the goal of increasing the survival rate during the acclimation phase. Somatic embryos from three embryogenic cell lines (L14-66, L16-18, and L17-B4) were used, and the acclimating substrates were commercial soils for Larix species (Larix-Soil) and horticultural corps (Hort-Soil), Elle-pot, and Peat-plug. The average initial shoot and root length was shortest in L14-66 and longest in L17-B4. The average survival rate by cell line was highest (87.0%) in L17-B4 and lowest (64.3%) for L14-66. Survival rates by substrate were highest in Elle-pot (88.5%) and Peat-plug (88.9%). The initial shoot length of the L14-66 plantlets was highly correlated with survival rates in the Larix-Soil (r = 0.852), Hort-Soil (r = 0.692), and Elle-pot (r = 0.867) substrates, but not in Peat-plug with high total nitrogen content. The initial shoot length of the L17-B4 plantlets was not correlated with the survival rate in any of the substrates. The initial root length of the L14-66 plantlets was highly related to survival rates in the Larix-Soil (r = 0.986), Elle-pot (r = 0.846), and Peat-plug (r = 0.802) substrates, and the survival rate of the plantlets was higher as the initial root length was longer. The initial root length of the L17-B4 plantlets was related to survival rate only in the Larix-Soil (r = 0.896) and Elle-pot (r = 0.696) substrates. In conclusion, to increase the survival rate of plantlets, root length should be considered over shoot length, and it is recommended to use substrates with high nitrogen content, such as Peat-plug, or to add fertilizer, during the acclimating process. In addition, in order to increase the survival rate, lines with high initial growth should be developed.

티크의 기내 줄기 생장 및 발근에 미치는 LED (light-emitting diode) 효과

이나념,김지아,김용욱,Lee, Na-Nyum,Kim, Ji-Ah,Kim, Yong-Wook 한국식물생명공학회 2019 식물생명공학회지 Vol.46 No.4

본 연구는 티크의 기내 생장 및 발근에 미치는 light-emitting diode (LED) 효과를 구명하기 위해 수행되었다. 티크의 정아 절편체의 줄기 신장은 LED 혼합광(BR, 50% 청색광 + 50% 적색광) 하에서 DKW 배지 배양 시 3.2 cm로 가장 좋았고, 줄기유도는 광원 및 배지(MS 및 DKW) 효과가 그리 크지 않은 것으로 나타났다. BA 농도 처리 시 최대 줄기 유도는 BR 광원 및 1.0 mg/L 처리구에서 2.4개, 최대 줄기 신장은 BR 및 0.5 mg/L 처리구에서 4.94 cm로 가장 효과적이었다. 발근에 영향하는 IBA 농도 효과는 정아 절편체를 BR 및 0.5 mg/L 처리구에서 배양 시 최대 발근율(93.8%)과 최대 뿌리 길이 신장(1.3 cm)을 보였고, 뿌리 수는 0.2 mg/L (3.1개/절편) 처리구에서 가장 좋은 것으로 나타났다. 발근 식물체는 상토로 이식 후 순화 과정을 거쳐 정상적은 생장을 보였다. This study was conducted to determine the effect of a light-emitting diode (LED) on in vitro shoot growth and rooting in teak (Tectona grandis L.). In the experiments with apical bud explants, the greatest shoot elongation (3.2 cm) occurred when they were cultured on DKW medium under 50% blue and 50% red LED mixture (BR), whereas no differences in growth were observed in different light sources (florescent light [F] or BR) or media (MS or DKW). The highest number of shoot multiplication (2.4/explant) or elongation (4.94 cm) was achieved with 0.5 or 1.0 mg/L 6-Benzyladenine (BA) treatment under BR. In addition, the best rooting rate (93.8%) or root length (1.3 cm) was recorded with 0.5 mg/L indole-3-butyric acid (IBA) treatment under BR, and the highest root induction (3.1/explant) was observed in 0.2 mg/L IBA under BR. The in vitro rooted plantlets were hardened and survived well on soil.

윤판나물(Disporum sessile) 아배양에 의한 식물체 재분화에 영향하는 배지 종류 및 생장조절물질 효과

이나념,김지아,김태동,김용욱,Lee, Na-Nyum,Kim, Ji-Ah,Kim, Tae Dong,Kim, Yong Wook 한국식물생명공학회 2017 식물생명공학회지 Vol.44 No.1

본 연구는 윤판나물의 효과적인 기내 증식을 위한 배양 최적 조건을 구명하기 위해 수행되었다. 신초 유도 및 생장에 영향하는 배지(MS, B5 및 WPM) 및 절편체 종류에 따른 실험에서 줄기 유도는 액아 절편을 식물생장조절물질 무첨가 MS 배지에 배양 시 절편 당 2.5개로 가장 좋았고, 줄기 신장은 정아 절편을 식물생장조절물질 무첨가 MS 배지 배양 시 7.2 cm로 가장 효과적이었다. BA 침지 처리 시 신초 유도 효과는 액아배양 유래 신초를 BA 침지 무처리구에서 신초 유도(2.29개/절편) 및 신초 신장(7.28 cm)이 가장 효과적이었으며, BA 침지 시간이 길어짐에 따라 신초 유도 및 길이 생장은 감소하였다. 발근에 영향하는 IBA 농도 효과는 정아유래 신초를 1.0 mg/L 처리구에서 배양 시 최대 발근율(100%) 및 뿌리 유도수(21.3개/절편)를 보였으며, 정아 혹은 액아 유래 신초지 종류에 따른 발근율의 차이는 크지 않은 것으로 관찰되었다. 발근된 식물체는 상토로 이식하여 형태적인 변이 없이 정상적인 생장을 보였다. This study was conducted to establish the optimal condition for in vitro propagation of Disporum sessile. In the experiments with different kinds of media (MS, B5 and WPM) and explant types for shoot induction and elongation, the highest number of shoot inductions (2.5/explant) was shown when the axillary bud explants were cultured on MS medium without plant growth regulators (PGRs). The best shoot elongation (7.2 cm) was also obtained when the apical buds were cultured on MS medium. The effect of BA pulse treatments with in vitro shoots was also examined. The highest in shoot induction (2.29/explant) and elongation (7.28 cm) was observed when the shoots from axillary buds were cultured on the media without PGRs. However, it decreased with increasing duration of BA pulse-treatment. The highest rooting rate (100%) and number of root inductions (21.3/explant) were achieved with 1.0 mg/L indole-3-butyric acid (IBA) treatment, whereas no differences were observed by different shoot types. The regenerated in vitro plantlets were hardened and successfully established in soil.

Multiple shoot induction and plant regeneration from axillary buds of Magnolia ‘Vulcan’

Tae-Dong Kim,Ji-Ah Kim,Na-Nyum Lee,Chang-Ho Choi (사)한국식물생명공학회 2020 JOURNAL OF PLANT BIOTECHNOLOGY Vol.47 No.1

An efficient protocol for multiple shoot induction and plant regeneration from axillary bud culture of Magnolia ‘Vulcan’ was developed in the present study. Primary shoots were obtained from axillary bud explants cultured on Murashige and Skoog (MS) medium containing 1.0 mg/L 6-benzylaminopurine (BA). To induce multiple shoots effectively, primary shoot tips were cultured on MS medium supplemented with different concentrations of BA and zeatin at 0, 0.2, 0.5, and 1.0 mg/L. Of these treatments, the MS medium with 0.5 mg/L BA resulted in the highest number of shoots per explant with an average value of 5.9, and it produced the greatest shoot height at 4.8 cm after 12 weeks of culturing. In the rooting of in vitro produced shoots, the greatest percentage of explants forming roots (91.3%), number of roots per explant (9.7), and root length (2.8 cm) were obtained in half-strength MS medium supplemented with 6.0 mg/L indole-3-butyric acid (IBA). Regenerated plantlets were successfully acclimatized and hardened off inside the culture room with 87.5% survival rate. Plants were transferred to a greenhouse with a 97.2% survival rate. The highly efficient shoot multiplication and plant regeneration system reported herein can be used for large-scale clonal propagation of valuable Magnolia species or cultivars.

쉽싸리의 기내 마디배양 시 탄소급원의 종류와 농도가 식물체 재분화 및 증식에 미치는 영향

이나념,김지아,김용욱,김태동,Lee, Na-Nyum,Kim, Ji Ah,Kim, Yong Wook,Kim, Tae Dong 한국식물생명공학회 2018 식물생명공학회지 Vol.45 No.2

본 연구에서는 쉽싸리의 재분화와 순화에 영향을 주는 탄소급원의 효과에 관한 연구를 수행하였다. 식물체 재분화는 1/2MS 배지에 3 ~ 10%의 sucrose와 glucose를 각각 다른 농도로 처리하여 재분화를 유도하였다. 그 결과 3%와 5% sucrose 처리에서 줄기증식과 발근이 증가되었으며, 고농도 처리에서는 줄기와 뿌리 길이 등의 생육이 억제되었다. 기외 순화 중 3% 혹은 5% sucrose 처리 배지에서 생육한 식물체의 뿌리비대, 뿌리길이, 줄기 길이 및 생존율은 glucose 및 다른 농도처리와 비교하여 가장 높게 나타났다. 따라서 3% 및 5% sucrose 처리는 쉽싸리의 기내 생육과 순화를 위해 가장 적정한 농도로 확인되었다. We examined the effect of carbon sources on the regeneration and ex vitro acclimatization of Lycopus lucidus Turcz. ex Benth. Plantlets were regenerated on the 1/2MS medium supplemented with different concentrations (3 ~ 10%) of sucrose and glucose. The sucrose concentrations of 3% and 5% that were supplied enhanced shoot multiplication and rooting but hampered high concentration growth (including the length of the shoot and root). During ex vitro acclimatization, the tuberization of the root, the root length, the shoot length and the survival rate of Lycopus lucidus plantlets grown using 3% and 5% sucrose were found to be better than the other carbon sources and concentrations. Thus a sucrose concentration of 3% and 5% in the 1/2MS medium appeared to be better for both in vitro growth and ex vitro acclimatization of Lycopus lucidus.

Multiple shoot induction and plant regeneration from axillary buds of Magnolia 'Vulcan'

Kim, Tae-Dong,Kim, Ji-Ah,Lee, Na-Nyum,Choi, Chang-Ho The Korean Society of Plant Biotechnology 2020 식물생명공학회지 Vol.47 No.1

An efficient protocol for multiple shoot induction and plant regeneration from axillary bud culture of Magnolia 'Vulcan' was developed in the present study. Primary shoots were obtained from axillary bud explants cultured on Murashige and Skoog (MS) medium containing 1.0 mg/L 6-benzylaminopurine (BA). To induce multiple shoots effectively, primary shoot tips were cultured on MS medium supplemented with different concentrations of BA and zeatin at 0, 0.2, 0.5, and 1.0 mg/L. Of these treatments, the MS medium with 0.5 mg/L BA resulted in the highest number of shoots per explant with an average value of 5.9, and it produced the greatest shoot height at 4.8 cm after 12 weeks of culturing. In the rooting of in vitro produced shoots, the greatest percentage of explants forming roots (91.3%), number of roots per explant (9.7), and root length (2.8 cm) were obtained in half-strength MS medium supplemented with 6.0 mg/L indole-3-butyric acid (IBA). Regenerated plantlets were successfully acclimatized and hardened off inside the culture room with 87.5% survival rate. Plants were transferred to a greenhouse with a 97.2% survival rate. The highly efficient shoot multiplication and plant regeneration system reported herein can be used for large-scale clonal propagation of valuable Magnolia species or cultivars.

편백의 다신초 유도 및 발근을 통한 식물체 재분화

김지아,이나념,김용욱,Kim, Ji Ah,Lee, Na-Nyum,Kim, Yong Wook 한국식물생명공학회 2019 식물생명공학회지 Vol.46 No.4

본 연구에서는 편백의 다신초 유도를 통한 기내 식물체 재분화기술을 개발하고자 본 연구를 수행하였다. 이를 위해 먼저 다신초 유도를 위한 적정 싸이토키닌의 종류 및 농도별 처리에서 1/2LM배지에 0.2 mg/L zeatin의 처리구에서 가장 높은 73%였으며, 절편체 당 평균 3.9개 다신초가 유도되었다. 그리고 싸이토키닌의 고농도→저농도의 순차적 처리의 경우 5.0 mg/L→0.5 mg/L zeatin 처리구에서 100%의 신초 유도율과 절편체 당 17.1개의 가장 많은 다신초가 유도되어 다신초 유도를 위한 최적 조건으로 확인되었다. 신초의 길이신장을 위한 배지는 LM배지에서 배양할 경우 2.62 cm로 가장 좋은 길이생장의 결과를 얻었다. 신초정단부의 갈변화 현상을 억제하기 위해 항산화제 처리 결과 0.5% MES처리구에서 37.5%의 갈변화증상만이 보여 무처구에 비해 2.5배 이상 갈변화 억제 효과가 있었으며, 그에 따른 줄기 길이 신장 또한 1.3 cm로 가장 좋게 나타났다. 신초의 발근은 0.2 mg/L IBA처리구에서 65% 발근율과 3.9개 뿌리가 유도되어 가장 효과적인 처리구였으며, 이렇게 재분화된 기내식물체는 순화를 거쳐 현재 생육포지에서 활발히 생육 중에 있다. A protocol for the in vitro propagation of Chamaecyparis obtusa was established in the present study. Multi-shoots were initiated from apical shoot explants from germinants after 10 weeks of culture on Litvay medium (LM) supplemented with different concentrations of cytokinin. The effects of pre-treatment with high concentrations of cytokinin and varying concentrations (0.2 to 5.0 mg/L) of zeatin on in vitro shoot elongation and shoot multiplication were investigated. Optimal shoot growth was achieved on LM medium, with over 10-mm shoots after 10 weeks of culture. In the anti-browning tests, ethanesulfonic acid triggered the least browning in the shoot tips. The highest multi-shoot induction was observed in the 0.5-mg/L zeatin treatments, which yielded 80% induction of shoots after 10 weeks of culture, and maximum shoot elongation was observed in the LM basal medium without the hormone. The highest rooting rates were 65% under 0.2 mg/L indole-3-butyric acid.

탄소급원에 의한 얌의 기내 비대근 형성과 순화

이나념,김지아,김용욱,김태동,Lee, Na Nyum,Kim, Ji Ah,Kim, Yong Wook,Kim, Tae Dong 한국식물생명공학회 2018 식물생명공학회지 Vol.45 No.2

이 실험에서 우리는 얌의 기내 비대근 유도와 순화에 영향을 미치는 탄소급원의 효과를 연구하였다. 첫 번째로 기내 비대근 유도를 위한 탄소원의 종류와 농도의 효과를 실험하였다. 7% sucrose처리에서 기내 비대근 유도 효율이 가장 높았으며, glucose처리에서는 기내비대근이 형성되지 않았다. 두 번째로 기내식물체의 순화 후 생존율과 비대근 형성율을 실험하였다. 순화 후 형성된 비대근의 직경(6.1 mm)과 생중량(0.5 g)은 7% sucrose에서 생육한 기내식물체에서 가장 높았다. 저농도(3%)의 sucrose처리에서 생육한 기내식물체의 순화 후 생존율은 100%였으나 식물체의 생육과 발달이 억제되었다. 이러한 결과는 얌의 기내 비대근 형성과 순화를 위해 7% sucrose처리가 적정하며 이러한 기술은 얌의 바이러스 및 병 저항성 클론과 우수 개체의 번식을 위해 적용이 가능하다. In this experiment, we investigated the effects of various carbon sources and concentrations on the microtuber induction and acclimatization of the yam (Dioscorea cayenensis). First, the effects of the in vitro carbon sources and concentrations on the microtuber induction were examined. The highest efficiency of the microtuber induction was obtained in the 7% sucrose treatment, whereas the glucose treatment shows no effect on the microtuber formation. Secondly, the effects of the survival rate and the microtuber formation rate after the acclimatization were examined. The diameter (6.1 mm) and fresh weight (0.5g) of the tuberous root are the highest in the pretreatment of the 7% sucrose. Although the survival rate of the pretreatment of the low concentration sucrose (3% sucrose) is 100 %, the growth and development were inhibited. These results suggest the 7% sucrose treatment is appropriate for the yam microtuber formation and acclimatization. In addition, this protocol could be used for the propagation of virus- or disease-free clones and the multiplication of elite yam cultivars.

Incidence and pattern of implant fractures: A long-term follow-up multicenter study

Lee, Jae-Hong,Kim, Yeon-Tae,Jeong, Seong-Nyum,Kim, Na-Hong,Lee, Dong-Woon Decker 2018 CLINICAL IMPLANT DENTISTRY AND RELATED RESEARCH Vol.20 No.4

<P>ConclusionPeri-implantitis-induced marginal and vertical bone loss and manufacturing-induced defects were considered to be major factors in IF. Therefore, using clinically verified implant systems and striving to minimize bone loss by preventing and actively treating peri-implantitis is essential to reduce IFs.</P>

High-frequency regeneration of plants in vitro from seedling-derived apical bud explants of Tilia mandshurica Rupr. & Maxim

Tae-Dong Kim,Nam-Ho Kim,Eung-Jun Park,Na-Nyum Lee 한국식물생명공학회 2021 JOURNAL OF PLANT BIOTECHNOLOGY Vol.48 No.1

This work describe an efficient method for the shoot induction and plant regeneration of seedling-derived apical bud explants of Tilia mandshurica Rupr. & Maxim. The highest rate of shoot induction (82.2%) was obtained when apical bud explants from juvenile seedlings (5 months old) were cultured on Murashige and Skoog (MS) medium containing 1.0 mg/L 6-benzylaminopurine (BAP). However, apical bud explants obtained from mature trees (12 years old) did not produce any shoots, even with BAP supplementation. Among the three cytokinins tested for shoot multiplication (BAP, zeatin, and kinetin), BAP was the most effective; the highest number of shoots per explant (2.1) was observed on MS medium supplemented with 1.0 mg/L BAP. In contrast, the longest average shoot length (3.0 cm) was observed after growth on MS medium with 2.0 mg/L zeatin. No multiplication occurred when apical bud explants were cultured with kinetin-supplemented media. During rooting of in vitro-elongated shoots, the highest rooting rate (100%) was observed in half-strength MS medium supplemented with 0.5 ~ 1.0 mg/L indole-3-butyric acid (IBA) or 3.0 mg/L 1-naphthaleneacetic acid (NAA). During the acclimatization process, plantlets that were rooted on the IBA (0.5 mg/L)- supplemented medium had the highest survival rate (100%) and maximum root length (18.5 cm). These findings suggest that a low concentration (0.5 mg/L) of IBA is appropriate for the rooting and acclimatization of T. mandshurica. Plants were successfully transferred to the greenhouse with a 100% survival rate. This protocol will be useful for the large-scale propagation of Tilia species.