• Journal of Plant Biotechnology
• /
• 제30권1호
• /
• pp.53-58
• /
• 2003

여러가지 생장조절제를 첨가한 MS배지에 참나리의 기내배양 소인편과 실생인편을 치상하여 재분화 과정을 알아보았다. 소인편의 부정아 발생은 BAP 1.0mg/L와 NAA 0.1 mg/L을 혼합 처리한 MS배지에서 4주간 배양했을 때 가장 왕성하게 일어났으며, 실생인편의 부정아 발생률은 BAP 0.5mg/L와 NAA 0.1 mg/L에서 4주간 암배양했을 때 가장 높았다. 소인편의 캘러스 발생은 BAP 0.1 mg/L와 2,4-D 1.0 mg/L 혼합 처리구에서 가장 높았다. 또한. 캘러스의 재분화는 NAA 0.5 mg/L와 BAP 0.1 mg/L를 혼합 처리한 MS배지에서 8주간 명배양했을 때 가장 높았다. 뿌리는 MS배지에서 쉽게 발달되었으며, 8주 후 굵고 긴 뿌리를 갖춘 완전한 재분화 식물체로 성장하여 화분에 이식하였다. 참나리의 재분화 전, 후의 염색체 수는 모두 2n=24로서 조직배양에 의한 염색체수의 변화가 없음을 확인하였다.

• Journal of Plant Biotechnology
• /
• 제43권1호
• /
• pp.110-118
• /
• 2016

We compared germination efficiency for somatic embryos (SE) of Liriodendron tulipifera using semi-solid (SS), temporary immersion bioreactors (TIB), and continuous immersion bioreactors (CIB) to produce vigorous plants. The bioreactors were designed to be immersed in liquid media with plantlets with an adjustable immersion time. TIB and CIB improved germination rates up to 80.86% and 95.21%, respectively, however, CIB produced more hyperhydric plantlets than TIB. The height of plantlets in TIB was significantly higher than for those in CIB. Fresh weights of plantlets grown in CIB of were significantly lower than for those grown in TIB. The lowest chlorophyll concentration was found in in vitro plantlets from CIB. We examined abnormally developed leaves, stems, and apical zones of in vitro plantlets that were produced in CIB. Among the three types, SS showed the highest stomatal density and the shortest stomatal length in in vitro plantlets. After acclimatization, plants from CIB exhibited the lowest values in biomass, such as height, root collar diameter, leaf fresh weight, leaf length, leaf width, petiole length, petiole diameter, and leaf area. Photosynthesis and transpiration rates of ex vitro plants were not significantly different among the three culture types, but stomatal conductance was higher in TIB than in the SS and CIB. Therefore, the results suggest that TIB is the preferable bioreactor to improve in vitro plantlet regeneration of L. tulipifera. TIB-originated plants showed higher growth rate than SS and CIB after transferring to soil.

• Plant Biotechnology Reports
• /
• 제2권4호
• /
• pp.245-252
• /
• 2008

The genetic status of somatic embryo-derived plantlets of Cymbopogon flexuosus was examined by randomly amplified polymorphic DNA (RAPD) analysis. Auxins such as 2, 4-dichlorophenoxyacetic acid (2, 4-D) (1-4 mg/l) were used in Murashige and Skoog (MS) medium for induction of calli from rhizomatous explants of Cymbopogon flexuosus. Optimum calli were induced on MS medium supplemented with 2, 4-dichlorophenoxyacetic acid (2, 4-D) (3.5 mg/l) alone or in combination with $N^6-benzyladenine$ (2 mg/l). Somatic embryogenesis was achieved from long term calli when cultured on MS medium containing 2, 4-dichlorophenoxyacetic acid (2, 4-D) (2 mg/l) along with $N^6-benzyladenine$ (BA) (1-2 mg/l). Regeneration was achieved when freshly induced embryogenic calli were sub-cultured on MS medium supplemented with $N^6-benzyladenine$ (3 mg/l) alone. Long-term cultured embryos showed profuse minute rooting on regeneration medium supplemented with N6 -benzyladenine (3 mg/l). Microshoots were rooted in the presence of indole-butyric acid (IBA) (2 mg/l). DNA samples from the mother plant and 18 randomly selected regenerated plants from a single callus were subjected to RAPD analysis with 6 arbitrary decamer primers for the selection of putative somaclones. A total of 64 band positions were scored, out of which 19 RAPD bands were polymorphic. From genetic similarity coefficient based on RAPD band data sharing, it was found that the majority of the clones were almost identical or more than 92% similar to the mother plant, except CL2 and CL9 (66%) which showed highest degree of genetic change with CL2 and CL9 showing presence of two non-parental bands each.

• Plant Biotechnology Reports
• /
• 제2권4호
• /
• pp.259-265
• /
• 2008

Picea koraiensis, called Korean spruce, is an evergreen tree and found mostly in northeast Asia. In this study, plant regeneration via somatic embryogenesis from open-pollinated immature zygotic embryos of nine geno-types of elite trees was established. Immature zygotic embryos were cultured onto RJW medium modified from 505 medium with $21.48{\mu}M$ NAA, $2.22{\mu}M$ BA, and $2.32{\mu}M$ KT. The average frequency for all nine genotypes was 74.2%. Embryogenic calluses of the nine genotypes of elite trees were subcultured on RJW basal medium containing $8.06{\mu}M$ NAA, $1.11{\mu}M$ BA, and $1.16{\mu}M$ kinetin. The calluses of three lines, $3^{\sharp}$, $9^{\sharp}$, and $2^{\sharp}$, were actively proliferated but others were not. Somatic embryogenesis was induced from the embryogenic callus in genotypes of $3^{\sharp}$, $9^{\sharp}$, and $2^{\sharp}$ on RJW medium with ABA and $60g\;l^{-1}$ sucrose. Cotyledonary somatic embryos were subjected to a drying process. The drying of embryos by uncapping the culture bottle for 5 days on a clean bench resulted in a high frequency of germination of somatic embryos (87% in RJW medium). However, plantlet conversion from germinated embryos was greatly reduced and the optimal medium for plant conversion was 1/2 WPM or 1/2 BMI medium. In conclusion, we have, for the first time, established a plant regeneration system via somatic embryogenesis in the Korean spruce, which can be applied for rapid micropropagation of elite trees.

• 한국자원식물학회:학술대회논문집
• /
• 한국자원식물학회 2019년도 추계학술대회
• /
• pp.15-15
• /
• 2019

The walnut (Juglans regia L.), a member of the Juglandaceae, is native to the mountain ranges of central Asia. This species of walnut is valued commercially for its nuts and in some areas for its timber. The seeds of walnut are recalcitrant and it has strong integument dormancy and their germination is irregular, making its natural propagation difficult. Low percentage of seed germination and long propagation cycle are the main problems of propagation. This study was conducted medium composition on in vitro plantlet regeneration from axillary buds of walnut. It has proved to be the most generally applicable and reliable method of in vitro propagation. Micropropagation culture that axillary buds are excised aseptically enables faster multiplication of plants. The axillary buds of walnut new cultivar "Sinlyeong" were cultured on two basal media which contained the different plant growth regulators depending on the respective shooting and rooting stage. After 12 weeks, the shoot generation rate was 85.3%, the shoot number and its length were 1.9/explant and 2.7 cm in the most favorable medium composition. The percentage of rooting was 25.4%. From these results, it was found the optimum basal medium and plant growth regulator for in vitro plant regeneration from axillary buds of walnut new cultivar "Sinlyeong". However, we have continued to search the other medium additives to enhance the rate of walnut root.

• Journal of Plant Biotechnology
• /
• 제40권3호
• /
• pp.141-146
• /
• 2013

본 연구는 낙엽송 배발생세포의 배양기간에 따른 현탁세포의 생장, 체세포배 유도 및 발아를 위한 다양한 삼투압제 종류 및 농도 효과를 조사하기 위해 수행되었다. 배발생 현탁세포의 생장은 배양일수가 증가함에 따라 settled cells volume (SCV)가 지속적으로 증가하는 경향을 보여 배양 18일 경에는 최대 10.1 ml의 SCV를 보였고, 그 이후로는 세포생장이 감소하였다. 현탁세포로부터 체세포배 발생을 위한 삼투압제 종류 및 농도에 따른 효과 비교에서 최대의 체세포배 유도 수는 0.2 M sucrose 처리구에서 352.3개(/g FW)로 나타났으며, 0.2 M maltose (203.7) 및 0.3 M maltose (193.7) 처리구에서 또한 효과가 있었다. 그러나 7.5% polyethylene glycol (PEG)이 첨가된 0.15 M sucrose 혹은 maltose 각각의 처리구에서는 체세포배 발생이 전혀 이루어지지 않았다. 다양한 삼투압제 종류 및 농도에서 발생된 체세포배의 발아율 비교에서는 최대 자엽 형성율(25.2%) 0.3 M maltose, 하배축 형성률(39%), 뿌리 발생율(30.3%) 및 식물체 재분화율(3.5%)은 0.2 M sucrose 처리구에서 각각 나타났다.

• 한국산림과학회지
• /
• 제79권2호
• /
• pp.109-114
• /
• 1990

피나무의 조직배양(組織培養)에 의(依)한 대량증식(大量增殖) 방법(方法)을 확립(確立)하기 위하여, 15년생(年生) 성숙목(成熟木)의 액아(腋芽)를 IS, MS 및 WPM배지(培地)에 배양(培養)하였다. 5수절(水凖)의 BAP농도(濃度)가 검정(檢定)되어, IS와 WPM배지(培地)에서는 BAP 1.0mg/l 첨가(添加)가 줄기 형성(形成)과 생장(生長)을 촉진(促進)한 반면(反面), MS배지(培地)에서는 BP 0.5mg/l 첨가(添加)가 효과적(效果的)이었다. 최적조건(最適條件)은 BAP 1.0mg/l 첨가(添加)의 WPM과 0.5mg/l 첨가(添加)의 MS배지(培地)이었다. 형성(形成)된 줄기는 다경유도(多莖誘導)를 위하여 BAP 0.2mg/l 첨가(添加) 배지(培地)에 다식(多植)하여, 한달안에 약(約) 2배(倍)로 증식(增殖)되었다. 발근(發根)은 고농도(高濃度)의 IBA에서 촉진(促進)되어, IBA 3.0mg/l 첨가(添加)의 1/2MS에서 83.3%의 발근율(發根率)을 얻었다. 재분화(再分化)된 유치물체(幼植物體)는 환경순화(環境馴化)될 수 있었다. 액아배양(腋芽培養)에 의(依)한 줄기 형성(形成)과 생장면(生長面)에서 클론간(間) 변이(變異)를 조사(調査)하기 위하여 수형목(秀型木) 7클론이 검정(檢定)되었다. 조직배양능력(組織培養能力)에 있어서 수형목(秀型木) 클론간(間) 변이(變異)는 Duncan의 다중검정(多重檢定) 5% 유의수준(有意水準)에서 유의성(有意性)이 인정(認定)되었으며, 강원(江原) 12호(號)가 BAP 1.0mg/l 첨가(添加) WPM에서 가장 좋은 반응(反應)을 나타냈다.

• 한국자원식물학회지
• /
• 제8권3호
• /
• pp.237-246
• /
• 1995

본 연구에서는 유자의 미성숙 배로부터 캘러스 유도와 재분화 및 발생단게별 미세구조적 특성에 관하여 연구하였다. 유자에서는 배양 5-6주 경에 담황색의 캘러스가 유도되었으며, 배양 6주경의 캘러스 세포에서는 인접하고 있는 작은 액포들이 융합하여 큰 액포를 형성하는 융합현상을 관찰하였다. 배양 12주 경에는 배발생 캘러스의 미분화 세포에서 특이하게 구형의 핵인을 가지고 있는 큰핵과 녹말로 가득차 있는 녹말체를 다수 관찰하였다. 배양 14-16주 경의 배발생 캘러스 세포에서는 활발하게 exocytosis가 일어나므로서 다수의 분배체들이 세포벽쪽에서 관찰되었으며 세포질로부터 작은 입자들이 세포 간극으로 빠져나가는 것을 볼수 있었다. 또한 배양 24주경의 배발생 캘러스에서는 발달된 커다란 구형의 엽록체들이 이중막으로 세포질과 경계되어 있었으며 기질중에 일정하게 전형적인 그라나가 분포되어 있었다. 배양 26주경의 어린 정상 식물체의 미성숙잎에서는 다수의 도관과 반세포들이 관찰되었다. 배양 30주경의 정상식물체의 미성숙잎에는 다수의 작은 반세포들과 사관 그리고 중심액포들이 나타났으며 또한 작은 제2차 액포들이 커다란 중심 액포 속으로 끼어들어가는 현상을 볼 수 있었으며 원형질막 근처에는 신장된 엽록체들이 분포되어 있었다.

• 한국약용작물학회지
• /
• 제5권1호
• /
• pp.21-27
• /
• 1997

반하의 조직배양을 통한 대량증식방법 확립의 일환으로 실시한 기내배양에서의 소괴경형성 및 식물체 분화에 영향을 미치는 배지 NAA와 Thidiazuron의 최적조건을 구명하고져 실시한 실험의 결과는 다음과 같다. 1, 식물생장조절물질로 NAA와 TDZ을 단독처리 할 때 TDZ $0.5\;{\mu}M$에서 shoot수가 45개로 가장 양호하였으며 root분화는 NAA 2.0 mg/l 에서 가장 양호하였다. 2. NAA 0.1 mg/l +TDZ $2.0\;{\mu}M$ 조합처리에서 shoot분화가 가장 양호하였으며 NAA 2.0 mg/l +TDZ $2.0\;{\mu}M$ 처리시에 가장 저조하였다. 3. 반하의 소괴경 형성은 MS배지에서는 TDZ $5.0\;{\mu}M$ 단독처리와 NAA 0.1 mg/l +TDZ $2.0\;{\mu}M$ 처리에서 소괴경 형성이 가장 양호하였다. B5배지에서는 TDZ $1.0\;{\mu}M$ 단독처리와 NAA 1.0 mg/l +TDZ $5.0\;{\mu}M$ 처리에서 소괴경 형성이 가장 양호하였으나 생체중은 NAA 0.1 mg/l 와 TDZ $5.0\;{\mu}M$의 단독처리에서 생체중이 가장 무거웠다. 4. MS, MG, B5배지조성에 따른 소괴경형성은 MG배지에서 30일배양후 가장 양호하였으며 생체중도 좋았다. 5. 분화된 식물체 뿌리분화에는 IAA보다 NAA가, MS 보다 1/2MS가 더 효과적인 결과를 보여 1/2MS배지에 NAA 2 mg/l 를 처리하였을 때 23.3개의 뿌리가 유도되어 가장 높은 결과를 보였다. 완전분화된 식물체를 vermiculite가 담긴 포트에 이식하여 온실에서 순화시킨 결과 80%정도의 생존율을 보였다.

• Journal of Plant Biotechnology
• /
• 제7권4호
• /
• pp.247-257
• /
• 2005

An efficient, rapid and large-scale in vitro clonal propagation of agronomically important Indian cereal crop genotypes (NSH27 & K5) of Sorghum bicolor (L.) Moench. by enhanced shoot proliferation in shoot tip segments was designed. MS medium fortified with plant growth regulators and coconut water markedly influenced in vitro propagation of Sorghum bicolor. In vitro plantlet production system has been investigated on Murashige and Skoog (MS) medium with the synergistic combination of 6-benzyladenine ($22.2\;{\mu}M$), kinetin ($4.6\;{\mu}M$), adenine sulphate ($2.8\;{\mu}M$), 5% coconut water and 3% sucrose which promoted the maximum number of shoots as well as beneficial shoot length. Subculturing of shoot tip segments on a similar medium enabled continuous production of more than 100 healthy shoots with similar frequency. When the healthy shoot clumps were cultured on MS medium fortified with 6-benzyladenine ($22.2\;{\mu}M$), kinetin ($4.6\;{\mu}M$), adenine sulphate ($2.8\;{\mu}M$), ${\alpha}$-naphthaleneacetic acid ($2.7\;{\mu}M$), ascorbic acid ($30.0\;{\mu}M$) and 5% coconut water, a rapid production of axillary and adventitious buds was developed after 8 wk culture. More than 300 shoots were produced 10 wk after culture. Rooting was highest (100%) on half strength MS medium containing 22.8 mM IAA. Micropropagated plants established in garden soil, farmyard soil and sand (2:1:1) were uniform and identical to the donor plant with respect to growth characteristics. These plants grew normally without showing any traits.