• 생명과학회지
• /
• 제11권3호
• /
• pp.254-258
• /
• 2001

The biochemical change during regeneration of perilla callus were investigated by comparing total protein and lipid contents, protein band pattern in SDS-PAGE, and fatty acid composition in the calli cultured for various period(0, 1, 3, 5 and 6 weeks) Calli were induced from cotyledon and hypocotyl explants of peplants of perilla on perilla on MS medium containing BA(0.5 mg/L) and NAA(0.5mg/L). The protein contents reached the peak at 3 weeks after induction of calli, and then was decreased. Total lipid contents was decreased as the culture period increased. The band pattern of polypeptides showed that 30KD and 45KD polypeptides and 22KD and 45KD polypetides were major proteins in the cotyledon and hypocotyl explants, respectively. However increase of culture period only 30KD protein was highly accumulated.

• Journal of Ginseng Research
• /
• 제6권2호
• /
• pp.162-167
• /
• 1982

Calli and leaflets of ginseng (Panax ginseng C.A. Meyer) were cultured on 1/2MS media supplement. with kinetin, 2 iP, NAA, 2,4-D and IBA to assess their capacity to regenerate embryoids and organs. Root calli produced numerous embryoids and shoots in 1/2MS medium supplemented with 2 mg/l NAA and 2mg/s 2iP, and the combination of 2 iP and NAA was more effective than the combination of kinetin and NAA in induction of embryoid and shoot from root calli. Culture of leaflet in the medium supplemented with IBA resulted in profuse root regeneration.

• 고려인삼학회:학술대회논문집
• /
• 고려인삼학회 1993년도 학술대회지
• /
• pp.143-149
• /
• 1993

Ginseng root explants and calli induced from selected cell lines were cultured on modified Murashige and Skoog's media supplemented with different concentrations of organic or inorganic compounds and plant growth requlators to clarify the effects of chemical composition and plant growth regulators in the medium on the growth of ginseng calli and the production of ginseng saponin. For optimum growth of calli, the concentrations of 2, 4-D and sucrose were the range of 1 to 3 mg/${\ell}$l and 1 to $3\%,$ respectively. And it was clarified that sucrose, nitrogen, phosphate, calcium, magmesian plant growth regulators and their concentrations influcenced the relative biosynthesis of saponin in tissue cultures of Panax ginseng. The patterns of ginsenosides, pharmacologically useful component, were different among the cell lines and contents of ginsenosides were much higher in selected cell lines than in original cell line.

• 한국산림과학회지
• /
• 제86권3호
• /
• pp.261-269
• /
• 1997

미류나무의 미성숙 ovule과 줄기로부터 유기된 캘러스에 plasmid pBI221 유전자를 유전자총을 이용하여 인위적으로 삽입하였다. Plasmid pBI221은 CaMV-35S 유전자에 의하여 발현되는 ${\beta}$-glucuronidase(GUS) reporter 유전자를 포함하고 있다. Plasmid pBI221이 물리적으로 삽입된 후 GUS 유전자의 발현정도는 5-bromo-4-chloro-3-indolyl-${\beta}$-gluconide(X-glue)의 반응에 의해 분석되었고, 유전자의 일시적 발현현상은 ovule, 캘러스 시료에 X-glue substrate의 반응에 의하여 나타나는 뚜렷한 점(spot)의 수에 따라서 조사하였다. 본 실험에서 particle bombardment후 GUS유전자의 발현검정에 있어 가장 중요한 요인은 bombardment 횟수와 X-glue substrate에 노출된 시간이었다. X-glue substrate와의 반응결과, 캘러스와 ovule에서 각각 56.8%, 75.9%의 반응 점들을 나타냈다. 여러 처리중 두번의 연속적인 shot와 bombardment 이후, X-glue과 sample을 48시간 반응시킨 후 24시간 동안 alcohol로의 침지가 가장 많은 수의 점을 유기시켰고, 이들 반응으로부터 평균 $25.75{\pm}2.77$(ovule), $11.43{\pm}1.22$(calli)개의 반응 점을 보였다. 유전자총에 의한 외래 유전자 도입에 관한 본 연구는 두종류의 시료로부터 빠른 시간 내에 유전자 발현을 볼 수 있을 뿐만 아니라, 지금까지 Agrobacterium을 이용한 형질전환이 보고되지 않은 미류나무(Populus deltoides)의 형질전환 연구에 대체 방법을 제공하리라 생각된다.

• Journal of Plant Biotechnology
• /
• 제30권1호
• /
• pp.73-80
• /
• 2003

패랭이꽃속의 분포와 개발을 위한 기초자료로 강원도에서 자생 2종과 수집가능한 7 외래종에 대한 기내배양체의 특성을 조사하였다. 캘러스의 유도를 위해 조직부위별 다양한 절편체들을 2,4-D. NAA, BA가 농도별로 처리된 MS 배지에 치상한 후, 27$^{\circ}C$ 광조건과 2.0mg/L 2,4-D와 0.5 mg/L BA의 조합배지에서 배양했을 때 3주 후 절단면에서 callus가 형성되기 시작하였으며, 패랭이꽃 (Dianthus chinensis)의 잎절편체에서 가장 양호하였다. Organogenic 캘러스의 선발은 기내에서 증식된 adventitious shoot의 절편체 유래 캘러스로부터 가능했으며 이들 캘러스를 1.0 mg/L BA와 0.1 mg/L NAA가 조합된 배지에 치상하였을 때 약 30% 정도의 shoot 형성률을 보였고 multiple shoot의 분화를 관찰할 수 있었다. In vitro에서 증식된 adventitious shoot 절편체를 1.0 mg/L BA와 0.1 mg/L NAA가 포함된 N6 배지에 치상했을 때, 캘러스 단계를 거치지 않고 직접 shoot이 형성되었고 D. gratianopol 유래 adventitious shoot 절편체는 52%의 높은 shoot 형성률을 보였다. 이들 multiple shoot로부터 재분화된 소식물체는 0.1 mg/L NAA가 포함된 MS 배지에서 뿌리가 유도되고 vermiculite가 담긴 pot에 이식 후 85% 습도하에서 활착되어 정상적인 개화된 개체를 획득할 수 있었다. 0.03 M EMS 처리된 패랭이꽃의 adventitious, shoot 절편체 유래 캘러스로부터 힌꽃의 패랭이꽃의 도연변체식물 (M23)이 분화되었으며 종자 형성이 가능하였다.

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

• Asian-Australasian Journal of Animal Sciences
• /
• 제17권10호
• /
• pp.1390-1394
• /
• 2004

We have achieved efficient transformation system for forage-type tall fescue plants by Agrobacterium tumefaciens. Mature seed-derived embryogenic calli were infected and co-cultivated with each of three A. tumefaciens strains, all of which harbored a standard binary vector pIG121Hm encoding the neomycin phosphotransferase II (NPTII), hygromycin phosphotransferase (HPT) and intron-containing $\beta$-glucuronidase (intron-GUS) genes in the T-DNA region. Transformation efficiency was influenced by the A. tumefaciens strain, addition of the phenolic compound acetosyringone and duration of vacuum treatment. Of the three A. tumefaciens strains tested, EHA101/pIG121Hm was found to be most effective followed by GV3101/pIG121Hm and LBA4404/pIG121Hm for transient GUS expression after 3 days co-cultivation. Inclusion of 100 $\mu$M acetosyringone in both the inoculation and co-cultivation media lead to an improvement in transient GUS expression observed in targeted calli. Vacuum treatment during infection of calli with A. tumefaciens strains increased transformation efficiency. The highest stable transformation efficiency of transgenic plants was obtained when mature seed-derived calli infected with A. tumefaciens EHA101/pIG121Hm in the presence of 100 $\mu$M acetosyringone and vacuum treatment for 30 min. Southern blot analysis indicated integration of the transgene into the genome of tall fescue. The transformation system developed in this study would be useful for Agrobacterium-mediated genetic transformation of tall fescue plants with genes of agronomic importance.

• 식물조직배양학회지
• /
• 제22권5호
• /
• pp.283-290
• /
• 1995

미나리의 체세포 배발생 조건 및 기작을 구명하고자 배발생 캘러스의 획득에 적합한 시료와 배지를 구명하고 배발달의 과정을 관찰하여 본 결과, 기내 식물체 확보를 위한 미나리의 경정배양은 발생한 줄기 수와 생육상태로 보아 BA가 0.1 - 0.5 mg/L 첨가된 MS배지가 적합하였으며 배발생 캘러스의 발생은 배유와 함께 채취ㆍ배양한 유배를 2,4- D가 1 mg/L 첨가된 배지에서 2 개월간 배양하였을 때 발생하였다. 기내에서 생장한 줄기와 엽병을 배양하였을 경우는 치상후 2과에 엽병의 절단 부위에서 발생한 캘러스는 모두 배발생 캘러스가 아니였으나 치상 후 6 - 8주에 줄기와 엽병의 표피부위에서 발생한 캘러스는 배발생 캘러스이었다. 배발생 캘러스는 계대배양에 의하여 대양 증식이 가능하며 이들은 배발생 캘러스와 비배발생 캘러스를 동시에 형성하였다. 비배발생 캘러스는 다양한 생장조절제를 첨가한 액체 및 고체배지에 반복 및 지체배양하여도 배발생 캘러스로의 분화는 일어나지 않았다. 비배발생 캘러스는 고체배지에 배양하면 쉽게 증식되어 뿌리를 분화하며, 액체배지 에 현탁배양할 경우에는 단세포로 단리되고, 다시 이들은 작은 원형세포괴를 형성한 후 단지 뿌리만을 발생하였다.

• Journal of Plant Biotechnology
• /
• 제5권4호
• /
• pp.233-238
• /
• 2003

Sweet potato cells derived from Yulmi were isolated from embryogenic callus and irradiated with 50 Gy dose. Resistant cells were selected on a MS medium containing 1.0 mM S-aminoethyl-L-cysteine (AEC). This level of AEC approximately inhibits non-selected wild type cells. The callus resistant to this analog of lysine was subcultured for 30 days in absence of AEC to proliferate. The three resistant calli (AR-1, AR-2 and AR-3) with better growth were divvied into 0.5~1mm diameter and placed on MS medium with 0, 0.4, 0.6, 0.8 and 1.0 mM AEC. There are considerable growth difference between control callus and AEC resistant callus on the AEC-medium. The selected calli were placed on the hormone-free medium for regeneration. Three plantlets, five plantlets and six plantlets were recovered from AR-1, AR-2 and AR-3 calli, respectively. Each two regenerants in AR-1, AR-2 and AR3 were randomly selected for RAPD and SDS PAGE analysis. RAPD polymorph isms between Yulmi and AEC resistant plant from irradiated calli were detected in several Wako primers. Also, it was identified that two AEC resistant plants had higher protein than the original variety Yulmi.

• Journal of Plant Biotechnology
• /
• 제30권3호
• /
• pp.233-239
• /
• 2003

We have developed a reliable and high-frequency genetic transformation and regeneration system via somatic embryogensis of Eleutherococcus sessiliflorus. Embryogenic callus obtained from seed were co- cultivated with Agrobacterium tumefaciens strain EHA101/pIG121Hm harboring genes for intron-$\beta$-glucoronidase(GUS), kanamycin and hygromycin resistance. Following co-cultivation, two types of samples(fine embrogenic calli and early globular embryo clusters) were cultivated on Murashige and Skoog(MS) medium containing 1 mg/L2.4-D for 3day in dark. Transient expression of GUS gene was found to be higher in the early globular embryo clusters than in the embryogenic calli. Also, co-cultivated period affected expression of GUS gene; the best result was obtained when globular embryo clusters were co-cultivated with Agrobacterium for 3 days. Subsequently, this callus transferred to selective MS medium containing 1mg/L2.4-D, 50mg/L kanamycin or/and 30mg/L hygromycin and 300mg/L cefortaxime. These embryogenic calls were subcultured to the same selection medium at every 2 weeks intervals. Approximately 24.5% of the early globular embryos co-cultivated with Agrobacterium for 3days produced kanamycin or/and hygromycin-resistant calli. Transgenic somatic embryos were converted into plantlets in half strength MS medium supplemented with 3mg/L GA$_3$ kanamycin and were confirmed by GUS histochemical assay and polymerase chain reaction analysis. Genomic Southem blot hybridization confirmed the incorporation of NPT II gene into the host genome.