To investigate the effects of ozonated water concentration and soaking time on adventitious root formation of willow, we studied the efficiency of root cuttings in the revegetation technology of biological engineering of willows. The ozonated water concentrations were used for 5 minutes and 2 hours at 1, 5, 10, 15, 20 ppm by soaking method and then the shoot characteristics were observed. The number and length of adventitious roots were determined. The results indicated that 20 ppm of ozonated water for 2 hours and 1 ppm of ozonated water for 5 minutes resulted in leaves turned wither away and no adventitious root production. Considering the appearance, number and length of the adventitious root, soaking willow cuttings into the ozonated water with dissolved ozone concentration, 5 ppm for 2 hours and 10-15 ppm for 5 minutes were suitable for generating adventitious roots.
In the present study, the plantlets in vitro of Paeonia suffruticosa ‘Wu Long Peng Sheng’ were used as laboratory materials. The proteome during adventitious root induction process was investigated to sift the related proteins by two-dimensional electrophoresis and mass spectrometry. The results indicated that the protein spots were concentrated in the acidity gel region (pH 4 - 7) and the spots number had a dynamic change ranged from 373 to 462 at the process of root induction (0 – 7 d). 8 spots significantly changed were analyzed with a mass spectrometer and identified using associated software and databases. The peptide information of the 8 spots was similar to the ATP synthase β-subunit of P. suffruticosa (Spots 1 - 4 and 8), P. tenuifolia (Spots 5), P. californica (Spot 6) and P. brownie ( Spots 7) r espectiv ely. T he expression levels of protein spots 1, 4, 5, 6 and 7 was dramatically downregulated, and that of protein spots 2 and 3 had a slightly opposite tendency on the 3rd day. The obviously decreased period is particularly interesting as it was consistent with the induction period of adventitious root primordial of tree peony plantlet in vitro. The ATP synthase β-subunit could be consumed for assembling the ATP synthase in order to supply energy to the rooting process. Therefore, we speculated that the ATP synthase β-subunit was involved in adventitious root initiation of tree peony plantlets in vitro and we expect that further studies should be carried out in order to export its action mechanism.
In this study, we examined immuno-modulatory activities of crude polysaccharides from wild ginseng adventitious roots (WGAR). The crude polysaccharide (WGAR-CP) was isolated from WGAR by hot water extraction, ethanol precipitation, and dialysis. The major constituents in WGAR-CP were neutral sugar (64.77%), and uronic acid (34.32%). WGAR-CP demonstrated anti-complementary activity dose-dependently. The immuno-modulatory effects of WGAR-CP were also analyzed by measuring nitric oxide and cytokines in the supernatants of mouse peritoneal macrophages. Mouse peritoneal macrophages stimulated with WGAR-CP produced nitric oxide and various cytokines such as interleukin (IL)-6 and IL-12 in a dosedependent manner. In conclusion, WGAR-CP may have immuno-modulatory activities by activating a complementary system and macrophages, which produces cytokines.
에크네시아의 뿌리로부터 유도된 캘러스에서 부정근 을 유도하고 이를 액체배양을 통해 효율적으로 증식하 는 배지조건을 조사하였다. 실생뿌리로부터 캘러스를 유도하는 조건은 NAA 0.1mg·L-1가 함유된 MS 기본 배지에 sucrose 30g·L-1, 한천 10g·L-1를 처리 한 것 이 가장 양호하였다. 캘러스로부터 부정근의 유도에는 NAA 0.01mg·L-1를 단독처리 한 배지에서 가장 양호 하였으며, NAA에 BA를 첨가하면 오히려 NAA의 부 정근 유도 효과를 감소시켰다. NAA처리는 동일한 농 도의 IBA보다 부정근의 수와 생체중의 증가에 효과적이었으며, 액체배양 후 약 6주후에 생체중이 최고에 달하는 것으로 나타났다. 무기염류는 SH배지 보다 MS배지가 효과적이었는데, 농도는 기본배지의 50% 성분량을 처리하였을 때 가장 양호하였다. 탄소원으로 는 glucose보다 sucrose가 효과적이며 60g·L-1를 처리 하였을 때 가장 효과적이었다. 질소원으로 KNO3와 NH4NO3의 첨가비를 비교한 결과 KNO3와 100%, NH4NO3는 50%를 첨가한 배지가 부정근의 생체중 증 가에 효과적이었다.
Background : A series of studies were conducted to optimize adventitious root induction in vitro from explants of Astragalus membranaceus using various nutrient media supplemented with plant hormones.
Methods and Results : Levels of active components were analyzed from adventitious roots induced under different media conditions. Among the different media conditions, Murashige and Skoog medium supplemented with 1.0㎎• ℓ −1 indole-3-butyric acid resulted in the greatest adventitious root induction rate. The amount of the major active component of the adventitious roots of Ama1, calycosin-7-O-β-D-glucopyranoside was higher than that of other adventitious root samples.
Conclusions : These results suggest that the adventitious roots of A. membranaceus could be used for the commercial production of medicines.
홍경천의 잎, 줄기, 뿌리 절편을 이용하여 기내 부정근의 생산 체계를 확립하였다. 먼저 홍경천의 잎, 줄기, 뿌리 절편을 0.1, 1.0, 3.0 및 5,0 mg/L의 IBA와 sucrose가 10, 30, 50 및 100 g/L가 첨가된 MS 배지위에 치상하여 부정근의 유도율을 조사하였다. 부정근의 유도는 잎, 줄기 절편에서 IBA의 농도가 5.0 mg/L 일때 가장 높은 유도율을 보였으며, 뿌리 절편은 IBA 3.0 mg/L 첨가된 배지에서 부정근 유도율이 가장 높았다. Sucrose의 농도는 30 g/L가 첨가되었을 때 잎, 줄기, 뿌리 절편에서 높은 유도율을 나타났다. 고체배지 조건에서 부정근의 유도율이 가장 우수한 조건을 기본으로 액체배양을 실시하였으며, 염의 농도에 따른 부정근의 증식조건을 조사하였다. 1/3MS 배지에서 홍경천의 부정근을 배양하였을 때 1/2MS, MS 액체 배지조건 보다 약 2배, 2.5배의 부정근 생장량을 보였다.
An adventitious root formation protocol from Smilax china L. was established for in vitro production of dioscin, a steroidal saponin having various bioactivities such as anticancer, antifungal, antiviral, and antiobesity. Optimal medium for root initiation from leaf explant was MS medium containing 30 g·L-1 of sucrose supplemented with 1.0 mg·L-1 kinetin + 2.0 mg·L-1 NAA. The induction of adventitious roots from in vitro initiated root segments was most favorable to MS liquid medium with 0.1 mg·L-1 kinetin + 2.0 mg·L-1 NAA. Among the 20 different adventitious roots originated from different plants, strain No. 10 was selected based on production ability of dioscin, and its stability through the successive suspension culture. The maximum growth stage of adventitious roots was noticed at 5 weeks after subculture while that of dioscin production in the adventitious root was at 7 weeks after subculture in suspension culture system. These results provide that suspension culture of adventitious roots of Smilax china L. have a potential for in vitro mass production of dioscin.
Echinacea purpurea is an important medicinal plant native of North America, which contains caffeic acid derivatives, alkamides, glycoproteins, and polysaccharides. For commercial roduction of bioactive compounds from Echinacea purpurea, adventitious roots were induced and proliferated in 5 and 20-liter bioreactors under various cultural conditions (Medium composition, inoculum density, air volume, light, temperature et al.). In a 5-liter bioreactor, more than 10-fold of biomass increment was achieved after 5 weeks of culture. Increases of root biomass and bioactive compounds were correlated with increased consumption of sucrose and macro/micro nutrients in the medium. Elicitations have been conducted with UV-C, NO, and SA to increase the accumulation of total polyphenolics, flavonoids and caffeic acid derivatives in the adventitious root cultures, which resulted in 1.2-1.6 times of increment. Based on the previous experimental results, 500-liter and 1000-liter scale bioreactors were applied for large scale cultures of Echinacea purpurea adventitious roots. Total root biomass of 3.62 kg and 5.05kg (dry weight) were achieved in a 500 L and a 1000 L bioreactor, respectively. The accumulation of 22.55 mg/g DW cichoric acid, 4.92 mg/g DW chlorogenic acid and 3.99 mg/g DW caftaric acids were also achieved from the adventitious root in a 1000-liter scale bioreactor.
식물생장조절제의 종류와 농도를 모색하여 할미꽃의 발근 최적 조건을 조사하여 정상적인 재분화체를 획득하고자 하였다. 지금까지의 선행연구를 토대로 최적조건에서 shoot를 유도시킨 후 발근 유도 생장조절제에 바로 옮기면 shoot가 너무 작아 재분화체가 성장하지 못하고 고사되었다. 1/2 MS에서 키운 shoot를 3.0 mg/L NAA를 첨가한 배지에서 3일간 생장시킨 후, 1/3 MS로 옮겨 치상 한 경우, 약 10일 전후에 뿌리의 유도가 관찰되었다. 이렇게 뿌리가 유도된 재분화체는 염의 농도와 설탕의 농도를 동시에 순차적으로 낮추어 순화처리되었다.
Scopolamine and hyoscyamine are important anticholinergic compounds obtained from Hyoscyamus niger. Adventitious roots induced from rhizome of H. niger and roots were cultured in SH medium supplemented with 3% (w/v) sucrose and 0.5 mg/L IBA. Roots were grown rapidly after 10 days of cultures. Scopolamine production was increased 7 times and hyoscyamine production was increased 12 times after 10 day of cultures. SH medium was best in root growth. But, highest scopolamine productivity was observed in WPM medium, followed White medium and best hyoscyamine productivity was resulted in MS medium. Sucrose was increased scopolamine and hyoscyamine production were increased the medium supplemented by sucrose comparing to than those by other carbon sources.
Scopolia parviflora Nakai, a rare and endangered species, is the sole plant producing tropane alkaloids (TA) among the Korean native species. In order to enhance TA productivity the SP72 root line was selected by screening 100 of root line, and the optimal culture media for root growth and TA production were investigated with the SP72 roots. Based on the several media, SH and 2B5 medium were determined as growth medium and White and NN medium as production medium. Among the four combinations of two-stage culture, 2BN (2B5 as growth medium plus NN as production medium) showed more enhanced root growth and TA production as compared with production media of White and NN medium and growth media of SH and 2B5 medium, respectively. However, bubble column bioreactor (BCB) cultures applying two-stage culture did not reveal the effective results despite of the each successful operation of two-stage culture in conical flasks and BCB cultures.
The effect of boron and aluminum on the development of adventitious roots was studied in sunflower cuttings. Three-day-old seedlings were de-rooted and grown in nutrient solutions with or without boron and supplemented with different concentrations (from 50 to 700 μM) of aluminum. The number and length of the adventitious roots and proline content in adventitious roots in response to insufficient boron and aluminum stress were determined periodically. The micronutrient boron caused the development of numerous roots in the lower parts of the hypocotyl. A dose-response of boron-induced rooting yielded an optimum concentration of 0.1 mM boron. In the absence of boron, in the majority of the adventitious roots, a significant inhibition was observed with or without aluminum, indicating that the most apparent symptom of boron deficiency is the cessation of root growth. Increasing concentrations of aluminum caused progressive inhibition of growth and rooting of the hypocotyls, and a parallel increase in proline levels of adventitious roots. Supplemental boron ameliorated the inhibitory effect of aluminum, suggesting that aluminum could inhibit root growth by inducing boron deficiency. Ascorbate added to medium in the absence of boron improved root growth and induced a significant decrease in proline levels. These findings suggest that adventitious root growth inhibition resulting from either boron deficiency or aluminum toxicity may be a result of impaired ascorbate metabolism.
새 번식방법 개발을 위해 둥굴레 뿌리는 눈이 없어도 부정아가 발생된다는 점에 착안하여 뿌리길이와 굵기에 대한 부정아 발생에 관한 시험 연구를 한 결과는 다음과 같다. 1년차는 뿌리길이가 5cm 경우 66.5%지만 15cm에서는 113% 부정아 발생되었고, 뿌리의 굵기 5mm이하는 98.3%지만 10mm이상은 113.3%로 46.7% 차이를 보였다. 2년차는 부정아 생성률은 15cm에서는 1년차의 3배, 시험구 설치 당시 전혀 눈이 없는 상태에서 4배의 증식을 보였다. 용토는 일반토양 2.6배 보다 모래나 펄라이트가 3.1~3.3배 증식 되었다. 대량번식을 위한 가장 좋은 방법은 10mm 둥굴레 뿌리를 5cm로 절단하여 모래에 식재하는 방법으로 판단된다.