• Proceedings of the Korean Society of Plant Biotechnology Conference
• /
• 2002.04b
• /
• pp.131-131
• /
• 2002

• Proceedings of the Korean Society of Plant Biotechnology Conference
• /
• 2005.11a
• /
• pp.436-436
• /
• 2005

• Journal of Forest and Environmental Science
• /
• v.25 no.2
• /
• pp.111-118
• /
• 2009

Multiple shoot induction and plant regeneration using apical bud and nodal explants of 100 year old tree of Anthocephalus cadamba, an important sacred and medicinal tree in India was achieved for the first time. Aseptic explants cultured in Murashige and Skoog (MS) medium augmented with different concentrations of BAP (0.1, 0.5, 1.0, 2.5, 5.0 and 10 mg/l), when maintained for 60 days, healthy shoots were induced in presence of BAP (1 mg/l). Lower concentrations of BAP (0.1 - 0.5 mg/l) induced only one shoot per explant. Increase in number of shoots per explant was observed in presence of higher concentrations of BAP (2.5, 5.0 and 10 mg/l). However, elongation of shoots was completely inhibited. Bud break and shoot regeneration was largely associated with seasonal factors. Apical buds cultured during June to August exhibited early bud break within two weeks of initial culture. In rest of the months, bud break and shoot regeneration was very slow irrespective of the various concentrations of BAP used in the medium. Explants sourced from three different maturity levels of shoots indicated that actively growing shoots from the mother plant with 1 - 2 nodal segments was more suitable for culture initiation than the explants collected from mature shoots at dormant stage. Regenerated shoots with 2 - 3 pairs of leaves when transferred to half strength MS medium fortified with IBA (1 mg/l), 60% of the shoots induced healthy roots, indicating the possibility of large scale micropropagation.

• Journal of Life Science
• /
• v.16 no.3 s.76
• /
• pp.459-463
• /
• 2006

Cultivated garlic, Allium sativum L. is economically important for leaves and bulbs, which historically were used in Korea for spices and condiments of Korean food as well as medicine crops. This experiment was carried out to investigate the effect of development and differentiation on culture of A. sativum (cv: white 6) by explant position, hormone composition and sucrose concentration in culture media. Culture method was investigated to induce shoot primordium. Culture efficiency was better with lower tissue of foliage leaf in explant position and on the medium with NAA 0.02 + BAP 1.0 mg/l in hormone composition than any other. Precocious shoot and callus were induced from shoot apex. Shoot was efficiently differentiated on 4,000 mg/l sucrose with increasing concentration of BAP. Shoot primordium was also induced with liquid rotary culture by histological observation. Rhizoid was induced from callus tissue cluster on medium with NAA 0.02 + BAP 2.0 mg/l.

• Korean Journal of Plant Tissue Culture
• /
• v.21 no.4
• /
• pp.233-237
• /
• 1994

Plant regeneration system via somatic embryogenesis in leaf explant cultures of Gentiana scabra var. buergeri has been established. Leaf segments formed calli when cultured on MS medium supplemented with 0.5 mg/L 2,4-D and 2 mg/L BAP After transferred to SH medium supplemented with 0.5 mg/L 2,4-D, 2 mg/L CPA and 0.5 mg/L kinetin, the callus became embryogenic. The embryogenic callus was subcultured every 3 to 4 weeks. Upon transfer onto SH basal medium the embryogenic callus gave rise to numerous somatic embryos, which subsequently developed into plantlets. The regenerated plants were potted in an artificial soil with mixture (peatmoss : pearlite : vermiculite : 2 : 1 : 1) and transplanted to the soil after kept under a high humidity for two weeks. A total of 78 plants out of 105 regenerated plants survived in the soil. Phenotypic variations in height, number of stems and the flowering time were observed in tile regenerated plants. Cytogenetical analyses showed no chromosomal variation.

• Plant Biotechnology Reports
• /
• v.3 no.1
• /
• pp.67-74
• /
• 2009

An efficient micropropagation protocol based on multiple shoot induction and callus regeneration has been standardized in Sarcostemma brevistigma, a rare medicinal plant. The nodal cuttings were cultured on MS medium supplemented with BA ($0.5-8{\mu}M$) or Kn ($0.5-8{\mu}M$) alone or in combination with NAA ($0.5-1.5{\mu}M$). Maximum multiple shoot induction was observed on MS medium supplemented with $4{\mu}M$ BA. On this medium, 100% cultures responded with an average number of 11.3 shoots per explant. However, the average shoot length was limited to only 0.9 cm on this medium. The addition of $1{\mu}M$ NAA along with $4{\mu}M$ BA gave rise to an average number of 10.9 shoots with an average shoot length of 1.8 cm. Luxuriantly growing callus was obtained on MS medium supplemented with BA ($5{\mu}M$) and 2,4-D ($2{\mu}M$). The callus was subcultured on MS medium supplemented with BA ($2-15{\mu}M$) or Kn ($2-15{\mu}M$) alone or in combination with NAA ($0.5-2{\mu}M$) for shoot organogenesis. Optimum callus regeneration was obtained on MS medium supplemented with $10{\mu}M$ BA and $1{\mu}M$ NAA. On this medium, 100% cultures responded with an average number of 13.4 shoots per culture. The shoots obtained via multiple shoot induction and organogenesis were rooted on half-strength MS medium supplemented with NAA ($1-7{\mu}M$) or IBA ($1-7{\mu}M$). IBA was better than NAA in terms of both the percentage of cultures that responded and the average number of roots per explant. The rooted shoots were successfully transplanted to soil with 86% success. This standardized protocol will help to conserve this rare medicinal plant.

• Plant Biotechnology Reports
• /
• v.4 no.2
• /
• pp.139-148
• /
• 2010

Jatropha curcas L. (Physic nut) is a commercially important non-edible oil seed crop known for its use as an alternate source of biodiesel. In order to investigate the morphogenic potential of immature embryo, explants from four developmental stages were cultured on medium supplemented with combinations of auxins and cytokinins. It was found that the size of embryo is critical for the establishment of callus. Immature embryos (1.1-1.5 cm) obtained from the fruits 6 weeks after pollination showed a good response of morphogenic callus induction (85.7%) and subsequent plant regeneration (70%) with the maximum number of plantlets (4.7/explant) on Murashige and Skoog's (MS) medium supplemented with IBA (0.5 $mg\;l^{-1}$) and BA (1.0 $mg\;l^{-1}$). The above medium when supplemented with growth adjuvants such as 100 $mg\;l^{-1}$ casein hydrolysate + 200 $mg\;l^{-1}$ L-glutamine + 8.0 $mg\;l^{-1}$ $CuSO_4$ resulted in an even higher frequency of callus induction (100%). Plant regeneration (90%) with the maximum number of plantlets (10/explant) was achieved on MS medium supplemented with 500 $mg\;l^{-1}$ polyvinyl pyrrolidone + 30 $mg\;l^{-1}$ citric acid + 1 $mg\;l^{-1}$ BA + 0.5 $mg\;l^{-1}$ Kn + 0.25 $mg\;l^{-1}$ IBA. It was observed that plantlet regeneration could occur either through organogenesis of morphogenic callus or via multiplication of pre-existing meristem in immature embryos. The age of immature embryos and addition of a combination of growth adjuvants to the culture medium appear to be critical for obtaining high regeneration rates. Well-developed shoots rooted on half-halfstrength MS medium supplemented with 0.5 $mg\;l^{-1}$ IBA and 342 $mg\;l^{-1}$ trehalose. The rooted plants after acclimatization were successfully transferred to the field in different agro-climatic zones in India. This protocol has been successfully evaluated on five elite lines of J. curcas.

• Journal of Plant Biotechnology
• /
• v.42 no.4
• /
• pp.380-387
• /
• 2015

Aronia (Aronia melanocarpa, Black chokeberry) is an important cash crop in domestic agriculture. We investigated the effects of plant growth regulators on shoot proliferation and rooting using in vitro tissue culture. The most effective shoot multiplication was observed on WPM (woody plant medium) supplemented with 1.0 mg/L zeatin ($8.3{\pm}1.0$ shoots/explant), while the highest rooting rate was obtained from half-strength WPM with 3.0 mg/L IBA (8.8 roots/explant). The rooted plantlets all survived in the artificial soil mixture (with a mixture of peat moss : perlite : vermiculite, 1:1:1, v/v/v) and grew up relatively uniform, ranging from 14 to 16 leaves, 8 to 10 cm in stem height, and 2.3 to 2.8 mm in stem diameter. While experimenting with 5 different varieties of Aronia, we found out that each variety had different characteristics of shoot proliferation and rooting. The total numbers of proliferated shoots per variety is as follows: $17.4{\pm}0.8$ for Nero, 14 to 15 for Purple and Mackenzie, and 10 for both Viking and Odamamachiko. Rooting rates were also various depending on the variety: 88% of Odamamachiko, 80% of Viking and Purple, and 76% of Nero and 60% of Mackenzie shoots rooted. The survival rate of the rooted plantlets was from 92% to 100%, varying by type. Further growth appeared to be better in auxin-treated plantlets, compared to untreated ones. Our results showed the possibility of establishing an effective in vitro micropropagation system for Aronia melanocarpa.

• Journal of Plant Biotechnology
• /
• v.34 no.3
• /
• pp.189-195
• /
• 2007

Helicteres isora is medicinally important plant effective against asthma, diabetes, hypolipidemia, HIV, besides a good source of diosgenin. Seed dormancy and low rate of natural fruit production make this plant a perfect candidate for developing an in vitro method useful for its clonal propagation and further biotechnological developments. This is the first report on in vitro production of this plant. Nodal explants obtained from aseptically germinated seedlings were cultured on MS medium (Murashige and Skoog 1962) fortified with indole-3-acetic acid (IAA) ($0.57-22.83\;{\mu}M$), indole-3-butyric acid (IBA) ($0.41-16.58\;{\mu}M$), 6-benzylaminopurine (BA) ($0.44-17.75\;{\mu}M$) and kinetin (Kin) ($0.46-13.94\;{\mu}M$) either singly or in combinations of IAA + BA, IAA + Kin and BA + Kin. Combinations of cytokinins (BA and Kin) were most suitable for multiple shoot induction and $13.94\;{\mu}M\;Kin\;+\;13.31\;{\mu}M\;BA$ was optimum (79% frequency) associated with high number of microshoots (7.1 shoots per explant) after 20 days of culture. Maximum shoot elongation and proliferation (10 shoots per explant with 4.8 cm average height) was achieved on MS media containing $2.32\;{\mu}M\;Kin\;+\;2.22\;{\mu}M\;BA\;+\;2.85\;{\mu}M\;IAA$. High rooting frequency (70%) was achieved on MS medium (1/2 basal strength) fortified with $4.14\;{\mu}M$ IBA, while activated charcoal showed inhibitory effects on rooting. Hardening was done with 76% survival rate and these plants were growing without any visual defects and morphologically mimicking the naturally growing plants.

• Journal of Plant Biotechnology
• /
• v.45 no.2
• /
• pp.140-145
• /
• 2018

Inducing genetic and morphological variation through conventional method is very difficult. Therefore, mutation induction through in vitro technology brings numerous advantages over the conventional breeding. Thus, the individual shoots (1 ~ 2 cm) were irradiated with gamma rays (10 ~ 70 Gy). The result revealed that the explants treated with higher doses (40, 50, 60, and 70 Gy) showed deleterious effects of ionizing radiation. The highest survival rate among ${\gamma}$ treated explants recorded was 71% in 10 Gy treatments while the lowest survivality was 15% in 70 Gy. Lethal dose 50% ($LD_{50}$) dose was found to be 33 Gy. In the in vitro condition, rooting reponse showed that increase in gamma irradiation dose resulted in the inhibition of root growth. Meanwhile, non-treated explants had the best rooting ability with the maximum number of root per explant (20) within a short period of time (6 days), with the highest root length of (15.1 cm). The longer period in rooting (12 days) and lowest number of root per explant (8) with shortest root length (10.1 cm) were recorded at 30 Gy treatment. The highest shoot length (13.6 cm) was observed at control treatment and the shortest shoot length (10.4 cm) was observed at 30 Gy. In the nursery, lowest leaf number (5) was observed at 30 Gy compared with other treatments. The highest chlorophyll content (49.8) was recorded at 10 Gy treated seedling. Irradiated explants with 10 Gy found to be superior over the control treatment and had positive effects in main growth parameters such as chlorophyll content.