• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.10a
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• pp.16-16
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• 2019

The Cassava (Manihot esculenta Crantz) is a perennial woody shrub cultivated mainly in the tropics for its starchy tuberous roots. It belongs to the family Euphorbiaceae which also includes rubber (Hevea brasiliensis) and castor bean (Ricinus communis). Among tropical crops, rice, sugarcane, maize and cassava are the most important sources of calories for human consumption. Problems in the propagation of cassava are virus diseases and low rates of seed germination. Thus, a study was undertaken to develop an efficient in vitro mass propagation protocol of Manihot esculenta Crantz. Young and actively growing stem segments were excised from adult plants of cassava. Samples were cut into a 3~4 cm nodal segments with single node after sterilization, and cultivated in the different medium supplemented with various plant growth regulators for 4 weeks. For shoot multiplication, single-node stem segments, approximately 1 cm in length, were taken from in vitro derived shoots and subcultured. After 4~6 weeks, the shoot generation rate was 55.6%, the shoot number and its length were 1.0/explant and 2.3 cm in the most favorable medium composition. Our experiments confirmed that in vitro growth and multiplication of plantlets could depend on its reaction to the different medium composition, and this micropropagation techniques could be a useful system for healthy and vigorous plant production.

• Journal of Plant Biotechnology
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• v.33 no.2
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• pp.117-122
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• 2006

Comparative studies on medium supply in bioreactors (raft, immersion and ebb and flood) have revealed that multiplication and growth of Alocasia Amazonica were greatest in the raft system, while lowest in ebb and flood system. In the raft system, the basal part of the shoots was continuously in contact with medium, which enabled a constant uptake of nutrients as well as aeration to the explants. The number and the size of leaf stomata were higher in the raft system compared with immersion and ebb&flood system. In the immersion system, plantlets were deformed and epidermal cells in leaves were irregular with a large intercellular space. The results suggested that the medium supply should be controlled properly to maintain normal and healthy plantlets during liquid cultures in bioreactors Which affects morphology and physiology Of the plantlets.

• Journal of Plant Biotechnology
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• v.47 no.1
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• pp.53-65
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• 2020

Here, we describe an efficient and rapid protocol for the micropropagation of Thymus pallidus Cosson ex Batt., a very rare medicinal and aromatic plant in Morocco. After seed germination, we tested the effect of different macronutrients, cytokinins alone or in combination with gibberellic acid (GA₃) or auxins, on T. pallidus plantlet growth. We found that Margara macronutrients (N₃₀K) had the best effect on the in vitro development of the plantlets. The addition of 0.93 μM/L 1,3-diphenylurea (DPU), 0.46 μM/L adenine (Ad), and 0.46 and 0.93 μM/L kinetin (Kin) resulted in the best shoot multiplication and elongation. In addition, the combination of 0.46 μM/L Kin, DPU, or Ad with gibberellic acid, in particular, 0.46 μM/L Ad + 0.58 μM/L GA₃ and 0.46 μM/L Kin + 1.15 μM/L GA₃, led to better bud and shoot multiplication. Moreover, the integration of the combinations of 0.46 μM/L Kin and auxins, namely 0.46 μM/L Kin + 2.85 μM/L indole-3-acetic acid (IAA), 0.46 μM/L Kin + 2.85 or 5.71 μM/L indole-3-butyric acid (IBA), and 0.46 μM/L Kin + 0.3 or 0.57 μM/L 1-naphthaleneacetic acid (NAA), in the culture medium led to better root development and optimized aerial growth. Finally, the in vitro plants from the medium containing N₃₀K + 0.46 μM/L Kin + 2.85 μM/L IAA were successfully acclimatized; these plants served as a source for repeating in vitro culture.

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

• KOREAN JOURNAL OF CROP SCIENCE
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• v.42 no.6
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• pp.678-686
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• 1997

This experiment was carried out to develop the mass propagation system for producing plug plantlets using stem cuttings of virus-tree microtubers in potato. Cocopeat, vermiculite, perlite and peatmoss were combined and used as plug nursery media to find out the best combination suitable for the growth of seedlings derived from microtubers. Seedling growth was favored in high temperature (above 2$0^{\circ}C$) and a long-day photoperiod(above 16 hours) condition, while stolons and microtubers formed in outdoor condition. Shoot and root multiplication was not affected by NAA 10mg /1 or IAA 10mg /1 treatment. At the early growth stage of plug plantlets, the number of leaves and roots and the length of root increased significantly when nodes from the upper (near to apex) part of shoots rather than from basal part were taken. But after transplanting, these differences among these characters were not observed. At ninety days after transplanting the plug plantlets in spring time, plant was around 70 to 80cm in height, and the number of stolons and tubers were ten and seven, respectively.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.10a
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• pp.15-15
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• 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.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2002.11a
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• pp.29-30
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• 2002

The goal of this research was to develop the effectiveness of in vitro culture method for A. formosanus and study the environment in vitro conditions affecting on growth. The first series of experiments were examined to investigate the response of three different basal media, MS (Murashige and Skoog, 1962), Knudson (KC; Knudson, 1946) and modified hyponex on growth and multiplication during in vitro culture. Multiple shoot proliferation was induced in shoot tip explants on Hyponex (H3) media supplemented with BA (1 mg1$\^$-1/) or TDZ (1-2 mg1$\^$-1/). Addition of activated charcoal (1%) to the TDZ containing medium promoted rapid shoot tip proliferation (11.1 shoots per explant) but the same medium had an opposite effect resulting in poor proliferation in the nodal explants. However, the regenerated shoots had slow growth rate and failed to elongate. This problem was overcome by transferring the shoot clumps to a hormone free H3 media supplemented with 2% sucrose and 0.5% activated charcoal. Using bioreactor culture for scaling up was also shown the best way for multiple shoot induction and growth of this plant. The second series of experiments was studied to investigate the effect of physical environment factors on growth of in vitro plantlets. The Anoectochilus formosanus plantlets were cultured under different air exchange rate (0.1, 0.9, 1.2h$\^$-1/), without sucrose or supplement 20g.1$\^$-1/ (photoautotrophic or photomixotrophic, respectively), and different photosynthesis photon flux (40, 80, 120 ,${\mu}$mol.m$^2$.s$\^$-1/- PPF). Under non-enrichment CO$_2$ treatment, slow growth was observed in photoautotrophical condition as compared with photomixotrophical condition on shoot height, fresh weigh and dry weight parameters; High air exchange (1.2.h-l) was found to be inadequate for plant growth in photomixotrophical condition. On the contrary, under CO$_2$, enrichment treatment, the plant growth parameters were sharply (visibly) improved on photoautotrophic treatments, especially on the treatment with air exchange rate of 0.9.h-1. The growth of plant in photoautotrophic condition was not inferior compared with photomixotrophic, and the best growth of plantlet was observed in treatment with low air exchange rate (0.9.h-1). Raising the PPF level from 80 to 120${\mu}$mol.m$\^$-2/.s$\^$-1/ decreased the plant height, particularly at 120${\mu}$mol.m$\^$-2/.s$\^$-1/ in photoautotrophic condition, fresh weight and dry weight declined noticeably. At the PPF of 120${\mu}$mol.m$\^$-2/,s$\^$-1/, chlorophyll contents lowed compared to those grown under low PPF but time courses of net photosynthesis rate was decreased noticeably. Light quality mainly affected morphological variables, changes of light quality also positively affected biomass production via changes in leaf area, stem elongation, chlorophyll content. Plant biomass was reduced when A. formosanus were grown under red LEDs in the absence of blue wavelengths compare to plants grown under supplemental blue light or under fluorescent light. Stem elongation was observed under red and blue light in the present experiment. Smaller leaf area has found under blue light than with other lighting treatments. Chlorophyll degradation was more pronounced in red and blue light compared with white light or red plus blue light which consequent affected the photosynthetic capacity of the plant. The third series of experiment were studied to investigate the effect of physical environment factors on growth of ex vitro plants including photosynthesis photon flux (PPF), light quality, growing substrates, electrical conductivity (EC) and humidity conditions. In the present experiments, response of plant on PPF and light quality was similar in vitro plants under photosynthesis photon flux 40${\mu}$mol.m,$\^$-2/.s$\^$-1/ and white light or blue plus red lights were the best growth. Substrates testing results were indicated cocopeat or peat moss were good substrates for A. formosanus growth under the greenhouse conditions. In case of A. formosanus plants, EC is generally maintained in the range 0.7 to 1.5 dS.m-1 was shown best results in growth of this plant. Keeping high humidity over 70% under low radiation enhanced growth rate and mass production.