• Journal of Forest and Environmental Science
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• v.25 no.2
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• pp.111-118
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• 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 Plant Biotechnology
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• v.45 no.2
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• pp.140-145
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• 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.

• Journal of Plant Biotechnology
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• v.6 no.4
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• pp.235-239
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• 2004

We studied the effect of genotype, explant, age of explant, medium (plant growth regulators and gelling agents), and standardized an efficient regeneration protocol for inbred lines of Chinese cabbage (Brassica rapa ssp. pekinensis). Of the different concentrations of NAA and BA tested, the combination of $5\;\cal{mg/L}\;BA\;and\;0.5\;\cal{mg/L}$ NAA gave the highest frequency of shoot regeneration. The cotyledonary explants had more shoot regeneration frequency ($\ge40\%$) than the hypocotyl ex-plants. Besides, the cotyledonary explants, excised from the 4-day old seedlings, showed higher shoot regene-ration ($56.7\%$). Of the three gelling agents and their concentrations used, 16 g/L agar was found to be the best for shoot regeneration. Shoot regeneration frequency in-creased significantly by supplementing the medium with $4\;\cal{mg/L}\;of\;AgNO_3$ MS medium devoid of NAA showed higher frequency of rooting in the regenerated shoots than the ones supplemented with NAA. Our improved regeneration protocol will be especially useful for the genetic transformation of Chinese cabbage inbred lines to develop transgenic hybrids.

• Journal of Plant Biotechnology
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• v.7 no.1
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• pp.57-65
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• 2005

The $F_1$ hybrid Red Coat is one of the most highly sought after cultivars of tomato in Australia and yields up to 7.5 $\cal{kg/plant}$. An experiment was conducted to de-termine the optimal strength and type of growth medium and sucrose concentration for shoot organogenesis of the Red Coat cultivar using cotyledonary explants. Two basal growth media, viz. MS and Gamborg' s $B_5$ at 0, 1/4, 1/2, full or double strength along with sucrose concentrations of 0, 0.5, 1.5, 3 or $5\%$, were evaluated using 25 replications. The main effects of treatment and their mutual interactions were evaluated for the proportion of explants that produced callus and/or shoots, number of shoots produced per explant, callus diameter and shoot height. The explants failed to produce shoots in the absence of mineral nutrient. Only a small proportion of the explants ($6\%$ with $B_5\;and\;3\%$ with MS) regenerated shoots in the absence of sucrose. Lower sucrose concentrations ($0.5-1.5\%$) along with full strength media were optimal for most of the traits studied. The $B_5$ medium outperformed MS medium for shoot organogenesis. For all the traits examined, significant differences in main effects (P < 0.05) and two-way interactions were detected, but no three-way interactions (medium type $\times$ medium concentration $\times$ sucrose concentration) were observed. Sucrose was found essential for the development of chlorophyll. Chlorophyll content increased with an increase in sucrose concentration up to $3\%$ and decreased at $5\%$ sucrose.

• Journal of Plant Biotechnology
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• v.5 no.4
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• pp.225-231
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• 2003

The hormonal requirement suiting micropropagation of Morus alba during any season throughout the year was studied. Sprouting frequency from axillary buds of M. alba was greatly influenced by the time of explant collection, the highest value was achieved when nodal explants were collected at the end of bud dormancy period (late in March) and cultured on Murashige and Skoog (MS) medium supplemented with low concentration (0.5 mg/L) of BAP, kinetin or IBA (85-68%). In addition, they showed higher axillary bud sprouting on growth-regulators-free medium (49%) than others collected in autumn or winter and cultured on medium supplemented with various growth regulators (47-48%). Regardless of that period, young explants with greenish buds collected in summer exhibiting high sprouting frequency (66%) on MS medium supplemented with 0.5 mg/L kinetin and 0.5 mg/L GA3. Shoot multiplication via adventitious bud formation was achieved when the nodal explants were cultured on MS medium supplemented with 2 mg/L BAP and 0.2 mg/L IBA. Further multiplication via nodal explants of in vitro grown shoots was obtained on MS medium supplemented with 0.5 mglL BAP and 0.5 mg/L GA3. While half strength MS medium supplemented with low concentration (0.5 mg/L) of IBA, IAA or 2,4-D stimulated adventitious root formation, IBA was the best. After transfer the plantlets to the soil, acclimatization for three weeks was essential prerequisite for survival in high frequency (92%). Peroxidase activity is related to break of bud dormancy where maximum enzyme activity was detected when the lateral buds were induced to commence growth under field condition (early in spring) or in vitro.

• Korean Journal of Plant Tissue Culture
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• v.28 no.3
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• pp.171-178
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• 2001

This study was investigated to establish a regeneration system of plant via adventitious bud formation from leaf explant cultures of strawberry (Fragaria ananassa Duch). Effects of plant growth regulators (2,4-D, BAP), agar sucrose and myo-inositol on adventitious bud formation were investigated. When the leaf explants were cultured on MS medium supplemented with 0.1 mg/L 2,4-D and 3 mg/L BAP, the adventitious bud formation was most promoted. The adventitious bud formation was not induced from leaf explants cultured on MS medium containing 2,4-D alone. Adventitious bud formation was enhanced to almost 3 times on medium with low level of agar concentration (0.4%) in comparison with those on the medium with high level of agar (1%), but almost of shoot was vitrificated on the medium. Therefore, the normal adventitious bud formation from leaf explants was most effective on the medium containing 0.05 mg/L 2,4-D,1 mg/L BAP, 0.8% agar, 30 g/L sucrose and 100 mg/L myo-inositol. Thus, the mass propagation of healthy strawberry could be established using leaf explants.

• Journal of Plant Biotechnology
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• v.3 no.2
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• pp.95-100
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• 2001

To select the section with shoot formation ability, the calli and shoot formation from three sections (first leaf including cotyledonary node, hypocotyl and cotyledon explants) of 5-days-seedlings of soybean were induced on MS medium supplemented with 1.0 mg/L BAP, 3% sucrose, and 0.3% gelrite for one month. The first leaf section exhibited the highest shoot formation rate (51%), followed the hypocotyl section (10%) and the cotyledon section (0%). The shoot formation rates and shoot number of the four excised sections (whole first leaf, a half of the first leaf, a third of the first leaf and only node) of the first leaf were also investigated on the same medium. A half of the first leaf explant and the third of the first leaf explant had higher shoot formation rates (76-80%) and numbers (3-4 / explants) than those in other two explants. Effects of six cytokinins (kinetin, zeatin, BAP, 2iP, PBA, and TDZ) on shoot formation were determined, using the half of the first leaf explants. Zeatin (1.0 mg/L) exhibited the highest in shoot formation rate (94%) and numbers (8 / explant). In addition, the combined effects of three cytokinins (zeatin, BAP, and TDZ; 0.5, 1.0, 2.0 mg/L, respectively) and an auxin (IAA; 0.0, 0.5, 1.0, 2.0 mg/L) were determined. The combination (1:1, v/v) of zeatin (1.0 mg/L) and IAA (1.0 mg/L) exhibited the highest in shoot formation rate (96%) and numbers (16 / explant), twice more than zeatin (1.0 mg/L) alone. The shoot cuttings were transferred and cultivated on the rooting media supplemented with only auxin, IBA at various concentrations. The highest root formation (8 / shoot) was achieved on the medium supplemented with 1.5 mg/L. After 4 weeks of cultivation, the plantlets with an extensive root system were transplanted in pots with a soil mixture of vermiculite and fine sand. Transferred to field, about 75% of the plantlets survived.

• Journal of Plant Biotechnology
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• v.30 no.1
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• pp.41-45
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• 2003

A study was under taken to investigate the appropriate explant sources of flower organ and suitable cultural conditions such as light, temperature, and sucrose in plant regeneration of Iris ensata culture. Explants of perianth, ovary, pedicel, and peduncle of Iris ensata were cultured at different daylength (0, 8, 16, 24 hour), different temperatures (10, 15, 25, 3$0^{\circ}C$), and sucrose concentrations (1, 3, 6, 9%) on MS medium. Formation of adventitious roots from explants of Iris ensata was effective in the dark, while that of adventitous shoots was effective in the light. The optimum daylength for young plant regeneration was 16 hours. The optimum temperature for shoot formation of Iris ensata explants was $25^{\circ}C$ but the formation at 10 and 15$^{\circ}C$ was ineffective. Especi-ally, perianth and ovary was effective in shoot formation from flower organ expants. T-he optimum concentration of sucrose for shoots and roots formation of Iris ensata explants was 3 and 6%, respectively.

• Journal of Life Science
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• v.9 no.1
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• pp.29-34
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• 1999

Cotyledon and hypocotyl explants of perilla were cultured on MS medium containing a combined concentration of BA(0.5, 1.0 and 1.5mg/$\ell$) and NAA(0.1, 0.5 and 2.0mg/$\ell$) in order to regenerate the explant and induce the callus. The best regeneration of the explant and induction of the callus were observed in a combined concenteration of 0.5mg/$\ell$ of BA and 0.5mg/$\ell$ NAA both in cotyledon and hypocotyl explants. In cotyledon explants, rooting was achieved upon transferring shoots to MS medium containing 0.5mg/$\ell$ of BA and 0.1mg/$\ell$ of NAA. We also investigated the change of protein and RNA content on developmental stage of callus and plant regeneration of perilla. Protein content was increased but RNA content was decreased as the culture period increases. The banding pattern of polypeptide revealed that both 30KD and 45KD polypeptides were obvious in cotyledon obtained from pre-culture explants, but only 30KD polypeptide was further getting obvious as the culture period increases.

• BMB Reports
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• v.47 no.12
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• pp.673-678
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• 2014

During Xenopus early development, FGF signaling is involved in mesoderm formation and neurogenesis by modulating various signaling cascades. FGF-MAPK signaling induces Xbra expression, which maintains mesodermal fate through an autocatalytic-loop. Interestingly, previous reports have demonstrated that basic FGF (bFGF) treatment alone does not induce neurogenesis in ectodermal explants, even though FGF signaling inhibits BMP signaling via phosphorylation in Smad1 linker region. In addition, the overexpression of dominantnegative Xbra induces neurogenesis in ectodermal explants. However, the detailed mechanism underlying these phenomena has not yet been clarified. In this work, we showed that bFGF-Xbra signaling increased the PV.1 expression. DN-Xbra was found to decrease PV.1 expression, and the co-injection of PV.1 with DN-Xbra reduced neurogenesis in ectodermal explants. Furthermore, the knockdown of PV.1 induced neurogenesis in bFGF-treated ectodermal explants. Taken together, our results demonstrate that FGF-Xbra signaling induces PV.1 expression and that PV.1 functions as a neural repressor in the FGF-treated ectoderm.