• Journal of Plant Biotechnology
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• v.37 no.4
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• pp.511-516
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• 2010

The process to acquire intron-GUS gene-expressed transformants from somatic embryos (including embryogenic calli) of Rosa hybrida cv. 'Sweet Yellow' using Agrobacterium-meditated transformation method was reported in this study. Somatic embryos including embryogenic calluses were infected with Agrobacterium tumefaciens AGL1 strain (O.D = 0.7~1.6) including intron-GUS gene for 30 min, and were co-cultured for 3 days. After co-cultivation, they were cultured on embryo germination medium (EGM) supplemented with $250\;mg{\cdot}L^{-1}$ cefotaxim at $4^{\circ}C$ for 7 days. Then, transient GUS gene expression was observed. Shoots were regenerated from the shoot primodia induced from the intron-GUS gene-transferred either somatic embryos or embryogenic calli cultured on EGM supplemented with both cefotaxim $250\;mg{\cdot}L^{-1}$ and ppt $2\;mg{\cdot}L^{-1}$. Before induction of rooting from shoots cultured on shoot growing medium supplemented with both cefotaxim $250\;mg{\cdot}L^{-1}$ and ppt $2\;mg{\cdot}L^{-1}$, the shoots were cultured on multi-shoot induction medium supplemented with both cefotaxim $250\;mg{\cdot}L^{-1}$ and ppt $2\;mg{\cdot}L^{-1}$ to induce multi-shoots. When expression of the gene from a part of the multi-shoots was identified by GUS transient assay, the putative transgenic multishoots were transferred to rooting medium supplemented with cefotaxim $250\;mg{\cdot}L^{-1}$. After the formation of healthy roots, transgenic plantlets were transferred to the greenhouse after acclimatization. The expression rate of the intron-GUS gene in the multi-shoots was 100%.

• Journal of Plant Biotechnology
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• v.40 no.2
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• pp.65-71
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• 2013

Lilium cernum Komarvo. is an important endangered plant belonging to the family Liliaceae. A method was developed for the rapid micropropagation of L. cernum through plant regeneration from bulb scales explant-derived calli. The bulb scales segments were cultured on Murashige and Skoog (MS) medium supplemented with 0, 0.5, 1.0, $3.0mg{\cdot}L^{-1}$ kinetin and 0, 0.1, 0.5, $1.0mg{\cdot}L^{-1}$ NAA or 2,4-D for callus induction. In media with $0.5{\sim}3.0mg{\cdot}L^{-1}$ kinetin and $0.1{\sim}1.0mg{\cdot}L^{-1}$ NAA and 2,4-D, 95~100% of explants produced callus. They were then transferred to MS medium supplemented with various concentrations of NAA (0, 0.01, 0.05 and $1.0mg{\cdot}L^{-1}$) in combination with BA (0, 1.0 and $2.0mg{\cdot}L^{-1}$) for bulbet formation. Bulbet induction (78%), weight (468 mg) and size (15.5 mm) were obtained the highest on MS medium containing $2.0mg{\cdot}L^{-1}$ BA and $1.0mg{\cdot}L^{-1}$ NAA. In vitro frequency of plant regeneration was not significantly treated in strength of MS and sucrose concentration. Chlorophyll contents in 1/2MS with $50g{\cdot}L^{-1}$ sucrose treatments were higher than those in control and another treatment. This in vitro propagation protocol will be useful for conservation and mass propagation of this endangered plant.

• Molecules and Cells
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• v.23 no.3
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• pp.323-330
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• 2007

The mature wheat embryo is arguably one of the best explants for genetic transformation because of its unlimited availability and lack of growth season restriction. However, an efficient regeneration system using mature wheat embryos (Triticum aestivum L.) is still not available. To identify genes related to the tissue culture response (TCR) of wheat, QTLs for callus induction from mature embryos and callus regeneration were mapped using an RIL population derived from the cross of 'Wangshuibai' with 'Nanda2419', which has a good TCR. By whole genome scanning we identified five, four and four chromosome regions conditioning, respectively, percent embryos forming a callus (PEFC), percent calli regenerating plantlets (PCRP), and number of plantlets per regenerating callus (NPRC). The major QTLs QPefc.nau-2A and QPcrp.nau-2A were mapped to the long arm of chromosome 2A, explaining up to 22.8% and 17.6% of the respective phenotypic variance. Moreover, two major QTLs for NPRC were detected on chromosomes 2D and 5D; these together explained 51.6% of the phenotypic variance. We found that chromosomes 2A, 2D, 5A, 5B and 5D were associated via different intervals with at least two of the three TCR indexes used. Based on this study and other reports, the TCRs of different explant types of wheat may be under the control of shared or tightly linked genes, while different genes or gene combinations may govern the stages from callus induction to plantlet regeneration. The importance of group 2 and 5 chromosomes in controlling the TCRs of Triticeae crops and the likely conservation of the corresponding genes in cereals are discussed.

• Journal of Korean Society of Forest Science
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• v.78 no.4
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• pp.401-411
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• 1989

The embryos of Pinus taeda, P. rigida, and P. taeda ${\times}$ rigida were cultured for adventitious shoot regeneration in vitro. Culture media were modified from Gresshoff and Doy (MGD), Murashige and Skoog (MMS), Lloyd and McCown (MLM), and Schenk and Hildebrandt (MSH). NAA was added to initiation media at a concentration of 0.1 or 0.01 mg/l. BAP was used at the concentrations of 0.1. 0.5, 1, 2, or 5mg/l. Each explant was induced for 3-4 weeks on solid medium. All explants were cultured up to 16 weeks. Illumination was about $1506{\pm}540lux$ at the level of the tissues in the growth room with a temperature of $25{\pm}2^{\circ}C$. A 16-hour photoperiod per 24 hours was used. Half-strength medium was used for all the subcultures. For shoot production by loblolly pine, MMS, MLM, or MSH is preferred with 5 mg/l BAP with either 0.1 or 0.01 mg/l NAA. For shoot production by pitch pine, MMS, MLM, or MSH is recommended with 2 or 5 mg/l BAP with 0.1 mg/l NAA. For shoot production by the hybrid pine, MMS or MLM is more effective with 1, 2 or 5 mg/l BAP with 0.1 mg/l NAA. There were no differences recognized among the species tried in the patterns of bud formation and shoot development. Different composition of media, in major and minor salts or possibly in vitamins, should be tested for the two developmental stages of adventitious shoots ; the induction of shoot buds and the elongation of them into shoots.

• FLOWER RESEARCH JOURNAL
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• v.16 no.1
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• pp.12-16
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• 2008

This study was carried out to produce haploid plants to verify a systematic breeding program and genetic analysis. Effect of developmental stage of pollen grains and pre-treatment temperature on production of haploid plants was investigated. Microscopic investigation of the explants (Lilium Asiatic hybrid 'reamland' revealed that the length of flower bud at 23.0-24.9, 25.0-26.9, and 27.0-28.9 mm long coincided with tetrad, uninucleate, and binucleate, respectively. When the efficiency of the anther culture from microgametogenetic stages was tested, late uninucleate to early binucleate stage, having the length of 23.0 to 28.0 mm long flower bud, was the best. The frequencies of the callus induction and plant regeneration from the stage mentioned above were 17.8 and 6.7%, respectively. When calli were cultured on the MS medium containing picloram and zeatin at $25^{\circ}C$, shoots were obtained. Roots of regenerated plantlets were confirmed as haploid through an microscopic observation.

• FLOWER RESEARCH JOURNAL
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• v.18 no.1
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• pp.9-14
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• 2010

This study was conducted to examine the effects of IAA and kinetin on induction of gemmae and gametophytes in Hypnum plumaeforme moss during tissue culture. Explants were obtained from sterilized gametophyte tips cultured on solid basal medium containing Knop's major salts and Nitsch and Nitsch's trace elements. After culture, inoculated gametophyte tip produced protonema firstly, changed to new gametophytes after four weeks. Aseptic gametophytes were chopped and inoculated on the same media containing 0.01, 0.1, 1 and $10{\mu}M$ of IAA and kinetin. As a result, secondary protonemata as well as protonemal gemmae were formed from gametophytes. But protonemal gemmae formation was varied according to the concentration of IAA and kinetin. Lower concentration of IAA and kinetin promoted gemma formation and bud induction. Especially, $0.01{\mu}M$ of kinetin showed the highest frequency of bud and gemma production. All the materials, obtained from $0.01{\mu}M$ kinetin medium, were subcultured to media supplemented with 0.01, 0.1, 1 and $10{\mu}M$ of IAA and kinetin again to induce gametophytes. After subculture, protonema and calli were developed from secondary protonema, and induced gametophytes finally. IAA regulated induction and growth of gametophytes, and kinetin influenced gemma formation and gametophyte induction also. All aspects of development of this moss species were governed by the external growth regulators.

• Journal of Plant Biotechnology
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• v.46 no.1
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• pp.32-36
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• 2019

Calla lilies (Zantedeschia spp.) are monocotyledonous ornamental plants which belongs to the Araceae family. After the release of elite calla cultivar, an efficient propagation system is needed for commercial use. Despite the use of conventional propagation methods such as splitting of tubers and rhizomes of calla, rapid and efficient propagation system should be developed. In order to achieve this goal, stem segments contained apical meristems derived from calla lily cultivar (cv. Gag-si) were cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of cytokinin and auxin. This was aimed at inducing embryogenic calluses, shoots and multiple shoots. As a result, about 25% of induction rates of yellow embryogenic calluses were observed with MS medium containing both $0.5mg{\cdot}L^{-1}\;NAA$ and $1.5mg{\cdot}L^{-1}\;BA$ as growth regulators. In the experiments involving the regeneration from embryogenic calluses through shoot formation, MS medium supplemented with $0.5mg{\cdot}L^{-1}\;IAA$ and $2.0mg{\cdot}L^{-1}\;BA$ showed the highest rates at approximately 85 ~ 90% with regard to the formation of shoots in calla. Moreover, multiple shoots needed for rapid propagation were generated when explants were cultured on MS medium supplemented with $0.5mg{\cdot}L^{-1}\;IAA$ and $2.0mg{\cdot}L^{-1}\;BA$ with 40% of formation rate. In this study, the combination of auxin and cytokinin showed positive effects on both the induction of embryogenic calluses, the formation of shoots as well as multiple shoots in calla. The regeneration system described here can contribute to the development of breeding programs of calla in the future.

• Journal of Plant Biotechnology
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• v.49 no.1
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• pp.82-89
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• 2022

In vitro techniques were developed for propagating Vaccinium oldhamii using shoots with apical buds. Explants having an apical bud were cultured on Murashige and Skoog (MS) medium supplemented with 1.0, 2.0, and 5.0 mg/L of each zeatin, thidiazuron, 6-benzylaminopurine (BA), and 6-(γ,γ-dimethylallylamino)purine (2-iP) in order to induce multiple shoots. Among the tested treatments, the 2.0 mg/L of 2-iP proved to be most suited for the multiplication and growth of shoots; the multiple shoot induction rate was 100.0%, the average number of shoots was 7.4 per explant, and the average shoot length was 51.7 mm. The in vitro elongated shoots were rooted on half-strength MS medium containing various concentrations of indole-3-butyric acid (IBA) and 1-naphthaleneacetic acid (NAA). However, overall callus overgrowth was observed in all treatments and resulted in necrosis and abnormal shoot growth in root formation. A low concentration (0.5 mg/L) of IBA was appropriate for normal root development and the in vitro rooting rate was 30%. Ex vitro treatments on root formation using various concentrations of IBA with Talc powder and two types of rooting substrates (Flexi-Plugs or Horticultural soil) were examined. The ex vitro rooting rate (80%) and length of roots (32.9 mm) were obtained when the cut ends of the shoots were treated with 1.0 mg/L IBA and cultivated in Horticultural soil for 2 months. These findings suggest that ex vitro rooting is the more effective method for improving root formation in Vaccinium oldhamii than in vitro rooting.

• Journal of Plant Biotechnology
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• v.49 no.3
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• pp.207-212
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• 2022

The optimal culture conditions for root organogenesis from the callus of peonies (Paeonia lactiflora Pall.) were investigated. Root explants with vascular bundles were cultured in Murashige and Skoog (MS) medium combined with 0.5-4.0 mg/L auxins (indole acetic acid [IAA], naphthalene acetic acid [NAA], indolebutyric acid [IBA], and 2,4-dichlorophenoxyacetic acid [2,4-D]) and 0.0-2.0 mg/L cytokinins (kinetin, zeatin, and benzylaminopurine [BAP]) to induce callus formation. The callus was then cultured in MS medium combined with three concentrations (0.1, 0.5, and 1.0 mg/L) of IAA, NAA, IBA, kinetin, zeatin, and BAP in the dark for 6 weeks. Based on the results, the effects of dark and light conditions on the callus cultured in MS medium with combinations of 0.1-1.0 mg/L IBA and zeatin for 6 weeks were studied. Callus formation was most effective (>+++) in the medium with a combination of 1.0 mg/L NAA and 1.0 mg/L zeatin. A high number of long adventitious roots were observed in the mediums with 0.1 mg/L IBA (6.66 and 4.82 cm) and 0.5 mg/L zeatin (2.32 and 0.72 cm) among auxins and cytokinins, respectively. The highest number (14.06) of adventitious roots were formed from the callus cultured in light in the MS medium combined with 0.1 mg/L IBA and 0.5 mg/L zeatin. This same medium induced the formation of the longest adventitious root (5.45 cm) in the dark. Thus, optimization of in vitro culture conditions may be possible for the mass propagation of adventitious roots in peonies.

• Korean Journal of Plant Resources
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• v.35 no.4
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• pp.502-507
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• 2022

In the present study, plant regeneration through adventitious shoot formation from hypocotyl segments of in vitro seedlings of groundcherry (Physalis angulata L.) was investigated to determine the optimum culture conditions for highly efficient regeneration of the species. Adventitious shoots in hypocotyl segments were efficiently induced on MS media with low concentrations of BAP, specifically, with 0.5-1.0 mg/L BAP singly or in combination with 0.1-0.5 mg/L NAA. The 1.0 mg/L BAP single treatment was most effective for forming multiple adventitious shoots. When the induced shoots were transferred to the root induction media, low concentrations of NAA, IBA, and IAA enhanced the development of adventitious roots from adventitious shoots, suggesting that low concentrations of auxins were optimal for producing regenerated plantlets. The number of roots per shoot was large (> 2.0), and the root length exceeded 8.0 cm. In particular, the development and the overall shape of the roots were ideal. Furthermore, the number and length of shoots exceeded 2 and 6.0 cm, respectively. When the regenerated plantlets were transferred to compost soil, the root and shoot systems had developed well to the point that all of the regenerated plantlets acclimated successfully, resulting in normal morphology and growth characteristics, similar to those of the mother plant. Therefore, plant regeneration via adventitious shoot formation is expected to be one of the main methods for producing groundcherry on a large scale for a stable supply of the raw materials.