• Korean Journal of Plant Resources
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• v.21 no.3
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• pp.184-191
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• 2008

This experiments were carried out to find out the effects of different explant materials, kinds and concentration of plant growth regulators, and total nitrogen and sucrose contents on the in vitro regeneration of Abeliophyllum distichum Nakai. The effects of growth regulators on regeneration from 3 explant sources (leaf, internode and node) were more or less same. Leaf explants produced only callus with 2ip (Isopentenyladenine) and NAA (Naphthaleneacetic acid) treatment and other regulators had no effects. Test with internode explants yielded about same results but callus was obtained with 2,4-D (2,4-Dichlorophenoxyacetic acid). Node explants resulted in shoot regeneration by all regulator treatment except NAA and 2,4-D, but control also showed similar results. Callus formation from internode and node explants was vigorous by 2ip, zeatin, and 2,4-D treatments and high NAA concentration resulted in higher callus formation. In this experiment, various mixed treatment of growth regulators were also employed, using node as explant material. Shoot regeneration was obtained with BA (Benzyl adenine) + NAA treatments but the results were comparable with control. Generally shoot and root regeneration was poor with all combined treatment except 2ip + NAA and 2,4-D + NAA. However, callus was formed readily with all treatments. In this experiment, combined treatments of regulators were applied on the callus derived from singular regulator treatment. The results showed no shoot and root regeneration with any combination of 2,4-D, IAA (Indoleacetic acid) and NAA, but soft milky white callus was formed in all the treatments. No shoot and root regeneration was observed with any combination of 2iP, NAA and IAA, but somewhat hard, light green callus was formed in all the treatments. Callus formation decreased with high kinetin concentration in case of kinetin + NAA treatment. The experiments with total nitrogen content of media showed that low concentrations of 15 and 30mM were effective for the shoot and root regeneration. Sucrose experiment demonstrated shoot regeneration with 1${\sim}$4% concentration, and root and callus formation with 2${\sim}$4%. No root and callus formation was observed with 0 and 1% sucrose.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2003.04a
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• pp.131-132
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• 2003

Leaf discs and apical meristems were cultured in Murashige and Skoog (MS) medium supplemented with cytokinin and auxin at different concentrations. Callus production was observed in all tested media after six days of incubation. Callus produced in the presence of high concentration of NAA (2.0mg/1) was fragile in texture and yellow in colour. Highest callus formation was observed from leaf discs in the medium supplemented with 1.0mg/1 NAA and 0.5 mg/l BAP in dark at $25{\pm}1{\circ}C$. Percentage of callus formation was 95% and mean callus fresh weight was 654.88 43.53 mg. Shoots were induced from the callus after 4 weeks in 1/2MS medium supplemented with BAP and kinetin both at 0.5mg/1. When elongated shoots were separated and transferred into multiplication medium (MS+0.5mg/1 BAP+0.5mg/1 kinetin) multiplication rate was 6.4 after 6 weeks. Higher concentrations of BAP caused callus production at the base. Direct shoot induction was observed from apical meristems in MS medium in the presence of 0.175 mg/1 IAA + 2.25mg/1 BAP and 0.175 mg/1 IAA + 3.0 mg/1 BAP in 16 hour day at $25{\pm}1{\circ}C$. Explants (apical meristems) elongated to form a single shoot forming a callus at the base. Adventitious buds were sprouted out from the base. Percentage explants which producing shoots was 28.57 and 65.5 respectively. Multiple shoot induction was also observed in the same media. Highest multiple shoot production was observed in the presence of 0.175 mg/l IAA and 3.0mg/l BAP, Mean number of shoots per explant was 5.36 and the mean shoot length was $16.66{\pm}4.15$mm. Shoots (20 30m length) were tested for root induction. Excised shoots were transferred into rooting media, which contains different concentrations of NAA and IAA. Best rooting performance was observed in 1/2MS medium supplemented with 0.1mg/1 NAA after 10 days of incubation in 16 hr photoperiod at $25{\pm}1{\circ}C$. Mean number of roots per shoot was 6 and the mean root length was 252mm. Rooted plantlets were transferred into sterile coir dust:sand (1:4) mixture and maintained in a humid chamber for two weeks, They were gradually exposed to the natural environment. After three weeks they were transferred to pots containing coir dust:sand (1:2) mixture for further development where the 90% survival was observed.

• Korean Journal of Plant Resources
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• v.36 no.5
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• pp.527-539
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• 2023

Pepper (Capsicum annuum L.) is an important vegetable and spice crop that has been cultivated worldwide. Pepper fruits have unique taste and aroma, providing a variety of antioxidants and compounds important for human health, which makes a high economic value. In addition, there is a high demand for new pepper varieties, according to consumer's preference. However, pepper is a recalcitrant plant for in vitro tissue and organ differentiation and plant regeneration, which makes it difficult to develop demanded varieties using newly developed technologies such as genetic engineering and gene editing. In this study, tissue culture and regeneration conditions were investigated using seven pepper varieties that were obtained from the core-collection of Seoul National University. We observed callus and bud induction and shoot formation using several media composition composed of different cytokinins and auxin concentrations. As a result, it was found that there were differences in callus induction and shoot formation of each variety depending on the hormone composition, and the highest regeneration was shown when the medium containing Zeatin Riboside and the petioles of seedlings were used. In particular, out of seven pepper varieties, CMV980 exhibited a higher regeneration efficiency (approximately 48%) than other varieties, followed by Yuwolcho. Therefore, this study provides CMV980 and Yuwolcho as good candidates that can be used for pepper transformation, which might contribute to the development of various varieties through gene editing technology in the future.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2021.04a
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• pp.24-24
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• 2021

The Cassava (Manihot esculenta Crantz) is one of the major food crops in the tropical or subtropical regions. Recently, clean planting materials of improved cassava cultivars are in high demand. Problems in the propagation of cassava are virus vulnerable and low rates of seed germination. Thus, the study was undertaken to develop an efficient in vitro mass propagation protocol of Manihot esculenta Crantz. So we tried to optimize protocols for mass production from axillary buds of Cassava. Young and actively growing stem segments were excised from adult plants of cassava. Samples were cut into a 3~4 cm nodal segments with axillary buds, and cultivated in the different medium supplemented with various plant growth regulators for 4 weeks. For shoot multiplication, axillary buds approximately 1 cm in length were taken from in vitro derived shoots and subcultured. After 4~6 weeks, the shoot generation rate showed 55.6%. The shoot number and its length was 1.0/explant and 2.3 cm in the most favorable medium composition. The auxin β-indolebutyric acid(IBA) 0~2.0 mg/L was proved to be effective on root development. Plantlets with fibrous roots easily generated tuberous roots in vitro. The tuberous roots were induced only when both kinetin and IBA were used in combination. after 8 weeks, the root generation rate showed 100%. The root number and its length was 17.2/explant and 2.2 cm in the most promising 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.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2021.04a
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• pp.37-37
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• 2021

Calystegia soldanella L. is a perennial herbaceous halophyte belonging to the convolvulaceae family, which mainly grows in coastal sand dunes in Korea. Shoots and rhizomes are edible, and roots called 'Hyoseon Chogeun' are known to have medicinal effects such as antipyretic, sterilization, and diuretic. In addition, physiological activities of antioxidant, anti-inflammatory, antiviral, antifungal and PTP-1B (protein tyrosine phosphate-1B) inhibition have been reported. In this study, in vitro induction cell lines of C. soldanella L. collected from the coastal sand dunes in Jeju island was redifferentiated into adventitious roots that can be used as medicinal resources. Also the biomass of mass-proliferated adventitious roots using a bioreactor were evaluated. Plants of C. soldanella L. were collected from the crevice of the seashore in the coastal area of Taeheung 2-ri, Namwon-eup, Seogwipo-si. Then, it was separated into leaves, stems, rhizomes, and roots, and surface sterilized with 70% ethyl alcohol and 2% NaOCl (sodium hypochlorite). After washing with sterilized water, each organ section was cultured in Hormone-free MS medium (Murashige & Skoog Medium). As a result, the induction response rates were evaluated at 85% and 55%, respectively, in terms of callus formation and shoot generation in the rhizome segment. In the case of the adventitious roots morphological characteristics induced by single-use treatment of auxin-based plant growth regulators IBA and NAA from redifferentiated shoots were compared. Most efficient adventitious root culture method as a rooting rate, number, length, and biomass proliferation in the bioreactor system was confirmed when treated by culturing in MS salts, Sucrose 30 g·L^-1, and IBA 1mg·L^-1 for 4 weeks. In this study, the medium composition and culture period were confirmed using a bioreactor system to mass-proliferate adventitious roots derived from C. soldanella L. in Jeju island. Also this adventitious root line developed a new medicinal material could increase value of the bio-industry ingredient through quantitative and qualitative screening of phyto-bioactive compounds.

• Horticultural Science & Technology
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• v.31 no.3
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• pp.359-365
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• 2013

Chrysanthemum is one of the most popular ornamental plants worldwide. Recently, lots of new and novel chrysanthemum varieties have been developed using mutagenesis. However, there was no study for comparison of tissue culture condition among the mutant varieties derived from one original variety, until now. This study was conducted to compare the efficient regeneration condition of the two chrysanthemum mutant varieties, 'ARTI-purple' and 'ARTI-queen'. Two different flower parts (disk and ray florets) at the unopened and early blooming stages were used for comparison of regeneration condition on MS medium supplemented with combinations of three growth regulators (BA, NAA, and IAA). The highest regeneration rate was identified on the NAA and BA combination when the disk florets at unopened blooming stage are used. The best optimum combinations of growth regulators were identified as NAA $1.0mg{\cdot}L^{-1}$ and BA $0.5mg{\cdot}L^{-1}$ at 'ARTI-purple', which displayed 47.9% regeneration. However, regeneration of 'ARTI-queen' was the highest as 25.6% at NAA $2.0mg{\cdot}L^{-1}$ and BA $1.0mg{\cdot}L^{-1}$. There results indicate that there is a difference for the optimum regeneration condition between the mutant varieties derived from one original variety. These results will be useful for construction of efficient regeneration system of diverse chrysanthemum mutants developed by mutation breeding.

• Journal of Bio-Environment Control
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• v.29 no.1
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• pp.80-88
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• 2020

This study was conducted to examine the effect by application method and concentration of the indole-3-butyric acid (IBA), which is auxin-based plant growth regulator, on the growth and runner plants production of strawberry in the greenhouse. The seedlings of strawberry were transplanted in the pot (150 ×135 × 90 mm) filled with coir medium on April 12, 2019. The IBA was applied with a foliar spray or drench as 50, 100, 150, and 200 mg·L^-1 (50 mL per plant), respectively. The treatment was started on April 29, 2019. The foliar spray and drench treatment of IBA were repeated at 2-week intervals for 9 weeks from the start date of treatment. At 9 weeks after treatment, the petiole length of mother plants was the shortest in the control. The number of runner plants showed a tendency to decreased in the foliar spray. The number of lateral buds showed a tendency to decreased in the IBA treatment, and the least in the foliar with 100 mg·L^-1. There was not significantly difference in the fresh and dry weights of the first and second runner plants. However, in the third runner plants, the fresh and dry weights were the greatest in the drench with 100 mg·L^-1. Therefore, when considering the growth of third runner plants and lateral bud suppression, the drench with the 100 mg·L^-1 could be better application method and concentration of IBA treatment for growth of the third runner plants and runner plants production of strawberry, and the results can be used as a basic research of plant growth regulator application to save the labor force and enhance the seedling quality in strawberry seedling stage.

• Journal of Plant Biotechnology
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• v.42 no.3
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• pp.204-214
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• 2015

SHI-RELATED SEQUENCE (SRS) genes are plant-specific transcription factors that contain a zinc-binding RING finger motif, which play a critical role in plant growth and development. Among Brassica rapa SRS genes, BrSRS7 and BrLRP1 genes, isolated from shoot apical regions are important regulators of plant growth and development. In order to explore the function of BrSRS genes in horticultural plant growth and development, two constructs containing BrSRS7 and BrLRP1 under the control of a cauliflower mosaic virus 35S promoter were introduced into petunia by Agrobacterium-mediated transformation. The resulting transgenic plants were dwarf and compact plants with reduced plant height and diameter. Additionally, these transgenic plants had upward-curled leaves of narrow width and short internodes. Interestingly, the flower shapes of petunia were different among transgenic plants harboring different kinds of SRS genes. These phenotypes were stably inherited through generations $T_2$ and $T_3$. Semi-quantitative RT-PCR analyses of transgenic plants revealed that BrSRS7 and BrLRP1 regulate expression of gibberellin (GA)- and auxinrelated genes, PtAGL15- and PtIAMT1-related, involved in shoot morphogenesis. These results indicate that the overexpression of BrSRS7 and BrLRP1 genes suppressed the growth and development of petunia by regulating expression of GA- and auxin-related genes. From these data, we deduce that BrSRS7 and BrLRP1 genes play an important role in the regulation of plant growth and development in petunia. These findings suggest that transformation with the BrSRS genes can be applied to other species as a tool for growth retardation and modification of plant forms.

• Korean Journal of Agricultural Science
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• v.9 no.2
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• pp.467-483
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• 1982

Leaflets of Allium sativum L. (garlic) and leaf scales Allium cepa L. (onion) were cultured on the Murashige and Skoog(MS) medium to study the effects of plant growth regulators, temperature and light on the formation of shoot and root. And also, the potentiality of shoot or root formation of tissues in accordance with the leaf age and the region was discussed. In the experiment with garlic, the formation of shoot and root was more effective on the medium containing $BA\;2mg/{\ell}$ and $NAA\;1mg/{\ell}$ and it occured on the basal region of outer leaflets. The shoot formation was more effective at $20^{\circ}C$ than $28^{\circ}C$ under 16-hour daylength, and it was more effective in darkness than light condition at $28^{\circ}C$. The results of shoot and root formation on the new cloves of garlic harvested in 1982 were similar to those the old cloves of garlic harvested in 1981. In the experiment with leaf seales of onion, when the tissues of dormant leaf scales of onions harvested in 1981 were explanted on the MS medium, a few shoots were formed on the medium containing $BA\;2mg/{\ell}$ and $NAA\;1mg/{\ell}$. In this case,there were no differences on the shoot formation among the age and region of leaflets. On the culture of leaf scales, the shoot was not formed on the leaf scales located at outer or middle region, but it was formed on the medium containing BA accompanied with NAA at inner scales. On the medium complemented with BA alone, the tissues were not differentiated, but on the medium complemented with NAA, the callus was formed from tissues and then the root was formed through the callus.

• Journal of Plant Biology
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• v.35 no.2
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• pp.107-116
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• 1992

The roles of RNA- and protein-synthesis and $H^{+}$ excretion in 1AA ($10\;\mu\textrm{M}$)-induced elongation were investigated using abraded hypocotyl segments of sunflower (Helianthus annuus L.). The response of elongation initiated about 13 min after IAA treatment. Removal of cuticle, acting as diffusion barrier for inhibitors, by mechanical abrasion of hypocotyl segments enhanced the effect of inhibitors markedly, but the degree of abrasion for the saturated effect of inhibition was different among inhibitors. The elongation induced by 1M was completely inhibited when cycloheximide ($10\;\mu\textrm{M}$) was applied to abraded hypocotyl segments as shortly as 4 min before the onset of the growth response (= 10 min after administration of IAA). Cordycepin ($200\;\mu\textrm{M}$) prevented completely 1AA-induced elongation when applied as shortly as 19 min before the onset of the growth response (=5 min before administration of 1AA). Vanadate (1 mM) inhibited both lAA-induced elongation and medium acidification via lAA-induced $H^{+}$ excretion to apoplast. Cycloheximide and cordycepin also prevented lAA-induced $H^{+}$ excretion strongly. However, inhibition by cycloheximide of lAA-induced elongation was not alleviated by acidifying the cell wall to pH 4.5. The results indicate that, a few minutes before the initiation of growih, protein synthesis is demanded for the initiation of 1AA-induced elongation and the $H^{+}$ excretion to cell wall, and that the H+ excretion, even though it may be necessary for elongation, does not seem to bring about acid growth simply through acidifying cell wall.l wall.