• 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.

• The Korean Journal of Pesticide Science
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• v.18 no.3
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• pp.202-209
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• 2014

RNA interference (RNAi) is the method which controls phenotypes of gene in live cells. Chitinase is the enzyme helping digestion and absorption of old cuticles during the ecdysis of insects. In order to investigate molting-inhibition effect with the chitinase related gene in Spodoptera litura, RNA was extracted from the $5^{th}$ instars. cDNA was synthesized and then we obtained about 700 bp size chitinase. After PCR products were cloned into a pGEM T-easy vector, colonies were picked. DNA was extracted from the colony cultures. EcoR I enzyme was used to check whether PCR products were inserted or not. And then we confirmed vector band of about 3 kb and insert band of about 700 bp. To synthesize the dsRNA, each DNA was cut with Spe I and Nco I enzymes (Circular DNA became lineared DNA). After synthesis of dsRNA, approximately 5 ul dsRNA was injected into the $3^{rd}$ abdominal segment of S. litura $4^{th}$ larvae. The concentration of dsRNA was about $10{\mu}g/{\mu}l$. We confirmed larval-larval molting : there were phenotypically abnormal individuals - for instance malformation, molting inhibition and change of integument color. Pupaadult molting : there were phenotypically abnormal individuals - for instance molting inhibition, change of wings and malformation. Also we could investigate the pupation, emergence and variation about noninjection, treated with DW and dsRNA. Each pupation was non-injection 83.3%, DW 78.3% and dsRNA 66.7%. Each emergence was non-injection 90.0%, DW 72.3% and dsRNA 65.0%. So we considered that chitinase dsRNA induced molting inhibition effect. But each variation was non-injection 8.9%, DW 2.9% and dsRNA 19.2%. Therefore dsRNA group showed the highest variation value. When 18 hours after injecting dsRNA, we could obtain abnormal individual.

• Korean Journal of Weed Science
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• v.16 no.3
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• pp.221-229
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• 1996

This study was carried out to determine the effects of growth regulators, carbon sources and silver nitrate on callus formation and plant regeneration of turfgrass. The results were summarized as fallows : Callus from Korean lawngrass (Zoysia japonica Steud.) and pencross creeping bentgrass (Agrostis palustrir Huds.) induced better in MS medium than in N6 medium and by addition of 2,4-D than by that of NAA. Callus formation from Korean lawngrass and penncross creeping bentgrass was very effective at MS medium adding 1mg/L 2,4-D and at the medium adding 2mg/L 2,4-D, repectively. Growth of callus was good at MS medium containing 2mg/L 2,4-D+0.2mg/L NAA. Callus growth of Korean lawngrass and penncross creeping bentgrass was good when kinetin was added 0.2mg/L and 0.3mg/L, individually, to MS medium containg 2mg/L 2,4-D+0.2mg/L NAA. Regeneration rate from leaf and stock callus of Korean lawngrass was 44% at MS medium adding 2,4-D 2mg/L+NAA 0.2mg/L+kinetin 0.3mg/L and 32% at the medium containing 2,4-D 2mg/L+NAA 0.2mg/L+kinetin 0.3mg/L, each and that from leaf and stock callus of penncross creeping bentgrass was 80% and 67%, each, at the medium adding 2,4-D 2mg/l+NAA 0.2mg/L+kinetin 0.3mg/L. Regeneration rate of Korean lawngrass and penneross creeping bentgrass increased by 3 to 4% and by 10 to 16%, respectively, when added $AgNO_3$ 1~2mg/L to the above-mentioned regeneration medium.

• Journal of Plant Biotechnology
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• v.50
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• pp.248-254
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• 2023

This study was conducted to establish an efficient plant regeneration system in 'Dayu', a Korean variety of Perilla frutescens developed for seed oil production, in conjunction with the previously studied variety 'Namcheon'. The healthiest callus was formed on the hypocotyl explants cultured on a medium containing 0.1 mg/L NAA and 0.5 mg/L BA, outperforming the leaf and cotyledon samples. In both dark and long-day conditions, Dayu consistently exhibited significantly higher shoot regeneration rates compared with Namcheon. The highest shoot regeneration rates in Dayu were observed from the hypocotyl explants cultured on 0.1 mg/L NAA and 0.5 mg/L BA media, with shoot regeneration rates of 84.4% and 86.7% under dark and long-day conditions, respectively. Various combinations of plant growth regulators were tested to establish the optimal shoot regeneration conditions for Dayu hypocotyl explants. The results demonstrated that the highest shoot regeneration rate (90%) was achieved when 0.5 mg/L of BA was added to the medium without NAA. Among the regenerated shoots, 70.5% were normal plants, while 19.3% were abnormal. The addition of NAA or an increase in its concentration led to a higher occurrence of abnormal plants. After the regenerated shoots were transferred to 1/2 MS medium, roots were observed within 10-15 days. By day 30, they had developed into complete plants. The results obtained from the regeneration experiments with the perilla variety Dayu can valuably inform molecular breeding reliant on transformation techniques such as genome-editing and genetic modification technology.

• Reproductive and Developmental Biology
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• v.28 no.3
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• pp.181-185
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• 2004

This study was conducted to examine whether activation treatments, source of oocytes and culture conditions affect in vitro developmental ability of caprine oocytes. Mature Korean native goats were pretreated with intravaginal CIDR for 10 days. The goats were then treated with a single intramuscular injection of 1,000 IU PMSG on Day 8 or twice daily injection of a total of 70 mg FSH for 3 days from Day 8 of CIDR insertion for superovulation. All the goats were injected with 10 mg PGF/sub 2a/ on Day 8 and 400 IU hCG on Day 10 of CIDR. Oocytes were surgically collected by oviduct flushing(in vivo maturation) or direct follicle aspiration(in vitro maturation) through mid-ventral incision at 35 h after hCG injection. Fifteen to twenty oocytes were placed in TCM-199 medium containing 25 mM Hepes and hormones under mineral oil at 39℃ in a humudified atmosphere of 5% CO₂ in air for 22 to 24 h. After maturation, the oocytes were activated by electric stimulation or ionomycin + 6-DMAP. The activated oocytes were then cultured in M16, TCM-199 and mSOF media supplemented with proteins at 39℃ for 6 to 7 days. Activation treatments did not affect cleavage of the oocytes. The cleavage rates were 64.1% (41/64) in oocytes activated by electric stimulation and 76.5% (218/285) in oocytes activated by ionomycin + 6-DMAP. The proportion of development to blastocyst was 15.6% (34/218) in oocytes activated by ionomycin + 6-DMAP, but activation by electric stimulation did not support embryos developed beyond morula stage. There were no differences in the cleavage rates of activated oocytes experiencing in vivo (86.8%, 66/76) and in vitro maturation (69.0%, 127/184). However, the development rate to blastocyst stage was significantly (P<0.05) higher for oocytes matured in vivo (50.0%, 33/66) compared to in vitro (0.8%, 1/127). Culture conditions did not affect the cleavage of -activated oocytes. The cleavage rates were 51.6% (49/95) in M16, 64.3% (18/28) in TCM-199 and 81.0% (145/179) in mSOF, respectively. By contrast, the development rate of activated oocytes to stage was greater (P<0.05) for oocytes cultured in mSOF medium (23.4%, 34/145) than in M16 or TCM-199 (0.0%). Our results suggest that source of oocytes and culture conditions are major factors affecting in vitro development of caprine parthenogenetic oocytes.

• Journal of Plant Biotechnology
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• v.32 no.1
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• pp.37-44
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• 2005

Microspores were isolated from pepper (Capsicum annuum L.) anthers by using a micro-blender and cultured in modified NLN medium at $25^{\circ}C$. The influence of pretreatment period at $32^{\circ}C$, adding the 2-hydroxynicotinic acid to a pretreatment medium, and co-pretreatment anthers with microscopes on the induction of embryo were examined. Globular and torpedo embryos were observed from 3 weeks after culture. Embryo development was not synchronized within culture. After 4 weeks in culture, in addition to globular and torpedo embryos, cotyledonary embryos were observed. Normal cotylodonary embryos developed into plantlets when transferred to a solid hormone free B5 medium containing $2\%$ sucrose. Embryo yields were significantly higher after 1- and 2-day pretreatment at $32^{\circ}C$. However the development of embryo ceased at the globular or heart stage. In contrast, embryo yields were lower after 3- to 6-day pretreatment at $32^{\circ}C$ and embryo developed at the cotyledonary stage. After adding the 2-hydroxynicotinic acid to anther pretreatment solution, embryo yields were slightly increased. However most embryos occurred were at the globular or heart stage. Co-pretreatment of microspores with anthers was deleterious for embryo induction and development. AS far as we know, this is the first report of success in obtaining high frequency of embryogenesis and plantlets formation from isolated microspores of pepper. Although the culture conditions have to be optimized further, this promising microspore culture system can be used for genetic transformation, selection for dominant and recessive traits as well as for the production of homozygous doubled haploid plants.

• Nuclear Medicine and Molecular Imaging
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• v.43 no.5
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• pp.487-494
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• 2009

Purpose: Small size of recombinant scFv antibody has many advantages such as rapid blood clearances and improved targeting antibodies to tumor region. On the other hand owing to small size, number of amino group is insufficient in conjugation with chelator and fluorescence labeling. This study is to introduce poly lysine tag to the C-terminal end of scFv lym-1 sequence for fluorescence chelator conjugation. Materials and Methods: Poly lysine scFv lym-1 gene, cloned into pET-22b (+) vector, was expressed in E. coli BL21 (DE3) strain. Antibody purification was performed with Ni-NTA column and then size exclusion column chromatography. Expression and purification levels of poly lysine tagged scFv lym-1 antibody were confirmed by western blot analysis. I-124, I-125, I-131 and Tc-99m were used for radiolabeling of purified poly lysine scFv lym-1. Flow cytometry analysis of FIT( conjugated poly lysine scFv lym-1 was performed for confirmation of immunoreactivity of human Burkitt's lymphoma cells. Results: Poly lysine scFv lym-1 antibody was purified through two steps and identified as molecular weight of 48 KDa. Radiolabeling yields of I-124, I-125, I-131 and Tc-99m into poly lysine scFv lym-1 were >99%, >99%, >95% and >99%, respectively. Flow cytometry analysis of poly lysine scFv and scFv lym-1 was showed similar immunoreactivity to human Burkitt's lymphoma cells. Conclusion: Poly lysine tag was useful for the sufficient number of amino groups to scFv lym-1 antibody for chelator conjugation with minimizing loss of immunoreactivity.

• Korean Journal of Poultry Science
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• v.8 no.1
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• pp.1-5
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• 1981

In trying to predict the effect of genetics on the broiler in the year 2000, this is a relatively short period of time as far as broiler genetics in concerned. Modern broiler genetics started around 1945 and tremendous gains when made in past 35 years. Futher improvements on broiler will depend on the evolution and revolution: 1. Evolution: (1) Growth rate has been made 4-5% per year. (2) Feed conversion has improved approximately 1% per year. (3) Abdominal fat is becoming a major complaint in broiler. (4) Because of the changing life-style, broiler meat sales in the future will be more and more in cut-up form. (5) Breeding for stress resistance and selection for docile temperament can be important in order to funker improve fled efficiency. (6) In female parent stock, reproduction characteristics are in many can negatively correlated with the desired broiler traits. (7) Egg production and hatchability in moot commercial parent nod m at a fairly high level. (8) In male parent stock, the heavier and mon super-meat-type male lines are desired to Product better broilers. 2. Revolution: Trying to forecast revolutionary change in broiler genetics is highly speculative, as sudden change are aften unpredictable. (1) Species hybridization, such as a turkey-chicken cross (2) Biochemical tools, such as blood typing. (3) Mutation breeding by radiation or chemical mutagentia. (4) Broiler breeding would be to change the phenotypic appearance by single gene, such as naked, wingless. (5) Changes in production techniques. such as growing in cage or growing in filtered air positive pressure houses.

• Journal of The Korean Society of Grassland and Forage Science
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• v.29 no.1
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• pp.1-6
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• 2009

In order to optimize tissue culture conditions for genetic transformation of zoysiagrass (Zoysia japonica Stued.), the effect of plant growth regulators and culture medium supplements on embryogenic callus induction from mature seeds of a cultivar 'Zenith' were investigated. The optimal concentration and treatment period of NaOCl is 30% (v/v) for 60 minutes. Cultivation of mature seed on the callus Induction medium containing 3 mg/L 2,4-D and 3 mg/L dicamba showed 17.5% of embryogenic callus formation frequency. Supplementation of 1 g/L casein hydrolysate and 500 mg/L L-proline improved frequency of embryogenic callus induction. Audition of the medium with 5 mg/L $AgNO_3$ and 20 mg/L cysteine enhanced frequencies of embryogenic callus induction. Efficient callus induction system established in this study will be useful for molecular breeding of Boysiagrass through genetic transformation.