• Journal of Plant Biotechnology
• /
• 제2권3호
• /
• pp.135-142
• /
• 2000

The activity of promoter sequences was evaluated in garlic cells using the $\beta$-glucuronidase (GUS) gene as a reporter. Histochemical GUS assay indicated transient GUS activity in leaf, callus and root cells 48 hours after particle bombardment transformation. Quantitative fluorometric assays in extracts of transformed leaves demonstrated that the CsVMV promoter induced the highest level of gene expression, which was, on average, ten fold the level induced by CaMV35S and by the Arabidopsis Act2 promoters and two fold the level expression observed with a construct containing a double CaMV35S plus the untranslated leader sequence from AMV. No activity or very low levels were observed when cells were transformed with plasmids rontaining the typical monocot promoters, Actl, from rice or the Ubi-1, from maize. The green fluorescent protein (GFP) was also tested as a marker gene for garlic transformation. Intense fluorescence was observed in leaf, callus and root cells transformed with a construct containing the gfp gene under control of the CaMV35 Promoter. No fluorescence was detected when the gfp was under control of the Ubi-1 promoter.

• Biotechnology and Bioprocess Engineering:BBE
• /
• 제11권2호
• /
• pp.154-159
• /
• 2006

Interleukin-18 (IL-18), otherwise known as interferon-gamma-inducing factor (IGIF), is one of several well characterized and important cytokines that contribute to host defenses. The complementary DNA (cDNA) of mature human interleukin-18 gene (hIL-18) was fused with the signal peptide of the rice amylase 1A gene (Ramy1A) and introduced into the plant expression vector under the control of a duplicated CaMV 35S promoter. The recombinant plasmid was transformed into tobacco (Nicotiana tabacum L. cv Havana) using the Agrobacterium-mediated transformation method. The integration of the hlL-18 gene into the genome of transgenic tobacco plants was confirmed by polymerase chain reaction (PCR) amplification and its expression was observed in the suspension cells that were derived from the transgenic plant callus by using Northern blot analysis. The hlL-18 protein was detected in the extracts of the transgenic callus and in the medium of the transgenic tobacco suspension culture by using immunoblot analysis. Based upon enzyme-linked immunosorbant assay (ELISA) results, the expression level of the hlL-18 protein approximated $166{\mu}g/L$ in the suspension culture medium. Bioassay results from the induction of $interferon-{\gamma}$ from a KG-1 cell line indicated that the hlL-18 secreted into the suspension culture medium was bioactive.

• 한국잠사곤충학회지
• /
• 제28권2호
• /
• pp.9-14
• /
• 1986

고조삽목의 실용화를 위해, 삽상의 종류, 상토, 삽목시기, 발근촉진제의 효과, 삽목전처리, 삼수채취부위 등을 연구한 결과 다음과 같은 결과를 얻었다. 1. 삽목방법 중 활착률이 가장 높은 구는 사토에서 유합조직을 형성시킨 후 삽목하는 구였으며, 보온못자리구에서는 완전히 고사하여 가장 낮았다. 2. 삽목적기는 3월 26일로 이때에는 수상자, 사토상토를 제외하고 활착률 100%를 보였다. 3. 삽목채취 부위별 활착률은 상<중<하부 순으로 높았으나, 유합조직구에서는 중부에서 70-100%의 높은 활착률을 보였다. 4. 발근촉진제 Rootone-F 포도는 지조상·중부, 그리고 온상 20cm구에서 포도하지 않은 것보다 활착률을 높였으나, 다른 구에서는 뚜렷한 효과가 없었다. 5. 상토의 토성은 활착률에 일정한 경향을 보이지 않는다. 6. 삽목전 처리 중 3월 15일 채취, 빙고에 저장하였다가 삽목하거나, 유합조직의 형성후 삽목하는 경우 타처리에 비해 현저히 활착률이 높았으며, 채취후 방치하였다 삽목하는 전혀 활착되지 않았다.

• 한국자원식물학회:학술대회논문집
• /
• 한국자원식물학회 2020년도 춘계학술대회
• /
• pp.19-19
• /
• 2020

The Cassava(Manihot esculenta Crantz) is a tropical root crop, originally from Amazonia, that provides the staple food of an estimated 800 million people worldwide. 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. So we tried to optimize protocols for mass production of somatic embryo amenable to large-scale vegetative propagation of Cassava. After in vitro eight-week culture of leaves of Cassava, the medium which contained the 2,4-D, BAP and IBA showed the highest callus induction rate, embryogenesis callus formation rate and somatic embryo formation in Cassava culture. In the medium with GA₃ and myo-inositol, shoots were most vigorously regenerated from somatic embryos of Cassava. 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.

• 한국작물학회지
• /
• 제47권1호
• /
• pp.62-67
• /
• 2002

An efficient system of rice microspore culture could contribute to the production of genetically modified rice. The microspores were isolated by mechanical or shed methods. The number of microspores per 100 anthers isolated at uninucleate stage was higher than (or similar to) those at binucleate stage in isolation method with pestle or spatular, but microspore divisions were not easily observed on both stages. On the other hand, pollen division in shed pollen culture was observed more frequently at uninuclear than at binuclear stage. Cold pretreatment at 1$0^{\circ}C$ for 10 days resulted in the best multicellular division to produce microcalli at 12.5% efficiency in shed microspores. Heat shock at 33$^{\circ}C$ for one hour before or after pollen shedding enhanced cell division and callus formation. Out of twelve green regenerants, two were haploids and ten were diploids based on the chromosome analysis of root tips. The size of stoma was 12$^{m}$ m in haploids and 15 ${\mu}{\textrm}{m}$ in diploids determined by scanning electron microscope (SEM).

• KSBB Journal
• /
• 제8권1호
• /
• pp.62-68
• /
• 1993

Transformed rice plantlet were recovered from protoplasts by electroporation with the plasmld pB 1121, which contain the plant expressible NPT-II and GUS genes. Embryonic cell suspension culture was established with embryonic callus induced from mature seeds of rice (Oryza sativa L. cv. Dong-jin) on the MS medium supplemented with 2.0 mg/l 2,4-D, 0.5 mg/l kinetin, 3% sucrose. Protoplasts isolated from embryonic cell suspensions were electroplated and then poterltialty-transformed tissues were selected by growth on the medium containing 200 mg/l kanamycin sulfate. When subjected to GUS assay, they stained blue, indicating the expression of the inserted GUS genes. Plantlets were regenerated from electroplated protoplasts on the hormone free MS medium. Transferred foreign genes in the plants were confirmed by southern hybridization. These results support use of electroporation for transformation of these important cereal plants.

• Plant Breeding and Biotechnology
• /
• 제6권4호
• /
• pp.426-433
• /
• 2018

Plant tissue culture is a technique that has invariably been used for various purposes such as obtaining transgenic plants for crop improvement or functional analysis of genes. However, this process can be associated with a variety of genetic and epigenetic instabilities in regenerated plants, termed as somaclonal variation. In this study, we investigated mutation spectrum, chromosomal distributions of nucleotide substitution types of single-nucleotide polymorphisms (SNPs) and insertions/deletions (InDels) by whole genome re-sequencing between Dongjin and Nipponbare along with regenerated plants of Dongjin from different induction periods. Results indicated that molecular spectrum of mutations in regenerated rice against Dongjin genome ranged from $9.14{\times}10^{-5}$ to $1.37{\times}10^{-4}$ during one- to three-month callus inductions, while natural mutation rate between Dongjin and Nipponbare genomes was $6.97{\times}10^{-4}$. Non-random chromosome distribution of SNP and InDel was observed in both regenerants and Dongjin genomes, with the highest densities on chromosome 11. The transition to transversion ratio was 2.25 in common SNPs of regenerants against Dongjin genome with the highest C/T transition frequency, which was similar to that of Dongjin against Nipponbare genome.

• Journal of Plant Biotechnology
• /
• 제47권3호
• /
• pp.227-234
• /
• 2020

벼(Oryza sativa L.)에서 세포 현탁 배양을 통한 miraculin 단백질의 생산을 위해 miraculin 유전자(AB512278)가 도입된 Agrobacterium tumefacience EHA105를 매개로 벼 캘러스에 형질전환하였다. 현탁배양세포주는 형질전환 캘러스를 이용하여 몇번의 선발과정 및 계대배양을 통해 선발하였고, 게놈 PCR 분석을 통해 miraculin 유전자가 벼 염색체에 안정적으로 도입된 것을 확인하였다. 또한, RT-PCR 분석을 통해 형질전환 세포주에서 도입된 miraculin 유전자가 과발현 되었다. 재조합 miraculin은 형질전환 현탁배양 HK-2 세포주에서 가장 높게 발현되어 total soluble protein (TSP) 대비 2.0%를 보였다. 이러한 결과는 형질전환 현탁세포배양이 miraculin과 같은 미각 수식 단백질의 대량생산 시스템을 구축하는데 이용 가능 할 것으로 사료된다.

• 식물조직배양학회지
• /
• 제27권3호
• /
• pp.163-167
• /
• 2000

벼 약배양에서 식물체 재분화 능력의 품종간 차이와 유전 양식에 대해 연구한 결과는 다음과 같다. ‘일미벼’ 외 46품종의 녹색체 재분화율은 자포니카형이 통일형이나 인디카형보다 높게 나타났고, 동일 품종 생태형간에도 배양 효율은 다양한 변이폭을 나타내었다. 자포니카 품종중에서 ‘일미벼’와 ‘향남벼’는 각각 19.4%와 18.1%의 높은 재분화 능력을 나타내었다. ‘밀양 23호’와 ‘추청벼’가 교배된 F$_2$집단의 녹색체 재분화율은 평균 6.6%로 나타났고 0~33.3%까지 넓은 변이폭을 가지는 연속적인 빈도 분포양상을 나타내었다. 캘러스 형성률과 식물체 재분화 능력에 대해 추정된 광의의 유전력은 각각 72.8%와 82.7%로 추정되었다.

• 한국연초학회지
• /
• 제27권2호
• /
• pp.153-162
• /
• 2005

Tobacco (Nicotiana tabacum L.) cells were transformed with rice expansin genes, OsEXPA4, OsEXPB3, OsEXPB4, and OsEXPB6, to elucidate the function of the genes in tobacco cells. The transformation increased the mass of the callus by $36\%-65 \%$, and the cell length by $12\%-28\%$. The cell width was decreased by $3\%$ for OsEXPB3, not changed for OsEXPB4, increased by $25\%\;and\;20\%$ for OsEXPA4 and OsEXPB6, respectively. From database search, seven expansin genes were found and six of them belong to EXPA group and one of them belongs to EXPB group. EXLA and EXLB were not found. All tobacco expansin genes were evenly distributed in the phylogenetic tree of rice and Arabidopsis expansin genes.