• Journal of Plant Biotechnology
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• v.3 no.2
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• pp.89-94
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• 2001

In vitro plant regeneration of inbred breeding line of hot pepper (Capsicum annuum L.) was established using leaf and petiole segments as explants. About 28 days old plants were excised and cultured on MS medium supplemented with TDZ and NAA or in combination with Zeatin. In all of the media compositions tested, combination of TDZ 0.5 mg/L, Zeatin 0.5 mg/L, and NAA 0.1 mg/L was found to be the best medium for shoot bud initiation. Young petiole was the most appropriate explant type for the plant regeneration as well as genetic transformation in hot pepper. In this study, HpMADS1 gene isolated from hot pepper was introduced using Agrobacterium-mediated transformation system. Based on the analysis of Southern blot and RT-PCR, HpMADS1 gene was integrated in the hot pepper genome. It has been known that floral organ development is controlled by a group of regulatory factors containing the MADS domain. Morphological characteristics in these transgenic plants, especially flowering habit, however, were not significantly altered, indicating this MADS gene, HpMADS1 may be non-functional in this case.

• The Plant Pathology Journal
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• v.32 no.1
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• pp.25-32
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• 2016

Potato is one of the most important crops worldwide. Its commercial cultivars are highly susceptible to many fungal and bacterial diseases. Among these, bacterial wilt caused by Ralstonia solanacearum causes significant yield loss. In the present study, integrated proteomics and genomics approaches were used in order to identify bacterial wilt resistant genes from Rs resistance potato cultivar CT-206-10. 2-DE and MALDI-TOF/TOF-MS analysis identified eight differentially abundant proteins including glycine-rich RNA binding protein (GRP), tomato stress induced-1 (TSI-1) protein, pathogenesis-related (STH-2) protein and pentatricopeptide repeat containing (PPR) protein in response to Rs infection. Further, semi-quantitative RT-PCR identified up-regulation in transcript levels of all these genes upon Rs infection. Taken together, our results showed the involvement of the identified proteins in the Rs stress tolerance in potato. In the future, it would be interesting to raise the transgenic plants to further validate their involvement in resistance against Rs in potato.

• Journal of Plant Biotechnology
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• v.37 no.4
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• pp.388-393
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• 2010

Recently, a number of successful research reports are accumulated to increase the carotenoid level in potato tuber such as $\beta$-carotene, precursor of vitamin A and keto-carotenoid like astaxanthin in which is not synthesized in most plants tissue since it does not contain a specific enzyme to add keto-ring in carotenoid molecule. In particular, keto-carotenoids are more interested due to their strong antioxidant activity. Currently, the content of $\beta$-carotene was increased up to 3,600-fold ($47\;{\mu}g/g$ dry weight) when compared to the control potato tuber, parental cultivar for genetic modification. In addition, astaxanthin, one of the major keto-carotenoid was accumulated up to $14\;{\mu}g/g$ dry weight in potato tuber with red color by over expressing the gene encoding $\beta$-carotene ketolase isolated from marine microorganisms. In this article, we summarized carotenogenesis-related genes that have been used for metabolic engineering of carotenoid biosynthetic pathway in potato. Furthermore, strategies for the accumulation of carotenoids and ketocarotenoids in specific potato tuber, bottle necks, and future works are discussed.

• Journal of Plant Biotechnology
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• v.29 no.2
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• pp.93-98
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• 2002

A VP6 fragments was subcloned with BamHI in the binary pMBP-1 vector under Califlower Mosaic Virus (CaMV) 355 promoter and neomycin phosphotransferase II (npt II) gene. The recombinant binary vector was mobilized into Agrobacterium-tumefaciens LBA4404 by the freeze-thaw method and potato (Solanum tubensum L. cv Desiree) was transformed by modified leaf-disc cocultivation. Shoots were induced on MS medium with 0.01 mg/L NAA, 0.1 mg/L GA$_3$, 2.0 mg/L Zeatin, 100.0 mg/L kanamycin, 500.0 mg/L carbenicillin. In order to identify the copy number of VP6 into potato plant, total genomic DNA was isolated from transgenic potato and analysed by Southern blotting. Genomic DNA and total mRNA analysis demonstrated the incorporation of the foreign gene into the potato genome, as well as their transcription.

• Journal of Plant Biology
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• v.37 no.1
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• pp.17-25
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• 1994

Two potato (Solanum tuberosum L.) cultivars were transformed by Agrobacterium tumefaciens harboring cauliflower mosaic virus (CaMV) 35S promoter and $\beta$-glucuronidase (GUS) gene. Expression patterns of the CaMV 35S promoter according to tissue types and developmental stages, and genetic stability of GUS gene were investigated in the clonal progenies of transgenic potatoes. Kanamycin-resistant shoot emerged from tuber disc after 4 weeks of culture, and root was induced 6 weeks after culture on the selection medium. Shooting frequency of cvs. Superior and Dejima were 43% and 27%, respectively. Mature transformants and their clonal progenies showed no phenotypical abnormality. GUS activity was expressed primarily at parenchymatous cells of phloem tissue around the vascular cambium in the stem and root, and higher activity was found at the apical meristem of shoot, root and adventious shoot bud. GUS activity was higher at tubers of young explants than at stored tubers. These facts indicate that expression level of the CaMV 35S promoter differed according to tissue types and developmental stages of the organs. The GUS gene was stably inherited to each clonal progeny and normally expressed.

• Journal of Plant Biotechnology
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• v.30 no.1
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• pp.13-18
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• 2003

This study was carried out to investigate the expression patterns of foreign gene that controlled by tuber-specific patatin promoter in transgenic potatoes. Potato leaf disc cultured in vitro were transformed by the Agrobacterium strain LBA4404 containing pBl121 or pATGUS from potato cv. Desiree. In order to select the transgenic lines, gene-specific primers deduced from the NPTII were synthesized and used for polymerase chain reaction. The down part of the putative transgenic potatoes was transplanted weekly onto sucrose-enriched medium to accelerate the microtuber formation. RNA gel blot analysis was performed on the total RNAs obtained from tuber that had been harvested at a week interval. Also, histochemical assay was observed in the explants transformed with either pBI121 or pATGUS. Results showed that the transgenic plant containing pATGUS expressed GUS transcripts mainly at the tuber, not in stem, with the highest expression level in 5 weeks-grown microtubers. In contrast to pATGUS plants, the transformed plants with pBI121 showed an equal expression pattern throughout the whole developing stages. Consistent with RNA gel blot analysis, histochemical GUS staining and enzyme activity exhibited pATGUS transcripts were at the highest level in 5 weeks cultures. From these results, we suggest that the best stage to analyze the foreign gene introduced by patatin promoter into potato plants is at 5 weeks cultures after tuber formation.

• Journal of the Korean Society of Tobacco Science
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• v.19 no.2
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• pp.117-123
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• 1997

Viral coat protein (CP) encoding cDNA with artificial start and stop codons was synthesized by reverse-transcriptase polymerase chain reaction (RT-PCR) from the Korean isolate of potato virus Y-vein nectrosis strain (pVY-VN). To make PVY CP cDNA to untranslatable form, three stop codons were inserted near the start codon by "megaprimer-PCR" method. The untranslatable CP cDNA was subcloned to plant expression vector and transferred to N. tabacum cv. NC82 by Agrobacterium-mediated transformation. Highly resistant plants to PVY infection were screened, based on symptom development after mechanical virus inoculation. By genomic PCR and Southern blot analysis, one or more copies of the untranslatable CP gene were found in all transformants. From northern blot analysis, highly resistant transgenic lines had very low level of CP transcript but susceptible lines had high level, suggesting resistance to PVY infection should be related to RNA-mediated mechanism.mechanism.

• Journal of Photoscience
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• v.7 no.2
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• pp.39-44
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• 2000

To examine the chilling tolerance lipids, we compared the chilling susceptibility of photosystem II of wild type tobacco plants with that of transgenic tobacco plants, in which the sensitivity to chilling had been enhanced by genetic modification of fatty acid unsaturation of chloroplast membrane lipids. The transgenic tobacco plants were found to contain reduced levels of unsaturated membrane fatty acids by being tansformed with cDNA for glycerol-3-phosphate acyltransferase from squash. For the purpose of studying on the functional integrity of photosystem II during low-temperature photoinhibition, the photochemical efficiency was measured as the ration of the maximun fluorescence of chlorophyll (Fv/Fm) of photosystem II. In parallel with an investigation on the transgenic plants, susceptibility of chilling-resistant species, such as spinah and pea, and of chilling-sensitive ones, such as squash and sweet potato, to low-temperature photoinhibition was also compared in terms of room temperature-induced chlorophyll fluorescence from photosystem II. When leaf disks from the two genotypes of tobacco plants were exposed to light at 5$^{\circ}C$, the transgenic plants showed more rapid decline in photochemical activity of photosysytme II than wild-type plants. When they were pretreated with lincomycin, an inhibitor of chloroplast-encoded protein synthesis, the extent of photoinhibition was even more accelerated. More impottantly, they showed a comparable extent of photoinhibition in the presence of lincomycin, making a clear contrast to the discrepancy observed in the discrepancy observed in the absence of lincomycin. Restoration of Fv/Fm during recovery from low-temperature photoinhibition occurred more slowly in the transgenic tobacco plants than the wild-type. These findings are discussed in relation to fatty acid unsaturation of membrane phosphatidylglycerol. It appears that the ability of plants to rapidly regenerate the active photosystem II complex from might explain, in part, why chilling-resistant plants can toleratlow-temperature photoinhibition.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2003.10a
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• pp.65-65
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• 2003

• Proceedings of the Zoological Society Korea Conference
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• 2001.10a
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• pp.226.1-226
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• 2001

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