• Journal of the Korean Society of Tobacco Science
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• v.21 no.1
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• pp.70-76
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• 1999

KF 116 was TMV resistant tobacco plant and KB 301 was PVY resistant plant transformed with TMV CP gene and PVY CP gene, respectively. These resistant plants were cross-fertilized and the 4 lines of the TMV-PVY resistant plants were selected from F1 hybrid plants. The rate of PVY-resistant plant in these hybrids was 100 percent and that of TMV-resistant plants including delay type was 90-98 percent at 4 weeks after virus inoculation. It was confirmed that the TMV and PVY CP genes were integrated into the genome of hybrid plants by genomic PCR, and Southern blot hybridization. The genome of F1 hybrid plants had one copy and 4 copies of PVY-CP gene and TMV-CP gene, respectively, and CaMV 35S promoters were not methylated, regardless of the difference symptom development to TMV.

• Journal of the Korean Society of Tobacco Science
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• v.19 no.2
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• pp.83-91
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• 1997

The vein-necrosis strain or potato virus Y (PVY-Vff) and black shank (Phytophlhora parasitica roar. nicotianae) causes severe damage on burley tobacco(Wicotiana tabacum L.) in Korea, A new burley tobacco resistance to PVY and black shank, KB 110, was developed by Korea Ginseng and Tobacco Research Institute. It was developed from the cross of Burley 21 with TC 591 in 1990, and was backrossed to Burley 21 in the following season. TC 591 has resistance to PVY and moderate resistance to race 0 of black shank, but it is susceptible to tobacco mosaic vim (TMV). KB 110 was evaluated for its resistance to PVY, TMV and black shank in the greenhouse and at fields for preliminary and performance trials. KB 110 which has secreting glandular trichomes was resistant to PVY-VN, TW and black shank. It had an erect growth habit and two more leaves per plant than that of Burley 21, and matures two to three days later. It yielded approximately 3 percent more cured leaf than the standard cultivar Burley 21, but other plant characteristics were very similar to those of Burley 21. It had acceptable standards for chemical and physical characteristics of lured leaf on regional farm test in 1995-1997. KB 110 produced average yields of good quality tobaccos and was appeared to be resistant to PVY inwhere occurrence of the virus are severe chronic at burley growing area.

• Journal of the Korean Society of Tobacco Science
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• v.16 no.2
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• pp.134-138
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• 1994

Potato Virus Y(PVY), vein necrosis strain, in Korea causes severe symptoms on burley tobacco(Nicotiana tabacum L.). As the results, programs to incorporate PVY resistance into commercial cultivars were initiatEd. But the development of the homozygous fertile line resistant to PVY is time consumming. This study was conducted whether the Fl hybrid could be used to reduce the yield losses caused by PVY. Four F1 hybrids were made between male - sterile(ms) NC 107 and KB 107 as maternal parent, and TC 612 and TC 613 as Pollen donor, respectively, and were evaluated for their PVY resistance and negatively associated traits. (ms NC 107 X TC 612) F1, named as KB 109, Ivas applied to yield trial and compared with commercial cultivars for the level of disease resistance, agronomic characteristics, chemical contents and physical properties. All Fl hybrid could be used commercially as the PVY resistant cultivar. Especially KB 109 have the resistance against PVY, tobacco mosaic virus and black shank(Phytophthora parasitica var. nicotianae). It had wider leaves, flowered one day later, and yield of acceptable quality was higher than that of Burley 21, standard cultivar in Korea.

• Journal of the Korean Society of Tobacco Science
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• v.20 no.1
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• pp.57-65
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• 1998

This study was conducted to develop a resistant tobarro against Potato virus Y (PVY) by transformation of the plants with genetically engineered viral genes. The complimentary DNAs (cDNAS) of potato virus Y-necrosis strain (PVY-Vn) replicase mutant genes (3'-deleted, 5'-deleted and ADD-mutant Nlbs) were synthesized through RT-PCR by using purified PVY-VN RNA and synthesized primers, and cloned in the sense orientation into a plant expression vector (pMBPI), The cDNAS of the genes were transferred into Agrobacterium tumefaciens LBA 4404, and then transformed into tobacco (Nicotiana tabacum cv. Burley 21) plants. Regenerated plants were tested for PVY resistance by inoculation test; 13 transgenic plants including 7 for 3'-deleted Nlb, 3 for 5'-deleted Nlb, and 3 for ADD-mutant Nlb appeared to be resistant at 4 weeks after inoculation with PVY-VN. Among the 13 transgenic tobacco plants, 8 plants had no symptom up to 14 weeks after inoculation. The progenies ($T_1$) from self-fertilization of the transgenic lines varied 0.0% to 81.2% in their resistance (% of resistant plants). The analysis of Nlb-31deleted, -5'deleted and -ADD mutant in the $T_1$ plants by polymerase chain reaction (PCR) showed that Nlb-3'deleted, -5'deleted and -ADD mutants were detected in all of the resistant plants. These results suggest that the PVY resistance was inherited in the $T_1$ generation.

• Journal of the Korean Society of Tobacco Science
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• v.20 no.1
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• pp.50-56
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• 1998

The complementary DNA (cDNA) of potato virus Y- vein necrosis strain (PVY-VN) replicase gene (Nlb) was transformed into tobacco (Nicotiana tabacum cv. Burley 21) plants. Out of 25 putative transformants regenerated, 3 were resistant to PVY-VN, one highly resistant plant with no symptom until seed harvest time and the other two with mild chlorotic spot symptoms at late stages after infection. No symptom was observed in the highly resistant plant, while mild vein necrotic symptoms were developed on suckers of the moderately resistant plants after seed harvest time, In the first generation (T1) via self fertilization, resistance to susceptibility frequency in transgenic plants from the highly resistant transformant was about 3 : 1, while it was lowered much (about 1:2 and 1:19) in T1 of the moderately resistant transformants. In the second generation (T2) of the highly resistant plant, resistance frequencies were similar to T1, but resistance levels varied greatly and appeared to be decreased. Key words : potato virus Y, viral replicate gene, transgenic tobacco plants, resistance.

• Proceedings of the Korean Society of Tobacco Science Conference
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• 1997.10a
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• pp.134-152
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• 1997

Plant viruses of tobacco including tobacco mosaic virus (TMV) and potato virus Y (PVY) cause severe economic losses in leaf-tobacco production. Cultural practices do not provide sufficient control against the viruses. Use of valuable resistant cultivars is most recommendable for the control of the viruses. However, conventional breeding programs are not always proper for the development of virus-resistant plants mostly owing to the frequent lack of genetic sources and introduction of their unwanted properties. Therefore, we tried to develop virus-resistant tobacco plants by transforming commercial tobacco cultivars, NC 82 and Burley 21, with coat protein (CP) or replicase (Nlb) genes of TMV and PVY necrosis strain (PVY-VN) with or without untranslated region (UTR) and with or without mutation. Each cDNA was cloned and inserted in plant expression vectors with 1 or 2 CaMV 35S promotors, and introduced into tobacco leaf tissues by Agrobacterium tumefaciens LBA 4404. Plants were regenerated in kanamycin-containing MS media. Regenerated plants were tested for resistance to TMV and PVY In these studies, we could obtain a TMV-resistant transgenic line transformed with TMV CP and 6 genetic lines with PVY-VN cDNAs out of 8 CP and replicase genes. In this presentation, resistance rates, verification of gene introduction in resistant plants, stability of resistance through generations, characteristics of viral multiplication and translocation in resistant plants, and resistance responses relative to inoculum potential and to various PVY strains will be shown. Yield and quality of leaf tobacco of a promising resistant tobacco line will be presented.

• Journal of the Korean Society of Tobacco Science
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• v.20 no.2
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• pp.153-159
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• 1998

The vein necrotic strain of Potato Virus Y (PVY - VN) and black shank (Phyto-phthora parasitica var. nicotianae) are the two major diseases causing severe damages especially in burley tobacco (N. tabacum L.) area in Korea. A new tobacco variety, KB 111, resistant to PVY and black shank disease, was developed by Korea Ginseng & Tobacco Research Institute in 1997. It is a male sterile(MS) F$_1$ hybrid of the cross between MS TC 613 and KB 108. KB 111 was compared to Burley 21 on the agronomic characteristics and disease resistances in performance tests: It possessed upright growth habit and flowered two days later than Burley 21. It was resistant to both PVY and black shank and yielded about 3% more cured leaf than Burley 21, but other characteristics are very simiar to those of Burley 21. The chemical composition and physical properties of the cured leaf of KB 111 were as much acceptable as those of Burley 21 while it produced average yield of good quality leaf and appeared to resistant to PVY and black shank disease on regional farm test in 1998.

• Research in Plant Disease
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• v.12 no.1
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• pp.28-31
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• 2006

One of major potato viruses is Potato virus Y (PVY) in Korea. In the southern part of Korea, potatoes have been grown as double crops in a year by using cv. 'Dejima' and 'Chubak' due to very short dormancy. However, they have caused a serious problem such as a rapid degeneration. It has been thought that the degeneration is affected by the high incidence of PVY in neighboring potato fields. Therefore, the investigation of factors causing the degeneration is very important in the production of healthy seed potato. In this study, the PVY reinfection rates of several potato varieties and the different seed sources of cv. 'Chubak' have been investigated. Results show that the lowest infection rate of PVY among four potato cultivars derived from minitubers is cv. 'Superior'. The others are in order of 'Dejima', 'Atlantic' and 'Chubak'. Also, the incidences of PVY differ significantly when several seed sources are examined. When the seed potatoes (G2, the progeny of microtuber) as spring potato crops are planted in area without potato field nearby, the infection rate of PVY is as low as that of microtubers. However, PVY incidence in the progenies of minitubers as fall potato crops largely increases. Therefore, the best way of potato production under double cropping system is to use the healthy seed potato produced in area without potato field and plant relatively resistant cultivar such as Dejima.

• 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 the Korean Society of Tobacco Science
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• v.23 no.2
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• pp.123-132
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• 2001

In order to transform pokeweed antiviral protein cDNA to tobacco plant, total RNA was extracted from Phytolacca americana. PAP-II cDNA was synthesized from purified total RNA via RT-PCR and subcloned to recombinant vector pBluescript II SK-. 10 deletion mutant PAP-II cDNA fragments which were sequentially deleted from N-terminal by 90bp were synthesized from PAP-II cDNA except leading frame by PCR with primers designed in our laboratory. To select non-cytotoxic clone, pAc55M was constructed with yeast expression vector pAc55 and multicloning site(MCS). Sequentially deleted mutant PAP-II cDNAs were cloned on downstream of gall promoter of pAc55M. 6 non-cytotoxic deletion mutant PAP-II cDNA were selected. Selected cDNAs were cloned into plant expression vector pKGT101BH for transformation of these clones to plant through Agrobacterium tumefacience. After cloning, recombinant pKGT101BH carrying deleted mutant PAP-IIcDNA were transformed to Nicotiana tabacum cv. NC567. Transformed tobacco plants cultured on shooting and rooting media were transfered to green-house. About four weeks later, these plants were infected with physically infection using carborandum with PVY-VN strain. After 4 weeks, plants resistant to virus were selected , and seeds of these plants were gathered. Southern blot hybridization showed deleted fragments by 220bp and 420bp, so resistant ability of these plants is due to mutant PAP-II cDNA.