• Title/Summary/Keyword: Expression vectors

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Transformation of Populus nigra × P. maximowiczii Using Agrobacterium tumefaciens vectors (Agrobacterium tumefaciens vector를 이용(利用)한 양황철의 형질전환(形質轉換))

  • Son, Suk Gyu;Hyu, Jung Oh
    • Journal of Korean Society of Forest Science
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    • v.87 no.2
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    • pp.164-172
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    • 1998
  • This study was conducted to find the optimum transformation condition using Agrobacterium harboring promoterless GUS gene. The optimal medium for shoot induction from leaves of Populus nigra${\times}$P. maximowiczii was MS medium supplemented with $0.1mg/{\ell}$ NAA, $0.5mg/{\ell}$ BAP(94% regeneration frequency and 11.5 average number of shoot) According to the test using pBI121, the concentration of antibiotics for selection marker gene was $100mg/{\ell}$ kanamycin or $60mg/{\ell}$ geneticin in the SIM(shoot inducing medium) 3. Two weeks later, callus was induced in the SIM 3 and this callus grew up to 0.5-1cm shoots after 6 weeks in the new SIM 3. And the treatment with methylation inhibitor(5-azacytidine) led to a dramatic increase in foreign gene expression rate from 5.7% to 26.7%. The vector systems showed. different transformation efficiencies based on the fluorometric and histochemical GUS assay. In this study the vector systems used for transformation seemed to affect transformation frequency, in which pEHA101 yielded more transformants(35.9%) than LBA4404/pBI121 did(5.7%). This result indicated that pEHA101 was effective to insert the promoterless foreign gene into a poplar genome.

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Molecular Breeding of Tobacco Plants Resistant to TMV and PVY (분자생물학적 TMV 및 PVY 저항성 연초 육종)

  • E.K. Pank;Kim, Y.H.;Kim, S.S.;Park, S.W.;Lee, C.H.;K.H.Paik
    • 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.

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Studies on the Occurrence, Host Range, Transmission, and Control of Rice Stripe Disease in Korea (한국에서의 벼 줄무늬잎마름병의 발생, 피해, 기주범위, 전염 및 방제에 관한 연구)

  • Chung Bong Jo
    • Korean journal of applied entomology
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    • v.13 no.4 s.21
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    • pp.181-204
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    • 1974
  • The study has been carried out to investigate the occurrence, damage, host range, transmission and control of rice stripe virus in Korea since 1965. 1 Disease occur「once and damage : The virus infection during the seedling stage ranged from 1.3 to $8\%$. More symptom expression was found in regrowth of clipped rice than infected intact plants, and the greater infection took place in early seasonal culture than in ordinary seasonal culture. A higher incidence of the disease was found on the rows close to the bank, and gradually decreased toward the centre of the rice paddy. Disease occurrence and plant maturity was highly correlated in that the most japonica rice types were diseased when they were inoculated within 3 to 7 leaf stage, and$50\%$, $20\%$ and no diseaseb were found if they were inoculated at 9, 11 and 13 leaf stages, respectively. Symptom expression required 7-15 days when the plants were inoculated during 3-7 leaf stages, while it was 15-30days in the plants inoculated during 9-15 leaf stages. On Tongil variety the per cent disease was relatively higher when the plants were infected within 1.5-5 leaf stages than those at 9 leaf stage, and no disease was found on the plants infected after 15 leaf stage. The disease resulted in lowered growth rates, maturity and sterility of Tongil variety although the variety is known as tolerant to the virus. 2. Host range: Thirty five species of crops, pasture grasses and weeds were tested for their susceptibility to the virus. Twenty one out of 35 species tested were found to be susceptible. and 3 of them, Cyperus amuricus Maximowics var. laxus, Purcereus sanguinolentus Nees and Eriocaulon robustius Makino, were found as new hosts of the virus. 3. Transmission: The vector of the virus, Laodelphax striatellus, produces 5 generations a year. The peak of second generation adults occurred at June 20th and those of third was at about July 30th in Suweon area. In Jinju area the peak of second generation adult proceeded the peak at Suweon by 5-7 days. The peaak of third generation adult was higher than the second at Jinju, but at Suweon the reverse was true. The occurrence of viruliferous Laodelphax striatellus was 10-15, 9, 17, 8 and about $10\%$ from overwintered nymph, 1st generation nymph, 2nd generation adult, End generation nymph and the remaining generations, respectively. More viruliferous L. striatellus were found in the southern area than in the central area of Korea. The occurrence of viruliferous L. striatellus depended on the circumstances of the year. The per cent viruliferous vectors gin 2nd and 3rd generation adult, however, was consistantly higher than that of other generations. Matings of viruliferous L. striatellus resulted in $90\%$ viruliferous progenies, and the 3rd, 4th and 5th instars of the vector had higher infectiviey than the rest of the vector stages. The virus acquisition rate of non-viruliferous L. striatellus was $7-9\%$, These viruliferous L. striatellus, however, could not transmit the virus for more than 3 serial times. The optimum temperature for the transmission of the viru3 was $25-30^{\circ}C$, while rare transmission occurred when the temperature was below $15^{\circ}C$. The per cent of L. striatellus parasitization by Haplogonatopus atratus were $5-48\%$ during the period from June to the end of August, and the maximum parasitization was $32-48\%$ at around July 10. 4. Control: 1) Cultural practices; The deeper the depth of transplanting more the disease occurrence was found. The higher infection rate, $1.5-3.5\%$, was observed during the late stages of seedling beds, and the rate became lower, $1.0-2.0\%$, in the early period of paddy field in southern area. Early transplanting resulted in more infection than early seasonal culture, and the ordinary seasonal culture showed the lowest infection. The disease also was favored by earlier transplanting even under tile ordinary seasonal culture. The higher the nitrogen fertilizer level the more the disease occurrence was found in the paddy field. 2) Resistant varieties; Tongil varieties shelved the resistant reaction to the virus in greenhouse tests. In the tests for resistance on 955 varieties most japonica types shelved susceptible reactions, while the resistant varieties were found mostly from introduced varietal groups. 3) Chemical control; Earlier applications of chemicals, Disyston and Diazinon, showed better results when the test was made 4 days after inoculation in the greenhouse even though none of the insecticides shelved the complete control of the disease. Three serial applications of chemicals on June 14, June 20 and June 28 showed bettor results than one or two applications at any other dates under field conditions.

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