• Title/Summary/Keyword: Transgenic

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Differentiation and Apoptosis of the Mammalian Embryo and Embryonic Stem Cells(ESC): I. Establishment of Mouse ESC and Induction of Differentiation by Reproductive Hormones (포유동물의 배아 및 기간세포의 분화와 세포사멸 기작: I. 생쥐 배아줄기세포의 확립과 분화유도에 미치는 생식호르몬의 영향)

  • 성지혜;윤현수;이종수;김철근;김문규;윤용달
    • Development and Reproduction
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    • v.6 no.1
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    • pp.55-66
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    • 2002
  • Embryonic stem cells(ES cells) are derived from the inner cell mass(ICM) of blastocysts, which have the potentials to remain undifferentiated, to proliferate indefinitely in vitro, to differentiate into the derivates of three embryonic germ layers. ES cells are an attractive model system for studying the initial developmental decisions and their molecular mechanisms during embryogenesis. Additionally, ES cells of significant interest to those characterizing the various gene functions utilizing transgenic and gene targeting techniques. We investigated the effects of reproductive hormones, gonadotropins(GTH) and steroids on the induction of differentiation and expressions of their receptor genes using the newly established mouse ES cells. We collected the matured blastocysts of inbred mice C57BL/6J after superovulation and co-cultured with mitotically inactivated STO feeder cells. After 5 passages, we confirmed the expression alkaline phosphatase(Alk P) activity and SSEA-1, 3, 4 expressions. The protocol devised for inducing ES differentiation consisted of an aggregation steps, after 5 days as EBs in hormone treatments(FSH, LH, E$_2$, P$_4$, T) that allows complex signaling to occur between the cells and a dissociation step, induced differentiation through attachment culture during 7 days in hormone treatments. Hormone receptors were not increased in dose-dependent manner. All hormone receptors in ES cells treated reproductive hormones were expressed lower than those of undifferentiated ES cell except for LHR expression in E$_2$-treated ES cells group. After hormone induced differentiation, at least some of the cells are not terminally differentiated, as is evident from the expression of Oct-4, a marker of undifferentiated. To assess their differentiation by gene expression, we analyzed the expression of 7 tissue-specific markers from all three germ layers. Most of hormone-treated group increased in the expression of gata-4 and $\alpha$ -fetoprotein, suggesting reproductive hormone allowed or induced differentiation of endoderm.

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Analgesic Effects on the Oral Cavity by Electroacupuncture in Dogs (개에서 전침에 의한 구강 수술 시 진통효과)

  • Choi, Kyeong-Ha;Lee, Jae-Yeon;Park, Chang-Sik;Jeong, Seong-Mok;Kim, Duck-Hwan;Kim, Myung-Cheol
    • Journal of Veterinary Clinics
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    • v.26 no.6
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    • pp.568-573
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    • 2009
  • The purpose of this study is to evaluate the analgesic effects on the oral cavity in dogs which are treated with xylazine and electroacupuncture (EA). Furthermore, this study aims to find out its effects on glucose (GLU), serum alanine aminotransferase (ALT), and blood urea nitrogen (BUN) variation values, vital signs (rectal temperature, heart rate, respiratory rate) and pain responses to the noxious stimuli. Eight healthy dogs were randomly assigned to receive either xylazine or EA. Xylazine group dogs with weight of 3.6${\pm}$1.0 kg received 1.5 mg/kg of xylazine intramuscularly. EA group dogs with weight of 3.9${\pm}$1.0 kg received 1 volt (10-15 hz) for 5 minutes, and then 1-9 Volts (25-30 hz) for 60 minutes totally. The acupoints used were LI-3 (San Jian), LI-4 (He Gu) and ST-7 (Xia Guan). All dogs were examined before and 10, 25, 40, 55 and 120 minutes after administration of xylazine or EA. The mean rectal temperatures of the EA group were significantly higher than those of xylazine group after 25, 40 and 55 minutes (p < 0.05). The mean heart rates of the EA group were significantly higher than those of xylazine group after 10, 25, 40 and 55 minutes (p < 0.05). The mean respiratory rates of the EA group were significantly higher than those of xylazine group after 55 and 120 minutes (p < 0.05). The mean GLU concentration of the EA group were significantly lower than those of xylazine group after 55 and 120 minutes (p < 0.05). The sum of mean pain scores (SMPS) of the EA group were significantly higher than those of xylazine group after 10, 25 40 and 55 minutes (p < 0.05). In this study, the pain control of the EA group was shown to be better than that of the xylazine group. Also, there do not appear to be any negative physiologic effects associated with acupuncture-induced surgical analgesia. So, it was considered that these acupoints of EA analgesia might be useful for minor oral surgery in weak patients.

Attenuation of Renal Ischemia-Reperfusion (I/R) Injury by Ascorbic Acid in the Canine Nephrotomy (개의 신장에 있어서 Ascorbic Acid에 의한 허혈/재관류 손상의 감소에 관한 연구)

  • Kim, Jong-Man;Lee, Jae-Yeon;Jeong, Seong-Mok;Park, Chang-Sik;Kim, Myung-Cheol
    • Journal of Veterinary Clinics
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    • v.27 no.5
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    • pp.553-558
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    • 2010
  • The purpose of this study was to investigate the effects of premedicated ascorbic acid and hepa-saline irrigation/aspiration on attenuation of ischemia-reperfusion (I/R) injury and recovery of renal function in canine nephrotomy model. In the canine model, nine mixed dogs were subjected to renal nephrotomy with premedicated ascorbic acid and hepa-saline irrigation-aspiration (treatment group 2), and only hepa-saline irrigation-aspiration (treatment group 1). The level of renal function and antioxidant enzymes after nephrotomy were measured. And the expression pattern of TNF-${\alpha}$ and INF-${\gamma}$ was examined in the renal tissue at $7^{th}$ day after nephrotomy. BUN and creatinine levels significantly decreased in the treatment group 1 and 2 compared to that of control group at the $3^{rd}$, 5th and $7^{th}$ day after reperfusion (p < 0.05). And, there was significant difference between treatment group 1 and 2 at the $3^{rd}$ day after reperfusion (p < 0.05). The activities of antioxidant enzymes in plasma was significantly increased in the treatment group 1 and 2 compared to that of control group at the $3^{rd}$, $5^{th}$ and $7^{th}$ day after reperfusion (p < 0.05). And, there was significant difference between treatment group 1 and 2 at the $3^{rd}$ day after reperfusion (p < 0.05). TNF-${\alpha}$ was decreased and INF-${\gamma}$ was increased in treatment groups. The result of this study suggested that irrigation-aspiration has effects on attenuation of renal ischemia-reperfusion injury, and the exogenous ascorbic acid has a role in the attenuation of renal ischemia-reperfusion injury and recovery of renal function in canine nephrotomy model.

Current Status of the Research on the Postharvest Technology of Melon(Cucumis melo L.) (멜론(Cucumis melo L.) 수확 후 관리기술 최근 연구 동향)

  • Oh, Su-Hwan;Bae, Ro-Na;Lee, Seung-Koo
    • Food Science and Preservation
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    • v.18 no.4
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    • pp.442-458
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    • 2011
  • Among Cucubitaceae, melon (Cucumis melo) is one of the most diversified fruits, with various forms, sizes, pulps, and peel colors, In addition, it is a commercially important crop because of its high sweetness, deep flavor, and abundant juice. In the species, there are both climacteric and non-climacteric melons depending on the respiration and ethylene production patterns after harvest. Ethylene is also considered a crucial hormone for determining sex expression, Phytohormones other than ethylene interact and regulate ripening, There are some indices that can be used to evaluate the optimum harvest maturity. The harvest time can be estimated after the pollination time, which is the most commonly used method of determining the harvest maturity of the fruit. Besides the physiological aspects, the biochemical alterations, including those of sweetness, firmness, flavor, color, and rind, contribute to the overall fruit quality. These changes can be categorized based on the ethylene-dependent and ethylene-independent phenomena due to the ethylene-suppressed transgenic melon. After harvest, the fruits are precooled to $10^{\circ}C$ to reduce the field heat, after which they are sized and packed. The fruits can be treated with hot water ($60^{\circ}C$ for 60 min) to prevent the softening of the enzyme activity and microorganisms, and with calcium to maintain their firmness. 1-methylenecyclopropene (1-MCP) treatment also maintains their storability by inhibiting respiration and ethylene production. The shelf life of melon is very short even under cold storage, like other cucurbits, and it is prone to obtaining chilling injury under $10^{\circ}C$. In South Korea, low-temperature ($10^{\circ}C$) storage is known to be the best storage condition for the fruit. For long-time transport, CA storage is a good method of maintaining the quality of the fruit by reducing the respiration and ethylene. For fresh-cut processing, washing with a sanitizing agent and packing with plastic-film processing are needed, and low-temperature storage is necessary. The consumer need and demand for fresh-cut melon are growing, but preserving the quality of fresh-cut melon is more challenging than preserving the quality of the whole fruit.

Comparison of Anesthetic Effects Induced by Tiletamine-Zolazepam and Azaperone Plus Tiletamine-Zolazepam in Growing Pigs (돼지에서 Tiletamine-Zolazepam 단독과 Azaperone, Tiletamine-Zolazepam 합제의 마취 효과에 대한 비교)

  • Kim, Young-Suk;Kim, Myung-Jin;Lee, Soo-Jin;Lee, Jae-Il;Jun, Moo-Hyung;Park, Chang-Sik;Kim, Myung-Cheol
    • Journal of Veterinary Clinics
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    • v.24 no.3
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    • pp.300-304
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    • 2007
  • The purpose of this study was to determine the anesthetic effects of tiletamine-zolazepam (TZ) alone and azaperone plus tiletamine-zolazepam in growing pigs, and to compare the various physiological parameters in both treatments. Cross experiment was accomplished at 2-week interval. Group 1 (TZ group): six pigs ($31.4{\pm}4.83$ kg) received 4.4 mg/kg of TZ alone. Group 2 (ATZ group); the same six pigs ($43.6{\pm}4.31$ kg) received 4.4 mg/kg of TZ twenty minutes after receiving 2 mg/kg of azaperone. All of the anesthetic drugs were injected into the trapezius muscles. The pigs were fasted for 24 hours before the experiments. Induction and recovery values were determined. Heart rate, respiratory rate, rectal temperature, $pO_2,\;pCO_2$ and pH were determined before administration and 5, 25, 45, 65 and 85 minutes after administration. Induction time of ATZ group was more rapid than that of TZ group (p<0.01). During recovery, sternal recumbency time, standing time and walking time of ATZ group were longer than those of TZ group (p<0.01). Heart rate, respiratory rate, $pO_2,\;pCO_2$, and pH did not show especial differences between the two groups. However, rectal temperature was significantly different between the TZ and ATZ group (p<0.05). As a result, ATZ group had a faster induction and a longer duration of anesthesia than TZ group did. Thus, it was concluded that ATZ combination could be usefully used for chemical restraint in pigs.

Influences of Insect-Resistant Genetically Modified Rice (Bt-T) on the Diversity of Non-Target Insects in an LMO Quarantine Field (LMO 격리 포장에서 해충저항성벼(Bt-T)가 비표적 곤충다양성에 미치는 영향)

  • Oh, Sung-Dug;Park, Soo-Yun;Chang, Ancheol;Lim, Myung-ho;Park, Soon Ki;Suh, Sang Jae
    • Korean Journal of Breeding Science
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    • v.50 no.4
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    • pp.406-414
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    • 2018
  • This study was conducted to develop environmental risk assessments and biosafety guides for insect-resistant genetically modified rice in an LMO (Living Modified Organism) isolation field. In the LMO quarantine area of Kyungpook National University, the species diversities and population densities of non-target insects found on insect-resistant genetically modified rice (Bt-T), rice resistant to Cnaphalocrocis medinalis, and non-GM rice (Dongjin-byeo and Ilmi-byeo) were investigated. The Bt-T plants were, therefore, evaluated under field conditions to detect possible impacts on above ground insects and spiders. In 2016 and 2017, the study compared transgenic rice and two non-GM reference rice, namely Dongjin-byeo and Ilmi-byeo, at Gunwi. A total of 9,552 individuals from 51 families and 11 orders were collected from the LMO isolation field. From the three types of rice fields, a total of 3,042; 3,212; and 3,297 individuals from the Bt-T, Dongjin-byeo, and Ilmi-byeo were collected, respectively. There was no difference between the population densities of the non-target insect pests, natural enemies, and other insects on the Bt-T compared to non-GM rice. The data on insect species population densities were subjected to principal component analysis (PCA) without distinguishing between the three varieties, namely GM, non-GM, and reference cultivar, in all cultivation years. However, the PCA clearly separated the samples based on the cultivation years. These results suggest that insect species diversities and population densities during plant cultivation are determined by environmental factors (growing condition and seasons) rather than by genetic factors.

Comparative Transcriptome Analysis of the Response of Two Lines of Rapeseed (Brassica napus L.) to Cold Stress (유채 두 계통에서 저온 스트레스에 반응하는 전사체 발현 비교 분석)

  • Lee, Ji-Eun;Kim, Kwang-Soo;Cha, Young-Lok;An, Da-Hee;Byun, Jong-Won;Kang, Yong-Ku
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.66 no.1
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    • pp.37-71
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    • 2021
  • Rapeseed is a typical winter crop, and its freezing stress tolerance is a major feature for winter survival. Therefore, it is important to comprehend clearly the physical and molecular mechanisms of rapeseed under freezing stress conditions. This study investigates the physical and transcriptome changes of two rapeseed lines, 'J8634-B-30' and 'EMS26', under cold acclimation and freezing temperature treatments. The proline content of 'J8634-B-30' at 5 ℃ increased 8.7-fold compared to that before treatment, and there was no significant change in that of 'EMS26' RNA-sequencing analysis revealed 5,083 differentially expressed genes (DEGs) of 'J8634-B-30' under cold acclimation condition. Among the genes, 2,784 (54.8%) were up-regulated and 2,299 (45.2%) were down-regulated. The DEGs of 'EMS26' under cold acclimation condition were 5,831 genes, and contained 2,199 up-regulated genes (37.7%) and 3,632 down-regulated genes (62.3%). Among them, only DEGs annotated in the cold response-related signaling pathways were selected, and their expression in the two rapeseed lines was compared. Comparative DEGs analysis indicated that cold response related signaling pathways are proline metabolism and ABA (Abscisic acid) signaling. And ICE (Inducer of CBF expression) - CBF (C-repeat-binding factor) - COR (Cold-regulated) signaling were the significantly differentially expressed transcripts in the two rapeseed lines. The major induced transcripts of 'J8634-B-30' induced P5CS (Δ'-pyrroline-5-carboxylate synthetase), which is related to proline biosynthesis, PYL (pyrabactin resistance-like protein, ABA receptor) and COR413 (cold-regulated 413 plasma membrane 1). In conclusion, these result provide a foundation for understanding the mechanisms of freezing stress tolerance in rapeseeds. Further functional studies should be performed on the freezing stress-related genes identified in this study, which can contribute to the transgenic and molecular breeding for freezing stress tolerance in rapeseed.

Temporal Transcriptome Analysis of SARS-CoV-2-Infected Lung and Spleen in Human ACE2-Transgenic Mice

  • Jung Ah, Kim;Sung-Hee, Kim;Jung Seon, Seo;Hyuna, Noh;Haengdueng, Jeong;Jiseon, Kim;Donghun, Jeon;Jeong Jin, Kim;Dain, On;Suhyeon, Yoon;Sang Gyu, Lee;Youn Woo, Lee;Hui Jeong, Jang;In Ho, Park;Jooyeon, Oh;Sang-Hyuk, Seok;Yu Jin, Lee;Seung-Min, Hong;Se-Hee, An;Joon-Yong, Bae;Jung-ah, Choi;Seo Yeon, Kim;Young Been, Kim;Ji-Yeon, Hwang;Hyo-Jung, Lee;Hong Bin, Kim;Dae Gwin, Jeong;Daesub, Song;Manki, Song;Man-Seong, Park;Kang-Seuk, Choi;Jun Won, Park;Jun-Won, Yun;Jeon-Soo, Shin;Ho-Young, Lee;Jun-Young, Seo;Ki Taek, Nam;Heon Yung, Gee;Je Kyung, Seong
    • Molecules and Cells
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    • v.45 no.12
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    • pp.896-910
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    • 2022
  • Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a highly transmissible and potentially fatal virus. So far, most comprehensive analyses encompassing clinical and transcriptional manifestation have concentrated on the lungs. Here, we confirmed evident signs of viral infection in the lungs and spleen of SARS-CoV-2-infected K18-hACE2 mice, which replicate the phenotype and infection symptoms in hospitalized humans. Seven days post viral detection in organs, infected mice showed decreased vital signs, leading to death. Bronchopneumonia due to infiltration of leukocytes in the lungs and reduction in the spleen lymphocyte region were observed. Transcriptome profiling implicated the meticulous regulation of distress and recovery from cytokine-mediated immunity by distinct immune cell types in a time-dependent manner. In lungs, the chemokine-driven response to viral invasion was highly elevated at 2 days post infection (dpi). In late infection, diseased lungs, post the innate immune process, showed recovery signs. The spleen established an even more immediate line of defense than the lungs, and the cytokine expression profile dropped at 7 dpi. At 5 dpi, spleen samples diverged into two distinct groups with different transcriptome profile and pathophysiology. Inhibition of consecutive host cell viral entry and massive immunoglobulin production and proteolysis inhibition seemed that one group endeavored to survive, while the other group struggled with developmental regeneration against consistent viral intrusion through the replication cycle. Our results may contribute to improved understanding of the longitudinal response to viral infection and development of potential therapeutics for hospitalized patients affected by SARS-CoV-2.

Tumorigenesis after Injection of Lung Cancer Cell Line (SW-900 G IV) into the Pleural Cavity of Nude Mice (누드마우스의 흉강에 폐암세포주의 주입에 의한 종양형성과 HER2/neu와 TGF-${\beta}_1$의 발현)

  • Park, Eok-Sung;Kim, Song-Myung;Kim, Jong-In
    • Journal of Chest Surgery
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    • v.43 no.6
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    • pp.588-595
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    • 2010
  • Background: Base on types of tumor, the types of expressed tumor is diverse and the difference in its expression rate is even more various. Due to such reasons an animal model is absolutely needed for a clinical research of lung cancer. The author attempted oncogenesis by cultivating a cell line of non-small cell carcinoma and then injecting it inside thoracic cavities of nude mice. The author conducted quantitative analyses of HER2/neu tumor gene - an epidermal growth factor receptor (EGFR) related to lung cancer, and TGF-${\beta}_1$, which acts as a resistance to cell growth inhibition and malignant degeneration. In order to investigate achievability of the oncogenesis, histological changes and the expression of cancer gene in case of orthotopic lung cancer is necessary. Material and Method: Among 20 immunity-free male BALB/c, five nude mice were selected as the control group and rest as the experimental group. Their weights ranged from 20 to 25 gm (Orient, Japan). After injection of lung cancer line (SW900 G IV) into the pleural cavity of nude mice, They were raised at aseptic room for 8 weeks. HER2/neu was quantitatively analyzed by separating serum from gathered blood via chemiluminiscent immunoassay (CLIA), and immunosandwitch method was applied to quantitatively analyze TGF-${\beta}_1$. SPSS statistical program (SPSS Version 10.0, USA) was implemented for statistical analysis. Student T test was done, and cases in which p-value is less than 0.05 were considered significant. Result: Even after lung cancer was formed in the normal control group or after intentionally injected lung cancer cell line, no amplification of HER2/neu gene showed reaction. However, the exact quantity of TGF-${\beta}_1$ was $28,490{\pm}8,549pg/mL$, and the quantity in the group injected with lung cancer cell was $42,362{\pm}14,449pg/mL$, meaning 1.48 times highly Significant (p<0.483). It proved that HER2/neu gene TGF-${\beta}_1$ had no meaningful interconnection. Conclusion: TGF-${\beta}_1$ gene expressed approximately 1.48 times amplification in comparison to the control group. The amplification of TGF-${\beta}_1$ meant somatic recuperation inhibition mechanism due to carcinogenesis in nude mice was definitely working. It may be implemented as a quantitative analysis that allows early detection of lung cancer in human body.

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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