Pot experiment was conducted to investigate the effects of fly ash on growth responses and on accumulation of the heavy metals in soil. Two kinds of fly ash, anthracite and bituminous coal, were treated with different levels of 0, 0.4, 0.8, and 1.2 kg/pot(20L). Tobacco growth was better by application of fly ash than that by the control. However, the early stage of growth by application of bituminous coal, 1.2 kg/pot, was decreased due to the boron toxicity occurred by fly ash. Generally, tobacco yield was significantly increased with applying fly ash, showing the highest yield at 1.2 kg/pot for anthracite and at 0.8 kg/pot for bituminous coal. The content of total nitrogen in leaves was higher with fly ash than that of the control, while the content of calcium in leaves was low, Contents of heavy metal and the other minerals were not significantly different between the control and the treatment of fly ash. Soil pH after experiment was linearly increased with application level of fly ash, indicating that the application of bituminous coal was more effective than that of anthracite. Contents of available phosphate, exchangeable $Ca^{2+}$+ and $Mg^{2+}$ in soil were increased by application of fly ash, especially with bituminous coal. Contents of Cu, Cr, and Ni were increased with application level of bituminous coati even if the contents were still lower than critical levels for farming land. The other heavy metals were similar between the control and the application of fly ash.

In the flue curing process, the wet bulb temperature is usually controlled at a constant level. To improve the quality of flue cured leaves, we studied the effect of the cyclic change of wet bulb temperature at the yellowing stage of flue curing on physical properties of cured leaves. The wet bulb temperature was automatically controlled between preset high (38℃) and low point (35℃) every one hour cyclically. As a result, the acceleration of the increase in the physical properties of cured leaves were observed. The leaves cured by this method were more or less orange in color, better bodied, and less brittle compared with the leaves produced by conventional curing. However, the leaves cured by this method had a little sharpness and harshness. As to the physical properties, there was decreased in occurrence of flat leaves than that of conventional ones. On the other hand, in case of cyclic curing method, increase of price per kg reached to 2-3% compared with those of conventional ones.

The bulk curing experiment to the improve the quality of flue-cured leaves were carried out to evaluate relationship between cyclic curing and conventional curing method. We studied the effect of the cyclic change of wet bulb temperature at the yellowing stage of flue curing on chemical properties of cured leaves. The wet bulb temperature was automatically controlled between preset high($38^{\circ}C$) and low point ($35^{\circ}C$) everyone hour cyclically during 12 hours after starting luring. As a result, the acceleration of the increase in the chemical properties of cured leaves were observed. As to the chemical properties, there was decreased in citric acid, increased in malic acid of the nonvolatile organic acids and all higher fatty acids content of leaves cured by this method compared with in that of conventional curing method, while a major chemical compounds in relation to aromatic essence of cigarette smoke in essential oil components of lured leaves was mostly higher in this method than those of conventional ones, and it was evaluated that there was decreased in CO, TPM, Tar, and $CO_2$ content of the cigarette smoke comparing to the conventional luring method.

This study was conducted to obtain the basic information on the burley tobacco (Nicotiana tabacum L.) curing methods. The effects of the three kinds of curing methods on the dry weight losses during curing, chemical contents and physical properties of cured leaves, and organoleptic and smoking qualities were observed. The dry weight losses during curing was the highest in the stalk curing, and the lowest in the conventional priming. The longer the curing periods was, the more the loss of dry weight. The percent of the excessive dried leaves was high and the price per kilogram was low in the conventional priming. The chemical contents and the physical properties of the cured leaves in the conventional priming were similar to those of yellowing or browning stage of curing. But, the chemical contents and the physical properties of the cured leaves in the stalk curing were comparable to those of the air curing. The smoking quality was the best in the stalk curing.

Survival, growth and reproduction of the cigarette beetle, Lasiodema sewicome F., were studied on flue-cured (NC82), air-cured (Br.21), aromatic (Izmir) tobacco and artificial diet (wheat feed 95% + yeast 5%). Cigarette beetle adults occurred from first generation in the flue-cured and aromatic tobacco, but no adults emerged in air-cured tobacco after the 2nd generations. The head capsule widths of larvae of the cigarette beetles L sewicome F.1 reared on different diets at 2$0^{\circ}C$, $25^{\circ}C$ and 3$0^{\circ}C$ were measured. The head capsule development was slower at 2$0^{\circ}C$ on the same diets than $25^{\circ}C$ and 3$0^{\circ}C$, and slower in the flue-cured and aromatic tobacco than the artificial diet at 2$0^{\circ}C$ and 2S$^{\circ}C$, At $25^{\circ}C$ the larval head capsule developed more rapidly in the aromatic tobacco than the flue-cured tobacco. However, no larval head development was noticed in the air-cured tobacco. The correlation between population densities of cigarette beetle,1 sewicome F., and sugar content of tobacco leaves was positive, but negative in nicotine contents. The regression between densities and sugar content was Y : 22.79 X + 57.29 (r=0.891**), and between nicotine content was Y : -141.31X + 321.40(r=-0.917**).

Protoplasts were isolated from mesophyll of tobacco(Nicotiana glauca) transformed with kanamycin-resistant gene (NPT II gene) and potato hairy root callus containing Ri plasmid of Agrobacterium rhiEogenes, and protoplasm fusion was made between the isolated protoplasts. The transgenic tobacco leaf tissue could grow on the media containing high concentrations of kanamycin, but not on the phytohormone-free media. On the other hand, the potato hairy root calli could be cultured on the phytohormone-free media but not on media containing more than 40 ㎍/ml kanamycin. In these conditions, the viability of both protoplasts were above 90%, These selection markers were used for the selection of protoplasts fused between the two, i.e. protoplast fusion was detected using selection media containing 100㎍/ml kanamycin and with no phytohormone. The mixture of 1.0% cellulase, 0.3% macerozyme, and 0.7M mannitol was best for the maximum protoplast production for tobacco, and that of 2.0% cellulase, 2.0% macerozyme, 1.0% dricelase, and 0.5M mannitol for potato. Both tobacco mesophyll and potato callus protoplasts were fused by using PEG solution on the selectable medium. Cell walls were regenerated after 5 days in this medium, and colonies were alive until 4 weeks after cultural, but died after 6 weeks.

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.

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.

TMV resistant lines (TRLs) originated from the Blo plant of Nicotiana tabacum cv. NC82 transformed with TMV coat protein cDNA which initially showed delayed disease symptom were selected for increased resistance in each subsequent generation. The result of field experiment of the transgenic tobacco lines in the fifth generation for TMV resistance and their response to other tobacco diseases (black shank, bacterial wilt, and powdery mildew) is described in this report. When fifteen TRLs of the fifth generation were tested for TMV resistance by mechanically inoculating the individual plants, over 95 percent of the plants of 6 lines showed complete resistance even 8 weeks after the inoculation. Average frequency of the resistant plants in TRLs of the fifth generation 8 weeks after the inoculation was 87%. Stable insertion and expression of TMV coat protein cDNA in the fifth generation of the transgenic tobacco plant were confirmed by PCR and immunoblot hybridization, respectively. All TRLs were resistant to the black shank but were susceptible to the bacterial wilt disease and the powdery mildew to the same degree as non-transgenic NC82 was. Therefore, it was indicated that the phenotypes related at least to disease resistance were not changed in the transgenic tobacco. Key words : TMV CP cDNA, TMV resistant tobacco plant, transformation.

Haemolyph carboxylesterases induced in diapausing pupae of Helicoverpa assulta Guenee were investigated. Increase in the activity of the electrophoresed isozyme bands were observed during the diapausing pupae. The isozymatic composition exhibited remarkable alterations represented as disappearance and induction of some isozyme bandsp which were identified as carboxylesterase (CE) on the basis of their specificities to inhibitors. Much higher activity of the induced CE was shown in reaction with $\beta$-naphthyl acetate ($\beta$-Na) than $\alpha$-naphthyl butyrate ($\alpha$-Nb), representing the high regioselectivity to $\beta$-naphthyl group. Optimal temperature for the enzyme activity was different to the substrates used 37$^{\circ}C$ in $\beta$-Na and 4$0^{\circ}C$ in $\alpha$-Nb, respectively. However, the optimal pH for the enzyme activity was the same as 7.5 regardless of the substrates used, and relatively high thermostability of the CE was demonstrated by showing the denaturation at high temperature (50~55$^{\circ}C$).

Changes in activity and classification of esterase isozymes during the tire cycle or Plodia inteipunctella (Hiibner) were investigated by the native polyacrylamide gel electrophoresis. The stage specificity in esterase activity and isozyme pattern was observed throughout the larvalpupal-adult transformation. The activity esterase was highest at the 3-day old adult stage, and the lowest level at the prepupal stage. A total of 12 esterase bands were identified throughout the development, and the bands showing high enzyme activity was observed in the middle part of gel. Twelve esterases on the basis of inhibition by the three types of inhibitors (organophosphates, eserine sulfate and sulfhydryl reagents) were classified into three class, namely, carboxylesterase (CE), arylesterase (ArE) and cholinesterase (ChE), and these classes contained 7, 3 and 2 isozymes, respectively.

After we made the computer source code with mathematical model of Muramatsu et al. that was expressed by the set of simultaneous first-order ordinary differential equations in evaporation-pyrolysis zone of cigarette, we simulated the distribution profiles of temperature and density of flue-cured tobacco. Those equations were solved numerically with the Runge-Kutta-Gill algorithm assuming step size of 0.025mm by Muramatsu at at,, but in this study the advanced algorithm of Runge-Kutta 4th Order assuming step size of 0.0005mm. The initial conditions and physical parameters of Muramatsu et at. were used for solving them. The calculated values corresponded well with results of Muramatsu et al., especially the gradient of the temperature profile increased with smoldering speed and the thickness of the evaporation-pyrolysis zone decreased with increasing of smoldering speed. On the other hand, the temperature gradient decreased with increasing of the effective thermal-conductivity value and the thickness of the evaporation-pyrolysis zone increased with the effective thermal-conductivity value.

Numerous studies have demonstrated a negative association between cigarette smoking and Parkinson's disease. The present study was undertaken to investigate whether chronic exposure of mice to cigarette smoke a(footed the metabolism of 1-methyl-1113,6-tetrahydro-pyridine (MPTP) by cytochrome P4SO (P-450) or flavin-containing monooxygenase (FMO) in the hepatic microsomes of C57BL6/J mice. Adult male C57BL6/J mice were exposed to mainstream smoke generated from 15 cigarettes for 10 min a day and 5 day per week for 6 weeks. MPTP (10 mg/kg body weight) was administered to mice by subcutaneous injection for 6 consecutive days. Microsolnal P-450 content was increased by MPTP, smoke exposure, or both, but NADPH cytochrome P-450 reductase activity was rather decreased by the same treatments. The activities of benzo(a)pyrene hydroxylase, 7-ethoxycoumarin O-deethylase and ethoxyresorufin O-deethylase were significantly increased by the exposure of cigarette smoke, but were not or little affected by MPTP treatment. Benzphetamine N-demethylase activity was not affected either by MPTP treatment or by cigarette smoke exposure, but it was significantly increased by the combined MPTP treatment with cigarette smoke exposure, showing their synergic effect for the induction of the enzyme activity. Interestingly, in vitro studies of hepatic FMO and P-450 system both O-oxygenation and N-demethylation of MPTP were increased in the smoke-exposed or in the MPTP-treated mice. These results suggest that the enhancement in the N-demethylation as well as O-deethylation of P-450 system and in the N-oxygenation of FMO activity by cigarette smoke exposure in mouse liver may contribute to attenuating the neurotoxic effects of MPTP on the nigrostriatal dopaminergic neurons.

This study was conducted to evaluate the techniques or analyzing tobacco smoke by statistical treatment method for the analytical data through Asia Collaborative Study V. In addition to five smoke components analysis, consisting of TPM, water, nicotine, NFDPM, and puff count of four cigarettes samples, statistical parameters such as mean, standard deviation, box-and-whisker plots, h plots, k plots, regression coefficients, reproducibility (R), and repeatability (r) were also calculated. Analysis of water content of cigarette smoke was the most difficult task, whereas puff count analysis was the easiest as well recognized by all laboratories. Analysis of nicotine and puff count accounted for both the lowest and the highest variation among four parameters. The water coefficients indicated more randomness or variation in the slops. The NFDPM data exhibited both types of deviations from linearity. Water content of sample D indicated the highest difference between two single results and between two interlaboratory test results. As a whole, KGTRI ranked higher in the analytical techniques for statistical evaluation of results when compared with the practices of 28 other laboratories.

The semi-volatile fraction of cigarette mainstream smoke contains the most important aroma and taste that enhance smoke flavor. The components in mainstream smoke were analysed for commercially available cigarettes. To compare the semi-volatile components delivered from mainstream smoke of domestic cigarette with imported cigarette, we analyzed three types of filter cigarettes such as cellulose acetate mono filter, carbon dual filter and slim filter cigarette. Mainstream smoke was collected with a combination of Cambridge filter pad and liquid trap containing a small amount of acetone cooled by dry ice-methanol. We have identified 43 components from mainstream smoke by GC and GC/MS and found that benzene, toluene, 4-hydroxy-4-methyl-2-pentanone, 5-methyl-2-furaldehyde, acetic acids nicotine and triacetin were the main peaks in mainstream smoke. Also we have compared the relative amounts of semi-volatile compounds delivered from the domestic and imported cigarettes with three different filters. From the results of this study, the relative amounts of limonene, acetic acid, 5-methyl-Lfuraldehyde delivered from the imported cigarettes were generally higher than those of the domestic one's.

This study was conducted to evaluate the ability of anion generating air cleaner to remove gases, vapor and particles from closed room contaminated with environmental tobacco smoke (ETS). The measurements covered particle sizes of 13.8-542.5nm, particle concentration, surface area, volumes UVPM, FPM, solanesol, and the following gases and vapor; carbon dioxide, carbon monoxide, nicotine, and 3-ethenylpyridine. Tobacco smoke was generated and mixed in a closed room in which the airflow rates were in the range of 0.00-0.04 m/s. The anion generating air cleaner was startedl and the decay rates for the gases, vapor and particles were measured, When the use of anion generating air cleaner, solid components of ETS, such as respirable suspended particle (RSP), utraviolet particulate matter (UVPM, fluorescent particulate matter (FPM) and solanesol was sharply decreased, and vapor phase components of ETS, such as nicotines 3-ethenylpyidine were modelately decreased by time elapse. Even the use of anion generation air cleaner, the decreasing rate of carbon dioxide concentration was similar with control, and the decreasing rate of carbon monoxide was slower than that of control. Our results indicated that the use of anion generting air cleaner had an effect on reduction of solid and vapor components from ETs, but it had no effect on gaseous components of ETS.

In order to minimize the trial frequency in the new filter cigarette design, we studied the relationship between smoke yield and tipping materials of cigarette. A three levels full factorial design involving filament denier (X1,2.5-3.3d), Porosity of the acetate filter plug wrap (X2, 3,500-16,000CU) and porosity of the tip paper (X3, 400-1,200CU) was used. Three independent factors (Xl, X2, X3) were chosen for their effects on the various responses and the function was expressed in terms of a quadratic polynomial equation, Y : $\beta$o + $\beta$1Xl + $\beta$2X2 + $\beta$3X3 + $\beta$11Xl2 + $\beta$22X22+ $\beta$33X32 + $\beta$12X1X2 + $\beta$13XIX3 $\beta$23X2X3 which measures the linear, quadratic, and interaction effects. Twenty-nine trial numbers were obtained as a results of using a three levels full factorial design and it was analyzed by the multiple regression analysis with backward stepwise in STATISTICA/pc under restricted conditions. Tar yields of the cigarette was affected by porosity of tip paper (0.66), filament denier (0.47) and porosity of plug wrap (0.28) in the decreasing order, and linear effect of tip paper porosity (B3) and filament denier (91) were significant at a level of 0.01($\alpha$). The filament denier and tipping paper porosity interaction F ratio among three factors had a P-value of 0,000041, indicating higher interaction between these factors. Based on the analysis of variance, the model fitted for Tar (Y1) was significant at 5% confidence level and the coefficient of determination (0.96) was the proportion of variability in the data fitted for by the model.