• Korean Journal of Weed Science
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• v.32 no.1
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• pp.25-34
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• 2012

Characteristics related to grain quality and physiochemical components such as mineral, total amino acid, free amino acid, and free sugar composition were investigated in Protox inhibitor resistanttransgenic rice (MX, PX, and AP37) and its nontransgenic counterpart (WT). Head rice, palatability, protein, and whiteness (except for MX and AP37) of milled transgenic rice were high or similar to those of the non-transgenic counterpart. Immature rice, unfilled grain, and cracked kernels (PX and AP37) of milled transgenic rice were lower than those of its non-transgenic counterpart. However, there were no significant differences in damaged grain between the transgenic rice lines and its counterpart. Potassium content in PX and copper contents in PX and AP37 were only low compared with their non-transgenic counterparts, but other mineral contents in transgenic rice lines were high or showed no significant differences compared with non-transgenic counterparts. Contents of most total amino acid composition in transgenic rice lines were high or similar to those in non-transgenic counterparts, but the content of isoleucine in AP37 was only low compared with its non-transgenic counterpart. On the other hand, free amino acid, leucine and tyrosine in PX and AP37, and total free amino acid in PX were low compared with their non-transgenic counterparts. However, the content of free amino acid in other kinds in transgenic rice lines were similar to those in their non-transgenic counterparts. Contents of sucrose in MX and PX were low compared with non-transgenic counterpars, but contents of fructose, glucose, and maltose in transgenic rice lines were high or similar compared with their non-transgenic counterparts. This results indicated that Protox genes had no negative affect on the nutritional composition of rice.

• Korean Journal of Weed Science
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• v.16 no.2
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• pp.150-159
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• 1996

This study was conducted to investigate the inhibition of protox activity and the PPIX accumulation of the oxyfluorfen-tolerant and-susceptible rice cultivars with barnyardgrass, a typical susceptible weed in accordance by oxyfluorfen treatment. The susceptible rice cultivars and barnyardgrass showed more inhibition of protox activity due to the treatment of oxyfluorfen than the tolerant rice cultivars. Especially in the concentration at $10^{-6}$M treatment, protox activity of the susceptible rice cultivars and barnyardgrass were the completely inhibited but the tolerant rice cultivars kept 32~59% of activity compared to the control. As the treatment concentration increased, the content of PPIX accumulation increased and it increased untill four hours of light exposure but it tended to decrease these after. The content of PPIX accumulation by the treatment of oxyfluorfen was more pronounced in the light condition than in the dark. Under the light and dark conditions, the susceptible rice cultivars and barnyardgrass showed more PPIX accumulation than the tolerant rice cultivate. Especialiy the susceptible barnyardgrass had more than the rice. With the treatment of GC and DA, tetrapyrrole biosynthesis inhibitor, the herbicidal activity by oxyfluorfen was inhibited, and the susceptible rice cultivars and barnyardgrass tended to have less effective than the tolerant rice cultivars and the content of chlorophyll or PPS accumulation tended to be similar.

• Korean Journal of Weed Science
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• v.31 no.2
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• pp.134-145
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• 2011

The objectives of this study were to investigate fitness difference in growth and rice yield in herbicide-transgenic rice overexpressing Myxococcus xanthus and Arabidopsis thaliana protoporphyrinogen oxidase (Protox) genes and non-transgenic rice. We also aimed to determine whether these fitness differences are related to ALA synthesizing capacity, accumulation of terapyrroles, reactive oxygen species, lipid peroxidation, and antioxidative enzymes at different growth stages of rice. Plant height of the transgenic rice overexpressing M. xanthus (MX) and A. thaliana (AP37) Protox genes at 43, 50, and 65 days after transplanting (DAT) was significantly lower than that of WT. Number of tiller of PX as well as MX and AP37 at 50 and 65 DAT was significantly lower than that of WT. At harvest time, culm length and yield of MX, PX and AP37 and rice straw weight of MX and AP37 were significantly low compared with WT. The reduction of yield in MX, PX, and AP37 was caused by spikelets per panicle and 1000 grain weight, ripened grain, spikelets per panicle, 1000 grain weight, and ripened grain, respectively. On the other hand, 135 the reduction of yield in MX, PX, and AP37 was also observed in another yearly variation experiment. The reduction of rice growth in MX, PX, and AP37 was observed in seedling stage as well as growth duration in field. There were no differences in tetrapyrrole intermediate Proto IX, Mg-Proto IX and Mg-Proto IX monomethyl ester, reactive oxygen species ($H_2O_2$ and ${O_2}^-$), MDA, antioxidative enzymes (SOD, CAT, POX, APX, and GR) and chlorophyll between transgenic lines and wild type, indicating that accumulated tetrapyrrole intermediate and other parameters were not related to growth reduction in transgenic rice. However, ALA synthesizing capacity in MX, PX, and AP37 at one day after exposure to light and 52 DAT was significantly lower than that of WT. Further study is required to elucidate the mechanisms underlying the growth and yield difference between transgenic and WT lines.

• Korean Journal of Weed Science
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• v.16 no.4
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• pp.337-345
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• 1996

This study was conducted to investigate the protox activity, the PPIX accumulation and the activity of antioxidative enzymes of the oxyfluorfen-tolerant and -susceptible rice cultivars by oxyfluorfen and oxytluorfen-similar herbicides treatment. When treated with acifluorfen, bifenox or oxadiazon, the oxyfluorfen-tolerant rice cultivars showed less decreased in fresh weight than the susceptible rice cultivars. The inhibition of protox activity was in the order of acifluorfen > oxyfluorfen > bifenox > oxadiazon, and the PPIX accumulation was in the sequence of oxadiazon > acifluorfen > oxyfluorfen > bifenox. The inhibition of protox activity and the PPIX accumulation by the herbicide was greater in the susceptible rice cultivars than in the tolerant rice cultivars. The effect inhibiting the decrease of chlorophyll content resulting from the treatment of GC, tetrapyrrole biosynthesis inhibitor, was in the order of oxyfluorfen > acifluorfen > bifenox > oxadiazon, and the tolerant rice cultivar had more than the susceptible rice cultivar. In the treatment of DPE and oxadiazon, the activities of MDAR, POX and GR was higher in the tolerant rice cultivar than in the susceptible rice cultivar, and in the case of POX isozyme the activities of main D and E bands increased in the tolerant rice cultivars at tested herbicides but they didn't in the susceptible rice cultivar.

• The Korean Journal of Pesticide Science
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• v.9 no.2
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• pp.181-184
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• 2005

The heterocyclic PPO inhibitor compounds have been studied due to their potent herbicidal effects without toxic to human and animals. We have designed and synthesized 4,5,6,7-tetrahydroindazole, maleimide, and tetrahydrophthalimide compounds carrying diverse epoxide substituents at 5- position of the phenyl group. Their herbicidal activities were evaluated under submerged paddy conditions. These results showed that 4,5,6,7-tetrahydroindazole compounds gave potent herbicidal activities especially to ECHOR, MOOVA, and CYPSE at a relatively low rate of 16 g/ha and improved tolerance on rice compared to S-275 as a standard herbicide in this experiment.

• Korean Journal of Weed Science
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• v.30 no.2
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• pp.122-134
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• 2010

We investigated the levels of resistance and accumulation of terapyrroles, reactive oxygen species, lipid peroxidation, and antioxidative enzymes for reasons of growth reduction in herbicide-transgenic rice overexpressing Myxococcus xanthus, Arabidopsis thaliana, and human protoporphyrinogen oxidase (Protox) genes. The transgenic rice overexpressing M. xanthus (MX, MX1, PX), A. thaliana (AP31, AP36, AP37), and human (H45, H48, H49) Protox genes showed 43~65, 41~72 and 17~70-fold more resistance to oxyfluorfen, respectively, than the wild type. Among transgenic rice lines overexpressing Protox genes, several lines showed normal growth compared with the wild type, but several lines showed in reduction of plant height and shoot fresh weight under different light conditions. However, reduction of plant height of AP37 was much higher than other lines for the experimental period. On the other hand, the reduction of plant height and shoot fresh weight in the transgenic rice was higher in high light condition than in low light condition. Enhanced levels of Proto IX were observed in transgenic lines AP31, AP37, and H48 at 7 days after seeding (DAS) and transgenic lines PX, AP37, and H48 at 14 DAS relative to wild type. There were no differences in Mg-Proto IX of transgenic lines except for H41 and H48 and Mg-Proto IX monomethyl ester of transgenic lines except for MX, MX1, and PX. Although accumulation of tetrapyrrole intermediates was observed in transgenic lines, their tetrapyrrole accumulation levels were not enough to inhibit growth of transgenic rice. There were no differences in reactive oxygen species, MDA, ALA synthesizing capacity, and chlorophyll between transgenic lines and wild type indicating that accumulated tetrapyrrole intermediate were apparently not high enough to inhibit growth of transgenic rice. Therefore, the growth reduction in certain transgenic lines may not be caused by a single factor such as Proto IX, but by interaction of many other factors.

• The Korean Journal of Pesticide Science
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• v.7 no.2
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• pp.100-107
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• 2003

To improve the growth inhibitory activity against the shoot and root of rice plant (Oryza sativa L) and barnyard grass (Echinochloa crus-galli), a series of N-substituted phenyl-3,4-dimethylmaleimdes derivatives as substrates were synthesized and then their the inhibitory activities of protoporphyrinogen oxidase (1.3.3.4), protox were measured. The quantitative structure-activity relationships (QSAR) between structures and the inhibitory activities were studied quantitatively using the 2D-QSAR method. And also, molecular sharp similarity between the substrate derivatives and protogen, substrare of protox enzyme were studied. The activities of the two plants indicated that barnyard grass had a higher activity than the rice plant and their correlation relationships have shown in proportion for each. Accordingly, the results of SARs suggest that the electron donating groups as $R_2=Sub.X$ group will bind to phenyl ring because the bigger surface area of negative charged atoms in the substrate molecule derivatives may increase to the higher the activity against barnyard grass. Based on the molecular shape similarity, when the derivatives and protogen, subsbrate of protox enzyme were superimposed by atom fitting, the similarity indices (S) were above 0.8 level but the correlation coefficients (r) between S values and the activities showed not good.

• Korean Journal of Weed Science
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• v.30 no.2
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• pp.111-121
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• 2010

Differential tolerance to protoporphyrinogen oxidase (Protox)-inhibiting herbicides, oxyfluorfen was observed between leaf ages in squash. Physiological responses to oxyfluorfen, including leaf injury, cellular leakage, accumulation of tetrapyrroles, and antioxidative enzymes activity, were investigated in leaf age classes of squash to identify mechanisms of oxyfluorfen tolerance. Leaf 1, 2, and 3 injuries for Joongangaehobak were >10,000, 1,286, and 1.6-fold higher than that of leaf 4, after treatment of oxyfluorfen. On the other hand, leaf 1, 2, and 3 injuries for Sintowjahobak were 725, 366, and >0.6-fold higher than that of leaf 4, after treatment of oxyfluorfen. However, in contrast to oxyfluorfen treatment results, leaf injury of squash leaf 4 treated with paraquat was much smaller than in leaves 1, 2 and 3. Electrolyte leakage from the tissues treated with oxyfluorfen was higher in the youngest leaf (Leaf 4) than in the older leaves 1, 2, and 3. Differential leaf response to oxyfluorfen of squash appears to be due in large part to differences in protoporphyrin IX (Proto IX), Mg-Proto IX, and Mg-Proto IX monomethyl ester accumulation in treated leaves. In contrast, leaf 4 had higher activities of superoxide dismutase, catalase, peroxidase, ascorbate peroxidase, and glutathione reductase than leaf 1 after treatment with oxyfluorfen. However, the induction in antioxidant activity in leaf 4 was not enough to overcome the toxic effects of a Protox inhibitor, oxyfluorfen, so the leaf eventually died.

• Korean Journal of Weed Science
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• v.32 no.1
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• pp.35-43
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• 2012

The objective of this research was to confirm the difference in physiological responses to environmental stresses such as chilling, high temperature, NaCl, and chemical stress (paraquat) in Protox inhibitor resistant-transgenic rice (MX, PX, and AP37) and its non-transgenic counterpart (WT). Transgenic and non-transgenic rice plants were exposed to a chilling temperature of $5^{\circ}C$ for 1 day or a high temperature of $45^{\circ}C$ for 4 days and allowed to recover at $25^{\circ}C$ for 6 days after the chilling treatment or 8 days after the high temperature treatment. Leaf injury, shoot fresh weight, porphyrin biosynthesis substances, and chlorophyll content were investigated in transgenic and non-transgenic rice at 6 days after 0.5% and 1% NaCl treatments or at 5 days after 0~300 ${\mu}M$ paraquat treatments. No significant difference in leaf injury and shoot fresh weight were observed between transgenic and non-transgenic rice during chilling and recovery. Plant height and shoot fresh weight were also similar between transgenic and non-transgenic rice during the high temperature and recovery period (0~5 days). However, plant height and shoot fresh weight in transgenic rice line MX and PX were lower than in non-transgenic rice at 6 days for recovery. Leaf injury, chlorophyll, and Mg-Proto IX ME contents had no significant difference between transgenic rice and non-transgenic rice after NaCl treatment, but Proto IX content for AP37 and shoot fresh weight for PX and AP37 in 0.5% NaCl treatment were significantly reduced compared with non-transgenic rice. There was no difference in leaf injury and shoot fresh weight when comparing transgenic rice and non-transgenic rice after paraquat treatment. Although transgenic rice and non-transgenic rice showed a little difference at a particular measurement period in certain environmental stresses, there was generally no difference in physiological responses between transgenic rice and non-transgenic rice.

• The Korean Journal of Pesticide Science
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• v.9 no.1
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• pp.23-25
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• 2005

3-Chloro-2-(4-chloro-2-fluoro-5-hydroxyphenyl)-4,5,6,7-tetrahydro-2H-indazole, which is the key intermediate of cyclic imide type compounds such as EK 5439 and S-275 series, were practically synthesized by the procedure of hydrolysis of by-products and were produced in the reaction of 2-(2-fluoro-4-chloro-5-hydroxyphenyl)-2,3a,4,5,6,7-hexahydroindazol-3-one with phosgene.