• The Plant Pathology Journal
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• v.38 no.6
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• pp.679-684
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• 2022

Rice blast is one of the most destructive diseases of rice worldwide, and the causative agent is the filamentous ascomycete Magnaporthe oryzae. With the successful cloning of more and more avirulence genes from M. oryzae, the direct extraction of M. oryzae genomic DNA from infected rice tissue would be useful alternative for rapid monitoring of changes of avirulence genes without isolation and cultivation of the pathogen. In this study, a fast, low-cost and reliable method for DNA preparation of M. oryzae from a small piece of infected single rice leaf or neck lesion was established. This single step method only required 10 min for DNA preparation and conventional chemical reagents commonly found in the laboratory. The AvrPik and AvrPi9 genes were successfully amplified with the prepared DNA. The expected DNA fragments from 570 bp to 1,139 bp could be amplified even three months after DNA preparation. This method was also suitable for DNA preparation from M. oryzae strains stored on the filter paper. All together these results indicate that the DNA preparation method established in this study is reliable, and could meet the basic needs for polymerase chain reaction-based analysis of M. oryzae.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.150.3-151
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• 2003

To analyze the genome of Cucumber mosaic virus(CMV) in pepper, we developed a new extraction method for double-stranded RNA(dsRNA). To isolate the dsRNA, 0.1g of pepper leaves homogenized with 1ml of 5${\times}$EXB extraction buffer[0.5M glycin, 0.5M NaCl, 5mM EDTA(pH9.0/NaOH), 10％ Sodium N-lauryl salcosinate(NLS), 10％ Sodium dodecylsulfate(SDS)] and purified with the 1/4 volume of phenol： chloroform： isoamylalcohol(25：24：1). dsRNAs from the aqueous phase was precipitated with isopropanol. This procedure was able to detect a minimal amount of dsRNA from CMV infected plant tissue and to distinguish different CMV satellite RNAs by polyacrylamide gel electrophoresis(PAGE). Moreover, this method can be applied CMV infected in pepper or Rice dwarf virus (RDV) infected rice.

• Korean journal of applied entomology
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• v.21 no.3 s.52
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• pp.113-122
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• 1982

Appressorial formation of Pyricularia oryzae on leaves showed no marked difference between highly resistant Tongil and susceptible Norm No. 6. Race N-2 of the blast fungus penetrated directly into motor cells of susceptible cultivar Norm No. 6, later extensively spreading hyphae were developed, while in the cultivar Tongil, after penetration, no further hyphal extension resulted. In discoloration of infected tissues, the highly resistant cultivar Tongil not only discolored rapidly, but also the percentage of discolored cells was higher than the susceptible cultivars, Jinheung and Norm No. 6. The respiratory rate, was generally higher in infected tissue than in healthy tissue. No significant difference in the respiration rate of resistant Suwon No. 180 was not found between the infected and healthy leaf tissue, whereas, in susceptible Jinheung, a marked increase in respiratory rate was caused by blast infection. The respiratory rate increased at the appearance of the first visible symptom in all cultivars resistant or susceptible. Higher peroxidase activity was found in infected tissues as compared with healthy tissue. Peroxidase activity increased in resistant and susceptible reactions. Particularly, in resistant reaction, the increase of the activity was more pronounced. In highly resistant reaction, there was no difference in peroxidase activity between healthy and infected tissues. Ascorbic acid oxidase, hydroquinone oxidase and catechol oxidase had the same trend as the peroxidase. In contrast, activity of catalase rather decreased in leaf tissues infected with compatible races of the fungus.

• Research in Plant Disease
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• v.21 no.2
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• pp.103-106
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• 2015

Isolates of the rice blast fungus show a range of tissue-specificities infecting leaves, nodes, neck and panicles. Although neck and panicle blast cause significantly greater yield losses than the leaf blast, virulence tests of the blast isolates have been performed only rice leaves instead of neck and panicles. In this study, we have developed a virulence test method for neck and panicle blast. We selected three representative isolates from each of leaf, neck, and panicle blast. We observed that severe disease lesions developed on the neck and the panicles when the infected rice plants were incubated in a dew chamber for 48 h instead of 24 h when tested on leaves. Unlike the leaf blast, a typical lesion on the neck and panicles appeared after 14 days post-infection as opposed to 7 days with leaf blast. This method will be applied to examine tissue-specificity of the rice blast fungus isolates.

• The Plant Pathology Journal
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• v.16 no.5
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• pp.257-263
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• 2000

Infection with rice blast fungus (Magnaporthe grisea) significantly reduced foliar net photosynthesis (A) of rice cultivars: Ilpoom, Hwasung, and Choochung in greenhouse experiments. By measuring the amount of diseased leaf area with a computer image analysis system, the relation between disease severity (DS) and net photosynthetic rate was curvilinearly correlated (r=0.679). Diseased leaves with 35% blast symptom can be predicted to have a 50% reduction of photosynthesis. The disease severity was linearly correlated (r=0.478) with total chlorophyll (chlorophyll a and chlorophyll b) per unit leaf area(TC). Light use efficiency was reduced by the fungal infection according to the light response curves. However, dark respiration (Rd) did not change after the fungal infection (p=0.526). Since the percent of reduction in photosynthesis greatly exceeded the percent of leaf area covered by blast lesions, loss of photosynthetic tissue on an area basis could not by itself account for the reduced photosynthesis. Quantitative photosynthetic reduction can be partially explained by decreasing TC, but cannot be explained by decreasing Rd. By photosynthesis (A)-internal CO$_2$ concentration (C$_i$ curve analysis, it was suggested that the fungal infection reduced ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activity, ribulose-1,5-bisphosphate (RuBP) regeneration, and inorganic phosphate regeneration. Thus, the reduction of photosynthesis by blast infection was associated with decreased TC and biochemical capacity, which comprises all carbon metabolism after CO$_2$ enters through the stomata.

• Korean Journal Plant Pathology
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• v.1 no.3
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• pp.178-183
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• 1985

The present researches were carried out to investigate the peroxidase activity in association with the reactions of the 4 cultivars of rice plant, Nagdong, Jinheung, Nongbaek and Taebaek to Pyricularia oryzae race KJ-I0l and KJ-301. Although the peroxidase activity was increased during the growth of the rice seedlings, the significant difference in the activity was not found among 4 cultivars. After inoculation of the fungus, the peroxidase activity was enhanced in diseased leaves, being considerably higher in the compatible than in the incompatible cultivars. The isozyme bands of peroxidases observed in mycelium of rice blast fungus were not found in the diseased leaves on the gel electrophoresis. The peroxidase activity was not affected by the increased application of nitrogenous fertilizer.

• Korean Journal of Agricultural Science
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• v.12 no.2
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• pp.206-241
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• 1985

Attempts to search infection period, infection speed in the tissue of neck blast of rice plant, location of inoculum source and effects of several conditions about the leaf sheath of rice plants for neck blast incidence have been made. 1. The most infectious period for neck blast incidence was the booting stage just before heading date, and most of necks have been infected during the booting stage and on heading date. But $Indica{\times}Japonica$ hybrid varieties had shown always high possibility for infection after booting stage. 2. Incubation period for neck blast of rice plants under natural conditions had rather a long period ranging from 10 to 22 days. Under artificial inoculation condition incubation period in the young panicle was shorter than in the old panicle. Panicles that emerged from the sheath of flag leaf had long incubation period, with a low infection rate and they also shown slow infection speed in the tissue. 3. Considering the incubation period of neck blast of rice plant, we assumed that the most effective application periods of chemicals are 5-10 days for immediate effective chemicals and 10-15 days for slow effective chemicals before heading. 4. Infiltration of conidia into the leaf sheath of rice plant carried out by saturation effect with water through the suture of the upper three leaves. The number of conidia observed in the leaf sheath during the booting stage were higher than those in the leaf sheath during other stages. Ligule had protected to infiltrate of conidia into the leaf sheath. 5. When conidia were infiltrated into the leaf sheath, the highest number of attached conidia was observed on the panicle base and panicle axis with hairs and degenerated panicle, which seemed to promote the infection of neck blast. 6. The lowest spore concentration for neck blast incidence was variable with rice varietal groups. $Indica{\times}Japonica$ hybrid varieties were infected easily compared to the Japonica type varieties, especially. The number of spores for neck blast incidence in $Indica{\times}Japonica$ hybrid varieties was less than 100 and disease index was higher also in $Indica{\times}Japonica$ hybrid than in Japonica type varieties. 7. Nitrogen content and silicate content were related with blast incidence in necks of rice plants in the different growing stage changed during growing period. Nitrogen content increased from booting stage to heading date and then decreased gradually as time passes. Silicate content increased from booting stage after heading with time. Change of these content promoted to increase neck blast infection. 8. Conidia moved to rice plant by ascending and desending dispersal and then attached on the rice plant. Conidia transfered horizontally was found very negligible. So we presumed that infection rate of neck blast was very low after emergence of panicle base from the leaf sheath. Also ascending air current by temperature difference between upper and lower side of rice plant seemed to increase the liberation of spores. 9. Conidial number of the blast fungus collected just before and after heading date was closely related with neck blast incidence. Lesions on three leaves from the top were closely related with neck blast incidence, because they had high potential for conidia formation of rice blast fungus and they were direct inoculum sources for neck blast. 10. The condition inside the leaf sheath was very favorable for the incidence of neck blast and the neck blast incidence in the leaf sheath increased as the level of fertilizer applied increased. Therefore, the infection rate of neck blast on the all panicle parts such as panicle base, panicle branches, spikelets, nodes, and internodes inside the leaf sheath didn't show differences due to varietal resistance or fertilizers applied. 11. Except for others among dominant species of fungi in the leaf sheath, only Gerlachia oryzae appeared to promote incidence of neck blast. It was assumed that days for heading of varieties were related with neck blast incidence.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.62 no.1
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• pp.1-8
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• 2017

We investigated the effect of silicate coating of rice seeds on bakanae disease incidence and the quality of seedlings raised in seedling boxes and transplanted into pots. The silicate-coated rice seed (SCS) was prepared as follows. Naturally infested rice seeds not previously subjected to any fungicidal treatment were dressed with a mixture of 25% silicic acid at pH 11 and 300-mesh zeolite powder at a ratio of 50 g dry seed - 9 mL silicic acid - 25 g zeolite powder. The following nursery conditions were provided : Early sowing, dense seeding in a glass house with mulching overnight and no artificial heating, which were the ideal conditions for determining the effect on the seed. The nursery plants were evaluated for Gibberella. fujikuroi infection or to determine the recovery to normal growth of infected nursery plants in the Wagner pot. Seedlings emerged 2-3 days earlier for the SCS than they did for the non-SCS control, while damping-off and bakanae disease incidence were remarkably reduced. Specifically, bakanae disease incidence in the SCS was limited to only 7.8% for 80 days after sowing, as compared to 91.6% of the non-SCS control. For the 45-days-old SCS nursery seedlings, the fresh weight was increased by 11% and was two times heavier, with only mild damage compared to that observed for non-SCS. Even after transplanting, SCS treatment contributed to a lower incidence of further infections and possibly to recovery of the seedlings to normal growth as compared to that observed in symptomatic plants in the pot. The active pathogenic macro-conidia and micro-conidia were considerably lower in the soil, root, and seedling sheath base of the SCS. In particular, the underdeveloped macro-conidia with straight oblong shape without intact septum were isolated in the SCS ; this phenotype is likely to be at a comparative etiological disadvantage when compared to that of typical active macro-conidia, which are slightly sickle-shaped with 3-7 intact septa. A active intact conidia with high inoculum potential were rarely observed in the tissue of the seedlings treated only in the SCS. We propose that promising result was likely achieved via inhibition of the development of intact pathogenic conidia, in concert with the aerobic, acidic conditions induced by the physiochemical characteristics associated with the air porosity of zeolite, alkalinity of silicate and the seed husk as a carbon source. In addition, the resistance of the healthy plants to pathogenic conidia was also important factor.