Ha-Kyoung Lee;Jong-Hwan Shin;Seong-Chan Lee;You-Kyoung Han
Research in Plant Disease
/
v.29
no.4
/
pp.390-398
/
2023
Apple white rot, caused by Botryosphaeria dothidea, is one of the important diseases in Korea. B. dothidea can cause pre- and postharvest decay on apple fruit as well as canker and dieback of apple trees. In this study, we isolated bacteria from the trunk of apple trees and tested their antagonistic activity against B. dothidea. Five bacterial isolates (23-168, 23-169, 23-170, 23-172, and 23-173) were selected that were most effective at inhibiting the mycelial growth of the pathogens. The isolate 23-172 was identified as Bacillus amyloliquefaciens and four isolates 23-168, 23-169, 23-170, and 23-173 were identified as Bacillus velezensis by RNA polymerase beta subunit (rpoB) and DNA gyraseA subunit (gyrA) gene sequencing. All isolates showed strong antagonistic activity against B. dothidiea as well as Colletotrichum fructicola and Diaporthe eres. All isolates exhibited cellulolytic, proteolytic and phosphate solubilizing activities. In particular, two isolates 23-168, 23-169 were shown to significantly reduce the size of white rot lesions in pretreated apple fruits. These results will provide the basis for the development of a fungicide alternative for the control of white rot of apple.
The antagonistic fluorescent pseudomonas, which was isolated from continuous cropping rhizosphere of pepper and cucumber, was identified as Pseudomonas fluorescens (P.f.). For further study, transformant was derived from the isolated P.f. after spontaneous mutation to give antibiotic resistance to nalidixic acid and rifampicin as marked strain. Both P.f. and transformant strains were used for this study and the results obtained were summarized as follows. 1. One of the most effective antagonistic strain, KR164, was selected against F. solani, F. oxysporum, R. solani and this strain was identified and classified as Pseudomonas fluorescens biotype IV. 2. Transformant, KR1641, was derived from strain KR164 and both strains had the same biological and biochemical characteristics. 3, Mycelial lysis and abnormal mycelia of plant pathogenic fungi were microscopically observed after simultaneous culture of fungus and given bacterial strain. 4. The length of chinese cabbage to the autolyzed became longer with given bacterial strain in dark culture. 5. Percentage of germination, number of leaves, length of height, and length of root in chinese cabbage in pot experiment were improved by inoculation of given bacterial strain. 6. The number of given bacterial strain kept generally stable until 34 days after inoculation of itself in pot experiment. Inoculation of given bacterial strain did affect the number of plant disease fungi to be decreased but did not affect the number of other bacteria, Bacillus, in pot experiment.
This experiment was carried out to examine physiological and cultural characteristics of two strains ASI 19003, 'Poplar field-cap mushroom' Agrocybe cylindracea, and ASI 19016, 'Chaxingo' A. chaxingu, at the bottle cultivation which have very similar morphological characteristics in genus Agrocybe. There was significant difference between the physiological and cultural characteristics of ASI19003 and ASI19016. The optimal temperature for the hyphal growth was $28^{\circ}C$ in the strain ASI19003 and $30^{\circ}C$ in ASI19016. The optimal pH was not different in two strains and these strains grew well at pH 5.5~7.0. But the optimal pH in the submerged culture was 5.5 in ASI19003 and 5.0 in ASI19016. Especially, hyphal growth of the strain ASI19016 was very poor at pH 6.0~7.3. The optimal carbon source for the growth was lactose in the strain ASI19003 and fructose in ASI19016, and nitrate sources were asparagine, alanine, and glycine in the strain ASI19003, and ammonium tartrate, asparagine, glycine, and alanine in ASI19016, respectively. The periods of incubation and fruiting body formation in the bottle cultivation during the spring were 27 and 13 days in the strain ASI19003, 29 and 17 days in ASI19016. The yields of fruit body were 114 g per bottle (850 $m{\ell}$ volume) in the strain ASI19003 and 100 g in ASI19016. In the summer, the periods of hyphal incubation and fruiting body formation were 29 and 11 days in the strain ASI19003, 30 and 12 days in ASI19016. The color of the cap in the ASI19003 strain according to temperature increase during the fruit body development become more pale, but the strain ASI19016 kept dark color relative to ASI19003. The fruiting body formation of the strain ASI19016 was faster than that of ASI19003. Accordingly, the cultivation of A. cylindracea ASI19003 during the spring, fall and winter, and A. chaxingu ASI19016 during the summer can keep high quality and stable supply all year round of these mushrooms.
The objectives of this study were to understand the characteristics of Cd and Pb accumulation in various tissues of poplar species and the effects of ectomycorrhizal inoculation on the accumulation of above two heavy metals in the tissues. The mycelial inoculum of ectomycorrizal fungus, Pisolithus tinctorius was produced in peatmoss and vermiculite mixtures, and inoculated into potted soil with fresh cuttings of four species of Populus, P. alba ${\times}$ glandulosa (Pag). P. koreana ${\times}$ nigra var. italica (Pkn), P. nigra ${\times}$ maximowiczii(Pnm), and P. euramericana(Pe). The potted soils were added with 0, 30, and 80 ppm Cd, and 0, 50, and 300 ppm Pb. The cuttings were grown outdoors for about five months until the plants were harvested for measurement of growth, mycorrhizal infection, and metal contents in leaves, stems, and roots. The total dry weight of Pe treated with Cd and Pb was increased by mycorrhizal inoculation, while that of three other species was not affected by the inoculation. Cd was accumulated in the highest concentration in Pag and its concentration was increased by four times by mycorrhizal inoculation. The Pag accumulated Cd in the highest concentration in the leaves, while three other species accumulated Cd most in the roots. Pb was accumulated in the highest concentration in the roots of all the four species, while Pkn accumulated Pb in the leaves as much as in the roots. Without mycorrhizal inoculation Pe accumulated Pb most among the four species, while Pkn with mycorrhizal inoculation accumulated Pb two times more than in the same species without mycorrhizal inoculation. It was concluded that Pag was the most effective species among the four poplar species in Cd absorption from contaminated soil, and that Pe instead effectively absorbed Pb. Mycorrhizal inoculation increased the Cd accumulation in the tissues by four times in Pag and also increased Pb accumulation by two times in Pkn, with leaves being the major sites of metal accumulation. It may be possible to use these two poplar species in remedying the metal in the soil through the raking and removing the litter out of the contaminate site.
The purposes of this study were to evaluate the effects of acid rain on tree growth and on the mycorrhizal formation and the effects of mycorrhizae on the host tolerance to acid rain. Simulated acid rain was applied for five months to Pinus $rigida{\times}taeda$ seedlings in pots inoculated with Pisolithus tinctorius (Pt) and Suillus luteus (Sl). Mycelial inocula of Pt and Sl were either mixed with entire pot soil (Mix) or casted as a band (Band) after soil sterilization. Three pH levels of acid rain (pH 3.0, 4.5 and 6.4 adjusted by 3 : 1 mixture of sulfuric and nitric acids) were tested. Pt-Mix was most effective in growth stimulation and resulted in 45-90% increase in tree height in sandy loam. Pt-Band was less effective than Pt-Mix in growth stimulation and mycorrhizal formation. Simulated acid rain at pH 4.5 stimulated height growth by 10-55%, while acid rain at pH 3.0 did not significantly affect the height growth. The top/root ratio was increased by pH 4.5 treatment, while pH 3.0 treatment reduced it. Mycorrhizal infection rate was not affected by acid rain. Pt inoculation reduced acid-induced leaf injury by 28-58% in both pH 3.0 and 4.5 compared with un-inoculated plants. Sl was also effective in growth enhancement, but was less effective than Pt in both mycorrhizal infection and reducing leaf injury.
Pinus koraiensis (Pk), P. rigida (Pr) and P. rigida ${\times}$ P. taeda (Pr. t) seedlings in a bare-rooted nursery were artificially inoculated with Pisolithus tinctorius (Pt) and Thelephora terrestris (Tt) to test long term effects of ectomycorrhizal inoculation on host growth. Mycelial inocula of Pt and Tt were mass-cultured in vermiculite-peatmoss mixture and introduced into fumigated nursery soil before seed sowing. Bare-rooted, inoculated seedlings at one to four years of age were outplanted to the field with $P_2O_5$ content of 25 ppm in soil. At the time of outplanting, Pk seedlings(4 years old), Pr seedlings(2 years old), and Pr.t seedlings(1 year old) all infected by Pt were significantly taller by 28%. 26%, and 77%, respectively, than controlled seedlings infected by natural population of mycorrhizal fungi in the non-fumigated plot. Ten years after inoculation or six to nine years after outplanting, Pk seedlings inoculated with Pt were significantly taller by 9% Pr.t seedlings significantly taller by 18%, and Pr slightly Caller by 2%(not significant) than controlled seedlings, suggesting that the stimulatory effect of Pt on host growth gradually declined or became minimal after outplanting. Tt failed to stimulate host growth either in the nursery or in the field, and the survival rate of outplanted seedlings was not different among fungal treatments. Considerable loss of the infected root system during lifting the seedlings for outplanting would be the primary cause of the reduced effect of Pt in the field. Pt infected more than 90% of the fine roots in the fumigated nursery during the first growing season, but Pt assumed to fail to compete successfully with natural population of ectomycorrhizal fungi in the field. It is necessary to select other mycorrhizal fungi which adapt well in both nursery and field.
Fluxapyroxad is classified as carboxamide fungicide that inhibits succinate dehydrogenase in complex II of mitochondrial respiratory chain, which results in inhibition of mycelial growth within the fungus target species. This study was carried out to assure the safety of fluxapyroxad residues in agricultural products by developing an official analytical method. A new, reliable analytical method was developed and validated using High Performance liquid Chromatograph-UV/visible detector (HPLC-UVD) for the determination of fluxapyroxad residues. The fluxapyroxad residues in samples were extracted with acetonitrile, partitioned with dichloromethane, and then purified with silica solid phase extraction (SPE) cartridge. Correlation coefficient($R^2$) of fluxapyroxad standard solution was 0.9999. The method was validated using apple, pear, peanut, pepper, hulled rice, potato, and soybean spiked with fluxapyroxad at 0.05 and 0.5 mg/kg. Average recoveries were 80.6~114.0% with relative standard deviation less than 10%, and limit of detection (LOD) and limit of quantification (LOQ) were 0.01 and 0.05 mg/kg, respectively. All validation parameters were followed with Codex guideline (CAC/GL 40). LC-MS (Liquid Chromatograph-Mass Spectrometer) was also applied to confirm the analytical method. Base on these results, this method was found to be appropriate fluxapyroxad residue determination and can be used as the official method of analysis.
This study was conducted to investigate the morphological characteristics and cultural conditions for artificial fruiting body(synnemata) production of Paecilomyces japonica. In the morphological characteristics of P. japonica, the size of it's conidia was ranged from $5.0{\sim}1.5\;to\;7.9{\sim}2.4\;{\mu}m$. The artificial fruiting body showed yellow in color, shape was confirmed ellipsoidal or obovoid type, and the length was $50.6{\sim}104.5\;mm$. The mycelial growth on the PDA medium treated with pH7, at $25^{\circ}C$ was superior to that of other treatments. The formation period of an artificial fruiting body of P. japonica treated with polypropylene and glass bottle culture was 30 days and 50 days, respectively. The length and number of fruiting body was longer and higher in the polypropylene bottle culture than those of the glass bottle culture. As the results, the artificial fruiting body production in the polypropylene bottle increased 1.2g per bottle compared to that of the glass bottle. It also increased in $100{\sim}400\;lx$ illumination, whereas the elongation of synnemata, pinheading and fruiting body growth were inhibited by continuous use of 900 lx illumination. The results of these experiment indicated that fruiting body formation seemed to be lower as the light intensity increased. The fruiting body formation was also dependent on the light color. There was a higher incidence in red color light and fluorescent light treatment than that of incandescent and blue color light. The fruiting body of the naked barley medium had so much better growth compared to other media that it would be able to use for it's production. The growth of fruiting body was affected by $CO_2$ concentration. It increased after putting the lid on the bottle.
This study was carried out in an attempt to select chemicals such as zeolite granules, chitosan, wood-vinegar, $Ca(OCl)_2$, NaOCl, against bacterial brown blotch caused by Pseudomonas tolaasii on Flammulina velutipes in laboratory and mushroom unit. Results obtained using these chemicals were summarized. Inhibitory effect on $Ca(OCl)_2$ and wood-vinegar with 0.5% concentration was shown to caual pathogen, P. tolaasii, by slight damage on the mycelial growth of F. velutipes. Those materials were recognized as promising one for control of bacterial brown bloth on F. velutipes. Disease incidence in control was the highest value as 43.8%; whereas chemical treatment was estimated as 14.6% with 0.5% of $Ca(OCl)_2$. Disease incidence was inhibited about 20% in chemical treatment with 0.5% of wood-vinegar, 1.0% of $Ca(OCl)_2$, and 26.1% with 1.0% of wood-vinegar. For the inhibition effects of chemical concentrations, it was effective at the low concentration which was 0.5% rather than that of 1.0%. Quality of mushrooms was significantly improved, and yields was also increased by 30% in the chemical treatment. In case of $Ca(OCl)_2$ treatment at 1.0% concentration, the yields was increased by 35.6% showed the highest value among tested chemicals. However, the overall effects including disease incidence, quality and yields suggests that $Ca(OCl)_2$ is relatively more effective than wood-vinegar, and the optimal concentration controlling the disease was $0.5{\sim}1.0%$ with $Ca(OCl)_2$ and 0.5% with wood-vinegar.
Sung, Jae-Mo;Yang, Kun-Joo;Kim, Soo-Ho;Harrington, Tom
The Korean Journal of Mycology
/
v.25
no.1
s.80
/
pp.46-67
/
1997
From 1985 to 1993, we collected 20 isolates throughout Kangwon and obtained 6 isolates from other sources. A. mellea formed rhizomorph actively, and some of A. osroyae were poor in the formation of rhizomorph and some without formation of rhizomorph. A. tabescens was active in the growth of aerial mycelium and poor in the development of rhizomorph. In A. gallica, the mycelium development among the isolates were variable greatly, and especially in isolate A8(KNU-250), the mycelial development was similar to that of A. osroyae, but A8(KNU-250) showed the feature of A. gallica to change medium into brown color. In PCR-RFLP analysis of the IGS region in rDNA, the homology between each isolate in the A. mellea and A. ostoyae showed 100% homology. A. tabescens showed $0.919{\sim}0.974$ homology, and A. gallica showed $0.619{\sim}1.000$ homology. A19 and A12 showed 100% homology as the same group, but compared with other subgroups they showed less than 10% homology as $0.051{\sim}0.108$ value. In RAPD analyses, the isolates of A. mellea showed high homology among themselves as $0.983{\sim}1.000$, and A. ostoyae also showed high similarity. The homology between isolates of A. tabescens showed $0.594{\sim}0.953$ value because A. gallica showed $0.280{\sim}0.733$ value, and the variations between isolates were greater than other species. Especially, A19 and A22 were identified as new novel group which were remoted from other groups, and the homology between these two isolates showed 0.921 value, and the genetic similarity between these groups and other 4 groups showed less than 7% as $0.012{\sim}0.069$ value. Of 5 species identified in this study, 4 species were identified as A. mellea, A. ostoyae, A. tabescens, and A. gallica that were already reported ones and 1 species was suggested as a new specie in Korea.
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