• Mycobiology
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• v.36 no.2
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• pp.81-87
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• 2008

The optimal conditions for mycelial growth of Phellinus linteus ATCC 26710 were determined to be a log length of 20 cm, temperature of $30^{\circ}C$ and pH of 6.0. Mycelial growth was excellent on the mushroom complete medium, and was optimal when sucrose, man nose and glucose were supplied as carbon sources. Potassium nitrate and sodium nitrate as nitrogen sources supported good mycelial growth. To evaluate P. linteus mycelial colonization on logs, sterilized short log inoculation, drilling inoculation and log-end sandwich inoculation techniques were used. Only sterilized short log inoculation produced good mycelial colonization. Initial mycelial growth and full mycelial colonization were best on 20 cm logs having 42% moisture content. The initial mycelial growth of P. linteus was accelerated over 12hr of sterilization. Basidiocarp formation was optimal using a burying method of logs after $5{\sim}6$ months, and fruiting body formation was superior in cultivation house conditions of $31{\sim}35^{\circ}C$ and in excess of 96% relative humidity.

• Research in Plant Disease
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• v.21 no.3
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• pp.201-207
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• 2015

This study was conducted to establish a simple mass-screening method for resistant melon to Fusarium wilt caused by Fusarium oxysporum f. sp. melonis (FOM). Root-dipping inoculation method has been used to investigate resistance of melon plants to Fusarium wilt. However, the inoculation method requires a lot of labor and time because of complicate procedure. To develop a simple screening method on melon Fusarium wilt, occurrence of Fusarium wilt on susceptible and resistant cultivars of melon according to inoculation method including root-dipping, soil-drenching, tip, and scalpel methods was investigated. Scalpel and tip methods showed more clear resistant and susceptible responses in the melon cultivars than root-dipping inoculation method, but tip method represented slightly variable disease severity. In contrast, in the case of soil-drenching inoculation method, disease severity of the susceptible cultivars was very low. Thus we selected scalpel method as inoculation method of a simple screening method for melon Fusarium wilt. By using the scalpel inoculation method, resistance degrees of the cultivars according to incubation temperature after inoculation (25 and $30^{\circ}C$) and inoculum concentration ($1{\times}10^6$ and $1{\times}10^7conidia/ml$) were measured. The resistance or susceptibility of the cultivars was hardly affected by all the tested conditions. To look into the effectiveness of scalpel inoculation methods, resistance of 22 commercial melon cultivars to FOM was compare with root-dipping inoculation method. When the melon cultivars were inoculated by scalpel method, resistance responses of all the tested cultivars were clearly distinguished as by root-dipping method. Taken together, we suggest that an efficient simple mass-screening method for resistant melon plant to Fusarium wilt is to sow the seeds of melon in a pot (70 ml of soil) and to grow the seedlings in a greenhouse ($25{\pm}5^{\circ}C$) for 7 days, to cut the root of seedlings with a scalpel and then pour a 10 ml-aliquot of the spore suspension of $1{\times}10^6conidia/ml$ on soil. The infected plants were cultivated in a growth room at 25 to $30^{\circ}C$ for about 3 weeks with 12-hr light a day.

• The Plant Pathology Journal
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• v.30 no.1
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• pp.25-32
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• 2014

Fusarium head blight (FHB; scab) caused mainly by Fusarium graminearum is a devastating disease of wheat and barley around the world. FHB causes yield reductions and contamination of grain with trichothecene mycotoxins such as deoxynivalenol (DON) which are a major health concern for humans and animals. The objective of this research was to develop an easy seed or seedling inoculation assay, and to compare these assays with whole plant resistance of twenty-nine Korean winter wheat cultivars to FHB. The clip-dipping assay consists of cutting off the coleoptiles apex, dipping the coleoptiles apex in conidial suspension, covering in plastic bag for 3 days, and measuring the lengths of lesions 7 days after inoculation. There were significant cultivar differences after inoculation with F. graminearum in seedling relative to the controls. Correlation coefficients between the lesion lengths of clip-dipping inoculation and FHB Type II resistance from adult plants were significant (r=0.45; P<0.05). Results from two other seedling inoculation methods, spraying and pin-point inoculation, were not correlated with adult FHB resistance. Single linear correlation was not significant between seed germination assays (soaking and soak-dry) and FHB resistance (Type I and Type II), respectively. These results showed that clip-dipping inoculation method using F. graminearum may offer a real possibility of simple, rapid, and reliable for the early screening of FHB resistance in wheat.

• Journal of Microbiology and Biotechnology
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• v.11 no.4
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• pp.700-703
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• 2001

The influence of process conditions on highly branched glucooligosaccharides production by mixed culture of Leuconostoc mesenteroides ATCC 13146 and Lipomyces starkeyi ATCC 74054 was studied. We divided the batch culture fermentations into two groups according to inoculation method. One-point inoculation was performed by coinoculation of L. mesenteroides and L. starkeyi at the ration of 10 to 1, and two-point inoculation by L. mesenteroides inoculation first and L. starkeyi inoculation after L. mesenteroides grew to the end of the log phase of growth. Two-point inoculation improved the yield of oligosaccharide by 1.5 to 20 fold more than one-point inoculation. In this process, the highest yield of oligosaccharides (48% of theoretical yield) and productivity (0.85 g/l/h) were obtained with starch as an initial substrate for L. starkeyi growth. The estimated composition of the end product consisted of 31.5% oligosaccharides, 17.6% dextran, and 46.5% mannitol.

• The Plant Pathology Journal
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• v.37 no.5
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• pp.455-464
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• 2021

Forty-eight spring barley genotypes were evaluated for deoxynivalenol (DON) concentration under natural infection across 5 years at Harrington, Prince Edward Island. These genotypes were also evaluated for Fusarium head blight (FHB) severity and DON concentration under field nurseries with artificial inoculation of Fusarium graminearum by the grain spawn method across 2 years at Ottawa, Ontario, and one year at Hangzhou, China. Additionally, these genotypes were also evaluated for FHB severity under greenhouse conditions with artificial inoculation of F. graminearum by conidial suspension spray method across 3 years at Ottawa, Ontario. The objective of the study was to investigate if reactions of barley genotypes to artificial FHB inoculation correlate with reactions to natural FHB infection. DON concentration under natural infection was positively correlated with DON concentration (r = 0.47, P < 0.01) and FHB incidence (r = 0.56, P < 0.01) in the artificially inoculated nursery with grain spawn method. Therefore, the grain spawn method can be used to effectively screen for low DON. FHB severity, generated from greenhouse spray, however, was not correlated with DON concentration (r = 0.12, P > 0.05) under natural infection and it was not correlated with DON concentration (r = -0.23, P > 0.05) and FHB incidence (r = 0.19, P > 0.05) in the artificially inoculated nursery with grain spawn method. FHB severity, DON concentration, and yield were affected by year, genotype, and the genotype × year interaction. The effectiveness of greenhouse spray inoculation for indirect selection for low DON concentration requires further studies. Nine of the 48 genotypes were found to contain low DON under natural infection. Island barley had low DON and also had high yield.

• Journal of Sericultural and Entomological Science
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• v.44 no.2
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• pp.74-81
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• 2002

The optimal conditions for mycelial growth of P. linteus ASI 26011 were 25-30$^{\circ}C$ and pH 6.0, respectively. The mycelial growth of P. linteus was excellent on MCM medium. In case of carbon sources, the mycelial growth of P. linteus was best on the culture media that were contained with sucrose, mannose and glucose. Potassium nitrate and sodium nitrate were good for the mycelial growth of P. linteus as a nitrogen source. For comparison of the mycelial colonization of P. linteus on logs, several techniques of inoculation were tested; the sterilized short log inoculation, drilling inoculation and log-end sandwich inoculation. The mycelial colonization of P. linteus on logs was good in the treatment of sterilized short log inoculation, but poor in the traditional methods such as drilling inoculation and log-end sandwich. The initial mycelial growth and the full mycelial colonization of P. linteus were the best on 20 cm logs under the condition of 42% of moisture content in log. Also the initial mycelial growth of P. linteus was accelerated over 12 hours of sterilization. Burying method of logs after 5-6 months of incubation was the best for formation of basidiocarp of P. linteus. The formation of fruiting body of P. linteus was quite good in the cultivation house at the 31-35$^{\circ}C$ and over 96% of relative humidity.

• The Korean Journal of Mycology
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• v.27 no.6 s.93
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• pp.378-382
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• 1999

This study was carried out to improve an artificial culture techniques and antioxidative activity of the crude extract isolated from sclerotia of Poria cocos(Fr.) Wolf. In the test of different spawns and inoculation method, the sclerotia formation, number of sclerotia and production yield were excellent in the both sides inoculation method of log spawn, whereas the both side inoculation method of sawdust spawn was poor in sclerotia formation and yield. The optimal spawn and inoculation method for the quality and productivity of P. cocos was in the order of log spawn (both sides inoculation > log spawn(cutting section inoculation) > sclerotia (both sides inoculation) > sawdust spawn (both sides inoculation). The physiological activity substance, crude extract content of P. cocos NIAST 13007 was about 83%. As the concentration of crude extracts increased, the relative viscosity tended to be increased. However, as the concentration of sodium chloride increased, the relative viscosity did not affected. In antioxidative activities, electron donating ability (EDA) of P. cocos was about 10% of butylated hydroxytoluene (BHT). Thiobarbituric acid (TBA) value was similar to that of the vitamin C, however the peroxide value (POV) was lower than those of BHT and vitamin C.

• The Plant Pathology Journal
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• v.39 no.2
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• pp.228-233
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• 2023

Two pear cultivars with different degrees of resistance to Venturia nashicola were evaluated on the basis of a disease severity rating for pear scab resistance under controlled environmental condition. Two inoculation techniques were tested: the procedure for inoculation by dropping conidia suspension of V. nashicola; the procedure by deposition of agar plug on the abaxial surface of pear leaves. All tested cultivars resulted in blight symptoms on the inoculated leaves and became spread to uninoculated region or other leaves. Although both methods provide satisfactory infection of V. nashicola on pear leaves, the mycelial plug method of inoculation was more reliable than the spray inoculation method for the evaluation of pear scab disease resistance. The incubation period of V. nashicola in the resistant pear cultivar, Greensis was longer than that in the susceptible cultivar, Hwasan.

• Journal of Plant Biotechnology
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• v.43 no.2
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• pp.240-247
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• 2016

In this study, routinely used transformation method, which includes transferring explants onto co-cultivation medium after inoculating them with bacterial solution for a while, was compared with 3 different inoculation methods. In every 3 methods, hypocotyl explants excised from 7-day-old sterile flax seedlings having cotyledon leaves and no root system dried under air flow in sterile cabin for 35 min were inoculated with different volumes of bacterial solution at different inoculation periods. GV2260 line of Agrobacterium tumefaciens having 'pBIN 19' plasmid containing npt II (neomycin phosphotransferase II) gene and GUS reporter gene was used in transformation studies. After inoculation, hypocotyl segments of seedlings (0.5 cm in length) - were excised and left to co-cultivation for 2 days. Then, explants were transferred to regeneration medium supplemented with different antibiotics. The presence of npt-II and GUS genes in transformants was confirmed by PCR and GUS analysis. The highest results in all characters examined in all cultivars were obtained from the 2 inoculation method in which hypocotyls excised from seedlings inoculated with $500{\mu}l$ of bacterial solution after drying in sterile cabin for 35 min were used.

• Journal of Microbiology and Biotechnology
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• v.22 no.7
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• pp.960-966
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• 2012

The diversity and composition of yeast populations may greatly impact wine quality. This study investigated the yeast microbiota in two different types of wine fermentations: direct inoculation of a commercial starter versus pied de cuve method at an industrial scale. The pied de cuve fermentation entailed growth of the commercial inoculum used in the direct inoculation fermentation for further inoculation of additional fermentations. Yeast isolates were collected from different stages of wine fermentation and identified to the species level using Wallersterin Laboratory nutrient (WLN) agar followed by analysis of the 26S rDNA D1/D2 domain. Genetic characteristics of the Saccharomyces cerevisiae strains were assessed by a rapid PCR-based method, relying on the amplification of interdelta sequences. A total of 412 yeast colonies were obtained from all fermentations and eight different WL morphotypes were observed. Non-Saccharomyces yeast mainly appeared in the grape must and at the early stages of wine fermentation. S. cerevisiae was the dominant yeast species using both fermentation techniques. Seven distinguishing interdelta sequence patterns were found among S. cerevisiae strains, and the inoculated commercial starter, AWRI 796, dominated all stages in both direct inoculation and pied de cuve fermentations. This study revealed that S. cerevisiae was the dominant species and an inoculated starter could dominate fermentations with the pied de cuve method under controlled conditions.