• Title/Summary/Keyword: Inoculation methods

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Effects of Seed Inoculation Methods on the Nodulation and the Growth of Alfalfa Seeding (근류균의 종자 접종방안의 차이가 근류형성 및 Alfalfa 유묘의 생장에 미치는 영향)

  • 이광회;이호진
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.26 no.2
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    • pp.192-197
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    • 1981
  • Alfalfa (Medicago sativa L. cv. Luna) seeded in agar was inoculated with two strains of Rhizobium meliloti isolated from root nodules of alfalfa for assessment of nodulation. The seedling growth after six weeks was remarkably increased by adding each rhizobia strains into agar media and also by nitrate application (70ug N/ml), but there was no significant difference among them. Nodulations started one week after inoculation and increased its numbers and sizes as seedling grew. Therefore, the two strains isolated from alfalfa root were concluded to be effective strains. For determining seed inoculation method the same cultivar was inoculated with both rhizobia strains using different inoculation methods such as broth-vacuum, peat-adhesive, peat & lime pelleting. They were seeded in pots of river sand and supplied with culture solution excluded nitrogen. The peat & lime pelleting was recognized the best method in both of nodulation and seedling growth after eight weeks growth. There were significant correlations between the weight of nodules and the shoot or root dry weight of alfalfa in both rhizobia strains.

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A New Method for Cultivation of Sclerotium of Grifola umbellata

  • Choi, Kyung-Dal;Lee, Kyung-Tae;Shim, Jae-Ouk;Lee, Youn-Su;Lee, Tae-Soo;Lee, Sang-Sun;Guo, Shun-Xing;Lee, Min-Woong
    • Mycobiology
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    • v.31 no.2
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    • pp.105-112
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    • 2003
  • Sclerotia of Grifola umbellata were cultivated by two methods such as burying and root inoculation methods. The sclerotia of G. umbellata produced by the burying method were $6.0{\sim}6.8{\times}3.4{\sim}4.6{\times}1.8{\sim}1.9cm$(Width$\times$Length$\times$Thickness) in size and $17.3{\sim}19.6g$ in weight, respectively. Their increase rate was $1.10{\times}1.12$ times. On the other hand, the sclerotia cultivated by the root inoculation method were $18.3{\sim}31.5{\times}12.5{\sim}26.4{\times}3.1{\sim}3.7cm(W{\times}L{\times}T)$ in size and $219.1{\sim}576.6g$ in weight, respectively. Their growth increment was $11.18{\sim}39.77$ times. The rhizomorphs of Armillaria mellea were developed with a high density under fallen leaves layer covering cultivation site, and distributed mainly between soil surface and soil depth of about 10 cm as well as colonized prominently on the inoculated wood logs. Fungal interaction between G. umbellata and A. mellea were observed mainly in the stage of white sclerotium of G. umbellata. The sclerotia of G. umbellata which were developed newly and harvested in the root inoculation method were twined with root hairs of host tree and rhizomorphs of A. mellea. The sclerotia of G. umbellata decomposing root hairs of host tree were confirmed through SEM examination. Physiochemical characteristics of soil in all cultivation sites had no significant differences. Soil pH were in the range of pH $3.98{\sim}4.40$. Organic matters were the range of $17.97{\sim}23.86%$ and moisture contents of soil were $12.00{\sim}18.20%$. Soil temperatures showed $12.9{\sim}13.8^{\circ}C$ in November and $22.0{\sim}23.9^{\circ}C$ in August, respectively. In conclusion, the root inoculation method seems to be a practical method for cultivating sclerotia of G. umbellata due to its many advantages such as simplicity of inoculation process, shortening of cultivation periods and facility of harvest.

Simple Mass-screening Methods for Resistance of Tomato to Fusarium oxysporum f. sp. lycopersici (토마토 시들음병에 대한 간편 대량 저항성 검정법)

  • Park, Myung Soo;Jang, Kyoung Soo;Choi, Yong Ho;Kim, Jin-Cheol;Choi, Gyung Ja
    • Horticultural Science & Technology
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    • v.31 no.1
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    • pp.110-116
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    • 2013
  • This study was carried out to establish the simple mass-screening methods for resistant tomato to Fusarium wilt caused by Fusarium oxysporum f. sp. lycopersici (FOL). Root dip inoculation method has been used in many studies on the resistance of tomato to disease. On the other hand, in mass-screening for resistant tomato to Fusarium wilt, the inoculation method is time-consuming and laborious procedure. Disease development of two FOL isolates on two cultivars of tomato according to inoculation method including root dip, tip and scalpel methods were investigated. In compatible interaction, tomato seedlings of each cultivar inoculated by tip method showed the lower and more variable disease severities than by root dip method. Whereas the seedlings by scalpel method represented clear resistant and susceptible responses to Fusarium wilt as root dip method. The resistance degree of each cultivar inoculated with FOL isolates by scalpel method was hardly affected by the tested incubation temperature and inoculum concentration. On the basis of the results, we suggest scalpel inoculation method as an efficient mass-screening method for resistant of tomato cultivars to Fusarium wilt. Roots of tomato seedlings at two-leaf stage grown in plastic cell tray were injured with scalpel and then spore suspension (more than $1{\times}10^7\;conidia{\cdot}mL^{-1}$) of FOL was poured directly on the roots. The infected plants were cultivated in a growth room at $25-30^{\circ}C$ for 4 weeks with 12-hours light a day.

Rhizobium meliloti Populations and Alfalfa Yields Due to Nitrogen Fertilization and Inoculation Methods at Cultivated Upland Soil (숙전(熟田)에서 질소시용(窒素施用)과 Rhizobium meliloti의 접종방법(接種方法)이 근류균(根瘤菌) 밀도(密度) 및 알팔파 수량(收量)에 미치는 영향(影響))

  • Kang, Ui-Gum;Jung, Yeun-Tae;Lee, Soo-Kwan;Ha, Ho-Sung
    • Korean Journal of Soil Science and Fertilizer
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    • v.25 no.3
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    • pp.275-280
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    • 1992
  • Inoculation responses of alfalfa[Medicago sativa(L.) Vernal] to Rhizobium meliloti with nitrogen fertilization were characterized by soil populations and plant yields from 1989 to 1991 at cultivated upland soil, which contained $3.1{\times}10$cells of R. meliloti/g.soil. The results obtained accordingly to fertilizing with 8Kg N/10a and differently inoculating the host such as initial and annual inoculation methods were as follows : 1. Soil populations of R. meliloti were increased more at no nitrogen fertilized condition compared to nitrogen fertilized condition to 2nd year experiment from 1st year that came up $1.6{\times}10^3$cells/g.soil, but at 3rd year the trend was reversed. Between inoculation methods, on the whole, the annual inoculation caused more populations, which were $2.0{\times}10^4$cells/g.soil in maximum number. And the populations declined after winter but recovered passing through summer season. 2. Alfalfa yields were mainly influenced by rhizobial populations rather than by fertilizing nitrogen showing a significant correlation(Y=0.36+0.287X, $r^2=0.58^{**}$) with the former. The increased extents of yields obtained by inoculation at no nitrogen and nitrogen fertilized conditions, respectively, were 66 and 10% in 1st : 13 and 20% in 2nd : 19 and 13% in 3rd year experiment with the initial inoculation, and were 66 and 10% in 1st ; 30 and 20% in 2nd : 35 and 36% in 3rd year experiment with the annual inoculation. 3. The results demonstrated the importance of inoculating, if possible, annual inoculating alfalfa to get much yields even at cultivated upland soil.

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Studies on the Varietal resistance to the rice stripe virus disease (줄무늬잎마름병(호엽고병)에 대한 벼품종의 저항성에 관한 연구)

  • Lee S. H.;Kim C. H.
    • Korean journal of applied entomology
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    • v.5_6
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    • pp.47-54
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    • 1968
  • In order to develop an effective control measure for the rice stripe disease, methods of testing for resistance and selection of resistant varieties among the leading varieties were investigated. For use as a parent in breeding for resistant variety to the disease, total of 410 rice varieties were tested. 1. Disease occurrence was higher at group inoculation than that of individual inoculation in comparing the inoculation methods. 2. In both methods, Lacrose responded susceptible; Zenith and St. No. 1 resistant, and the rest moderate. 3. Suseptible symptom type A was prevalant among the susceptible varieties, while resistant symptom type C was abundant among the resistant varieties, There was no difference between the inoculation methods. 4. 410 rice varieties tested could he divided into 3 groups as susceptible (21 varieties), moderate (377 varieties) and resistant (12 varieties). Resistant varieties wers St. No. 1 and 2, Shin-2, Gulfrose Bonnet, Arkrose, Sun Bonnet, Zenith, Yeechunchilichal, Norm-24, Opaikjoke, Yangchubatchal and Nonglimana-1, Nams-97,-149, -159, -216. -265, Iri-243, -265, Kanchuk -5, -7, -8, -10, -41, -43, -47, -50, Suwon-56, -77, Norin-22, Cod-4. Lacrose and Chukna were susceptible. 5. There was slight differance in the disease occurrence in regard to maturing period. However late varieties seem to be more resistant than early or medium varieties. The medium varieties seem to be susceptible. Most of the introduced varieties from foreign countries and the upland cultivated varieties were resistant. 6 Among the leading varieties, Shin-2 in Kangwon-Do was resistant, Kosi in Choongchung-Do, was susceptible, aad the others were moderate.

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Effect of the Mixed Culture of Bacillus subtilis and Lactobacillus plantarum on the Quality of Cheonggukjang (Bacillus subtilis와 Lactobacillus plantarum의 혼합배양이 청국장의 품질에 미치는 영향)

  • Ju, Kyung-Eun;Oh, Nam-Soon
    • Korean Journal of Food Science and Technology
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    • v.41 no.4
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    • pp.399-404
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    • 2009
  • The goal of this study was to improve the quality of cheonggukjang by the optimization of the inoculation methods of the Bacillus subtilis (B. subtilis) and Lactobacillus plantarum (L. plantarum) strains. In order to optimize the mixed cultivation of B. subtilis and L. plantarum, the B. subtilis strain was inoculated into steamed soybeans after cultivation of L. plantarum. Inoculation size of B. subtilis was changed to the simultaneous inoculation method in order to stimulate the growth of the L. plantarum in cheonggukjang. The viable cell count of L. plantarum increased from $2{\times}10^7$ CFU/g to $2-6{\times}10^8$ CFU/g and B. subtilis grew to $9{\times}10^8$ CFU/g. These results showed that 2 strains were successfully able to grow in the steamed soybean for good quality of cheonggukjang by optimization of the inoculation methods. The sensory evaluation indicated that a favorable aroma and overall acceptance of cheonggukjang by the optimized mixed cultivation of B. subtilis and L. plantarum, which was relatively higher than those of cheonggukjang by single strain inoculation of B. subtilis.

Seasonal Occurrence and Infection Site of Strawberry Anthracnose (딸기탄저병의 시기별 발생과 감염경로)

  • 김승한;최성용;임양숙;윤재탁;최부술
    • Research in Plant Disease
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    • v.8 no.1
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    • pp.45-49
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    • 2002
  • Seasonal occurrence of strawberry anthracnose in greenhouses caused by Colletotrichum sp. was examined from 1997 through 1999 at three locations, Kyeongju, Goryeong, and Cheongdo in Kyungbuk province, Korea. Also some factors related to the anthracnose infection such as initial infection sites, inoculation methods, and soil nature were studied through in vitro and field experiments. The anthracnose disease begun to occur from 15 days after transplanting in early October, and continued but gradually decreased thereafter for 2 months until December, After transplanting, initial infection mainly occurred through the runner of which the tissue was more susceptible to the anthracnose than those of the leaf and petiole when the fungal mycelial disk was inoculated. Postplanting inoculation by irrigation with spore suspension was much more effective in inducing the anthracnose disease than preplanting soil mix. However without inoculation, no or little anthracnose occurred regardless of commercial, non-cultivated or diseased field soils when healthy seedlings were planted. This suggests that occurrence of strawberry anthracnose in fields may be related to contamination of plant propagules with the anthracnose pathogen.

Virus-induced Gene Silencing as Tool for Functional Genomics in a Glycine max

  • Jeong, Rae-Dong;Hwang, Sung-Hyun;Kang, Sung-Hwan;Choi, Hong-Soo;Park, Jin-Woo;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • v.21 no.2
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    • pp.158-163
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    • 2005
  • Virus-induced gene silencing (VIGS) is a recently developed gene transcript suppression technique for characterizing the function of plant genes. However, efficient VIGS has only been studied in a few plant species. In order to extend the application of VIGS, we examined whether a VIGS vector based on TRV would produce recognizable phenotypes in soybean. Here, we report that VIGS using the Tobacco rattle virus (TRV) viral vector can be used in several soybean cultivars employing various agro-inoculation methods including leaf infiltration, spray inoculation, and agrodrench. cDNA fragments of the soybean phytoene desaturase(PDS) was inserted into TRV RNA-2 vector. By agrodrench, we successfully silenced the expression of PDS encoding gene in soybean. The silenced phenotype of PDS was invariably obvious 3 weeks after inoculation with the TRV-based vector. Real-time RT-PCR analyses showed that the endogenous level of GmPDS transcripts was dramatically reduced in the silenced leaf tissues. These observations confirm that the silenced phenotype is closely correlated with the pattern of tissue expression. The TRV-based VIGS using agrodrench can be applied to functional genomics in a soybean plants to study genes involved in a wide range of biological processes. To our knowledge, this is the first high frequency VIGS method in soybean plants.

Characteristics of sawdust cultivation of Lentinula edodes with different methods of spawn inoculation

  • Chang, Hyun You;Seo, Geum Hui;Lee, Yong Kuk;Jeon, Sung Woo
    • Journal of Mushroom
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    • v.16 no.2
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    • pp.61-64
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    • 2018
  • This study was carried out to investigate the management characteristics and growth performance of L. edodes from the cooling stage to incubation. Bags of different heights and weights are available for bagging. When the medium size of $17{\times}13cm$ was used and the size of the inoculation hole was changed from 1/3 to 2/3, the browning period was shortened to 30 days. Mycelial growth was evaluated according to the cooling temperature after sterilization. It was observed to be the highest at 122 mm/15 days at $10^{\circ}C$ and 114 mm/15 days and 117 mm/15 days at $15^{\circ}C$ and $20^{\circ}C$, respectively. The contamination rate of the sawdust media before inoculation was measured as 0, $4.5{\times}10$, $1.3{\times}10^2$, $4.0{\times}10^3cfu$ at $5^{\circ}C$, $10^{\circ}C$, $15^{\circ}C$, and $24^{\circ}C$ respectively. The average of $1.6{\times}10^8$ colony forming units (cfu) of microorganisms was observed in the sawdust that had been piled for six months outdoors. In summer, the sawdust has to be used immediately after mixing. The sterilized medium had an average of $4{\times}10^3cfu$ of microorganisms at $24^{\circ}C$ and $1.3{\times}10^2cfu$ at $15^{\circ}C$. After 15 days of inoculation in vitro, the growth conditions of the sawdust was the best at 132 mm, followed by grain and liquid. When inoculated with liquid spawn, the moisture content of the substrate should be adjusted between 50% and 55% in advance.

Development of an Efficient Mechanical Inoculation Technique to Screen Barley Genotypes for Resistance to Barley mild mosaic virus Disease and its Comparison to Natural Infection

  • Jonson, Gilda;Park, Jong-Chul;Noh, Tae-Hwan;Kim, Mi-Jung;Hyun, Jong-Nae;Kim, Jong-Gon
    • The Plant Pathology Journal
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    • v.22 no.4
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    • pp.348-352
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    • 2006
  • Barley mild mosaic virus(BaMMV) is a soilborne Bymovirus vectored by root-infecting fungus, Polymyxa graminis. Mechanism of cultivar's resistance to BaMMV in field tests are difficult to assess since resistance could be either due to the virus or to P. graminis, or both. Whereas, available mechanical inoculation methods for BaMMV and other related viruses are labor intensive, give inconsistent results and generally result in low infection rates. Inoculation method using stick with gauze(SWG) was developed for BaMMV. The improved method proved to be simple, efficient, and reliable. The infected leaf tissues were preserved by drying in a frozen state under high vaccum(freeze dried barley infected leaves) to circumvent reduction of virus infectivity during storage. Five Korean barley cultivars were mechanically inoculated with BaMMV-infected sap by the improved method. Infection rates obtained were compared with natural infection. Cultivar Naehanssalbori showed resistance to BaMMV in the field trials but was found highly susceptible in the greenhouse tests by mechanical inoculation, indicating that the field resistance may be possibly due to resistance to P. graminis.