• Title/Summary/Keyword: Liquid culture inoculation

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Effect of Sawdust Culture on Oak Mushroom, Lentinula edodes (Berk.) Pegler by Inoculation of the Liquid Spawn (액체종균(液體種菌) 접종(接種)에 의한 표고톱밥재배(栽培) 효과(效果))

  • Lee, Tai-Soo;Cho, Nam-Seok;Min, Du-Sik
    • Journal of the Korean Wood Science and Technology
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    • v.26 no.1
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    • pp.19-28
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    • 1998
  • For cultivation on sawdust-bed of oak-mushroom until present time, inoculation of spawn on sawdust bed has been performed by sawdust spawn. But, liquid spawn may have advantages for rapid mass production of spawn, and now, sawdust-cultivation by liquid spawn inoculation should be applied instead of sawdust spawn. Therefore, investigations were performed to evaluate the effect of sawdust-cultivation by liquid spawn inoculation. The results were as follows: 1. When 11 kinds of liquid media were applied, the oak-mushroom culture medium was the most excellent in growth. Most suitable temperature at PDA was $25^{\circ}C$, and $22.5\sim27.5^{\circ}C$ in range were optimal for liquid culture. In liquid culture, amount of mycelial growth increases rapidly up to 40 days of cultivation. Incubation at fermentor brought yield of 106mg dry mycelia per 40ml media after 17 days. 2. In 1l-spawn bottle, growth of mycelium by inoculation of 20ml-liquid spawns were faster than 6g-sawdust spawn in spread of mycelia. On 2kg-bag culture, inoculations of 10ml-, 20ml- and 30ml-liquid spawns were all slower than 20g-sawdust spawn in mycelial spread. So, amount increasement in ampunt of liquid spawn should be discussed. Yields of mushrooms until third sproutings of 2kg-bag culture were 580g in 30ml-liquid spawn inoculation, but 510g, 486g and 470g from 20g-sawdust spawn, 20ml-liquid spawn and 10ml-liquid spawn, respectively. Thus, 30ml-liquid spawn inoculation was highest in yield.

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Proliferation of Tricholoma matsutake Mycelial Mats in Pine Forest Using Mass Liquid Inoculum

  • Lee, Won-Ho;Han, Sang-Kuk;Kim, Beom-Seok;Shrestha, Bhushan;Lee, Soo-Yong;Ko, Cheol-Soon;Sung, Gi-Ho;Sung, Jae-Mo
    • Mycobiology
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    • v.35 no.2
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    • pp.54-61
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    • 2007
  • Two isolates of Tricholoma matsutake T-008 and T-034, preserved in Entomopathogenic Fungal Culture Collection (EFCC) of Korea, were used in the present study. The isolates had 100% Bootstrap homology with Tricholoma matsutake U62964 and T. matsutake AB188557 and AF309538 preserved in Gene Bank of NCBI. Mycelial growth of T. matsutake was highest in TMM and MYA at $25^{\circ}C$. The highest dry wt. of mycelium was obtained after 65 days of culture, when 6 mycelial discs were inoculated in 100 ml of broth in 250 ml shaking flask. Mycelial mats were observed in clumped condition at the inoculation sites of pine forest after two weeks of inoculation. After 5 months of inoculation, mycelia mats were observed growing inside soil and walls of a few inoculation sites, while mycelial mats growth up to $5{\sim}8$ cm were observed in the roots of pine tree after 6 months. The survival rate of the inoculum was about 40% of the total inoculation sites. The survival rate was found below 20% when the mycelium was inoculated in the summer. The reasons for low survival rates of the mycelium were mainly due to dry season and the soil-borne small animals such as earthworm and mole. After one year of inoculation, no external difference was observed between the artificially inoculated mycelia and the naturally existing mycelia of T. matsutake. The present study showed that fruiting bodies of T. matsutake could be produced by artificial inoculation under the appropriate environmental conditions.

Effects of Culture Type and Inoculation Quantity in Bioreactor on Production of Potato Plantlets

  • Choi Ki Young;Son Sung Ho;Lee Joo Hyun;Lee Yong-Beom;Bae Jong Hyang
    • Journal of Bio-Environment Control
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    • v.14 no.4
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    • pp.298-301
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    • 2005
  • Potato (Solamum tuberosum 'Dejima') plantlets were investigated on culture type and initial quantity of inoculation in bioreactor and survival rate by hydroponics for mass production. rode stems (1 to 1.5cm in length) of potato plantlets multiplied in vitro were grown for 3 weeks in liquid Murashige and Skoog (MS) medium with sucrose $30 g\; L^{-1}$. When plantlets (80-node inoculation) were raised in 10L balloon type bubble (BB) bioreactor, the healthiest growth of plantlets was obtained from explants cultured in ebb & flow culture with medium supplied periodically 12 times per day. The suitable inoculation quantity of 20L BB bioreactor was 120 pieces of stem segments (mean 2.2g fresh weight) in ebb & flow culture. Number of nodal shoot was eight on the average. In controlled culture room, survival rate of plantlets at 7 days after stem cutting was above $70\%$ when they were acclimatized by hydroponics grown in deep flow and solid medium culture. The highest survival rate of the stem cutting plantlets was in nutrient solution adjusted to EC $1.4dS{\cdot}m^{-1}$. Stem cutting plantlets through one culture could be obtained $670\~900$, when plantlets were grown in ebb & flow culture during 3 weeks using a 20L bioreactor with initial 120 pieces of nodal segments. 11 is possible In do mass production of seedlings cultured in bioreactor and hydroponics.

In Vitro Mycorrhization and Morphological Characterization of Xanthoconium affine with Pinus densiflora

  • Wang, Eun-Jin;Jang, Yeongseon;Ka, Kang-Hyeon
    • The Korean Journal of Mycology
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    • v.45 no.4
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    • pp.319-327
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    • 2017
  • In this study, we investigated the culture conditions of four ectomycorrhizal fungi, namely, Amanita spissacea NIFoS 2719, Pisolithus arhizus NIFoS 2784, Suillus spraguei NIFoS 2848, and Xanthoconium affine NIFoS 2716, in solid and liquid culture media. In addition, we attempted to induce in vitro mycorrhization of the fungi with Pinus densiflora. Prior to liquid culture, we determined the optimal culture conditions for each species in solid media. The results revealed that all species examined are capable of growth in potato dextrose agar (PDA), malt extract agar (MEA), and modified Melin-Norkran's medium (MMN), although their preferred growth media were different. Liquid culture experiments showed that inorganic nitrogen did not enhance the mycelial growth of all four species. Therefore, we used MMN-based liquid inocula to promote the growth of ectomycorrhizal fungi in our symbiosis culture system. Mycorrhization was observed in Xanthoconium affine NIFoS 2716. Morphological analysis revealed that fungi-inoculated roots of P. densiflora form simple and dichotomous lateral roots with dense mycelia. In addition, inoculation with X. affine NIFoS 2716 promoted root and shoot developments.

Occurrence of Phytophthora Rot of Strawberry Caused by Phytophthora nicotianae var. nicotianae (Phytophthora nicotianae var. nocotianae에 의한 딸기 역병의 발생)

  • 송주희;노성환;하주희;정연화;문병주
    • Korean Journal Plant Pathology
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    • v.14 no.5
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    • pp.445-451
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    • 1998
  • A severe Phytophthora rot of strawberry caused by a species of Phytophthora has been widely occurred at major cultivation areas of Kimhae on August in 1997. Incidence of the disease was obtained in the range of 69.2~83.6% in surveyed 4 fields and showed an average of 75.2%. A species of Phytophthora was mostly isolated from the crown of infected strawberry plants and all the isolates were identified as P. nicotianae var. nicotianae (=P. parasitica). The fungus showed strong pathogenicity on strawberry by inoculation test. As a result of the leaf inoculation using mycelial disks of the fungus, both leaves and petioles were darkly browned, and were finally blighted. As a result of the root inoculation of zoospore suspension, both roots and crowns were rotten with dark brown. Although the fungus produced sporangia either on V-8 juice agar medium or liquid medium, the sporangia observed on the liquid medium appeared to be broadly turbinate and noncaducous. Moreover the fungus cultured on the liquid medium often produced sporangia having two papilla. The number of zoospores in sporangia was found to be ranged from 3 or 4 to as many as 20 or 25. In addition, the released zoospore from the sporangium became the cystospore during the prolonged culture of the fungus. The sporangia were measured as av. 49$\times$35 ${\mu}{\textrm}{m}$ with l/b ratio of 1.43. All isolates from crowns were heterothallic and A1 mating type since oospores were abundantly formed on clarified V-8 juice agar by dual culture with P. capsici A2 mating type. Aplerotic oospores were sized 24-26 ${\mu}{\textrm}{m}$. Antheridia were always amphigynous and recoreded an average of 12$\times$10 ${\mu}{\textrm}{m}$. Hyphal swlling were easily observed, and terminal or intercalary chlamydospores were abundantly formed on V-8 juice agar as well as in C/Z solution and sized av. 28.2 ${\mu}{\textrm}{m}$. This is the first report of Phytophthora rot of strawberry in Korea.

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The effect of different culture conditions of liquid spawn on the quality characteristics of shiitake mushroom (Lentinula edodes) (표고의 액체종균 배양 조건 및 품질 특성)

  • Lee, Soo-Jung;Kim, Hun-Hwan;Kim, Seon-Ho;Kim, Sung-Hee;Sung, Nak-Ju
    • Journal of Mushroom
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    • v.17 no.3
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    • pp.99-106
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    • 2019
  • To improve the productivity of shiitake mushroom (Lentinula edodes), seven different types of media for liquid spawn (denoted as "A" to "G") were prepared with 0.3% soybean meal and varying sugar and glucose concentrations. During 14 days of incubation, the pH of the liquid culture gradually acidified with increasing incubation period. Additionally, there was a significant, but not prominent, difference in the degree of acidification depending on the sugar to glucose ratio. Liquid spawn culture "G," which had the highest sugar content was the most acidic on the last day of incubation. Mycelium dry weight increased significantly with increasing incubation period, and there was no significant difference in mycelium dry weight irrespective of the sugar to glucose ratio even after 14 days of culture. The inoculation of liquid spawn in sawdust medium with an inoculation volume ${\geq}45mL$ and incubation period of 15 to 18 days were the optimal culture conditions. Productivity of fruit bodies in sawdust medium and mushrooms treated with liquid spawn was significantly higher compared to solid spawn treatment. The mushrooms treated with liquid spawn had better chewiness, and the free amino acid content, which is associated with savory taste, was higher in these mushrooms compared to those treated with solid spawn.

Changes of Nutrients in Media and Mycelia on Liquid Spawn Culture of Lentinula edodes (표고 액체종균 배양시 배지와 균사체의 양분변화)

  • Shim, Kyu-Kwang;Yoo, Young-Jin;Koo, Chang-Duck;Kim, Myung-Kon
    • The Korean Journal of Mycology
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    • v.42 no.2
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    • pp.144-149
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    • 2014
  • Lentinula edodes liquid spawn growth under explosive aeration (supplying air with tiny bubbles) and soybean meal addition to liquid culture medium were investigated in terms of mycelial growth and residual free sugar content. The two treatments were effective for homogeneous culturing of mycelial spawn and for separating colonies during multiplication after an exponential growth period without limiting sustaining nitrogen nutrients. The mycelial growth and carbon dioxide concentration were greatest on the 13th day since the inoculation. At 12th day, however, free sugars were almost depleted in the upper part of the liquid medium. Total nitrogen content within precipitated mycelia was the highest at the 13th day. Chitin and sucrose contents in the mycelia were the highest at the 18th day, but ergosterol content became highest at 22 days. These results suggest that Lentinula edodes liquid spawn is ready in 18 days after inoculation.

Production of Pleurotus ostreatus and Flammulina velutipes Using Liquid Spawn Inoculation System (느타리 팽나무버섯 재배에서 액체종균 배양 및 접종시스템 적용방법의 구명)

  • Hong, Sung-Jun;Lee, Won-Ho;Shin, Beom-Soo;Sung, Jae-Mo
    • The Korean Journal of Mycology
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    • v.31 no.1
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    • pp.22-27
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    • 2003
  • This research aimed at developing the efficient method and device applicable to the inoculation of mushroom using (spawn was intended to utilize) liquid spawn for the stable production of mushroom. For the mass production of liquid culture, optimal inoculum volume and cultural period were $5{\sim}6%$ and 4 days for Flammulina velutipes and 4% and 5 days for Pleurotus ostreatus. Fruit body weight in 850 ml polythene bottle was highest at 6% or 15 ml liquid inoculum for P. ostreatus and $4{\sim}6%$ or $10{\sim}15ml$ for F. velutipes. Weight of fruit body by the application of liquid spawn inoculation system increased up to 33.7% for P. ostreatus and 32.8% for F. velutipes, respetively, compared to conventional spawn making system. The system of liquid spawn inoculation was successfully operated without malfunction in opening or closing the lid, and it took 26 min to inoculate 1200 bottles.

Growth Rate of Entomopathogenic Fungi in Mass Culture System (곤충병원성 진균의 대량 배양체계에서의 성장율)

  • 이인기;서종복
    • Journal of Sericultural and Entomological Science
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    • v.38 no.2
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    • pp.150-153
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    • 1996
  • To develope a microbial pesticide for the control of agricultural and forestal pests in Korea, the mass culture system of entomopathogenic fungi was studied. Previously, we have developed the mass culture system which was adaptable for the culture of Beauveria bassiana. In this study, we determined the efficacy of this mass culture system for other entomopathogenic fungi, B. bassiana, Beauveria brongniartii, Metarhizium anisopliae, and Verticillium lecanni. To determine the efficacy of mass culture system, we examined the growth rate of entomopathogenic fungi in this system which was composed of 1st liquid media for growth of blastospore and 2nd pellet media for growth of conidia. As the result, we obtained that the blastopore numbers increased 103-104 times in liquid media at 72 hrs post inoculation. The results showed that this mass culture system for the growth of entomopathogenic fungi was effective.

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Effect of Co-inoculation of Two Bacteria on Phosphate Solubilization

  • Lee, Yu-Jin;Lee, Heon-Hwak;Lee, Chan-Jung;Yoon, Min-Ho
    • Korean Journal of Soil Science and Fertilizer
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    • v.49 no.4
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    • pp.318-326
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    • 2016
  • Two phosphate solubilizing bacteria, Pantoea rodasii PSB-11and Enterobacter aerogenes PSB-12, were isolated from button mushroom compost and employed to assess their synergistic effect in liquid medium and on growth of green gram plants by single and co-inoculation of the strains. Co-inoculation of two strains was found to release the highest content of soluble phosphorus ($521{\mu}g\;ml^{-1}$) into the medium, followed by single inoculation of Pantoea strain ($485{\mu}g\;ml^{-1}$) and Enterobacter strain ($470{\mu}g\;ml^{-1}$). However, there was no significant difference between single inoculation of bacterial strain and co-inoculation of two bacterial strains in terms of phosphorous release. The highest pH reduction, organic acid production and glucose consumption was observed in the E. aerogenes PSB-12 single inoculated culture medium rather than those of co-inoculation. According to the plant growth promotion bioassay, co-inoculated mung bean seedlings recorded 10.6% and 10.7% higher shoot and root growth respectively compared to the control. Therefore, in concluding, co-inoculation of the strains P. rodasii and E. aerogenes displayed better performance in stimulating plant growth than inoculation of each strain alone. However, being short assessment period of the present study, we recommend in engaging further works under field conditions in order to test the suitability of the strains to be used as bio-inoculants.