• The Plant Pathology Journal
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• 제39권2호
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• pp.220-227
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• 2023

Rose crown gall caused by Agrobacterium tumefaciens is a major disease that damages the production of cutroses in Korea. The effective prevention methods for this disease include the use of resistant varieties. This study was conducted to evaluate the resistance of 58 Korean cultivars and six foreign cultivars to crown gall disease with nodal explants in vitro. Among 180 A. tumefaciens strains, pathogenic strain RC12 was selected as an inoculant strain. The strain RC12 was identified based on characteristics of some selective media, pathogenicity test, and polymerase chain reaction analysis. Forty rose cultivars formed tumors on explants inoculated with A. tumefaciens RC12. However, 24 cultivars, including 22 Korean cultivars and 2 foreign cultivars, showed resistance to A. tumefaciens RC12 without forming any tumors. Six cultivars with tumor formation rates of over 30% formed initial tumors within 23 days after inoculation. Six cultivars with low tumor formation rates of around 5% formed initial tumors after 28 days of inoculation. It was found that gall formation rate was highly correlated with the initial gall formation period. Thus, the relationship between the period of gall formation and the rate of gall formation could be useful for assessing resistance to crown gall disease. In vitro inoculation methods could be used to evaluate resistance of cut-rose cultivars to crown gall diseases.

• 한국산림과학회지
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• 제83권4호
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• pp.531-539
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• 1994

본 연구는 모래밭버섯 균근균(菌根菌)(Pisolithus tinctorius)의 인공접종(人工接種)이 30년생(年生) 상수리나무와 소나무 조직배양묘(組織培養苗)의 줄기생장과 발근(發根)에 미치는 영향(影響)을 조사(調査)하여, 조직배양(組織培養)을 통한 영양증식(營養增殖)에 균근균(菌根菌)의 잠재적(潛在的) 이용가능성(利用可能性)을 파악(把握)하고자 수행(遂行)하였다. 상수리나무의 아배양(芽培養)은 겨울철에 휴면상태(休眠狀態)에 있는 30년생(年生) 개체목(個體木)에서 채취(採取)한 휴면지(休眠枝)의 동아(冬芽)를 이용(利用)하여 실시(實施)하였고, 소나무의 기내삽목(器內揷木)은 종자(種子)에서 발아(發芽)한 실생묘(實生苗)를 이용(利用)하였다. 전반적(全般的)으로 상수리나무와 소나무 모두 모래밭버섯 균근균(菌根菌)의 접종(接種)으로 부정근(不定根)의 수(數), 길이, 발근율(發根率), 생존율(生存率) 등이 향상(向上)되었다. 소나무는 생존율(生存率)이 100%로 처리간(處理間)에 차이(差異)를 보이지 않았지만, 상수리나무에서는 접종구(接種區)가 100%인 반면(反面), 비접종구(非接種區)는 75%로 나타났다. 발근율(發根率)에 있어서도 소나무 기내삽목(器內揷木)에서는 발근율(發根率)이 비접종구(非接種區)에서 63%에 그친 반면(反面)에, 접종구(接種區)는 100%로 나타났고, 상수리나무는 비접종구(非接種區)가 20%로 매우 낮은 데 비(比)하여 접종구(接種區)가 70%로 유의적(有意的)인 균근균(菌根菌) 접종효과(接種效果)가 나타났다. 따라서 기내(器內) 삽목(揷木)과 조직배양(組織培養)에 균근균(菌根菌)을 응용(應用)함으로서, 발근유도(發根誘導)와 뿌리 발달(發達) 및 줄기 생장(生長)을 촉진(促進)시킬 수 있을 것으로 기대(期待)된다.

• Animal Bioscience
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• 제35권10호
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• pp.1592-1605
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• 2022

Objective: The objective of this study was to select an effective in vitro digestion-fermentation model to estimate the effect of decreasing dietary crude protein (CP) on odor emission during pig production and to suggest potential prediction markers through in vitro and in vivo experiments. Methods: In the in vitro experiment, three diet formulations with different CP contents (170 g/kg, 150 g/kg, and 130 g/kg) but containing the same standardized ileal digestible essential amino acids (SID-EAA) were assessed. Each diet was evaluated by two different in vitro gastric-intestinal phase digestion methods (flask and dialysis), combined with fresh pig feces-ferment inoculation. Eighteen growing barrows (31.9±1.6 kg) were divided into three groups: control diet (180 g CP/kg, without SID-EAA adjustment), 170 g CP/kg diet, and 150 g CP/kg diet for 4 weeks. Results: The in vitro digestion results indicated that in vitro digestibility was affected by the gastric-intestinal phase digestion method and dietary CP level. According to the gas kinetic and digestibility results, the dialysis method showed greater distinguishability for dietary CP level adjustment. Nitrogen-related odor compounds (NH₃-N, indole, p-cresol, and skatole) were highly correlated with urease and protease activity. The feeding study indicated that both EAA-adjusted diets resulted in a lower odor emission especially in p-cresol and skatole. Both protease and urease activity in feces were also closely related to odor emissions from nitrogen metabolism compounds. Conclusion: Dialysis digestion in the gastric-intestinal phase followed by fresh fecal inoculation fermentation is suitable for in vitro diet evaluation. The enzyme activity in the fermentation and the fecal samples might provide a simple and effective estimation tool for nitrogen-related odor emission prediction in both in vitro and in vivo experiments.

• 대한미생물학회지
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• 제7권1호
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• pp.29-41
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• 1972

조직배양(組織培養)을 이용(利用)한 마우스복강거식세포내 인나균증식실험(人癩菌增殖實驗)으로서 trypsin-정제인나균(精製人癩菌)을 사용하여 1) 인나균(人癩菌)의 마우스복강내접종(腹腔內接種)에 의(依)하여 야기(惹起)되는 복강거식세포내의 생체내(生體內) 인나균감염(人癩菌感染)에 대(對)한 동역학적양상(動力學的樣相), 2) in vivo infection-in vitro cultivation에 의(依)한 인나균(人癩菌)의 복강거식세포내 증식태도(增殖態度) 그리고 3) 인나균(人癩菌)의 복강내접종(腹腔內接種)으로 인(因)한 마우스비장조직(脾臟組織)의 병리학적변화(病理學的變化)를 구명(究明)코저 본연구(本硏究)를 실시(實施)하였으며 아래의 결론(結論)을 얻었다. 1. 인나균(人癩菌)의 마우스복강내접종후(腹腔內接種後) 복강거식세포배양실시까지의 기간(其間)이 연장(延長)됨에 따라 배양(培養)된 복강거식세포에 있어서 세포질내(細胞質內)에 식균된 항산균수(抗酸菌數)와 항산균(抗酸菌)을 식균한 거식세포수(細胞數)가 각각 계속적(繼續的)으로 현저(顯著)하게 감소(減小)되어감이 관찰(觀察)되었다. 2. 인나균(人癩菌)의 복강내접종후(腹腔內接種後) 5개월(個月)에 이르기까지 생체내(生體內)에 존재(存在)하는 마우스복강거식세포에 있어서의 인나균증식(人癩菌增殖)을 관찰(觀察)할 수 없었다. 3. 인나균(人癩菌)의 복강내접종후(腹腔內接種後) 24시간(時間) 및 1주(週)만에 실시(實施)된 복강거식세포배양을 2 내지(乃至) 3개월간(個月間) 그 배양상태(培養狀態)를 유지(維持)하였던바 배양(培養)된 거식세포(細胞)의 염색표본(染色標本)에서 거식세포내(細胞內) 항산균수(抗酸菌數)의 뚜렷한 증가양상(增加樣狀)을 관찰(觀察)할수 있었다. 4. in vivo infection-in vitro cultivation 수기(手技)로서 배양(培養)된 복강거식세포를 사용(使用)한 총항산균수측정실험(總抗酸菌數測定實驗)에서 조직배양개시(組織培養開始) 9 및 11주(週)에 이르러 배양(培養)된 거식세포내(細胞內) 항산균수증가(抗酸菌數增加)에 대한 양적증거(量的證據)를 얻을수 있었다. 5. 인나균(人癩菌)의 복강내접종(腹腔內接種)으로 야기되는 마우스비장조직(脾臟組織)의 병리학적소견(病理學的所見)은 주로 적수부위(赤髓部位)에 나타난 변성변화(變性變化)이었으며, 균접종후(菌接種後) 5개월(個月)에 이르기까지 마우스비장내(脾臟內)에 있어서의 인나균(人癩菌)의 증식양상(增殖樣狀)을 관찰(觀察)할 수 없었다.

• Radiation Oncology Journal
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• 제12권3호
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• pp.301-305
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• 1994

Purpose :. To investigate the effect of aqueous extract of Ganoderma lucidum(G.I.) on the surival of tumor cells in vitro and on the growth of tumors in vivo. Materials and Methods : Dried G.I. was made into powder, extracted with distilled water, filtered and diluted from a maximum concentration of 100 mg/ml in sequence. The cytotoxicity of G.I, in vitro was evaluated from its ability to reduce the clonogenicity of SCK tumor cells. For the tumor growth delay study, about $2{\times}10^5$$ of SCK tumor cells were subcutaneously inoculated in the legs of A/J mice. The first experimental group of mice were injected i.p. with 0.2ml of 250 mg/kg of G.I. from the first day after tomor inoculation for 10 days. The second experimental group of mice were injected i.p. with 0.2ml of 250 mg/kg of G.I. either once a day for 10 days or twice a day for 5 days beginining from the 7th day after tumor inoculation Results : 1. Cytotoxicity in vitro;survival fraction, as judged from the curve, at G.I. concentration of 0.5, 1,5, 10, 25, 50 and 100 mg/ml were 1.0, $0.74{\pm}0.03$, $0.18{\pm}0.03$, $0.15{\pm}0.02$, $0.006{\pm}0.002$, 0.015 and 0.0015, respectively. 2. Tumor growth delay in vivo; a) the time required for the mean tumor volume to grow to $1,000mm^3$ was 11 days in the control group and 14 days in the experimental group. b) the time required for tumor volume to increase 4 times was 11 days in the control group while it was 10.5 and 12 days in the groups injected with G.I. once a day and twice a day from the 7th day after tumor inoculation respectively. Conclusion : Aqueous extracts of G.I. showed a marked cytotoxicity on the SCK mammary cells in vitro. Tumor growth delay was statistically signiricant when G.I. in-jection was started soon after tumor inoculation, but it was not significant when injection was started after the tumors were firmly established.

• 한국환경농학회지
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• 제20권5호
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• pp.335-339
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• 2001

Two successive in vitro experiments were carried out to examine the effect of MPS bacterial inoculation on growth, and nitrogen and phosphorus accumulation of maize plants under greenhouse condition in the same soil. There were four treatments, uninoculated control and three phosphate solubilizing bacterial inoculations, viz., Pseudomonas striata, Burkholderia cepacia and Serratia marcescens. The inoculated plants showed the higher plant height, total dry mass, nitrogen and phosphorus accumulation when compared to uninoculated control plants in both experiments. In the combined data analysis from two experiments, the plants inoculated with P. striata and B. cepacia showed significantly higher plant height, total dry mass and P accumulation when compared to S. marcescens inoculated plant and uninoculated control plants. The P. striata and B. cepacia inoculation enhanced total dry matter accumulation by 14% and phosphorus accumulation by 25% over the uninoculated control plants. The nitrogen and phosphorus concentration of maize plants were also increased due to MPS bacterial inoculation, however, the effect was not significant.

• Animal Bioscience
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• 제35권9호
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• pp.1379-1389
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• 2022

Objective: This study identified the major lactic acid bacteria (LAB) strains from different fermented total mixed rations (FTMRs) via metataxonomic analysis and evaluated the ability of their standard strain as ensiling inoculants for corn stover silage. Methods: The bacterial composition of eight FTMRs were analyzed by 16S rDNA sequencing. Corn stover was ensiled without LAB inoculation (control) or with 1×10⁶ cfu/g LAB standard strain (Lactobacillus vaginalis, Lactobacillus reuteri, Lactobacillus helveticus, or Lactobacillus paralimentarius) selected from the FTMRs or 10 g/t commercial silage inoculant (CSI) around 25℃ for 56 days. For each inoculation, a portion of the silage was sampled to analyze ensiling characteristics at time intervals of 0, 1, 3, 7, 14, 28, and 56 days, gas production (GP), microbial crude protein and volatile fatty acids as the measurements of rumen fermentation characteristics were evaluated in vitro with the silages of 56 days after 72 h incubation. Results: Lactobacillus covered >85% relative abundance of all FTMRs, in which L. pontis, L. vaginalis, L. reuteri, L. helveticus, and L. paralimentarius showed >4% in specific FTMRs. CSI, L. helveticus, and L. paralimentarius accelerated the decline of silage pH. Silage inoculated with L. paralimentarius and CSI produced more lactic acid the early 14 days. Silage inoculated with L. paralimentarius produced less acetic acid and butyric acid. For the in vitro rumen fermentation, silage inoculated with CSI produced more potential GP, isobutyric acid, and isovaleric acid; silage inoculated with L. helveticus produced more potential GP and isovaleric acid, silage inoculated with L. paralimentarius or L. reuteri produced more potential GP only. Conclusion: The standard strain L. paralimentarius (DSM 13238) is a promising ensiling inoculant for corn stover silage. The findings provide clues on strategies to select LAB to improve the quality of silage.

• 식물병연구
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• 제29권1호
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• pp.82-87
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• 2023

Exogenous ferrous chloride (FeCl₂) suppressed in vitro growth of Ralstonia pseudosolanacearum, causing bacteria for tomato bacterial wilt. More than 50 μM of FeCl₂ reduced the in vitro bacterial growth in dosedependent manners. Two to 200 μM of FeCl₂ did not affect the fresh weight of detached tomato leaves at 3 and 5 days after the petiole dipping without the bacterial inoculation. The bacterial wilt of the detached tomato leaves was evaluated by inoculating two different inoculum densities of R. pseudosolanacearum (10⁵ and 10⁷ cfu/ml) in the presence of FeCl₂. Bacterial wilt in the detached leaves by 10⁵ cfu/ml was efficiently attenuated by 10-200 μM of FeCl₂ at 3 and 5 days post-inoculation (dpi), but bacterial wilt by 10⁷ cfu/ml was only reduced by 200 μM of FeCl₂ at 3 and 5 dpi. These results suggest that iron nutrients can be included in the integrated disease management of tomato bacterial wilt.

• BMB Reports
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• 제40권3호
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• pp.404-411
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• 2007

Transformation of cantaloupes with the coat protein (cp) gene of papaya ringspot virus type W (PRSV-W), Thai isolate, was used to introduce virus resistance. Binary vectors containing either the full length coat protein coding region under control of the 35S CaMV promoter(pSA1175), or the inverted-repeat of a coat protein coding region (pSA1304), were constructed and used for Agrobacteriummediated transformation of cotyledonary explants of the cantaloupe cultivar Sun Lady. Four independent transgenic lines were obtained using pSA1304 and one using pSA1175. Integration of the PRSV-W cp gene into the genome of these transgenic lines was verified by PCR amplification, GUS assays and Southern blot hybridization. In vitro inoculation of these lines with PRSV-W revealed that whereas the line containing pSA1175 remained sensitive, the four lines containing pSA1304 were resistant. The presence of small RNA species, presumably siRNA, corresponding to regions of the viral cp gene in transgenic lines resistant to PRSV-W supports the involvement of post-transcriptional gene silencing in the establishment of resistance.

• 대한수의학회지
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• 제31권4호
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• pp.441-447
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• 1991

In this study, effect of levamisole, selenium and tocopherol on the lymphocyte blastogenesis and antibody production in Korean native goat were evaluated in vitro and in vivo. Lymphocyte blastogenesis of goat blood increased significantly (p<0.01 and p<0.05) when the cells were treated in vitro with levamisole at the concentration of $50{\sim}500{\mu}g/ml$, with selenium at the concentration of $0.062{\sim}1.0{\mu}g$ and with tocopherol nt the concentration on $12.5{\mu}g$. Increased lymphocyte blastogenesis was detected from 2 to 24 hours after oral administration of levamisole (2.5mg/kg of body weight). After 7 days, increased mitogenic response of lymphocytes was not detected. Meanwhile increased blastogenesis of lymphocyte from goats given the selenium-tocopherol mixture (selenium $100{\mu}g$-tocopherol 200IU/head/day) was detected from 10 days after feeding, and the tendency continued throughout the entire experimental period. When immune responses of goats against PPD were subjected to test by ELISA, the mean IgG titers of levamisolc group (1 : 1,800) and selenium-tocopherol group (1 : 960) were higher than that of control group (1 : 600) at 2 weeks after lst inoculation. At 3 weeks after lst inoculation and 1 week after 2nd inoculation, the significant (p<0.05) differences in IgG titers were detected among the three groups. The mean IgG titers of levamisole group, selenium-tocopherol group and control at that time were 1 : 20,480, 1 : 5,120 and 1 : 2,640, respectively. The IgG production of levamisole group was significantly (p<0.01) higher than that of control group.