• Title/Summary/Keyword: Resistant screening

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An in vitro Actinidia Bioassay to Evaluate the Resistance to Pseudomonas syringae pv. actinidiae

  • Wang, Faming;Li, Jiewei;Ye, Kaiyu;Liu, Pingping;Gong, Hongjuan;Jiang, Qiaosheng;Qi, Beibei;Mo, Quanhui
    • The Plant Pathology Journal
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    • v.35 no.4
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    • pp.372-380
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    • 2019
  • Pseudomonas syringae pv. actinidiae (Psa) is by far the most important pathogen of kiwifruit. Sustainable expansion of the kiwifruit industry requires the use of Psa-tolerant or resistant genotypes for the breeding of tolerant cultivars. However, the resistance of most existing kiwifruit cultivars and wild genotypes is poorly understood, and suitable evaluation methods of Psa resistance in Actinidia have not been established. A unique in vitro method to evaluate Psa resistance has been developed with 18 selected Actinidia genotypes. The assay involved debarking and measuring the lesions of cane pieces inoculated with the bacterium in combination with the observation of symptoms such as callus formation, sprouting of buds, and the extent to which Psa invaded xylem. Relative Psa resistance or tolerance was divided into four categories. The division results were consistent with field observations. This is the first report of an in vitro assay capable of large-scale screening of Psa-resistance in Actinidia germplasm with high accuracy and reproducibility. The assay would considerably facilitate the breeding of Psa-resistant cultivars and provide a valuable reference and inspiration for the resistance evaluation of other plants to different pathogens.

Mycobacterium tuberculosis DNA Detection and Molecular Drug Susceptibility Test in AFB-stained Sputum Slides

  • Jung, Dongju;Lee, Hyeyoung;Park, Sangjung
    • Biomedical Science Letters
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    • v.22 no.1
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    • pp.24-28
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    • 2016
  • Tuberculosis (TB) remains an unsolved community health problem since identification of its causing microorganism called Mycobacterium tuberculosis (MTB) by Robert Koch in 1882. Annually, eight million TB cases are newly reported and 2~3 million patients die from TB. Pulmonary TB is highly infectious and untreated pulmonary TB patients are believed to infect >10 people in a year. The conventional methods for diagnosis of TB are chest X-ray and isolation of the causing microorganisms from patient specimens. Screening of TB is conducted with smeared sputum in slides, and TB is confirmed by identification of MTB in cultured specimens. One of the fatal pitfalls of screening detection for smeared sputum is that it is impossible to distinguish MTB and other acid-fast bacilli (AFB) because they are stained equally with Ziehl-Neelsen (ZN) stain. Culture of MTB is the most reliable method for diagnosis of TB but it takes 4~8 weeks. In this report, we suggest a fast and highly-reliable MTB detection method that distinguishes AFB in sputum samples. Purified DNA from the AFB stained slide samples offered by The Korean Institute of Tuberculosis were used to detect infected MTB in patients. PCR, real-time PCR and reverse blot hybridization assay (REBA) methods were applied to purified DNA. Conclusively, the real-time PCR method was confirmed to produce high sensitivity and we were able to further detect drug-resistant MTB with REBA.

Fungicidal Activities of 51 Fruit-Derived Extracts in vivo against Six Phytopathogenic Fungi

  • Lee, Hoi-Seon;Lee, Seon-Woo;Cho, Kwang-Yun;Kim, Moo-Key;Ahn, Young-Joon
    • Journal of Applied Biological Chemistry
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    • v.44 no.3
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    • pp.147-153
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    • 2001
  • Methanol extracts from 51 fruits were tested for their fungicidal activities against six phytopathogenic fungi in a greenhouse. The efficacy varied with both the plant pathogen and fruit species used. At 10 and 5 mg/pot, methanol extracts of Poncirus trifoliata peel and seed gave over 80% control values against Pyricularia grisea, and strong fungicidal activities against Rhizoctonia solani were showed from the extracts of Citrus paradisi peel and Punica granatum leaf. In a test with Botrytis cinerea at 5 mg/pot, the extracts of C. sinensis seed and D. kaki leaf produced potent fungicidal activities, and the extracts of C. crenata peel and leaf, Ch. sinensis seed, P. trifoliata peel, and Z. jujuba leaf had strong fungicidal activities. At 5 mg/pot, strong fungicidal activities were produced in the extracts of P. trifoliata peel and seed against Phytophthora infestans and in the extracts of P. ussriensis var. macrostipes fruit and seed, C. crenata peel, C. crenata leaf, C. paradisi peel, P. trifoliata peel, P. granatum peel, and Z. jujuba leaf against Puccinia recondita. In a test with E. graminis, potent activities at 10 mg/pot were produced from the extracts of Ch. sinensis seed, C. sinensis seed, P. trifoliata leaf, P. ussriensis var. macrostipes fruit and seed, and Vitis vinifera seed. In the control effect of seven extracts against B. cinerea strains resistant to carbendazim, procymidone, and diethofencarb, extracts of C. crenata peel and leaf, Ch. sinensis seed, and P. trifoliata peel were highly effective against all strains of B. cinerea. Furthermore, potent fungicidal activities were produced from the extracts of C. sinensis seed and D. kaki leaf against the SSR, SRR, and RRS, and Z. jujuba leaf against the SSR and RRS strains. As a naturally occurring fungicide, these fruit-derived materials could be useful as new fungicidal products against phytopathogenic fungi.

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Studies on the Relationship between Inoculum Concentration and Disease Development in Bacterial Leaf Blight of Rice (벼흰빛잎마름병에 있어서 병원균의 접종 농도가 병의 발전속도에 미치는 영향)

  • Cho Yong Sup
    • Korean journal of applied entomology
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    • v.14 no.1 s.22
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    • pp.1-5
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    • 1975
  • The study has been carried out for the development of an inoculation method in screening resistant varieties and/or lines to bacterial leaf blight of rice with special consideration on plant ages and inoculum concentrations. A higher incidence of the disease was found on younger plants than on the older ones when the plants were inoculated with the same concentration of inoculum by clipping method under the same circumstances. Applications of extremly high concentration of inoculum resulted the death of younger seedlings from all varieties within short period after inoculation, and the inoculum that was lower than optimum concentration failed in distinguishing varietal characteristics in regard of resistance. The proper inoculum size for the screening of resistance among the varieties and/or lines depended on the age of plants. The optimum concentration of inoculum for the plant age of 14, 37, 48 and 58days was $10^6,\;10^7,\;10^7-10^{-9}\;and\; 10^9 cells/ml.,$ respectively.

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Susceptibility to Calonectria ilicicola in Soybean Grown in Greenhouse and Field

  • Kim, K. D.;Russin, J. S.;Snow, J. P.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.43 no.4
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    • pp.239-244
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    • 1998
  • Susceptibility of soybean cultivars to Calonectria illicicola was evaluated in a greenhouse by inoculating seedlings with mycelium in agar discs placed on the stems at the soil line. A range of responses was detected among cultivars following inoculation with a virulent isolate of C.ilicicola. Rankings of cultivars between greenhouse tests 1 and 2 were similar for disease severity and areas under the disease progress curves (AUDPC). In addition, rankings of cultivars for Final disease severity were highly correlated with AUDPC in test 1 ($r_s$ =0.88, t =5.48, p<0.001), test 2 ($r_s$ =0.99, t =22.10, p<0.001), and when tests were combined ($r_s$=0.89, t=5.82, p<0. 001). Final disease severity and AUDPC consistently identified Asgrow 7986, Braxton, Cajun, and Forrest as soybean cultivars least susceptible to red crown rot. In 1993 and 1994 field tests, a range in disease susceptibility was observed for tested cultivars but none was completely resistant. Soybean cultivars Braxton, Cajun, and Forrest, which were least susceptible to red crown rot in greenhouse tests, also ranked among cultivars with the lowest disease incidence and AUDPC in field tests. Comparisons .between rankings of the eight cultivars common to greenhouse and field tests showed a correlation between final disease severity from combined greenhouse tests and both final disease incidence ($r_s$=0.63, t =1.99, p<0.1) and AUDPC ($r_s$=0.60, t =1.82, p < 0.2) from the combined field tests. However, AUDPC from greenhouse tests did not correlate with either final disease incidence or AUDPC from field tests. The green-house screening method provided consistent results between greenhouse and field tests and successfully identified the least susceptible cultivars Braxton, Cajun, and Forrest.

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Screening of Rice Germplasm for the Distribution of Rice Blast Resistance Genes and Identification of Resistant Sources

  • Ali, Asjad;Hyun, Do-Yoon;Choi, Yu-Mi;Lee, Sukyeung;Oh, Sejong;Park, Hong-Jae;Lee, Myung-Chul
    • Korean Journal of Plant Resources
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    • v.29 no.6
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    • pp.658-669
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    • 2016
  • Rice blast, caused by a fungus Magnaporthe oryzae, is one of the most devastating diseases of rice worldwide. Analyzing the valuable genetic resources is important in making progress towards blast resistance. Molecular screening of major rice blast resistance (R) genes was determined in 2,509 accessions of rice germplasm from different geographic regions of Asia and Europe using PCR based markers which showed linkage to twelve major blast R genes, Pik-p, Pi39, Pit, Pik-m, Pi-d(t)2, Pii, Pib, Pik, Pita, Pita/Pita-2, Pi5, and Piz-t. Out of 2,509 accessions, only two accessions had maximum nine blast resistance genes followed by eighteen accessions each with eight R genes. The polygenic combination of three genes was possessed by maximum number of accessions (824), while among others 48 accessions possessed seven genes, 119 accessions had six genes, 267 accessions had five genes, 487 accessions had four genes, 646 accessions had two genes, and 98 accessions had single R gene. The Pik-p gene appeared to be omnipresent and was detected in all germplasm. Furthermore, principal component analysis (PCA) indicated that Pita, Pita/Pita-2, Pi-d(t)2, Pib and Pit were the major genes responsible for resistance in the germplasm. The present investigation revealed that a set of 68 elite germplasm accessions would have a competitive edge over the current resistance donors being utilized in the breeding programs. Overall, these results might be useful to identify and incorporate the resistance genes from germplasm into elite cultivars through marker assisted selection in rice breeding.

Feasibility on Differentiation of Resistance of Rice Varieties to Whitebacked Planthopper (Sogatella furcifera) using Radioisotope (방사성동위원소를 이용한 흰등멸구(Sogatella furcifera)의 저항성 검정법에 관한 연구 (2))

  • Chung K.H.;Choi S.Y.
    • Korean journal of applied entomology
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    • v.22 no.1 s.54
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    • pp.41-44
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    • 1983
  • In order to evaluate the technique of P-32 labelling method for screening lines of rice to whitebacked planthopper, the relationship between the amount of ingestion and feeding preference of insects were observed with the resistant and susceptible lines where characteristics were predetermined by comparing their feeding preference and antibiosis method. The feeding preference of the insects was significantly correlated with the amount of P-32 ingestion. It was more manifest in adults than nymphs of the whitebacked planthopper. The density of nymph and radioactivity of P-32 were high in the susceptible line. This suggested that deforming the ingestion amount of P-32 in the insects seemed to be useful technique for accurate screening. For this technique, the uniform labelling of P-32 on the rice seedings was prerequisite and the uniformity was increased by triming roots and leaves in the length and number.

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Isolation and Characterization of Some Promoter Sequences from Leuconostoc mesenteroides SY2 Isolated from Kimchi

  • Park, Ji Yeong;Jeong, Seon-Ju;Kim, Jeong A;Kim, Jeong Hwan
    • Journal of Microbiology and Biotechnology
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    • v.27 no.9
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    • pp.1586-1592
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    • 2017
  • Some promoters were isolated and characterized from the genome of Leuconostoc mesenteroides SY2, an isolate from kimchi, a Korean traditional fermented vegetable. Chromosomal DNA of L. mesenteroides SY2 was digested with Sau3AI and ligated with BamHI-cut pBV5030, a promoter screening vector containing a promoterless cat-86. Among E. coli transformants (TFs) resistant against Cm (chloramphenicol), 17 were able to grow in the presence of $1,000{\mu}g/ml$ Cm and their inserts were sequenced. Transcription start sites were examined for three putative promoters (P04C, P25C, and P33C) by primer extension. Four putative promoters were inserted upstream of a promoterless ${\alpha}$-amylase reporter gene in $pJY15{\alpha}$. ${\alpha}$-Amylase activities of E. coli TFs containing $pJY15{\alpha}$ (control, no promoter), $pJY03{\alpha}$ ($pJY15{\alpha}$ with P03C), $pJY04{\alpha}$ (with P04C), $pJY25{\alpha}$ (with P25C), and $pJY33{\alpha}$ (with P33C) were 66.9, 78.7, 122.1, 70.8, and 99.3 U, respectively. Cells harboring $pJY04{\alpha}$ showed 1.8 times higher activity than the control. Some promoters characterized in this study might be useful for construction of food-grade expression vectors for Leuconostoc sp. and related lactic acid bacteria.

Screening of Antagonistic Actinomycetes for Potato Scab Control and Isolation of Antibiotic Compound (감자 더뎅이병원균에 대해 길항활성을 갖는 방선균 탐색 및 항균 활성물질의 분리)

  • Lee, Hyang-Burm;Cho, Jong-Wun;Lim, Chi-Hwan;Kim, Chang-Jin
    • Applied Biological Chemistry
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    • v.47 no.2
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    • pp.164-169
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    • 2004
  • In the course of our screening for biocontrol agent (BCA) against Streptomyces scabiei and S. turgidiscabies causing potato scab using 5,000 actinomtcete isolates, 9 antagonistic strains were selected as BCA candidates through in vitro and in vivo assay. An antagonistic strain, A020645 was highly resistant to some pesticides and antibiotics such as dazomet and mancozeb and showed high control value in vivo. Two bioactive compounds (compound A, B) were purified by anion exchange chromatography, solid phase (ODS) extraction, TLC and reverse phase HPLC. Their chemical structures are now thought to be nucleoside derivative as determined by $^1H-NMR$ data analysis. Their full chemical structures would be elucidated through $^{13}C-NMR$, HMQC and HMBC analyses. Further studies will be focused on fitness in soil and formulation of the BCA candidates.

Effect of Rice Downy Mildew (Sclerophthora macrospora) on Rice Growth and Screening of Disease Resistance of Cultivars (벼 누른오갈병(Sclerophthora macrospora) 발생이 벼 생육에 미치는 영향 및 병 저항성 품종 검정)

  • Lee, Young-Hwan;Cha, Kwang-Hong;Ko, Sug-Ju;Park, Ki-Beum;Kim, Young-Cheol
    • Research in Plant Disease
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    • v.9 no.1
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    • pp.52-56
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    • 2003
  • It was conducted to investigate the effect of rice downy mildew (RDM) infection to plant growth and yield components in water seeding stage, and to screen of varietal resistance to downy mildew. Being infected by rice downy mildew, chlorotic spot appeared in the leaf and leaf length was shortened. As the infected rice was growing, internode was not elongated properly and was deformed, and then panicle was not arised or mal-formed. Plant height of infected rice was shortened at all growth stage, and while the number of tillers of infected rice was more decreased than that of healthy plant before maximum tillering stage, and that of infected rice was more increased after heading stage. While the number of internode of infected tiller was much increased than that of healthy tiller internode length of infected tiller was shorter. As the rice infected by RDM severely, the number of panicles per square meter and ripening of rice was more decreased and yield of rice was extremely much decreased. As result of the varietal resistance screening with rice seedling, Geyh-wabyeo and Donjinbyeo were resistant varieties to downy mildew.