• Title/Summary/Keyword: plant diseases detection

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Simultaneous Detection of Three Bacterial Seed-Borne Diseases in Rice Using Multiplex Polymerase Chain Reaction

  • Kang, In Jeong;Kang, Mi-Hyung;Noh, Tae-Hwan;Shim, Hyeong Kwon;Shin, Dong Bum;Heu, Suggi
    • The Plant Pathology Journal
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    • v.32 no.6
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    • pp.575-579
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    • 2016
  • Burkholderia glumae (bacterial grain rot), Xanthomonas oryzae pv. oryzae (bacterial leaf blight), and Acidovorax avenae subsp. avenae (bacterial brown stripe) are major seedborne pathogens of rice. Based on the 16S and 23S rDNA sequences for A. avenae subsp. avenae and B. glumae, and transposase A gene sequence for X. oryzae pv. oryzae, three sets of primers had been designed to produce 402 bp for B. glumae, 490 bp for X. oryzae, and 290 bp for A. avenae subsp. avenae with the $63^{\circ}C$ as an optimum annealing temperature. Samples collected from naturally infected fields were detected with two bacteria, B. glumae and A. avenae subsp. avenae but X. oryzae pv. oryzae was not detected. This assay can be used to identify pathogens directly from infected seeds, and will be an effective tool for the identification of the three pathogens in rice plants.

Characteristics of the Infection of Tilletia laevis Kuhn (syn. Tilletia foetida (Wallr.) Liro.) in Compatible Wheat

  • Ren, Zhaoyu;Zhang, Wei;Wang, Mengke;Gao, Haifeng;Shen, Huimin;Wang, Chunping;Liu, Taiguo;Chen, Wanquan;Gao, Li
    • The Plant Pathology Journal
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    • v.37 no.5
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    • pp.437-445
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    • 2021
  • Tilletia laevis Kuhn (syn. Tilletia foetida (Wallr.) Liro.) causes wheat common bunt, which is one of the most devastating plant diseases in the world. Common bunt can result in a reduction of 80% or even a total loss of wheat production. In this study, the characteristics of T. laevis infection in compatible wheat plants were defined based on the combination of scanning electron microscopy, transmission electron microscopy and laser scanning confocal microscopy. We found T. laevis could lead to the abnormal growth of wheat tissues and cells, such as leakage of chloroplasts, deformities, disordered arrangements of mesophyll cells and also thickening of the cell wall of mesophyll cells in leaf tissue. What's more, T. laevis teliospores were found in the roots, stems, flag leaves, and glumes of infected wheat plants instead of just in the ovaries, as previously reported. The abnormal characteristics caused by T. laevis may be used for early detection of this pathogen instead of molecular markers in addition to providing theoretical insights into T. laevis and wheat interactions for breeding of common bunt resistance.

Multi-spectral Mueller Matrix Imaging for Wheat Stripe Rust

  • Yang Feng;Tianyu He;Wenyi Ren;Dan Wu;Rui Zhang;Yingge Xie
    • Current Optics and Photonics
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    • v.8 no.2
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    • pp.192-200
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    • 2024
  • Wheat stripe rust, caused by Puccinia striiformis, has reduced winter wheat yield globally for ages. In this work, multi-spectral Mueller matrix imaging with 37 measurements using the method of double rotatable quarter-wave plates was used to investigate wheat stripe rust. Individual Mueller matrix measurements were performed on incident monochromatic light with nine bands in the range of 430 to 690 nm. As a result, it was found that the infected area absorbed linearly polarized light and was sensitive to circularly polarized light in the spectral domain. Both linear depolarization and linear diattenuation images distinguished between wheat stripe rust and healthy tissue. The responsiveness of stripe rust to polarized light reveals the potential of using polarization imaging to detect plant diseases. This further suggests that the multi-spectral Mueller matrix imaging system provides us with an alternative approach to agricultural disease detection.

Simultaneous Detection of Barley Virus Diseases in Korea (국내 맥류에 발생하는 바이러스병 동시진단 방법)

  • Lee, Bong-Choon;Bae, Ju-Young;Kim, Sang-Min;Ra, Ji-Eun;Choi, Nak Jung;Choi, Man Young;Park, Ki Do
    • Research in Plant Disease
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    • v.23 no.4
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    • pp.363-366
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    • 2017
  • Barley mild mosaic virus (BaMMV), Barley yellow mosaic virus (BaYMV) and Barley yellow dwarf virus (BYDV) have been identified as an important causative agents for an economically important disease of winter barley in Korea. In this study, a multiplex reverse transcription polymerase chain reaction (mRT-PCR) method was used for the simultaneous detection. Three sets of virus-specific primers targeted to the capsid protein coding genes of BaMMV, BaYMV and BYDV were used to amplify fragments that were 594 bp, 461 bp, and 290 bp, respectively. Several sets of primers for each target virus were evaluated for their sensitivity and specificity by multiplex RT-PCR. The optimum primer concentrations and RT-PCR conditions were determined for the multiplex RT-PCR. The mRT-PCR assay was found to be a better and rapid virus diagnostic tool of specific barley diseases and potential for investigating the epidemiology of these viral diseases.

Improved Detection and Purification of Grapevine Leafroll-associated 3 Closterovirus Using Tissue Culture (포도 조직배양에 의한 Grapevine Leafroll-associated 3 Closterovirus의 증식과 검출효율 증대)

  • 김현란;정재동;정봉남;이봉춘;박진우;최용문
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.6
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    • pp.335-339
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    • 2001
  • Grapevine leafroll-associated 3 closterovirus (GLRaV-3) is phloem limited virus, and one of the most severe viral diseases found in Korea. However, nonhomogenous distribution and low concentration and seasonal variations of GLRaV-3 in grapevines remain as main problems which prevent the introduction and molecular biology or serology experiments. Virus-infected plantlet in vitro was obtained from node tissue cuttings, which was GLRaV-3 infected 'kyoho' vines. The amount of purified virus was highest in vitro plantlet. Moreover, viruses seem to be relatively homogeneously distributed in all organs including leaf, stem and callus derived from in vitro plantlets. RT-PCR detection using in vitro plantlet tissue as template was most effective. When comparing ELISA to RT-PCR, RT-PCR detection was 1,000 times as effective as ELISA. These results can be explained by improved quality such as tenderness or less tannins in plantlet in vitro. In conclusion, until infected herbaceous host will be available, tissue culture can be usefully adopted as a technique for a good source of GLRaV-3 closterovirus for further studies.

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PCR Detection Method for Rapid Diagnosis of Bacterial Canker Caused by Clavibacter michiganensis on Tomato (토마토 궤양병 신속 진단을 위한 Clavibacter michiganensis의 PCR 검출법)

  • Park, Mi-Jeong;Back, Chang-Gi;Park, Jong-Han
    • Research in Plant Disease
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    • v.24 no.4
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    • pp.342-347
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    • 2018
  • Bacterial canker caused by Clavibacter michiganensis is considered to be one of the most serious diseases, leading to economic damage to tomato worldwide. Diagnosis of the bacterial canker on tomato is known to be difficult because the causal pathogen is slow-growing on artificial media as well as causes latent infection in tomato. In this study, as a less time-consuming method, a specific primer set was newly designed for rapid detection of C. michiganensis. The method presented here is so simple, easy, and fast that it can be useful and practical in direct detection of the bacterial canker pathogen from tomato plants.

One-step Multiplex RT-PCR Method for Simultaneous Detection of Seed Transmissible Bacterium and Virus Occurring on Brassicaceae Crop Seeds (십자화과 작물 종자에서 종자전염 세균 및 바이러스 동시 검출을 위한 One-step Multiplex RT-PCR 방법)

  • Jeong, Kyu-Sik;Soh, Eun-Hee
    • Research in Plant Disease
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    • v.17 no.1
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    • pp.52-58
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    • 2011
  • The aim of this research was to develop specific and sensitive PCR-based procedures for simultaneous detection of economically important plant pathogenic bacteria and seed borne virus in commercial Brassicaceae crop seeds, Xanthomonns campestris pv. campestris (Xcc) and Lettuce Mosaic Virus (LMV). Bacterial and virus diseases of Brassicaceae leaves are responsible for heavy losses. PCR with arbitral primers: selection of specific primers, performance of PCR with specific primers and determination of the threshold level for pathogens detection. To detect simultaneously the Xcc and LMV in commercial Brassicaceae crop seeds (lettuce, kohlrabi, radish, chinese cabbage and cabbage), two pairs of specific primer (LMV-F/R, Xcc-F/R) were synthesized by using primer-blast program (http://www.ncbi.nlm.nih.gov/tools/primer-blast/). The multiplex PCR for the two pathogens in Brassicaceae crop seeds could detect specifically without interference among primers and/or cDNA of other plant pathogens. The pathogen detection limit was determined at 1 ng of RNA extracted from pathogens. In the total PCR results for pathogen detection using commercial kohlrabi (10 varieties), lettuce (50 varieties), radish (20 varieties), chinese cabbage (20 varieties) and cabbage (20 varieties), LMV and Xcc were detected from 39 and 2 varieties, respectively. In the PCR result of lettuce, LMV and Xcc were simultaneously detected in 8 varieties.

Detection, isolation, and characterization of the cucumber mosaic virus in Pseudostellaria heterophylla from Korea

  • Lee, Da Hyun;Kim, Jinki;Han, Jun Soo;Lee, Jae-Hyeon;Lee, ByulHaNa;Park, Chung Youl
    • Journal of Plant Biotechnology
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    • v.47 no.2
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    • pp.150-156
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    • 2020
  • Weeds play an important role in the survival of viruses and are potential inoculum sources of viral diseases for crop plants. In this study, specimens of Pseudostellaria heterophylla exhibiting symptoms of the cucumber mosaic virus (CMV) were collected in Bonghwa, Korea. The characteristics of the disease were described and leaf RNA was extracted and sequenced to identify the virus. Three CMV contigs were obtained and PCR was performed using specific primer pairs. RNA from positive samples exhibiting CMV leaf symptoms was amplified to determine the coat protein. A sequence comparison of the coat protein gene from the CMV BH isolate shared the highest nucleotide identity (99.2%) with the CMV ZM isolate. Phylogenetic analysis showed that CMV-BH belonged to subgroup IA and that the most closely-related isolate was CMV-ZM. All test plants used for the biological assay were successfully infected with CMV and exhibited CMV disease symptoms such as blistering, mosaic, and vein yellowing. To our knowledge, this is the first report of CMV infection in P. heterophylla from Korea.

One-step Multiplex RT-PCR Method for Simultaneous Detection of Seed Transmissible Bacteria and Viruses in Pepper and Tomato Seeds (고추와 토마토 종자에서 종자전염 세균 및 바이러스의 동시 검출을 위한 One-step Multiplex RT-PCR 방법)

  • Jeong, Kyu-Sik;Soh, Eun-Hee
    • Research in Plant Disease
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    • v.17 no.1
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    • pp.44-51
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    • 2011
  • The aim of this study was to develop specific and sensitive PCR-based procedures for simultaneous detection of economically important plant seed infection pathogenic bacteria and virus, Xanthomonns campestris pv. vesicatoria (Xcv), Clavibacter michiganensis subsp. michiganensis (Cmm), Erwinia carotovora subsp. carotovora (Ecc), Pepper mild mottle virus (PMMoV) and Tobacco mild green mosaic virus (TMGMV) in pepper and tomato seeds. Most of pepper and tomato bacterial and virus diseases are responsible for germination and growth obstruction. PCR with arbitral primers: selection of specific primers, performance of PCR with specific primers and determination of the threshold level for pathogens detection. To detect simultaneously the Xcv, Cmm, Ecc, PMMoV and TMGMV in pepper and tomato seeds, five pairs (Cmm-F/R, Ecc-F/R, Xcv-F/R, PMMoV-F/R, TMGMV-F/R) of specific primer were synthesized by primer-blast program. The multiplex PCR for the five pathogens in pepper and tomato seeds could detect specially without interference among primers and/or cDNA of plant seeds and other plant pathogens. The PCR result for pathogen detection using 20 commercial pepper and 10 tomato seed samples, Ecc was detected from 4 pepper and 2 tomato seed samples, PMMoV was detected from 1 pepper seed sample, and PMMoV and TMGMV were simultaneously detected from 1 pepper seed sample.

Use of Dienes' Stain in Diagnosis of Plant Mycoplasmal Diseases and Modification of Diagnostic Procedure (Dienes 염색법을 이용한 마이코플라스마성 식물병의 진단과 몇가지 염색방법의 개선)

  • Shin Hyeon Dong;La Yong J eon
    • Korean journal of applied entomology
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    • v.23 no.4 s.61
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    • pp.215-220
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    • 1984
  • Mulberry dwarf, paulownia witches' broom, jujube witches' broom, and sumach witches' broom are known to be associated with mycoplamalike organisms(MLO) in Korea. Simple microscopic detection of MLO infection in these plants was attempted. Periwinkle plant was also tested. Application of $0.2\%\;and\;0.4\%$ solution of Dienes' stain gave diagnoatic value for MLO-induced diseases of periwinkle and mulberry. Among the various plant parts examined, young herbaceous stem just below the apical part gave the best result. Density of staining reaction was proportional to disease severity. Longitudial sections were superior to transverse sections in confirming MLO infection by staining. Light source without blue filter was useful for increasing the color contrast between sieve tube and xylem vessel and for eliminating misinterpretation. Paulownia, jujube, and sumach samples gave no clear difference in staining reaction between healthy and diseased sections even when various modifications of Dienes' staining procedure were tried.

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