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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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The Plant Pathology Journal
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Journal DOI :
Korean Society of Plant Pathology
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Volume & Issues
Volume 27, Issue 4 - Dec 2011
Volume 27, Issue 3 - Sep 2011
Volume 27, Issue 2 - Jun 2011
Volume 27, Issue 1 - Mar 2011
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Evaluation of Potential Reference Genes for Quantitative RT-PCR Analysis in Fusarium graminearum under Different Culture Conditions
Kim, Hee-Kyoung ; Yun, Sung-Hwan ;
The Plant Pathology Journal, volume 27, issue 4, 2011, Pages 301~309
DOI : 10.5423/PPJ.2011.27.4.301
The filamentous fungus Fusarium graminearum is an important cereal pathogen. Although quantitative realtime PCR (qRT-PCR) is commonly used to analyze the expression of important fungal genes, no detailed validation of reference genes for the normalization of qRT-PCR data has been performed in this fungus. Here, we evaluated 15 candidate genes as references, including those previously described as housekeeping genes and those selected from the whole transcriptome sequencing data. By a combination of three statistical algorithms (BestKeeper, geNorm, and NormFinder), the variation in the expression of these genes was assessed under different culture conditions that favored mycelial growth, sexual development, and trichothecene mycotoxin production. When favoring mycelial growth, GzFLO and GzUBH expression were most stable in complete medium. Both EF1A and GzRPS16 expression were relatively stable under all conditions on carrot agar, including mycelial growth and the subsequent perithecial induction stage. These two genes were also most stable during trichothecene production. For the combined data set, GzUBH and EF1A were selected as the most stable. Thus, these genes are suitable reference genes for accurate normalization of qRT-PCR data for gene expression analyses of F. graminearum and other related fungi.
Effect of Rice stripe virus NS3 on Transient Gene Expression and Transgene Co-Silencing
Sohn, Seong-Han ; Huh, Sun-Mi ; Kim, Kook-Hyung ; Park, Jin-Woo ; Lomonossoff, George ;
The Plant Pathology Journal, volume 27, issue 4, 2011, Pages 310~314
DOI : 10.5423/PPJ.2011.27.4.310
Nonstructural protein 3 (NS3) encoded by RNA3 of Rice stripe virus (RSV), known to be a suppressor of gene silencing, was cloned and sequenced. The cloned NS3 gene is composed of 636 nucleotides encoding 211 deduced amino acids, and showed a high degree of similarity with the equivalent genes isolated from Korea, Japan and China. The NS3 gene promoted the enhancement of transient gene expression and suppressed transgene co-silencing. In the transient GFP expression via agroinfiltration, GFP expression was dramatically enhanced in terms of both protein yield and expression period in the presence of NS3. The highest accumulation of GFP protein reached to 6.8% of total soluble proteins, which corresponded to a two-fold increase compared to that obtained in the absence of NS3. In addition, NS3 significantly suppressed the initiation of GFP co-silencing induced by the additive GFP infiltration in GFP-transgenic Nicotiana benthamiana. The NS3 gene was also found to be a stronger suppressor than Cucumber mosaic virus 2b. These observations are believed to be derived from the strong suppressive effect of NS3 on gene silencing, and indicate that NS3 could be used as an effective enhancer for the rapid production of foreign proteins in plants.
Soybean mosaic virus Infection and Helper Component-protease Enhance Accumulation of Bean pod mottle virus-Specific siRNAs
Lim, Hyoun-Sub ; Jang, Chan-Yong ; Bae, Han-Hong ; Kim, Joon-Ki ; Lee, Cheol-Ho ; Hong, Jin-Sung ; Ju, Ho-Jong ; Kim, Hong-Gi ; Domier, Leslie L. ;
The Plant Pathology Journal, volume 27, issue 4, 2011, Pages 315~323
DOI : 10.5423/PPJ.2011.27.4.315
Soybean plants infected with Bean pod mottle virus (BPMV) develop acute symptoms that usually decrease in severity over time. In other plant-virus interactions, this type of symptom recovery has been associated with degradation of viral RNAs by RNA silencing, which is accompanied by the accumulation of virus-derived small interfering RNAs (siRNAs). In this study, changes in the accumulation of BPMV siRNAs were investigated in soybean plants infected with BPMV alone, or infected with both BPMV and Soybean mosaic virus (SMV) and in transgenic soybean plants expressing SMV helper component-protease (HC-Pro). In many potyviruses, HC-Pro is a potent suppressor of RNA silencing. In plants infected with BPMV alone, accumulation of siRNAs was positively correlated with symptom severity and accumulation of BPMV genomic RNAs. Plants infected with both BPMV and SMV and BPMV-infected transgenic soybean plants expressing SMV HC-Pro exhibited severe symptoms characteristic of BPMVSMV synergism, and showed enhanced accumulation of BPMV RNAs and siRNAs compared to plants infected with BPMV alone and nontransgenic plants. Likewise, SMV HC-Pro enhanced the accumulation of siRNAs produced from a silenced green fluorescent protein gene in transient expression assays, while the P19 silencing suppressor of Tomato bushy stunt virus did not. Consistent with the modes of action of HC-Pro in other systems, which have shown that HC-Pro suppresses RNA silencing by preventing the unwinding of duplex siRNAs and inhibiting siRNA methylation, these studies showed that SMV HC-Pro interfered with the activities of RNA-induced silencing complexes, but not the activities of Dicer-like enzymes in antiviral defenses.
Evaluation and Verification of Barley Genotypes with Known Genes for Resistance to Barley yellow mosaic virus and Barley mild mosaic virus Under Field Conditions in South Korea
Kim, Hong-Sik ; Baek, Seong-Bum ; Kim, Dea-Wook ; Hwang, Jong-Jin ; Kim, Si-Ju ;
The Plant Pathology Journal, volume 27, issue 4, 2011, Pages 324~332
DOI : 10.5423/PPJ.2011.27.4.324
Soil-borne barley yellow mosaic disease caused by Barley yellow mosaic virus (BaYMV) or Barley mild mosaic virus (BaMMV) gives a serious threat to the winter barley cultivated in the southern regions in Korea. It is important to develop resistant varieties for stable and high-yield production. The objectives of this study were to evaluate 22 genotypes of exotic barley germplasms carrying the resistance genes rym1 through rym12, with the exception of rym10, and to determine the genes that confer resistance to BaYMV or BaMMV in Korea. Using the traditional visual scoring of symptoms at 4 locations over 3 years, average disease rate values differed (P < 0.001) among the genotypes. ELISA test revealed the presence of both BaYMV and BaMMV in all of the field sites but Jinju and significantly different rates of infection among genotypes and years. Barley genotypes differed in how virus quantities and pathogen-induced symptoms were correlated, especially in response to BaYMV. Disease incidence was affected by the climatic conditions present during the early growing stage before overwintering. A Chinese landrace, 'Mokusekko 3', carrying rym1 and rym5 was comparatively resistant at all locations studied. The barley genotypes carrying either rym6 or rym9 were susceptible to the viral strains. The genotypes carrying rym5 were resistant in Jinju and Milyang but susceptible in Iksan and Naju. The resistance genes rym2 and rym3 were effective in local strains and would be potent contributors to disease resistance.
Biocontrol Efficacies of Bacillus Species Against Cylindrocarpon destructans Causing Ginseng Root Rot
Jang, Ye-Lim ; Kim, Sang-Gyu ; Kim, Young-Ho ;
The Plant Pathology Journal, volume 27, issue 4, 2011, Pages 333~341
DOI : 10.5423/PPJ.2011.27.4.333
Two antifungal bacteria were selected from forest soils during the screening of microorganisms antagonistic to Cylindrocarpon destructans, a cause of ginseng root rot. The antifungal bacteria were identified as Bacillus subtilis (I4) and B. amyloliquefaciens (yD16) based on physiological and cultural characteristics, the Biolog program, and 16S rRNA gene sequencing analyses. Antagonistic activity of both bacterial isolates to C. destructans increased with increasing temperature. More rapid starch hydrolytic activity of the bacteria was seen on starch agar at higher temperatures than at lower temperatures, and in the higher density inoculum treatment than in the lower density inoculum treatment. The bacterial isolates failed to colonize ginseng root the root tissues inoculated with the bacteria alone at an inoculum density of
cfu/ml, but succeeded in colonizing the root tissues co-inoculated with the bacteria and C. destructans. Scanning electron microscopy showed that the pathogen was damaged by the low-density inoculum treatment with the bacterial isolates as much as by the high-density inoculum treatment. Both bacterial isolates were more effective in reducing root rot when they were treated at a concentration of
cfu/ml than at
cfu/ml. Also, only the former treatment induced prominent wound periderm formation, related to structural defense against pathogen infection. The results suggest that the bacterial antagonists may have high potential as biocontrol agents against ginseng root rot at relatively low-inoculum concentrations.
Purification and Structural Analysis of Surfactin Produced by Endophytic Bacillus subtilis EBS05 and its Antagonistic Activity Against Rhizoctonia cerealis
Wen, Cai-Yi ; Yin, Zhi-Gang ; Wang, Kai-Xuan ; Chen, Jian-Guang ; Shen, Shun-Shan ;
The Plant Pathology Journal, volume 27, issue 4, 2011, Pages 342~348
DOI : 10.5423/PPJ.2011.27.4.342
Bacillus subtilis EBS05, an endophytic bacteria strain isolated from a medicinal plant Cinnamomum camphor, can produce antagonistic compounds that effectively inhibit plant pathogenic fungi. The greenhouse experiments showed that wheat sharp eyespot disease (WSED) was reduced by 91.2%, 88.2% and 43.0% after the treatment with fermentation broth, bacteria-free filter and a fungicide fludioxonil, respectively. The culture broth of strain EBS05 can more effectively control WSED than can fludioxonil. The fermentation broth and bacteria-free filter ability to suppress WSED was not significantly different, suggesting that an active secreted substance played a major role in controlling WSED. Separation and purification of the active compounds was carried out by serial processes, including hydrochloric acid (pH 2.0) treatment, methanol extraction and Sephadex LH-20 column chromatography, silica gel column chromatography and reverse-phase high-pressure liquid chromatography (HPLC), respectively. The purified compounds, one of active peaks in the HPLC spectrum, were obtained from the collection. Analysis of the chemical structures by time-of-flight mass spectrometry (TOF-MS) and electrospray ionization mass spectrometry/mass spectrometry (ESI-MS/MS) showed that the active substances produced by the endophytic bacteria EBS05 are mixture of the
surfactin A isomers with 1035.65 Da, 1021.64 Da, 1007.63 Da and 993.65 Da molecular weights, respectively.
Estrogenic Compounds Compatible with a Conditional Gene Expression System for the Phytopathogenic Fungus Fusarium graminearum
Lee, Jung-Kwan ; Son, Ho-Kyoung ; Lee, Yin-Won ;
The Plant Pathology Journal, volume 27, issue 4, 2011, Pages 349~353
DOI : 10.5423/PPJ.2011.27.4.349
The ascomycete fungus Fusarium graminearum is an important plant pathogen responsible for Fusarium head blight in small grains and ear rot on maize. This fungus also produces the estrogenic metabolite, zearalenone (ZEA) that causes estrogenic disorders in humans and animals. Previously, we developed a conditional gene expression system for this fungus using a ZEA-inducible promoter (Pzear). In the present study, four other estrogenic compounds, including
-estradiol, estriol, estrone, and secoisolariciresinol, were screened as possible substitutes for ZEA in this system. Among them,
-estradiol was able to successfully induce the expression of a gene controlled by Pzear, while estrone was only able to partially induce its expression but the other two compounds were not effective. In combination, these results demonstrate that
-estradiol can replace ZEA in this conditional gene expression system, thereby eliminating the need to use the more expensive reagent, ZEA, and facilitating high-throughput functional analyses of F. graminearum in future studies.
Difference in Chemotype Composition of Fusarium graminearum Populations Isolated from Durum Wheat in Adjacent Areas Separated by the Apennines in Northern-Central Italy
Prodi, A. ; Purahong, W. ; Tonti, S. ; Salomoni, D. ; Nipoti, P. ; Covarelli, L. ; Pisi, A. ;
The Plant Pathology Journal, volume 27, issue 4, 2011, Pages 354~359
DOI : 10.5423/PPJ.2011.27.4.354
Chemotype composition of Fusarium graminearum strains, isolated from durum wheat kernels from naturally FHB infected fields in Northern and Central Italy, was investigated by multiplex PCR. The different climatic and environmental conditions of the two examined areas separated by the Apennines affected the composition of chemotypes. 15Ac-DON chemotype was predominant in both the sub areas. Nivalneol chemotype was more frequent in the warmer sub area.
Reclassification of Xanthomonas Isolates Causing Bacterial Leaf Spot of Euphorbia pulcherrima
Li, Bin ; Yu, Rongrong ; Shi, Yu ; Su, Ting ; Wang, Fang ; Ibrahim, Muhammad ; Xie, Guanlin ; Wang, Yanli ; Sun, Guochang ;
The Plant Pathology Journal, volume 27, issue 4, 2011, Pages 360~366
DOI : 10.5423/PPJ.2011.27.4.360
Bacterial leaf spot of Euphorbia pulcherrima has been reported in many countries. Characterization by polyphasic approaches indicated that the isolates from India, USA and New Zealand could be distinguished based on rep-PCR profiles and gyrB phylogenies, while the Chinese isolates should be ascribed to Xanthomonas axonopodis pv. poinsettiicola.
Multiplex PCR Assay for Simultaneous Detection of Korean Quarantine Phytoplasmas
Kim, Young-Hwan ; Win, Nang Kyu ; Back, Chang-Gi ; Yea, Mi-Chi ; Yim, Kyu-Ock ; Jung, Hee-Young ;
The Plant Pathology Journal, volume 27, issue 4, 2011, Pages 367~371
DOI : 10.5423/PPJ.2011.27.4.367
Multiplex PCR assays were developed for the simultaneous detection of ten important Korean quarantine phytoplasmas. The species-specific primers were designed based on ribosomal protein, putative preprotein translocase Y, immunodominant protein, elongation factor TU, chaperonin protein and the 16S rRNA genes of 'Candidatus (Ca.) Phytoplasma' species. Three main primer sets were prepared from ten designed primer pairs to limit nonspecific amplification as much as possible. The multiplex PCR assay using the three primer sets successfully amplified the correct conserved genes for each 'Ca. Phytoplasma' species. In addition, ten important 'Ca. Phytoplasma' species could be easily determined by recognizing band patterns specific for each phytoplasma species from three primer sets. Moreover, a high sensitivity of multiplex PCR for each primer set was observed for samples containing a low DNA concentration (10 ng/
). This study provides the useful multiplex PCR assay as a convenient method to detect the presence of ten important quarantine phytoplasmas in Korea.
Characteristics of Cucumber mosaic virus isolated from Zea mays in Korea
Kim, Mi-Kyeong ; Kwak, Hae-Ryun ; Lee, Su-Heon ; Kim, Jeong-Soo ; Kim, Kook-Hyung ; Cha, Byeong-Jin ; Choi, Hong-Soo ;
The Plant Pathology Journal, volume 27, issue 4, 2011, Pages 372~377
DOI : 10.5423/PPJ.2011.27.4.372
A virus causing mottle and stunt symptom on Zea mays was observed around Ulleng-do, Korea and identified as Cucumber mosaic virus (CMV-ZM) based upon biological, serological, and molecular characteristics. In host range studies, the CMV-ZM isolate produced local lesions on Datura stramonium, Vigna unguiculata, Cucurbita moschata, Chenopodium amaranticolor, Ch. quinoa, whereas this isolate produced systemic mosaic on Nicotiana tabacum cv. 'Xanthi-nc', Capsicum annuum, Solanum lycopersicum, Solanum melongena, Cucurbita pepo, and Z. mays. In addition, chlorotic local rings on inoculated leaves along with severe mosaic, malformation, and fern leaf symptoms on upper systemic leaves were shown in N. glutinosa plants. Complete nucleotide sequences of each genomic RNA segment was determined and compared to those of the other CMV strains. Comparison of the nucleotide sequence of 1a open reading frame (ORF) revealed approximately 89.2-92.4% sequence identity with each CMV subgroup IA and IB strain, while showing only 78% sequence identity with CMV subgroup II. Nucleotide sequence analysis of RNA2 ORFs revealed 85.3-97.6% sequence identity with subgroup I. In ORFs of RNA3, levels of nucleotide sequence identities were higher than 92-99.2% with CMV subgroup I and lower than 82% with CMV isolates of subgroup II. These results suggest that CMV-ZM isolate is more closely related to subgroup I than subgroup II and therefore, CMV-ZM isolate might be classified into as CMV subgroup I based on biological and molecular analysis.
Molecular Evidence of Recombination on Korean Isolates of Tomato yellow leaf curl virus by Nucleotide Transversions and Transitions
Lee, Hye-Jung ; Park, Jung-An ; Auh, Chung-Kyoon ; Lee, Kyeong-Yeoll ; Kim, Chang-Seok ; Lee, Gwan-Seok ; Soh, Hyun-Cheol ; Choi, Hong-Soo ; Lee, Suk-Chan ;
The Plant Pathology Journal, volume 27, issue 4, 2011, Pages 378~384
DOI : 10.5423/PPJ.2011.27.4.378
Tomato yellow leaf curl virus (TYLCV), a member of genus Begomovirus, was isolated in Korea in 2008. We sequenced and analyzed the DNA-A of 51 TYLCV isolates from Korea, and 13 of the TYLCV isolates were selected as type representatives of TYLCV from six Korean provinces. The 13 TYLCV isolates were classified into Korea Group 1 (KG1, nine isolates) and Korea Group 2 (KG2, four isolates) based on the results of phylogenetic analysis and genome size (2774 and 2781 nucleotides, respectively). A recombination detection program 3 (RDP3) revealed two recombinations between the TYLCV Korea isolates and other TYLCV isolates [Thailand (AF206674), Iran (AJ132711), and Israel (X76319)]. TYLCV Jeju isolate was characterized by two recombination events (E1 and E2) caused by the presence of E1 in ORF V1 and C3, which may seem to be the mutations of the high nucleotide transversion and transition rate. Collectively, our results suggest that the occurrence of nucleotide transversions and transitions in TYLCV DNA-A might have induced novel recombination events within the TYLCV Korea isolates.
Simple and Rapid Detection of Potato leafroll virus by Reverse Transcription Loop-mediated Isothermal Amplification
Ju, Ho-Jong ;
The Plant Pathology Journal, volume 27, issue 4, 2011, Pages 385~389
DOI : 10.5423/PPJ.2011.27.4.385
A new reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for the Potato leafroll virus (PLRV) was developed and compared with conventional reverse transcription polymerase chain reaction (RT-PCR) to address its advantages over RTPCR. RT-LAMP primers were designed from the open reading frame 3 (ORF3) sequence of PLRV. The RT-LAMP reactions were conducted without or with a set of loop primers. By real-time monitoring using Turbimeter, the RT-LAMP (with loop primers) detects PLRV in less than 30 min, compared to 120 min of RT-PCR. By adding fluorescent reagent during the reaction, final products of the RT-LAMP were fluorescently visualized under UV light or could be differentiated by naked-eye inspection under normal light. The RT-LAMP was extremely sensitive, about 2000-fold more sensitive than RT-PCR. This study presents great potential of the RT-LAMP for diagnosis and PLRV epidemiology because RT-LAMP method is speedy, sensitive, inexpensive, and convenient.
Mannitol Amendment as a Carbon Source in a Bean-based Formulation Enhances Biocontrol Efficacy of a 2,4-diacetylphloroglucinol-producing Pseudomonas sp. NJ134 Against Tomato Fusarium Wilt
Kang, Beom-Ryong ;
The Plant Pathology Journal, volume 27, issue 4, 2011, Pages 390~395
DOI : 10.5423/PPJ.2011.27.4.390
Fusarium wilt caused by Fusarium oxysporum has become a serious problem world-wide and relies heavily on chemical fungicides. We selected Pseudomonas sp. NJ134 to develop an effective biocontrol strategy. This strain shows strong antagonistic activity against F. oxysporum. Biochemical analyses of ethyl-acetate extracts of NJ134 culture filtrates showed that 2,4-diacetylphloroglucinol (DAPG) was the major compound inhibiting in vitro growth of F. oxysporum. DAPG production was greatly enhanced in the NJ134 strain by adding mannitol to the growth media, and in vitro antagonistic activity against F. oxysporum increased. Bioformulations developed from growth of NJ134 in sterile bean media with mannitol as the carbon source under plastic bags resulted in effective biocontrol efficacy against Fusarium wilt. The efficacy of the bioformulated product depended on the carbon source and dose. Mannitol amendment in the bean-based formulation showed strong effective biocontrol against tomato Fusarium wilt through increased DAPG levels and a higher cell density compared to that in a glucose-amended formulation. These results suggest that this bioformulated product could be a new effective biocontrol system to control Fusarium wilt in the field.
Anthracnose Caused by Colletotrichum gloeosporioides on Sweet Crabapple in Korea
Choi, Ok-Hee ; Seo, Jong-Bum ; Kwon, Jin-Hyeuk ; Kim, Jin-Woo ;
The Plant Pathology Journal, volume 27, issue 4, 2011, Pages 396~396
DOI : 10.5423/PPJ.2011.27.4.396
Ophiostoma ips Isolated from Reddish Brown Stained Japanese Red Pine Wood
Kim, Jae-Jin ; Hyun, Min-Woo ; Suh, Dong-Yeon ; Kim, Seong-Hwan ; Shin, Sang-Chul ;
The Plant Pathology Journal, volume 27, issue 4, 2011, Pages 397~397
DOI : 10.5423/PPJ.2011.27.4.397
Occurrence of Podosphaera pannosa Teleomorph on Rosa rugosa from Korea
Lee, Sang-Hyun ; Han, Kyung-Sook ; Park, Ji-Hyun ; Shin, Hyeon-Dong ;
The Plant Pathology Journal, volume 27, issue 4, 2011, Pages 398~398
DOI : 10.5423/PPJ.2011.27.4.398