- Evaluation of Bakanae Disease Progression Caused by Fusarium fujikuroi in Oryza sativa L.
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In Sun Hwang , Woo-Ri Kang , Duk-Ju Hwang , Shin-Chul Bae , Sung-Hwan Yun , Il-Pyung Ahn
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J. Microbiol. 2013;51(6):858-865. Published online December 19, 2013
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DOI: https://doi.org/10.1007/s12275-013-3472-3
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Bakanae disease caused by Fusarium fujikuroi is an important
fungal disease in rice. Among the seven strains isolated from
symptomatic rice grains in this study, one strain, FfB14,
triggered severe root growth inhibition and decay in the
crown and root of rice seedlings. The remaining six strains
caused typical Bakanae symptoms such as etiolation and
abnormal succulent rice growth. To reveal the relationship
between mycelial growth in the infected tissues and Bakanae
disease progression, we have established a reliable quantification
method
using real time PCR that employs a primer
pair and dual-labeled probe specific to a unigene encoding
F. fujikuroi PNG1 (FfPNG1), which is located upstream of
the fumonisin biosynthesis gene cluster. Plotting the crossing
point (CP) values from the infected tissue DNAs on a standard
curve revealed the active fungal growth of FfB14 in
the root and crown of rice seedlings, while the growth rate
of FfB20 in rice was more than 4 times lower than FfB14.
Massive infective mycelial growth of FfB14 was evident in
rice stems and crown; however, FfB20 did not exhibit vigorous
growth. Our quantitative evaluation system is applicable
for the identification of fungal virulence factors other
than gibberellin.
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Citations
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In Sun Hwang, Il-Pyung Ahn
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Molecular Breeding.2015;[Epub] CrossRef
- Evaluation of Fusarium Head Blight in Barley Infected by Fusarium graminearum
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Woo-Ri Kang , Duk-Ju Hwang , Shin-Chul Bae , Theresa Lee , Soonok Kim , Il-Pyung Ahn
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J. Microbiol. 2013;51(4):540-543. Published online August 30, 2013
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DOI: https://doi.org/10.1007/s12275-013-3338-8
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607
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Abstract
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Fusarium head blight, which is primarily caused by Fusarium graminearum, is a devastating disease in the barley field. A real-time PCR protocol was developed to evaluate the growth of this pathogen in the host plant tissues. All four strains
harbored the gene encoding ATP-BINDING CASSETTE TRANSPORTER (FgABC; FGSG_00541) as a single copy within their genomes. Our Southern blot result was identical with the genomic data for F. graminearum strain PH-1. Based on the crossing point (CP) values obtained in our TaqMan real-time PCR analysis, two standard curves describing the relationship among the CP value, FgABC copy number, and amount of fungal DNA were constructed. Chronological enumeration of fungal growth was coincided with the symptom development.
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Citations
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Combination effect of wheat cultivars and antagonistic activity of
Trichoderma
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Houria Boulahouat, Sihem Touati-Hattab, Leila Mekliche, Houda Boureghda
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- Quantification of Rice Sheath Blight Progression Caused by Rhizoctonia solani
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Mukhamad Su’udi , Jong-Mi Park , Woo-Ri Kang , Duk-Ju Hwang , Soonok Kim , Il-Pyung Ahn
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J. Microbiol. 2013;51(3):380-388. Published online June 28, 2013
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DOI: https://doi.org/10.1007/s12275-013-3274-7
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671
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Rhizoctonia solani has a wide host range, including almost all cultivated crops and its subgroup anastomosis group (AG)-1 IA causes sheath blight in rice. An accurate measurement of pathogen’s biomass is a convincing tool for enumeration
of this disease. Mycological characteristics and molecular diagnosis simultaneously supported that all six strains in this study were R. solani AG-1 IA. Heterokaryons between strains Rs40104, Rs40105, and Rs45811 were stable and viable, whereas Rs40103 and Rs40106 did not form viable fused cells, except for the combination of Rs40106 and Rs40104. A primer pair was highly specific to RsAROM gene of R.
solani strains and the amplified fragment exists as double copies within fungal genome. The relationship between crossing point (CP) values and the amount of fungal DNA was reliable (R2>0.99). Based on these results, we determined R.
solani’s proliferation within infected stems through real time PCR using a primer pair and a Taqman probe specific to the RsAROM gene. The amount of fungal DNA within the 250 ng of tissue DNA from rice cv. Dongjin infected with Rs40104,
Rs40105, and Rs45811 were 7.436, 5.830, and 5.085 ng, respectively. In contrast, the fungal DNAs within the stems inoculated with Rs40103 and Rs40106 were 0.091 and 0.842 ng. The sheath blight symptom progression approximately coincided with the amount of fungal DNA within the symptoms. In summary, our quantitative evaluation method provided reliable and objective results reflecting the amount of fungal
biomass within the infected tissues and would be useful for evaluation of resistance germplasm or fungicides and estimation of inoculum potential.
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Citations
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Asmita Singh, Prashantha S. Tadasanahaller, Sapna Sharma, Gopala Krishnan Subbaiyan, Rashmi Aggarwal, V. Shanmugam, Mahender S. Saharan, Bishnu Maya Bashyal
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Chuping Luo, Huafei Zhou, Jincheng Zou, Xiaoyu Wang, Rongsheng Zhang, Yaping Xiang, Zhiyi Chen
Applied Microbiology and Biotechnology.2015; 99(4): 1897. CrossRef - Evaluation of bakanae disease progression caused by Fusarium fujikuroi in Oryza sativa L.
In Sun Hwang, Woo-Ri Kang, Duk-Ju Hwang, Shin-Chul Bae, Sung-Hwan Yun, Il-Pyung Ahn
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Gyula Oros, Zoltán Naár, Donát Magyar
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- Quantification of Rice Brown Leaf Spot through Taqman Real-Time PCR Specific to the Unigene Encoding Cochliobolus miyabeanus SCYTALONE DEHYDRATASE1 Involved in Fungal Melanin Biosynthesis
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Mukhamad Su’udi , Jong-Mi Park , Woo-Ri Kang , Sang-Ryeol Park , Duk-Ju Hwang , Il-Pyung Ahn
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J. Microbiol. 2012;50(6):947-954. Published online December 30, 2012
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DOI: https://doi.org/10.1007/s12275-012-2538-y
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Rice brown leaf spot is a major disease in the rice paddy field. The causal agent Cochliobolus miyabeanus is an ascomycete fungus and a representative necrotrophic pathogen in the investigation of rice-microbe interactions. The aims of this research were to identify a quantitative evaluation method to determine the amount of C. miyabeanus proliferation in planta and determine the method’s sensitivity. Real-time polymerase chain reaction (PCR) was employed in combination
with the primer pair and Taqman probe specific to CmSCD1, a C. miyabeanus unigene encoding SCYTALONE DEHYDRATASE, which is involved in fungal melanin biosynthesis. Comparative analysis of the nucleotide sequences of CmSCD1 from Korean strains with those from the Japanese and Taiwanese strains revealed some sequence differences. Based on the crossing point (CP) values from Taqman realtime PCR containing a series of increasing concentrations of cloned amplicon or fungal genomic DNA, linear regressions with a high level of reliability (R2>0.997) were constructed. This system was able to estimate fungal genomic DNA at the picogram level. The reliability of this equation was further confirmed using DNA samples from both resistant and susceptible cultivars infected with C. miyabeanus. In summary, our quantitative system is a powerful alternative in brown leaf spot forecasting and in the consistent evaluation of disease progression.
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In Sun Hwang, Woo-Ri Kang, Duk-Ju Hwang, Shin-Chul Bae, Sung-Hwan Yun, Il-Pyung Ahn
Journal of Microbiology.2013; 51(6): 858. CrossRef
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