| Construction and application of molecular detection system for adzuki bean rust |
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| DOI：10.7606/j.issn.1000-7601.2023.06.25 |
| Key Words: Vigna angularis rust disease Uromyces vignae molecular detection nested PCR |
| Author Name | Affiliation | | YIN Lihua | Heilongjiang Bayi Agricultural University / National Coarse Cereals Engineering Research Center/Heilongjiang Provincial Key Laboratory of Crop-Pest Interaction Biology and Ecological Control, Daqing, Heilongjiang 163319, China | | ZHANG Jinpeng | Keshan Branch of Heilongjiang Academy of Agricultural Sciences, Qiqihar, Heilongjiang 161005, China | | ZHANG Guoqing | Heilongjiang Bayi Agricultural University / National Coarse Cereals Engineering Research Center/Heilongjiang Provincial Key Laboratory of Crop-Pest Interaction Biology and Ecological Control, Daqing, Heilongjiang 163319, China | | LIU Guohui | Heilongjiang Agricultural Environment and Cultivated Land Protection Station, Harbin, Heilongjiang 150030, China | | KE Xiwang | Heilongjiang Bayi Agricultural University / National Coarse Cereals Engineering Research Center/Heilongjiang Provincial Key Laboratory of Crop-Pest Interaction Biology and Ecological Control, Daqing, Heilongjiang 163319, China | | ZUO Yuhu | Heilongjiang Bayi Agricultural University / National Coarse Cereals Engineering Research Center/Heilongjiang Provincial Key Laboratory of Crop-Pest Interaction Biology and Ecological Control, Daqing, Heilongjiang 163319, China |
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| Abstract: |
| Adzuki bean rust, caused by the fungus Uromyces vignae Barclay, is one of the most serious diseases in adzuki bean production. Estabishing early molecular detection system can play important roles in the disease early diagnosis and timely prevention. Primers designed based on the ITS sequence of U. vignae and the specific primers of the genus Uromyces were used for specific primer screening in this study. A specific primer pair UV-ITS of U. vignae was screened based on the normal PCR by amplifying the genomic DNA, in which the fungi such as Alternaria solani, Rhizoctonia solanii, Fusarium graminearum, Magnaporthe oryzae, Botryosphaeria dothidea, Fusarium semitectum, Alternaria tenuis, Fusarium oxysporum, and Gaeumanomyces graminis were used as control. Accordingly, a pair of nested PCR primers UV-MX was designed, and a high sensitivity molecular detection system for adzuki bean rust based on nested PCR was established. The nested PCR system still achieved effective amplification when the genomic DNA concentration was only 4×10-6 ng·μL-1, and the detection efficiency was 100,000 times higher than the normal PCR. The application effect of this detection system was tested for U. vignae detection in the leaves of susceptible cultivar ‘Baoqinghong’ that inoculated with U. vignae at different time points. The results showed that a specific band of U. vignae was amplified at 12 h after inoculation. The molecular detection system of adzuki bean rust established in this study has high specificity and sensitivity and has the potential for early diagnosis of latent disease. This nested PCR detection system can provide important technical support for early diagnosis and timely prevention of adzuki bean rust. |
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