A kind of detection method of apple mosaic disease
A technology of apple mosaic virus and detection method, which is applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of difficulty in taking into account virus mutation strains, heavy workload, high cost, etc., and expand the range of detection objects , easy to observe, easy to judge the effect
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Embodiment 1
[0031] Example 1 Establishment of a detection method for apple mosaic disease
[0032] 1.1 Main reagents
[0033] EASYspin Plant RNA Extraction Kit (Beijing Biomed Technology Development Co., Ltd.), Reverse Transcription cDNA Synthesis Kit (Bao Bioengineering Dalian Co., Ltd.), SYBR Reaction Solution (Bao Bioengineering Dalian Co., Ltd.), primer synthesis (Beijing Qingqing Co., Ltd.) Kexinye Biotechnology Co., Ltd.), cloning vector pMD19-T and other conventional reagents (Beijing Leichuang Biotechnology Co., Ltd.).
[0034] 1.2 Real-time fluorescent quantitative PCR reaction
[0035] (1) Extract the total RNA of the apple leaf, branch, flower or fruit sample to be tested, and reverse-transcribe it into cDNA;
[0036] (2) Reaction system: 5.0 μL of SYBR reaction solution, 0.5 μL of 10pM / μL primer solution of SEQ ID No.1 and 0.5 μL of SEQ ID No.2 primer solution, 1.0 μL of cDNA template, and 3.0 μL of RNase-free Water, a total of 10 μL;
[0037] (3) Reaction conditions: The r...
Embodiment 2
[0049] Example 2 Field application of apple mosaic disease detection
[0050] 2.1 Detection object
[0051] In June 2018, a total of 13 apple varieties or rootstock leaves in the apple orchards of the headquarters and experimental base of Beijing Forestry and Fruit Tree Research Institute were collected, and the method of the present invention was applied to detect apple mosaic disease.
[0052] In March 2019, high-temperature detoxification was performed on the above-mentioned apple varieties or rootstock mosaic disease-positive plants; from May to August 2019, the method of the present invention was applied to each detoxified seedling plant obtained by grafting of detoxified young shoots. Initial inspection and re-inspection of leaf disease.
[0053] 2.2 Test results
[0054] The test of 13 apple varieties or rootstocks in the field showed that apple plants with many years of age, such as Fuji, Gala, Wanglin, Hong Jonagold, and rootstock SH6, were positive regardless of wh...
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