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Multiple-PCR (polymerase chain reaction) detection and warning method for sweet potato chlorotic stunt virus (SPCSV) and sweet potato twin virus (Sweepoviruses) in sweet potato roots

A technology of chlorotic dwarf virus and gemini virus, applied in the field of bioengineering, can solve the problems of high cost, affecting the quality of nucleic acid extraction and virus detection efficiency, and low efficiency

Active Publication Date: 2017-05-31
INST OF PLANT PROTECTION HENAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A single PCR method requires multiple PCRs, which is costly and inefficient, especially because the starch and polysaccharide content in sweet potato tubers is high, which seriously affects the quality of nucleic acid extraction and the detection efficiency of viruses

Method used

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  • Multiple-PCR (polymerase chain reaction) detection and warning method for sweet potato chlorotic stunt virus (SPCSV) and sweet potato twin virus (Sweepoviruses) in sweet potato roots
  • Multiple-PCR (polymerase chain reaction) detection and warning method for sweet potato chlorotic stunt virus (SPCSV) and sweet potato twin virus (Sweepoviruses) in sweet potato roots
  • Multiple-PCR (polymerase chain reaction) detection and warning method for sweet potato chlorotic stunt virus (SPCSV) and sweet potato twin virus (Sweepoviruses) in sweet potato roots

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1 A kind of multiplex PCR method that simultaneously detects chlorotic dwarf virus (SPCSV) and geminivirus (Sweepoviruses) in sweet potato tuber root

[0038] (1) Materials and methods

[0039] 1. Virus material and sampling method

[0040] Potato tubers from diseased plants harvested and stored in the field were tuber tubers containing SPCSV alone, Sweepoviruses alone, SPCSV and Sweepoviruses at the same time, and sweet potato feather mottle virus (SPFMV) and / or latent virus (SPLV) and other mixed viruses potato chips. Before the test, use a sterilized scalpel blade to cross-cut the 2-3cm of the end of the sweet potato tuber, and dig out the potato flesh for testing.

[0041] 2. Synthesis of primers

[0042]

[0043] 3. Extraction of nucleic acid

[0044]The total RNA of sweet potato tubers was extracted with the Plant Total RNA Extraction Kit produced by Gene Mark Company, following the instructions of RNALysisSolution B. The total DNA extraction of...

Embodiment 2

[0072] Example 2 Effects of Poisonous Seed Potatoes on Symptoms and Yield of Sweet Potato Plants at Seedling Stage and Field Stage

[0073] 1. Materials and methods

[0074] 100 diseased field potato pieces were randomly selected from the laboratory potato cellar, and the variety was Shangshu 19.

[0075] Using the established detection method, 100 potato cubes were tested for virus respectively, and then these 100 potato cubes were respectively raised in flower pots equipped with vermiculite, and the symptoms at the seedling stage were observed, and the potatoes grown from each potato cube were The seedlings were transplanted to the field for planting. The potato seedlings of each potato block were planted in one plot, and the plots were randomly arranged, with a total of 100 plots. During the field period, the symptoms of the plants were regularly observed and recorded, and the severity of the disease in each plot was investigated. The yield was measured during the harvest p...

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Abstract

The invention discloses a multiple-PCR (polymerase chain reaction) detection and warning method for sweet potato chlorotic stunt virus (SPCSV) and sweet potato twin virus (Sweepoviruses) in sweet potato roots. For the two types of viruses of SPCSV and Sweepoviruses, specific primer pairs CP-F and CP-R and primer pairs of VF and VR are designed, the detection primers are good in specificity, and only a stripe of the size of specificity can be amplified in a targeted virus sample. The detection method for SPCSV and Sweepoviruses is established by the aid of multiple-PCR primers, primary PCR reaction can be passed, two kinds of sweet viruses of SPCSV and Sweepoviruses can be detected whether to exist in the sweet potato roots or not, and quality of potato seeds is further judged; relation of sweet potato root virus with sweet potato seedling, field period virus symptoms and yield loss is determined, basis is provided for judging whether the potato seeds are qualified or not by detecting whether the potato seeds carry the virus or not, and great importance is achieved in quality of detoxified sweet potatoes and disease epidemic warning.

Description

technical field [0001] The invention relates to multiple PCR detection primers and a method for detecting sweet potato chlorotic dwarf virus and sweet potato geminivirus in sweet potato tubers and a quality early warning method for virus-free seed potatoes, belonging to the technical field of bioengineering. Background technique [0002] Sweet potato is an important food crop in my country, and it is also an important food processing and industrial raw material. my country is the largest sweet potato producer in the world, and the sweet potato planting area accounts for about 45% of the world's planting area. Viral diseases are an important class of diseases that harm sweet potato, which can cause serious yield loss and seed degradation. At present, there are more than 30 kinds of viruses that have been reported to infect sweet potato in the world. Among them, sweet potato chlorotic dwarf virus (SPCSV) and sweet potato geminivirus (Sweepoviruses) are two types of important ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/94
CPCC12Q1/686C12Q1/701C12Q2600/16C12Q2537/143C12Q2565/125
Inventor 王爽张振臣田雨婷乔奇秦艳红王永江张德胜
Owner INST OF PLANT PROTECTION HENAN ACAD OF AGRI SCI
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