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Multiple RT-PCR detection method of three sweet potato viruses

A technology of RT-PCR and sweet potato virus, applied in the field of bioengineering, can solve problems such as quality deterioration, seed degeneration, and production hazards of sweet potatoes, and achieve the effects of reducing production costs, improving detection efficiency, ensuring quality and increasing production

Active Publication Date: 2010-06-09
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

The three viruses all belong to the genus Potyvirus, and they often infect together, resulting in reduced yield, poor quality and species degradation of sweet potatoes, causing serious damage to sweet potato production.
However, this method is not suitable for the detection of sweet potato virus

Method used

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  • Multiple RT-PCR detection method of three sweet potato viruses
  • Multiple RT-PCR detection method of three sweet potato viruses
  • Multiple RT-PCR detection method of three sweet potato viruses

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Experimental program
Comparison scheme
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Embodiment 1

[0034] Embodiment one: the multiplex RT-PCR detection method of sweet potato virus

[0035] (1) Materials and methods

[0036] 1. Design and synthesis of primers:

[0037] Three pairs of primers were designed according to the conserved regions of the nucleotide sequences of the coat protein (CP) genes of SPLV, SPVG and SPFMV, and the primers were synthesized by TaKaRa Company.

[0038] Primers for SPFMV are:

[0039] SEQ 1: 5'-CTTCAGTGACGTTGCTGAAGC-3'

[0040] SEQ 2: 5'-TGCACACCCCTCATTCCTAAG-3'

[0041] The primers for SPLV are:

[0042] SEQ 3: 5'-CCAAATATCAATGGTGATTGGGT-3'

[0043] SEQ 4: 5'-CCACGCATTCCAAGTAGTGTGTGT-3'

[0044] The primers for SPVG are:

[0045] SEQ 5: 5'-GCACCAATGGCAAATGGAAGAATAG-3'

[0046] SEQ 6: 5'-GAGGTTATGTATATTCCTTGTAAC-3'

[0047] The sizes of target fragments amplified by the above three pairs of primers for SPFMV, SPLV and SPVG three viruses are 300bp, 420bp and 600bp respectively.

[0048] 2. Virus material

[0049] The stem tips of pota...

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Abstract

The invention relates to the bioengineering technical field, in particular to relate to a multiplex RT-PCR detection process of three sweet potato viruses. The process comprises respectively synthesizing sweet potato latent virus primer, sweet potato G virus primer and sweet potato feathery mottle virus primer, then extracting complete RNA of virus in sample, conducting reverse transcription, respectively placing upstream primer and downstream primer of virus in a PCR reaction system after mixed, conducting PCR amplification using reverse transcription products as template after placing in thePCR reaction system, and finally detecting amplified products with agarose gel electrophoresis. The process builds a multiplex RT-PCR detection technology which can simultaneously detect three sweetpotato viruses, namely simultaneously detecting three sweet potato viruses in one-time PCR reaction, effectively increases detection efficiency, reduces production cost, and can guarantee the qualityof virus free sweet potato and the effect of increasing yield.

Description

1. Technical field: [0001] The invention relates to the technical field of bioengineering, in particular to a multiple RT-PCR detection method for three sweet potato viruses. 2. Background technology: [0002] Sweet potato latent virus (SPLV), sweet potato G virus (Sweet PotatoVirus G, SPVG) and sweet potato feathery mottle virus (Sweet potato feathery mottle virus, SPFMV) are the main viruses that harm sweet potato. The three viruses belong to the genus Potyvirus, and they often infect together, resulting in reduced yield, poor quality and species degradation of sweet potato, causing serious harm to sweet potato production. There is no particularly effective chemical control method for sweet potato virus disease at present. The most effective way to control virus disease is to cultivate virus-free sweet potato by culturing shoot apex meristem. In the process of cultivating virus-free sweet potatoes, it is necessary to detect viruses on sweet potato shoot-tip seedlings. The...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/68
Inventor 张振臣张业辉张德胜蒋士君乔奇王永江秦艳红
Owner INST OF PLANT PROTECTION HENAN ACAD OF AGRI SCI