Dual PCR primer for detecting two types of pathogens of grape canker at same time and application of dual PCR primer

A technology of grape canker and Vitis vinifera is applied in the directions of microorganisms, microorganism-based methods, and microbial determination/inspection. The effect of identification time

Active Publication Date: 2015-12-23
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because Botryosphaeria dothidea and Neofusiccum parvum are often mixed infestation, and the symptoms caused by infestation of grapes are similar, it is difficult to distinguish them, so traditional early diagnosis methods cannot quickly obtain accurate results

Method used

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  • Dual PCR primer for detecting two types of pathogens of grape canker at same time and application of dual PCR primer
  • Dual PCR primer for detecting two types of pathogens of grape canker at same time and application of dual PCR primer
  • Dual PCR primer for detecting two types of pathogens of grape canker at same time and application of dual PCR primer

Examples

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Embodiment 1

[0026] Example 1. A set of dual PCR primers for simultaneous detection of two kinds of grape canker bacteria and their detection methods

[0027] 1. Obtaining dual PCR primers for grape canker bacteria Neofusicoccumparvum and Botryosphaeriadothidea

[0028] Extract the EF elongation factor and β-tubulin sequence of 4 species (Botryosphaeriadothidea, Lasiodiplodiatheobromae, Neofusicoccumparvum and Diplodiaseriata) that have been reported to cause grape canker in my country from GeneBank (accession numbers are: JX462265; GU294727; KJ146834; KJ146833; JX462263; JX462263; JX462263) JX462280; HQ288289; JX462291; GU294713; KJ146836; KJ146835; JX462289; JX462299; JX462306; HQ288254; JX521848) for homology comparison. The specific primers Bd-F / Bd-R of Botryosphaeriadothidea were designed according to the differences in the sequence of Botryosphaeriadothidea and the other three EF elongation factors. The sequences are the upstream forward primer Bd-F sequence: 5'-GTCTGCATCATTCTCAGCGTGGG-3' ...

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Abstract

The invention discloses dual PCR primer for detecting two types of pathogens of grape canker at the same time and application of the dual PCR primer. The dual PCR primer is composed of nucleotides shown in the first sequence in the sequence list, nucleotides shown in the second sequence in the sequence list, nucleotides shown in the third sequence in the sequence list and nucleotides shown in the fourth sequence in the sequence list. The primer can detect existence of the two types of pathogens of the grape canker, namely, Neofusicoccum parvum and Botryosphaeria dothidea in a dual PCR reaction, it shows that the primer can rapidly, accurately and flexibly determine the two types of pathogens of the grape canker in grape nursery stock and field grape disease samples, and foundation is provided for nursery stock bacterium-carrying detection and specially preventing and controlling the grape canker caused by the two types of pathogens.

Description

Technical field [0001] The present invention belongs to the invention belongs to a new technology for molecular detection of plant fungal pathogens, and specifically relates to the establishment and application of a method for detecting two grape canker pathogens by adopting double PCR technology. Background technique [0002] Grape canker disease (grapeBotryosphaeriadieback) caused by Botryosphaeriaceae fungi has become more and more harmful in the world in recent years, and has become one of the important factors that threaten the quality and yield of grapes (PascoeI.Trunkdiseasesofgrapevines-perspectivefromatourofCaliforniaGrapegrower and Winemaker.1998 ,417:68-71.VanNiekerkJM,FouriePH,HallennF,etal.Botryosphaeriaspp.asgrapevinetrunkdiseasepathogens.PhytopathologiaMediterranea.2006,45(4):43-54.Urbez-TorresJR.Thefectusofvines.Phyteriantoea. Up to now, reports have shown that there are 17 species in the Botrytis family that can cause grape canker (AugerJ, EsterioM, RickeG, etal....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12R1/645
CPCC12Q1/6895C12Q2600/16
Inventor 张玮燕继晔李兴红郭飞飞刘梅周莹陈震
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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