Sugarcane smut bacteria nest type polymerase chain reaction (PCR) quick detection method

A sugarcane smut and detection method technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganism measurement/inspection, etc., can solve the problem of being easily interfered by other pathogens, unable to accurately detect, and poor detection specificity and other problems, to achieve the effect of fast detection, high accuracy and strong specificity

Inactive Publication Date: 2012-01-25
GUANGZHOU SUGARCANE IND RES INST +1
View PDF3 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this method can also detect whether sugarcane is infected with smut, but because it uses fungal ITS general primers instead of sugarcane smut specific primers, its detection specificity is extremely poor, and it is easy to be affected during the detection process. Interference from other pathogens; the sensitivity is limited, and it cannot be accurately detected when the infection amount of smut

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Sugarcane smut bacteria nest type polymerase chain reaction (PCR) quick detection method
  • Sugarcane smut bacteria nest type polymerase chain reaction (PCR) quick detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0026] The nested PCR rapid detection method of sugarcane smut comprises the following steps:

[0027] Step A, genomic DNA extraction:

[0028] In April 2011, 9 heart leaves of sugarcane variety Xintaitang No. 22 seedlings artificially inoculated with teliospores of sugarcane smut were randomly collected (without early symptoms of sugarcane smut). Genomic DNA was extracted, and the samples were obtained and numbered as samples 4-12 in sequence;

[0029] Step B, the first round of PCR amplification:

[0030] Using samples 4-12 as templates, the first round of PCR amplification products were obtained through PCR amplification using universal primers ITS4 and ITS5 of the fungal ribosomal gene transcription spacer ITS;

[0031] The primer sequence is: ITS4:5`-TCCTCCGCTTATTGATATGC-3`;

[0032] ITS5:5`-GGAAGTAAAAGTCGTAACAAGG-3`;

[0033] PCR reaction system: Genomic DNA 1 μL, 5 μmol / L ITS4 and ITS5 primers 1 μL each, Taq enzyme 0.2 μL (5U / μL), 10×PCR reaction buffer (containin...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention relates to the field of sugarcane pathogenic bacteria detection, and provides a sugarcane smut bacteria nest type polymerase chain reaction (PCR) quick detection method. The method comprises the steps of DNA extraction, primary PCR amplification, secondary PCR amplification and PCR amplification product detection, wherein universal primers ITS4 and ITS5 of a fungal ribosome genetic transcription spacer ITS are adopted in the primary PCR amplification; and specific primers Smut-L1 and Smut-R2 of a sugarcane smut ribosome genetic transcription spacer ITS are adopted in the secondary PCR amplification. The method has the substantial characteristics that: 1, the detection sensitivity is extremely high, and the smut bacteria can be accurately detected even if the sugarcane has nosmut symptom or is infected little by the smut bacteria; 2, the method is strong in specificity, low in interference of other bacteria and high in accuracy; and 3, the method has short time consumption and high detection speed, and is suitable for sugarcane variety introduction quarantine, toxin-free seedling quality detection, sugarcane smut resistance identification, early symptom-free leaf detection and the like.

Description

technical field [0001] The invention relates to the field of detection of sugarcane pathogenic bacteria, in particular to a method for detecting sugarcane smut. Background technique [0002] by Saccharomyces sativa ( Ustilago scitaminea ) caused by sugarcane smut is a worldwide important sugarcane disease, and it is also the most economically harmful sugarcane disease in my country. Powdery mildew, sugarcane smut spreads infection during storage and planting. Therefore, the detection method of sugarcane smut during the sugarcane introduction and quarantine process is an important way to prevent the widespread spread of sugarcane smut. [0003] The routine PCR detection method of sugarcane smut, such as the patented "Sugarcane smut rapid detection method" (application number: 201010230534), this conventional PCR detection method mainly includes the following steps: Step 1, using CTAB method to extract sugarcane smut genomic DNA ; Step 2, using the sugarcane smut genomic DNA...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12Q1/68C12R1/645C12Q1/04
Inventor 沈万宽姜子德邓海华习平根刘睿李敏慧陈健文杨湛端陈仲华
Owner GUANGZHOU SUGARCANE IND RES INST
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products