Nested PCR reagents and detection system for detecting pear pollen pear fire blight bacteria

A technology of fire blight bacteria and pink pears, which is applied to the nested PCR reagent and detection field for detecting pear pollen blight blight bacteria, can solve problems such as missing and lack of detection methods, and achieve the goal of ensuring safe and efficient production and sustainable development Effect

Pending Publication Date: 2021-09-03
TARIM UNIV +1
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the lack of inspection technology for pear pollen pathogens, any pollination method has a potential safety hazard of spreading pathogens during pollination
[0003] In order to block the spread of pathogens, it is necessary to detect the pathogens in pear pollen, so as to remove the pollen carrying the pathogens. At present, there is no effective detection method to detect the pear pollen branch blight bacteria, so it is urgent to develop a new one. System and method for detecting pear pollen branch blight

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
  • Nested PCR reagents and detection system for detecting pear pollen pear fire blight bacteria
  • Nested PCR reagents and detection system for detecting pear pollen pear fire blight bacteria
  • Nested PCR reagents and detection system for detecting pear pollen pear fire blight bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1, the nested PCR detection of Yali pollen Phytophthora amylovora

[0034] 1. Design of detection primers for Yali pollen Phytophthora amylovora

[0035]The primers used in this implementation test are three pairs of primers derived from the E. amylovora pathogen plasmid pEA29, the first primer pair AJ75 / AJ76, the second primer pair PEANT1 / PEANT2 and the third primer pair p29A / p29B, the sequences of the three pairs of primers, The loci and the size of the amplified bands are shown in Table 1. Primers were synthesized by Sangon Bioengineering (Shanghai) Co., Ltd. As shown in Table 1, the primer list for the detection of E. amylovora from Yali pollen.

[0036] Table 1

[0037]

[0038] 2. Culture of Erwinia amylovora

[0039] After the prepared LB medium was sterilized by steam at 121°C and 0.1MPa for 20 minutes, it was poured into a plate on an ultra-clean workbench, and allowed to cool for later use. Take the preserved Erwinia amylovora strain out of ...

Embodiment 2

[0065] Embodiment 2, the establishment of mock sample nested PCR detection system

[0066] 1. Take 10g of healthy Yali pollen and add it to 50ml of sterilized PBS solution. Shake at 180 rpm on a constant temperature shaker at room temperature for 1 hour. Filter with sterilized filter paper in a clean bench. Take 7 sterilized 1.5ml centrifuge tubes, and centrifuge each Add 1ml of filtrate to the tube, centrifuge at 13000rpm for 10min, discard the supernatant, pour out the supernatant as much as possible, add 900μL of sterilized water to suspend, and finally, add 100μL of 7 concentration gradient bacteria suspensions to the 7 centrifuge tubes, and vortex Mix well and set aside. Sensitivity detection of simulated samples According to the reaction system established above, 2 μL of simulated samples with the first 5 gradient concentrations were taken for sensitivity detection, and ddH 2 O is blank control, detected by 1% agarose gel electrophoresis.

[0067] 2. Sensitivity of moc...

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 discloses nested PCR reagents and a detection system for detecting pear pollen pear fire blight bacteria, the detection reagents comprise a first primer pair and a second primer pair; the nucleotide sequence of the upstream primer of the first primer pair is as shown in SEQ ID NO.1, and the nucleotide sequence of the downstream primer is as shown in SEQ ID NO. 2; and the nucleotide sequence of the upstream primer of the second primer pair is as shown in SEQ ID NO.3, and the nucleotide sequence of the downstream primer is as shown in the specification. According to the invention, the nested PCR system for high-sensitivity detection of the pear pollen branch blight bacteria is established, and the nested PCR molecular method is utilized to establish the high-sensitivity pear pollen blight bacteria detection system, so that the sterility and safety of the commercial pear pollen are ensured, and the path of propagation and diffusion of the bergamot pear branch blight bacteria through the pollen is cut off.

Description

technical field [0001] The invention relates to the technical field of microbial detection, in particular to a nested PCR reagent and a detection method for detecting P. amylovora amylovora from pear pollen. Background technique [0002] Pear is a fruit-bearing crop strictly cross-pollinated. In recent years, the situation of the prevention and control of branch blight of fragrant pears in Korla in Xinjiang is severe. Pollination by bees is restricted, and the fragrant pear industry is seriously threatened. UAV pollination is a highly efficient spray pollination technology that is currently popularized. However, due to the lack of pear pollen pathogen inspection technology, any pollination method has a potential safety hazard of spreading pathogens during pollination. [0003] In order to block the spread of pathogens, it is necessary to detect the pathogens in pear pollen, so as to remove the pollen carrying the pathogens. At present, there is no effective detection method...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/6848C12R1/01
CPCC12Q1/689C12Q1/6848C12Q2531/113C12Q2549/119
Inventor 郝海婷冯宏祖王兰羊坚谢伟
Owner TARIM UNIV
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

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap