Primer for detecting pseudomonas syringae pv.actinidae of kiwifruit as well as detection method and application thereof

A technology of kiwi fruit and pathogenic bacteria, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., to achieve the effect of easy implementation and simple detection process

Inactive Publication Date: 2018-07-24
绵阳市农业科学研究院 +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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  • Primer for detecting pseudomonas syringae pv.actinidae of kiwifruit as well as detection method and application thereof
  • Primer for detecting pseudomonas syringae pv.actinidae of kiwifruit as well as detection method and application thereof

Examples

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[0041] Example 1

[0042] (1) Extract DNA from kiwi fruit; the specific method is:

[0043] 1) Take an appropriate amount of kiwi fruit into a 1.5ml EP tube, add liquid nitrogen, and quickly grind it into powder with a grinding rod;

[0044] 2) Add 550μL of 65°C preheated CTAB extract, shake vigorously, and place it in a 65°C incubator for 1 hour with an interval of 10 minutes between shaking and mixing once.

[0045] 3) Add 550μL of chloroform-isoamyl alcohol (24:1v / v), shake vigorously and mix well, let stand for 10min, centrifuge at 13000r / min for 5min;

[0046] 4) Take the supernatant, add 600μL of ice ethanol, centrifuge at 13000r / min for 5min, discard the supernatant, and rinse the pellet with 75% ethanol; repeat this step at least once;

[0047] 5) Centrifuge at 7500r / min for 3min, suck up the excess liquid at the bottom with a pipette, and let it dry naturally at room temperature;

[0048] 6) After drying, add proper amount of sterilized ddH 2 O is fully dissolved, and then stored...

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Abstract

The invention provides a group of specific primers for high sensitively detecting pseudomonas syringae pv.actinidae of the kiwifruit. The specific primer consists of a forward primer and a reverse primer, the forward primer is 5'-AAAACCGCTGTGATACAATTCG-3', as shown in SEQ ID NO. 1, and the reverse primer is 5'-GATAGCCCTGAATGGTTTCCGAA-3', as shown in SEQ ID NO.2. The invention also provides a detection method and application based on the specific primer.

Description

technical field [0001] The invention belongs to the technical field of agriculture and plant quarantine, and in particular relates to a high-sensitivity specific primer for detecting kiwifruit canker pathogen Pseudomonas syringae pv. actinidae, a detection method and an application. Background technique [0002] Kiwi fruit is known as the king of fruits and the crown of vitamin C. It has high economic benefits. The main domestic planting areas are Sichuan, Shaanxi and Guizhou. The detection, prevention and control of kiwifruit diseases is a necessary means to improve its economic value and promote the economic development of the kiwifruit industry in the region. [0003] Canker is a fatal bacterial disease that seriously threatens the production of kiwifruit, and has been listed as a national forest plant quarantine object. Pseudomonas syringae is the main pathogenic bacteria of kiwifruit disease, and one of the main pathogenic bacteria is currently identified as Pseudomona...

Claims

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Application Information

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IPC IPC(8): C12Q1/689C12Q1/686C12Q1/04C12N15/11C12R1/38
CPCC12Q1/686C12Q1/689C12Q2565/125
Inventor 石军胡容平黄廷友刘定友项祖芬侍守佩彭涛李春财杨伟张杰
Owner 绵阳市农业科学研究院
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