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Primer for detecting botryosphaeria dothidea of kiwi fruit and detection method and application

A technology of kiwi fruit and pathogenic bacteria, applied in the field of high-sensitivity specific primers and detection, can solve the problems of unsatisfactory sensitivity, etc., and achieve the effect of easy implementation and simple detection process

Inactive Publication Date: 2018-07-24
INST OF PLANT PROTECTION SICHUAN ACAD OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] At present, although the Chinese patent with the application number 201310233134.3 has realized the specific detection of Botryosphaeria dothidea, the sensitivity in trace detection is not satisfactory

Method used

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  • Primer for detecting botryosphaeria dothidea of kiwi fruit and detection method and application
  • Primer for detecting botryosphaeria dothidea of kiwi fruit and detection method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] (1) DNA is extracted from kiwi fruit; the specific method is:

[0047] 1) Put 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;

[0048] 2) Add 550 μL of CTAB extract solution preheated at 65°C, shake vigorously, place in a 65°C incubator for 1 hour, shake and mix once every 10 minutes

[0049] 3) Add 550 μL of chloroform-isoamyl alcohol (24:1 v / v), shake vigorously and mix well, let stand for 10 min, and centrifuge at 13000 r / min for 5 min;

[0050] 4) Take the supernatant, add 600 μL ice ethanol, centrifuge at 13000 r / min for 5 min, pour off the supernatant, and wash the precipitate with 75% ethanol; repeat this step at least once;

[0051] 5) Empty centrifuge at 7500r / min for 3min, suck off excess liquid at the bottom with a pipette, and let it dry naturally at room temperature;

[0052] 6) After drying, add an appropriate amount of sterilized ddH 2 O was fully dissolved, and then s...

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Abstract

The invention provides a group of specific primers with high sensitive for detecting botryosphaeria dothidea of kiwi fruit. The group of specific primers comprise a forward primer and a reverse primer, wherein the forward primer is 5'-TCATTCGTGATCTCAAGCCAGA-3' and is as shown in SEQ ID No.1, the reverse primer is 5'-TGCTGGTGAAGATAGACATCTGC-3' and is as shown in SEQ ID No.2. The invention further 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 soft rot pathogen Botryosphaeria dothidea, 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] Soft rot is one of the main pathogens of kiwifruit diseases. The pathogen has high spore-producing ability from mid-June to the end of September. When kiwifruit young fruits are infected with this fungus, it will seriously affect the growth of the fruit. When infected in June, pre-harvest fruit drop is prone to occu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/686C12Q1/04C12N15/11C12R1/645
CPCC12Q1/686C12Q1/6895C12Q2565/125
Inventor 胡容平石军陈松叶慧丽陈庆东林立金
Owner INST OF PLANT PROTECTION SICHUAN ACAD OF AGRI SCI
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