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A Complete Set of Reagents and Its Application for Detecting Puccinia solani

A technology of Solanum solani mold and reagents, applied in the biological field, can solve the problems of tomato fruit quality decline, etc., and achieve the effects of increasing practicability, reducing detection costs, and high accuracy

Active Publication Date: 2020-05-19
INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Among the 30,000 greenhouses planted in Yutai County, Shandong Province in 2002, the rate of diseased greenhouses was 43%. In 2003, the rate of diseased greenhouses was 90%. If the control was not timely, the tomato production decreased by about 20%, and some even decreased by 80%. Decreased tomato fruit quality
But there is not yet a good method for detecting Stemphylium solani

Method used

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  • A Complete Set of Reagents and Its Application for Detecting Puccinia solani
  • A Complete Set of Reagents and Its Application for Detecting Puccinia solani
  • A Complete Set of Reagents and Its Application for Detecting Puccinia solani

Examples

Experimental program
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Effect test

Embodiment 1

[0052] Embodiment 1, the preparation of the complete set of reagents that detects Puccinia solani

[0053] The complete set of reagents for detecting P. solani provided by the present invention are LAMP primers, which are set reagent 1 and set reagent 2. The set of reagents 2 consists of Stem14-FIP, Stem14-BIP, Stem14-F3 and Stem14-B3 and single-stranded DNA named Stem14-LF25 and Stem14-LB25 respectively. The information of each single-stranded DNA is shown in Table 1 .

[0054] Wherein, Stem14-FIP is a single-stranded DNA whose nucleotide sequence is SEQ ID No.1 in the sequence listing;

[0055] The nucleotide sequence of Stem14-BIP is the single-stranded DNA of SEQ ID No.2 in the sequence listing;

[0056] The nucleotide sequence of Stem14-F3 is the single-stranded DNA of SEQ ID No.3 in the sequence listing;

[0057] The nucleotide sequence of Stem14-B3 is the single-stranded DNA of SEQ ID No.4 in the sequence listing.

[0058] The nucleotide sequence of Stem14-LF25 is t...

Embodiment 2

[0063] Embodiment 2, detect the specificity of the complete set of reagents of Puccinia solani

[0064] 1. Test strains

[0065] The selected strains for testing were: 17 strains of Puccinia solani, 1 strain of Fusarium solani, 1 strain of Botrytis cinerea, 3 strains of Alternaria solani, 2 strains of Corynesium multimata, 4 strains of Alternaria and 1 strain of Alternaria brassicae (Table 2), the tested strains are preserved in the Vegetable Disease Integrated Control Research Group of Vegetable and Flower Research Institute, Chinese Academy of Agricultural Sciences.

[0066] Table 2. Information on tested strains

[0067]

[0068] In Table 2, numbering is YQZ11020901 Solanum solani (Stemphylium solani) recorded in literature (Xieet al., First report of Stempylium solani causing leaf spot on wild eggplantin China, Can.J.Plant Pathol., 2016, Vol.38 , No.4,517–521);

[0069] Botrytis cinerea numbered HG09021201 is recorded in the literature (Tang Ming et al., Comprehensiv...

Embodiment 3

[0078] Example 3, Stem14-LF25 and Stem14-LB25 in the complete set of reagents 2 can improve the efficiency of LAMP reaction

[0079] The genomic DNA of Puccinia solani with the strain No. FQ11112407 in Example 2 was selected to detect the LAMP reaction efficiency of the set of reagents 1 and 2 in Example 1.

[0080] Wherein, the LAMP reaction system of the set of reagents 1 is shown in Example 2, and the reaction system of the set of reagents 2 is as follows:

[0081]

[0082]

[0083] Each LAMP reaction system was reacted at 65°C for 60 min in a real-time turbidity monitor LA-300.

[0084] The results show that the reaction time of the reaction system of the complete set of reagents 1 is about 40min; after adding the loop primers Stem14-LF25 and Stem14-LB25, the reaction time of the reaction system of the complete set of reagents 2 can be accelerated to about 27min (such as figure 2 ), the results showed that after adding the loop primers Stem14-LF25 and Stem14-LB25, ...

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Abstract

The invention discloses a complete set of reagents for detecting stemphylium solani and application. The complete set of reagents comprise single strand DNA named after Stem14-FIP, single strand DNA named after Stem14-BIP, single strand DNA named after Stem14-F3, single strand DNA named after Stem14-B3, single strand DNA named after Stem14-FL25, and single strand DNA named after Stem14-LB25, and the sequences are SEQ ID No. 1 to 6 separately in a sequence table. The complete set of reagents can perform specific detection on stemphylium solani, and the sensitivity for detecting stemphylium solani can reach 2.4*100 pg genome DNA / micro [mu]L.

Description

technical field [0001] The invention relates to a complete set of reagents for detecting Puccinia solani and its application in the field of biotechnology. Background technique [0002] Stemphylium fungi can infect a variety of crops around the world, and can cause a variety of vegetable diseases. The main damage vegetables include tomato, lettuce, pepper, cabbage, spinach, and garlic, and the damage to tomato is the most serious. Tomato leaf spot caused by Puccinia spp. has been reported in the United States, Israel, New Zealand and other places, which has caused huge economic losses to local tomato production. In recent years, the occurrence of gray leaf spot of tomato in 500,000 mu in Northeast, North, East and South China is particularly serious. Among them, Shouguang in Shandong, Haikou in Hainan, and Tianyang in Guangxi are the most severely affected. The total affected area exceeds 100,000 mu, and the loss exceeds 10 million yuan. [0003] Tomato is a very important...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/6844C12Q1/04C12N15/11C12R1/645
CPCC12Q1/6844C12Q1/6895C12Q2531/119
Inventor 谢学文李宝聚曹金强石延霞柴阿丽
Owner INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI