CN113604590A - Fluorescent quantitative PCR detection method and kit for plant pathogenic bacteria Pantoea ananatis - Google Patents

Fluorescent quantitative PCR detection method and kit for plant pathogenic bacteria Pantoea ananatis Download PDF

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CN113604590A
CN113604590A CN202110909246.0A CN202110909246A CN113604590A CN 113604590 A CN113604590 A CN 113604590A CN 202110909246 A CN202110909246 A CN 202110909246A CN 113604590 A CN113604590 A CN 113604590A
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pantoea ananatis
quantitative pcr
fluorescence quantitative
plant pathogenic
standard substance
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陈美娟
陈成成
陈冲
曹琴琴
杨敏
陆梦娇
王振球
章进军
贾俊才
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Shanghai Woji Gene Technology Co ltd
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Abstract

The invention discloses a fluorescence quantitative PCR detection method and a kit for plant pathogenic bacteria Pantoea ananatis. The invention also provides a method for carrying out fluorescence quantitative PCR detection on plant pathogenic bacteria Pantoea ananatis through the kit, which comprises the following steps: step 1, extracting genome DNA of a sample to be detected; and 2, setting a positive standard substance control and a negative control by taking the genomic DNA obtained in the step 1 as a template, preparing a PCR reaction system with the Pantoea ananatis PCR reaction solution and the specific primer, and carrying out fluorescence quantitative PCR reaction under optimized reaction conditions. The method has higher specificity, sensitivity and stability when detecting the Pantoea ananatis in crops and trees, and is a rapid and accurate detection method of the Pantoea ananatis.

Description

Fluorescent quantitative PCR detection method and kit for plant pathogenic bacteria Pantoea ananatis
Technical Field
The invention relates to a fluorescent quantitative PCR detection method and a kit in the field of molecular biology, in particular to a fluorescent quantitative PCR detection method and a kit for plant pathogenic bacteria Pantoea ananatis.
Background
The plant pathogenic bacterium Pantoea ananatis (Pantoea ananatis) is a gram-negative bacterium, a common epiphytic plant. Pantoea ananatis is distributed all over the world and can be separated from plants and the environment. As a plant pathogen, it causes a great amount of economic loss to crops and trees worldwide. It can infect monocotyledonous and dicotyledonous plants, and different infected hosts cause different symptoms, such as fruit decay caused by infecting melons such as pineapple and Hami melon; the corn infection can cause necrotic spots, stripes and the like; the infected rice can cause the rotting of the root stalk and the like. Infection of eucalyptus can cause leaf blight, withered bud tip, etc. Environmental factors have different effects on the severity of disease caused by infection of different hosts. It has been demonstrated that the pathogens of Pantoea ananatis (Pantoea ananatis) can be transmitted by pollen, plant surface wounds, and direct plant-to-plant contact under certain conditions. As a plant pathogen, the disease condition is serious due to the rapid onset of disease, and the production of commercial crops in partial areas is seriously damaged. At present, no method for quickly and accurately detecting Pantoea ananatis exists.
Disclosure of Invention
The invention aims to provide a fluorescent quantitative PCR detection method and a kit, which are used for quickly and accurately detecting the plant pathogenic bacteria Pantoea ananatis.
In order to achieve the aim, the invention provides a fluorescent quantitative PCR detection kit for plant pathogenic bacteria Pantoea ananatis, wherein the kit consists of PCR reaction enzyme premix, Pantoea ananatis specific upstream primer, downstream primer, positive control and negative control; the upstream primer is TATCCGCGCCTTTGTTGAGT; the downstream primer is ATGCCGTCTTTCTCGGTTGA.
The fluorescence quantitative PCR detection kit for the plant pathogenic bacteria Pantoea ananatis is characterized in that the positive control adopts a designed and synthesized positive standard substance sequence, and the positive standard substance sequence is as follows: 5-TATCCGCGCCTTTGTTGAGTACCTGAATAAAAACAAAACGCCTATTCA CCCAACCGTATTCTATTTCTCAACCGAGAAAGACGGCAT-3.
The fluorescence quantitative PCR detection kit for the plant pathogenic bacteria Pantoea ananatis is characterized in that the positive standard substance sequence is diluted by positive standard substance plasmids to prepare a positive standard substance gradient so as to achieve a quantitative effect.
The fluorescence quantitative PCR detection kit for the plant pathogenic bacteria Pantoea ananatis is characterized in that the preparation process of the positive standard substance gradient is as follows: taking 10 mu l of positive standard substance plasmid, diluting according to 10 times of gradient, and sequentially diluting for 4-7 gradients to obtain positive standard substance gradient.
The fluorescence quantitative PCR detection kit for the plant pathogenic bacteria Pantoea ananatis is characterized in that after the positive standard substance is prepared in a gradient manner, fluorescence quantitative PCR reaction is carried out on the gradient positive standard substance and sample DNA under the same condition, the logarithm value of the concentration of the gradient positive standard substance is taken as the ordinate, the Ct value of the experimental result is taken as the abscissa, a standard curve is drawn, and the concentration of the Pantoea ananatis in the sample is obtained through calculation according to the Ct value obtained by sample testing.
The invention also provides a method for carrying out fluorescence quantitative PCR detection on plant pathogenic bacteria Pantoea ananatis through the kit, wherein the method comprises the following steps: step 1, extracting genome DNA of a sample to be detected; and 2, setting a positive standard substance control and a negative control by taking the genomic DNA obtained in the step 1 as a template, preparing a PCR reaction system with the Pantoea ananatis PCR reaction solution and the specific primer, and carrying out fluorescence quantitative PCR reaction.
The method for carrying out the fluorescence quantitative PCR detection of the plant pathogenic bacterium Pantoea ananatis is characterized in that the concentration of the PCR reaction system in the step 2 comprises the following steps: 1XPCR buffer, upstream and downstream primers 0.2-1. mu.M each, and 1-10 ng/. mu.l of OD260/OD280 between 1.7-1.9.
The method for carrying out the fluorescent quantitative PCR detection on the plant pathogenic bacterium Pantoea ananatis is described above, wherein the PCR reaction conditions in the step 2 are as follows: pre-denaturation at 95 ℃ for 30s, further denaturation at 95 ℃ for 5s, and annealing extension at 60 ℃ for 30s for 40 cycles.
The fluorescence quantitative PCR detection method and the kit for the plant pathogenic bacteria Pantoea ananatis provided by the invention have the following advantages:
the invention designs specific primers aiming at the gene sequence of the gyrB of the Pantoea ananatis to develop a kit for detecting the Pantoea ananatis, wherein the sequence of the gyrB gene has conservation and variability and can be used as a target molecule in the research of bacterium classification and identification based on nucleotide sequence. The specific primer designed by taking the method as a detection target spot detects the Pantoea ananatis through a fluorescent quantitative PCR technology, has higher specificity, sensitivity and stability when detecting the Pantoea ananatis in crops and trees, and is a rapid and accurate detection method for the Pantoea ananatis.
Drawings
FIG. 1 is a graph of the amplification profile of a gradient standard of PCR amplification.
FIG. 2 is a melting curve diagram of PCR amplification.
FIG. 3 is a standard graph of PCR amplification.
FIG. 4 is a graph showing the results of gradient fluorescent quantitative PCR assay.
FIG. 5 is a drawing taken at 10^ a9Copy/. mu.l, 10^ l7Copy/. mu.l, 10^ l5Copy/. mu.l, 10^ l3Copy/. mu.l electrophoresis results.
Detailed Description
The following further describes embodiments of the present invention with reference to the drawings.
The kit consists of a PCR reaction enzyme premix, Pantoea ananatis (Pantoea ananatis) specific upstream primers, downstream primers, a positive control and a negative control.
Wherein, specific primers are designed and synthesized according to the gyrB gene sequence (GenBank accession number: MW981333.1) of Pantoea ananatis, and the sequences are as follows:
the forward primer was TATCCGCGCCTTTGTTGAGT (SEQ ID NO: 1).
The downstream primer is ATGCCGTCTTTCTCGGTTGA (SEQ ID NO: 2).
The positive control adopts a positive standard substance sequence which is designed and synthesized and is as follows: 5-TATCCGCGCCTTTGTTGAGTACCTGAATAAAAACAAAACGCCTATTCA CCCAACCGTATTCTATTTCTCAACCGAGAAAGACGGCAT-3 (SEQ ID NO: 3).
When the positive standard substance sequence is used, positive standard substance plasmids are diluted to prepare a positive standard substance gradient so as to achieve the quantitative effect.
The preparation process of the positive standard substance gradient comprises the following steps: taking 10 mu l of positive standard substance plasmid, diluting according to 10 times of gradient, and sequentially diluting for 4-7 gradients to obtain positive standard substance gradient.
And after the positive standard substance is prepared in a gradient manner, performing fluorescence quantitative PCR reaction on the gradient positive standard substance and the sample DNA under the same condition, drawing a standard curve by taking the concentration logarithm value of the gradient positive standard substance as a vertical coordinate and the Ct value of the experimental result as a horizontal coordinate, and calculating to obtain the concentration of the pantoea ananatis in the sample according to the Ct value obtained by the sample test.
The invention also provides a method for carrying out fluorescence quantitative PCR detection on plant pathogenic bacteria Pantoea ananatis through the kit, which comprises the following steps: step 1, extracting genome DNA of a sample to be detected; and 2, setting a positive standard substance control and a negative control by taking the genomic DNA obtained in the step 1 as a template, preparing a PCR reaction system with the Pantoea ananatis PCR reaction solution and the specific primer, and carrying out fluorescence quantitative PCR reaction under optimized reaction conditions.
The concentration composition of the PCR reaction system in the step 2 is as follows: 1XPCR buffer, upstream and downstream primers 0.2-1. mu.M each, and 1-10 ng/. mu.l of OD260/OD280 between 1.7-1.9.
The PCR conditions in step 2 were: pre-denaturation at 95 ℃ for 30s, further denaturation at 95 ℃ for 5s, and annealing extension at 60 ℃ for 30s for 40 cycles.
The present invention will be further described with reference to the following examples.
Example 1
1. Designing a characteristic primer for detecting Pantoea ananatis.
Specific primers are designed and synthesized aiming at the gyrB gene sequence (GenBank accession number: MW981333.1) of the Pantoea ananatis, and BLAST analysis is carried out on the designed specific primer sequence, and the result shows that the primers have higher specificity. The sequence is as follows:
an upstream primer: TATCCGCGCCTTTGTTGAGT (SEQ ID NO: 1).
A downstream primer: ATGCCGTCTTTCTCGGTTGA (SEQ ID NO: 2).
Design and synthesis of positive standard substance sequence:
5-TATCCGCGCCTTTGTTGAGTACCTGAATAAAAACAAAACGCCTAT TCACCCAACCGTATTCTATTTCTCAACCGAGAAAGACGGCAT-3 (SEQ ID NO: 3).
2. Extracting genome DNA of sample to be detected
3. And (3) setting a positive standard substance control and a negative control by taking the extracted genome DNA as a template, preparing a PCR reaction system with the Pantoea ananatis PCR reaction solution and the specific primer, and carrying out fluorescence quantitative PCR reaction under the optimized reaction condition.
The concentration composition of the PCR reaction system is as follows: 1XPCR buffer, upstream and downstream primers 0.2-1. mu.M each, and 1-10 ng/. mu.l of OD260/OD280 between 1.7-1.9.
The composition of the PCR reaction system is specifically shown in Table 1 below.
Table 1.PCR reaction System composition Table.
Figure BDA0003202865460000051
The PCR reaction procedure is shown in Table 2 below.
Table 2 PCR reaction schedule.
Figure BDA0003202865460000052
4. Establishment of a standard curve: by using the above reaction system and reaction conditions, qPCR amplification was performed on each gradient of the standard plasmid, and a gradient standard amplification curve (shown in fig. 1), a melting curve (shown in fig. 2), and a standard curve (shown in fig. 3) were obtained.
Example 2 evaluation of the Performance of the fluorescent quantitative PCR detection method.
1. And (4) evaluating the sensitivity of the detection method.
Plasmid standards were diluted 9 gradients 109/μl，108Copy/. mu.l, 107Copy/. mu.l, 106Copy/. mu.l, 105Copy/. mu.l, 104Copy/. mu.l, 103Copy/. mu.l, 102Copy/. mu.l, 101Mu.l. The results of the fluorescent quantitative PCR detection of these 9 gradients by the above detection method are shown in FIG. 4, and the lines marked 1, 2, 3, 4, 5, 6, 7, 8 and 9 are the concentrations: 109/μl，108Copy/. mu.l, 107Copy/. mu.l, 106Copy/. mu.l, 105Copy/. mu.l, 104Copy/. mu.l, 103Copy/. mu.l, 102Copy/. mu.l, 101Mu.l amplification Curve, 101Mu.l plasmid has no obvious exponential amplification curve, and the lowest detection line of the method is 102Copies/. mu.l.
2. Evaluation of the reproducibility of the detection method:
get 103Copy/. mu.l, 105Copy/. mu.l, 107Copy/. mu.l, 109Copies/. mu.l
The three independent detections are carried out on the 4 plasmid standard substance gradients, and the detection repeatability is better.
The results are shown in table 3 below.
Table 3. table of repeatability evaluation results of the test methods.
Figure BDA0003202865460000061
The kit is provided with a PCR reaction enzyme premix, a Pantoea ananatis specific upstream primer, a downstream primer, a positive control and a negative control, and is used for quickly and accurately detecting the Pantoea ananatis, a plant pathogen. The method has higher specificity, sensitivity and stability when detecting the Pantoea ananatis in crops and trees, and is a rapid and accurate detection method of the Pantoea ananatis.
While the present invention has been described in detail with reference to the preferred embodiments, it should be understood that the above description should not be taken as limiting the invention. Various modifications and alterations to this invention will become apparent to those skilled in the art upon reading the foregoing description. Accordingly, the scope of the invention should be determined from the following claims.
Ohio 110 Shanghai Voji Gene science and technology Co Ltd
Fluorescent quantitative PCR detection method and kit for plant pathogenic bacteria Pantoea ananatis
〈160〉3
〈210〉1
〈211〉20
〈212〉DNA
Artificial sequence of < 213 >
〈220〉
< 223 > upstream primer sequence for identifying Pantoea ananatis specificity
〈400〉1
tatccgcgcctttgttgagt 20
〈210〉2
〈211〉20
〈212〉DNA
Artificial sequence of < 213 >
〈220〉
< 223 > downstream primer sequence for identifying Pantoea ananatis specificity
〈400〉2
atgccgtctttctcggttga 20
〈210〉3
〈211〉87
〈212〉DNA
Artificial sequence of < 213 >
〈220〉
Sequence of positive standard substance of '223' pineapple fungus
〈400〉3
tatccgcgcctttgttgagtacctgaataaaaacaaaacgcctattcacccaaccgtatt 60
ctatttctcaaccgagaaagacggcat 87

Claims (8)

1. A fluorescence quantitative PCR detection kit for plant pathogenic bacteria Pantoea ananatis is characterized in that the kit consists of PCR reaction enzyme premix, Pantoea ananatis specific upstream primers, downstream primers, positive control and negative control;
the upstream primer is TATCCGCGCCTTTGTTGAGT;
the downstream primer is ATGCCGTCTTTCTCGGTTGA.
2. The fluorescence quantitative PCR detection kit for the plant pathogenic bacterium Pantoea ananatis according to claim 1, wherein the positive control adopts a designed and synthesized positive standard substance sequence which is: 5-TATCCGCGCCTTTGTTGAGTACCTGAATAAAAACAAAACGCCTATTCACCCAACCGTATTCTATTTCTCAACCGAGAAAGACGGCAT-3.
3. The fluorescence quantitative PCR detection kit for the plant pathogenic bacterium Pantoea ananatis as claimed in claim 2, wherein the positive standard substance sequence is diluted by positive standard substance plasmid to prepare positive standard substance gradient so as to achieve quantitative effect.
4. The fluorescence quantitative PCR detection kit for the plant pathogenic bacterium Pantoea ananatis according to claim 3, wherein the preparation process of the positive standard substance gradient is as follows: taking 10 mu l of positive standard substance plasmid, diluting according to 10 times of gradient, and sequentially diluting for 4-7 gradients to obtain positive standard substance gradient.
5. The fluorescence quantitative PCR detection kit for the plant pathogenic bacteria Pantoea ananatis according to claim 4, wherein after the gradient preparation of the positive standard is completed, the fluorescence quantitative PCR reaction is performed on the gradient positive standard and sample DNA under the same conditions, the logarithm value of the concentration of the gradient positive standard is taken as the ordinate, the Ct value of the experimental result is taken as the abscissa to draw a standard curve, and the concentration of Pantoea ananatis in the sample is obtained by calculation according to the Ct value obtained by the sample test.
6. A method for carrying out fluorescence quantitative PCR detection on plant pathogenic bacteria Pantoea ananatis through the kit according to any one of claims 1 to 5, wherein the method comprises the following steps:
step 1, extracting genome DNA of a sample to be detected;
and 2, setting a positive standard substance control and a negative control by taking the genomic DNA obtained in the step 1 as a template, preparing a PCR reaction system with the Pantoea ananatis PCR reaction solution and the specific primer, and carrying out fluorescence quantitative PCR reaction.
7. The method for carrying out the fluorescence quantitative PCR detection of the plant pathogenic bacterium Pantoea ananatis according to claim 6, wherein the concentration of the PCR reaction system in the step 2 comprises: 1XPCR buffer, upstream and downstream primers 0.2-1. mu.M each, and 1-10 ng/. mu.l of OD260/OD280 between 1.7-1.9.
8. The method for carrying out the fluorescence quantitative PCR detection of the plant pathogenic bacterium Pantoea ananatis according to claim 6, wherein the PCR reaction conditions in the step 2 are as follows: pre-denaturation at 95 ℃ for 30s, further denaturation at 95 ℃ for 5s, and annealing extension at 60 ℃ for 30s for 40 cycles.
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114350828A (en) * 2022-01-21 2022-04-15 安徽省农业科学院植物保护与农产品质量安全研究所 Specific primer for amplifying Pantoea ananatis and application thereof

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103966307A (en) * 2013-02-05 2014-08-06 中国农业科学院蔬菜花卉研究所 Fluorescent quantitative PCR detection technology of brassicaceous vegetable plasmodiophoromycetes, and its application

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103966307A (en) * 2013-02-05 2014-08-06 中国农业科学院蔬菜花卉研究所 Fluorescent quantitative PCR detection technology of brassicaceous vegetable plasmodiophoromycetes, and its application

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
丁正: "新型HARPIN泡腾片剂型的研制及其防治玉米细菌新病害的研究", 中国优秀硕士学位论文全文数据库 工程科技I辑, pages 2 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114350828A (en) * 2022-01-21 2022-04-15 安徽省农业科学院植物保护与农产品质量安全研究所 Specific primer for amplifying Pantoea ananatis and application thereof
CN114350828B (en) * 2022-01-21 2023-11-03 安徽省农业科学院植物保护与农产品质量安全研究所 Specific primer for amplifying Pantoea ananatis and application thereof

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