CN113584208A - Method for detecting Diaporthe novem by using PCR (polymerase chain reaction) primers - Google Patents
Method for detecting Diaporthe novem by using PCR (polymerase chain reaction) primers Download PDFInfo
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- 241000983238 Diaporthe novem Species 0.000 title description 5
- 238000003752 polymerase chain reaction Methods 0.000 title 2
- 238000001514 detection method Methods 0.000 claims abstract description 20
- 238000011895 specific detection Methods 0.000 claims abstract 2
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 5
- 230000003321 amplification Effects 0.000 claims description 4
- 239000012634 fragment Substances 0.000 claims description 4
- 238000012408 PCR amplification Methods 0.000 claims description 2
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- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
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Abstract
The present invention relates to the detection of plants and plant products using PCR technologyDiaporthe novemBelonging to the field of plant pathogenic fungi detection. The invention designs and synthesizesD.novemSpecific primers of (1), for the pairD.novemSpecific detection of genomic DNA. The invention establishes a method which is rapid, simple, convenient, strong in specificity, accurate and reliableD.novemThe molecular detection method of (1) completes the whole detection within one working day.
Description
Technical Field
The present invention relates to the detection of plants and plant products using PCR technologyDiaporthe novem J.M. Santos, Vrandečić & A.J.L. Phillips, sp. nov.Belonging to the field of plant pathogenic fungi detection.
Background
Diaporthe novemBelongs to the kingdom of Fungi (Fungi), phylum Ascomycota (Ascomycota), class Ascomycetes (Ascomycetes), order Sitophyllales (Diaporthales), family Sitophyllaceae (Diaporthaceae), genus Sitophyllum (A)Diaporthe) In asexual stages thereofPhomopsissp.9. It is associated with the pathogenic bacteria of northern tuber ulcer of soybean which seriously harms soybeanD. caulivoraSouthern Stem ulcer pathogen of soybeanD. aspalathiPhomopsis thalianaD. longicollaThe species are similar to each other, wherein the soybean northern stem canker pathogen and the soybean southern stem canker pathogen are listed in the entry plant quarantine pest directory of China.
D. novemOriginally officially named by Joge Manual Santos et al in 2011, the existing data show that the pathogen is distributed in areas such as Chilean, Crohn's disease, Italy, Portugal, south Africa and Australia. The method is characterized in that the method harms important economic crops such as soybean, sunflower, kiwi fruit, grape, kumquat, lime and the like, seeds and fruits are rotted, so that serious economic loss is caused, and the method also has separation reports on hydrangea, south African black tea and rubiaceae Raglania, and China has not reported yet.
The soybean is used as an important grain and oil crop and feed raw material, has great demand in China, is one of the most important imported agricultural products in China, and the imported total amount of the soybean in China in 2019 reaches 8851 ten thousand tons. Repeated detection of port in ChinaD. novemSince the pathogen is not in the quarantine directory, the harm is easy to be overlooked. The pathogen causes stalk ulcers and seed rot in soybeans, resulting in reduced yield and quality of soybeans. Soybean oil produced from the damaged seeds and other products such as feed, fertilizer and soybean cake obtained after oil extraction from soybean are low in quality and not suitable for use as feed. In addition, the host of the pathogen is wide, and once the pathogen is introduced into China along with the imported soybeans, the pathogen causes inestimable loss on the production and agricultural ecology of the soybeans and other crops in China.
In view ofD. novemTransmitted into ChinaRisk and severity of pathogen harm, and establishes a set of rapid, accurate and complete methods for protecting the production safety of agriculture and forestry in China, maintaining the benefits of China, further standardizing the quarantine of the important disease, improving the accuracy and the verification speed of port epidemic situation detection, preventing the pathogen from transmitting into ChinaD. novemThe detection method of (2) is imminent.
Disclosure of Invention
The invention collects the diaporthe 8 fungi (see table 1), establishes a rapid, simple, convenient, strong-specificity, accurate and reliable molecular detection method for the diaporthe fungi, and can detect the diaportheD. novemAccurately identified. The method is rapid in detection and reliable, the whole process is completed within one working day, and the method can be effectively popularized and applied to port detection.
The specific technical scheme is as follows:
the invention aims to provide detectionD. novemA method for PCR of genomic DNA comprising the steps of:
(1) extracting plant genome DNA;
（2）D. novemthe specific primer has the following sequence:
DnoF1：CTCACACTGGCTGGATTATG
DnoR1：AGCCGCTTATCTCCTATGC
the primer is used forD. novemThe genome DNA is specifically detected, and the amplified fragment is about 329 bp.
Compared with the prior art, the invention has the beneficial effects that: the invention establishes the characteristics of rapidness, simplicity, convenience, strong specificity, accuracy and reliabilityD. novemA molecular detection method ofD. novemIs distinguished from other species of the saddle shell. The method is rapid in detection and reliable, the whole process is completed within one working day, and the method can be effectively popularized and applied to port detection.
The present invention will be described in further detail with reference to the accompanying drawings and specific embodiments.
Drawings
FIG. 1 shows a schematic view of aD. novemAmplification of genomic DNA specific primers
In the figure, lanes 1 to 9 areD. novem, D. longicolla , D. caulivora, D. aspalathi, Diaporthe sp., D. eres , D.phaseolorum, D. vacciniiAnd a negative control, lane M is 2000 bp DNA marker.
Detailed Description
Example 1: extraction of genomic DNA from plant material
This experimentD. novemThe strain is from plant inspection laboratory of Tianjin customs animal food planting center. The total number of the strains is 8, and the related information is shown in the table 1.
TABLE 1 code of test materials, Latin name, Chinese name and year of Collection
| Serial number | Latin name | Name of Chinese | Year of |
| 1 | Diaporthe novem | - | 2019 |
| 2 | Diaporthe longicolla | Phomopsis longissima (L.) F.sp. | 2019 |
| 3 | Diaporthe caulivora | Bacterial canker of northern tuber of soybean | 2019 |
| 4 | Diaporthe aspalathi | Bacterial canker of south Stem of Soybean | 2019 |
| 5 | Diaporthesp. | Sunflower seat shell | 2020 |
| 6 | Diaporthe eres | Cherry house seat shell | 2020 |
| 7 | Diaporthephaseolorum | Kidney bean room seat shell | 2020 |
| 8 | Diaporthe vaccinii | Blueberry fruit rot fungi | 2020 |
Picking crimp bandSterilizing the diseased bean with lesion on the surface with 1% sodium hypochlorite for 5 min, washing with sterilized water for 3 times, maintaining moisture at 25 deg.C, culturing for 24 hr, freezing at-20 deg.C for 24 hr, culturing on PDA culture medium at 22 deg.C under 12 hr in dark. After 5d, the culture results were observed. The suspicious colony is separated and purified by PDA, a proper amount of mycelium blocks are taken, liquid nitrogen is fully ground, DNA is extracted by a Dneasy plant mini kit plant gene extraction kit of Germany QIAGEN, and the DNA is dissolved in 100μThe cells were stored at-20 ℃ in L1 XTE buffer.
Example 2: design of specific primers
Artificially synthesizedD. novemThe specific primer of (1) has the following primer sequence:
DnoF1：CTCACACTGGCTGGATTATG
DnoR1：AGCCGCTTATCTCCTATGC
example 3:D. novemPCR amplification of specific primers
1. Preparation of reaction mixture
The reaction system configuration is shown in Table 2.
TABLE 2D. novemSpecific amplification system
| Reagent | Sample addition amount/. mu.l |
| 2× Premix | 12.5 |
| ddH2O | 10.0 |
| DnoF1 | 0.5 |
| DnoR1 | 0.5 |
| Form panel | 1.5 |
| Total | 25.0 |
2. PCR reaction procedure
Pre-denaturation: 94 ℃ for 3min
Denaturation: 94 ℃ for 30s
Annealing: 60 ℃ for 20s
Extension: 72 ℃ and 20s
The number of cycles: 35 are provided with
Extension: 72 ℃ for 6min
2.3 analysis of results
Genomic DNA from 8 experimental materials was amplified with specific primers DnoF1/DnoR1, and sterile water was added as a template for negative control. The amplified product is electrophoresed through 1.5% agarose gel, and EB staining can observe a band with a specific target size, and the amplified fragment of the universal primer is about 329bp, and only the amplified fragment isD. novemShowing a band of interest of a specific size (see lane 1 in FIG),D. longicolla , D. caulivora, D. aspalathi, Diaporthe sp., D. eres , D.phaseolorum, D. vaccinii and negative control has no corresponding product (shown in lanes 2-9, and lanes 1-9 in the figure areD. novem, D. longicolla, D. caulivora, D. aspalathi, Diaporthe sp., D. eres, D.phaseolorum, D. vacciniiAnd negative control), lane M is 2000 bp DNA marker.
Sequence listing
<110> Tianjin customs animal, plant and food detection center
<120> a method for detecting Diaporthe novem by using PCR primer
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 20
<212> DNA
<213> Diaporthe novem
<400> 1
ctcacactgg ctggattatg 20
<210> 2
<211> 19
<212> DNA
<213> Diaporthe novem
<400> 2
agccgcttat ctcctatgc 19
Claims (3)
1. Detection by PCR primerDiaporthe novemIs characterized by being self-designedD. novem PCR specific primer pair with the following sequence:
DnoF1：CTCACACTGGCTGGATTATG
DnoR1：AGCCGCTTATCTCCTATGC
the amplified fragment was 329bp and used forD. novemAnd (3) specific amplification detection of genome DNA.
2. The method of claim 1, wherein the detection is performed using PCR primersD. novemThe method is characterized by comprising the following steps:
(1) extracting plant genome DNA;
(2) establishingD. novemA PCR amplification system of genome DNA, which is used for amplification according to a reaction program;
(3) and detecting the amplification product by agarose gel electrophoresis.
3. The assay as claimed in claim 1D. novemThe specific PCR primer set of (1) or the assay of claim 2D. novemIn the specific detectionD. novemThe application of (1).
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102094086A (en) * | 2010-12-01 | 2011-06-15 | 天津出入境检验检疫局动植物与食品检测中心 | Method for detecting Solanum rostratum Dunal by PCR (Polymerase Chain Reaction) primer |
| CN102373233A (en) * | 2010-08-23 | 2012-03-14 | 深圳出入境检验检疫局动植物检验检疫技术中心 | Positive standard molecule for detecting diaporthe phaseolorum (Cooke etEll) Sacc. var. caulivora Athow et Caldwell (DPC) and diaporthe phaseolorum (Cooke etEll) Sacc. var. meridionalis F.A.Fernandex (DPM) as well as preparation method and detection method thereof |
| CN105400869A (en) * | 2015-10-26 | 2016-03-16 | 天津出入境检验检疫局动植物与食品检测中心 | Method for detecting amaranthus palmeri through PCR primers |
| CN105543217A (en) * | 2015-12-08 | 2016-05-04 | 天津出入境检验检疫局动植物与食品检测中心 | Phomopsis(Sacc.)Bubak glyceraldehyde-3-phosphate dehydrogenase (GPDH) gene amplification primers and their designing method and use |
| CN108707687A (en) * | 2018-05-30 | 2018-10-26 | 河北省农林科学院谷子研究所 | A kind of PCR Testing and appraisal methods that can distinguish millet blast bacterium and Pyricularia oryzae |
| CN110016515A (en) * | 2019-03-26 | 2019-07-16 | 天津出入境检验检疫局动植物与食品检测中心 | A method of kiwi fruit maize ear rot bacterium is detected with PCR primer |
-
2021
- 2021-08-06 CN CN202110899407.2A patent/CN113584208B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102373233A (en) * | 2010-08-23 | 2012-03-14 | 深圳出入境检验检疫局动植物检验检疫技术中心 | Positive standard molecule for detecting diaporthe phaseolorum (Cooke etEll) Sacc. var. caulivora Athow et Caldwell (DPC) and diaporthe phaseolorum (Cooke etEll) Sacc. var. meridionalis F.A.Fernandex (DPM) as well as preparation method and detection method thereof |
| CN102094086A (en) * | 2010-12-01 | 2011-06-15 | 天津出入境检验检疫局动植物与食品检测中心 | Method for detecting Solanum rostratum Dunal by PCR (Polymerase Chain Reaction) primer |
| CN105400869A (en) * | 2015-10-26 | 2016-03-16 | 天津出入境检验检疫局动植物与食品检测中心 | Method for detecting amaranthus palmeri through PCR primers |
| CN105543217A (en) * | 2015-12-08 | 2016-05-04 | 天津出入境检验检疫局动植物与食品检测中心 | Phomopsis(Sacc.)Bubak glyceraldehyde-3-phosphate dehydrogenase (GPDH) gene amplification primers and their designing method and use |
| CN108707687A (en) * | 2018-05-30 | 2018-10-26 | 河北省农林科学院谷子研究所 | A kind of PCR Testing and appraisal methods that can distinguish millet blast bacterium and Pyricularia oryzae |
| CN110016515A (en) * | 2019-03-26 | 2019-07-16 | 天津出入境检验检疫局动植物与食品检测中心 | A method of kiwi fruit maize ear rot bacterium is detected with PCR primer |
Non-Patent Citations (3)
| Title |
|---|
| S. M. THOMPSON: ""Diaporthe novem isolated from sunflower (Helianthus annuus) and other crop and weed hosts in Australia"", 《EUR J PLANT PATHOL》, vol. 2018, no. 152, pages 823 - 831 * |
| 张淼 等: ""大豆新发病害Diaporthe novem 干热处理研究初报"", 《动植物检疫》, vol. 4, no. 9, pages 50 - 54 * |
| 张莹 等: ""进境美国大豆中值得关注的真菌Diaporthe novem"", 《菌物学报》, vol. 41, no. 3, pages 412 - 419 * |
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