Rapid detection method for gibberellic acid microspecies generated in fusarium moniliforme

A technology of rice baculum and detection method, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, and determination/inspection of microorganisms to achieve the effects of good specificity, good practicability and accurate identification

Pending Publication Date: 2020-11-03
CHINA NAT RICE RES INST
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no report on the detection of Fusarium fujikura based on the diff

Method used

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  • Rapid detection method for gibberellic acid microspecies generated in fusarium moniliforme
  • Rapid detection method for gibberellic acid microspecies generated in fusarium moniliforme
  • Rapid detection method for gibberellic acid microspecies generated in fusarium moniliforme

Examples

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example 1

[0043] Example 1, molecular identification of four Fusarium races of rice bakanae pathogen

[0044] The Fusarium isolated from rice bakanae disease-susceptible samples was extracted from the genomic DNA of the strain, and the Fusarium protein translation elongation factor EF Universal primers designed for gene conservation sequences for molecular identification.

[0045] Step 1: Strain Culture

[0046] Inoculate the strain on potato dextrose agar solid medium (potato 200g, glucose 18g, agar 20g, distilled water 1L, PDA), and take the 5d-grown bacteria block and transfer it into 100mL potato dextrose liquid medium (potato 200g, glucose 18g, distilled water 1L , PDB), after 5 days of shaking culture in a constant temperature shaker at 28°C and 120 r / min, the mycelia were collected.

[0047] Step 2: Rapid Extraction of Strain DNA

[0048](1) Sample pretreatment: put 20 mg of mycelia in a 2 mL centrifuge tube, add 500 µL of Extraction Buffer (1 M KCL, 100 mM TrisHCl, 10 mM EDT...

example 2

[0058] Example 2, Comparison of Four Fusarium Race Gibberellin Biosynthetic Gene Clusters

[0059] Wiemann et al. (2013) have published the sequence of the gibberellin synthesis gene cluster in Fusarium Fujikura [Wiemann P, Sieber C M, von Bargen K W, et al. Deciphering the cryptic genome: genome-wide analyzes of the rice pathogen Fusarium fujikuroi reveal complex regulation of secondary metabolism and novel metabolites. PLoS Pathog, 2013,9(6): e1003475]. According to the gibberellin biosynthetic gene cluster DES (SEQ ID NO.20), P450-4 (SEQ ID NO. 21), P450-1 (SEQ ID NO.22), P450-2 (SEQ ID NO.23), GGS2 (SEQ ID NO.24), CPS / KS (SEQ ID NO.25) and P450-3 (SEQ ID NO.26) gene sequence, respectively designed three pairs of specific primers (Table 1) at the front, middle and back of each gene, for the Fusarium lamina, Fusarium pseudovermitilis and F. andiyazi Genomic DNA was amplified by PCR. The PCR reaction system is: strain genomic DNA 50ng, forward and reverse primers ...

example 3

[0062] Example 3, the comparison of Fusarium Fujikura and the whole genome of Fusarium laminarum

[0063] In order to carry out the functional genome comparison of Fusarium fujikura and Fusarium laminarum, considering the availability, representativeness and importance of the data, the Fp9 strain of Fusarium fujikura (SRA accession: PRJNA517537) and Fusarium fujikura were downloaded from the NCBI database. The whole genome sequence of Fusarium 58289 strain (ANFV00000000), including gene, protein sequence and gene annotation information. Using the whole genome sequence alignment mGenomeSubtractor online analysis software (http: / / bioinfo-mml.sjtu.edu.cn / mGS / ), the pairwise comparison of the two Fusarium genome sequences was performed ( E set the value to 1e -5 ), screened and obtained the specific gene encoding GA20-oxidase in the late stage of gibberellin synthesis pathway GA20ox , the gene exists in the Fusarium fusarium genome (GenBank No. FFUJ_12912), but does not exist i...

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Abstract

The invention discloses a rapid detection method for gibberellic acid microspecies generated in fusarium moniliforme, and belongs to the technical field of plant disease control and plant pathogen detection. According to the method, specific primers are designed according to specific gene differences of fusarium graminearum, fusarium cladosporium, fusarium verticillatum and F.andiyazi, multiplex PCR amplification is performed by taking DNA of a to-be-detected sample as a template through the specific primers, and gibberellic acid microspecies generated in the fusarium moniliforme are judged through amplification results. The method provided by the invention is suitable for rapid and reliable detection and identification of rice bakanae diseases induced by the fusarium moniliforme, the fusarium cladosporium, the fusarium verticillatum and the F.andiyazi, and has important practical application value in prevention and treatment of the rice bakanae diseases induced by the fusarium graminearum, the fusarium cladosporium, the fusarium verticillatum and the F.andiyazi.

Description

technical field [0001] The invention belongs to the technical field of plant disease control and plant pathogen detection, and in particular relates to a rapid detection method for producing gibberellin races in rice bakanae pathogens. Background technique [0002] Gibberellin (Gibberellic acid, GA), as one of the five major plant hormones, is an efficient plant growth regulator. It has a variety of physiological functions for plant growth and development, and can regulate plant seed germination, stem and leaf elongation, and fruit development. . At present, 136 species of gibberellin substances have been found from plants, fungi and bacteria, most of which exist in higher plants, some exist in fungi or bacteria, and the other part is shared by fungi and plants. Gibberellins are tetracyclic diterpenoids whose basic backbone consists of four isoprene units. Due to the difference in the number and position of double bonds, hydroxyl groups, and the presence or absence of lact...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/686C12Q1/04C12N15/11C12R1/77
CPCC12Q1/6895C12Q1/686C12Q2600/16C12Q2537/143Y02A50/30
Inventor 王玲黄世文刘连盟
Owner CHINA NAT RICE RES INST
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