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
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[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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