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Three specific genes capable of generating fumonisins fusarium microspecies and application thereof

A specific gene, fumonisin technology, applied in the field of molecular biology

Active Publication Date: 2018-08-28
CHINA NAT RICE RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The present invention obtains the specific genes of each race and its detection method by comparing the whole genomes of Fusarium sp. There is no report on the differential detection of rice ear rot based on the whole genome of the pathogen

Method used

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  • Three specific genes capable of generating fumonisins fusarium microspecies and application thereof
  • Three specific genes capable of generating fumonisins fusarium microspecies and application thereof
  • Three specific genes capable of generating fumonisins fusarium microspecies and application thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0040] Example 1: Comparison of whole genomes of three Fusarium races

[0041] In order to perform functional genome comparisons of Fusarium races, considering the availability, representativeness and importance of the data, the strains of Fusarium sp. ET1 were downloaded from the NCBI database (https: / / www.ncbi.nlm.nih.gov . nih.gov / genome / 13188), including gene, protein sequence and gene annotation information. Using the whole genome sequence comparison mGenomeSubtractor online analysis software (http: / / bioinfo-mml.sjtu.edu.cn / mGS / ), the pairwise comparison of the three Fusarium genome sequences (E value is set to 1e -5 ), to screen out specific gene fragments in each race. When the coverage of the protein sequence Blastp of the gene is less than 30%, it is defined as a specific gene.

[0042] After screening, 8 genes were obtained that existed in Fusarium aeruginosa, but did not exist in Fusarium pseudovermitilis and Gibberella fujikura, and had clear gene function annot...

example 2

[0045] Example 2: Acquisition of Race-Specific Genes of Fusarium sp.

[0046] Step 1: Test strains

[0047]The Fusarium sp. used has a wide range of sources, and the strains were collected and isolated from different provinces in China, basically covering the areas where rice ear rot is currently prevalent.

[0048] Step 2: Morphological identification

[0049] According to the shape and growth mode of the megaspore and microspore of the strain, the characteristics of the conidiophores, the growth color of the colony, etc., it was determined to be Fusarium laminarum after morphological identification.

[0050] Step 3: Strain Culture

[0051] 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 7 days of shaking culture in a constant temperature shaker at 2...

example 3

[0067] Example 3: Acquisition of Fusarium pseudoverticillium race-specific genes

[0068] Step 1: Test strains

[0069] The Fusarium pseudoverticillium used has a wide range of sources, and the strains were collected and isolated from different provinces in China, basically covering the areas where rice ear rot is currently prevalent.

[0070] Steps 2-5: Same as Example 2, after morphological identification and molecular identification of EF gene, the screened Fusarium pseudoverticillium was used for race-specific gene analysis.

[0071] Step 6: Screening of specific genes of Fusarium pseudoverticillium

[0072] The 12 pairs of specific gene primers in Example 1 were used to design primers for PCR verification. The PCR reaction system is: strain genomic DNA 50ng, forward and reverse primers (10μM) 1μL each, dNTPs (2.5mM) 4μL, 10×PCR buffer 2μL, Taq polymerase (5U / μL) 0.3μL, add ddH 2 O to make up 20 μL. The PCR reaction program was as follows: pre-denaturation at 94°C for ...

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Abstract

The invention discloses three specific genes capable of generating fumonisins fusarium microspecies and application thereof and belongs to the field of molecular biology. On the one hand, three specific gene sequences for generation of the fumonisins fusarium microspecies are disclosed; on the other hand, the application of the three specific genes capable of generating the fumonisins fusarium microspecies in detection of rice spike monilinia fiucticola is also disclosed. The three specific genes have the advantages that specific genes of the microspecies are obtained from genomic sequence information of three kinds of fusarium capable of generating fumonisins, detection of pathogenic bacteria is achieved according to specific nucleotide sequences of the microspecies, detection primers have high specificity, and therefore bands with specificity can be amplified from a target strain. The specific genes, the primers and the detection method can be adopted for fast, conveniently and accurately distinguishing three kinds of bacterial strains which can generate fumonisins and trigger rice spike rot.

Description

technical field [0001] The invention belongs to the field of molecular biology, and specifically relates to three specific genes for producing fumonisin Fusarium races and applications thereof. Background technique [0002] Fusarium ( Fusarium spp .) Belongs to the subphylum Tumorellales. The microscopic features based on the identification of Fusarium species mainly include: 1. The shape of the megaspore, the characteristics of the apical cell and the basal cell; The characteristics of sporozoites and the types of phialides; 4. The presence and location of chlamydospores; 5. The growth rate and color of the colonies. Dai Fanglan's "Chinese Fungus Collection" divides Fusarium into 16 groups, 51 species and 45 varieties. The morphological identification of Fusarium is relatively difficult due to the rich variety of its species, the complexity of the classification organs, and the collection and classification of pure cultures that are prone to variation due to improper cul...

Claims

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Application Information

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IPC IPC(8): C12N15/31C12N15/11C12Q1/6895C12Q1/686C12Q1/04
CPCC07K14/37C12Q1/686C12Q1/6895C12Q2600/16C12Q2537/143
Inventor 王玲黄世文刘庆刘连盟
Owner CHINA NAT RICE RES INST
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