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The ribosomal RNA gene and application of erysiphe alphitoides pathogenic tussah powdery mildew

A powdery mildew and pathogenic bacteria technology, applied in the direction of DNA/RNA fragments, application, genetic engineering, etc., can solve the problems of uncertain pathogenic fungal species, high variability of ITS segments, fungal species research, etc., and achieve good application prospects Effect

Active Publication Date: 2021-02-19
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, based on the rDNA sequences of 5.8S, 18S, and 28S, it is difficult to study the classification of fungal species, and the species of pathogenic fungi cannot be determined. ITS sequences are often used to identify species (spieces) level
However, the high variability of the ITS segment presents many problems in application

Method used

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  • The ribosomal RNA gene and application of erysiphe alphitoides pathogenic tussah powdery mildew
  • The ribosomal RNA gene and application of erysiphe alphitoides pathogenic tussah powdery mildew
  • The ribosomal RNA gene and application of erysiphe alphitoides pathogenic tussah powdery mildew

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Embodiment 1 detects the acquisition of the nucleotide sequence of the ribosomal RNA gene of the oak powdery mildew pathogen Erysiphe alphitoides

[0057] 1. Experimental method

[0058] (1) High-throughput sequencing

[0059] The leaves with typical powdery mildew lesions were randomly found in the affected oak garden, collected, cut off the lesions in the oak leaves, and the material of the lesions was cut and fully ground with liquid nitrogen. The total DNA was extracted using Dingguo fungus DNA extraction kit, according to its operating instructions, the extracted total DNA is stored at -20°C; according to the Illumina library construction process, the total DNA is constructed as a paired-end high-throughput sequencing library with a fragment size of 500bp; Illumina Hiseq2500 is used for sequencing The instrument performed high-throughput sequencing on the constructed DNA library, and a total of 11.17M pairs of sequencing fragments were measured. The sequencing rea...

Embodiment 2

[0069] Verification experiment of embodiment 2 ribosome assembly result

[0070] 1. Experimental method for verification of ribosome assembly results

[0071] (1) PCR amplification reaction

[0072] The leaves with typical white diseased spots on the back were randomly found in the diseased oak garden, collected, and the diseased area in the oak leaves was cut off, and the material of the diseased spots was cut and fully ground with liquid nitrogen to extract the total DNA of the diseased leaves of the oak tree. Store at -80°C;

[0073] Design 4 sets of primers for verification of ribosome assembly results (respectively primer set oak1, primer set oak2, primer set oak3 and primer set oak4), the nucleotide sequences of the 4 sets of primers are shown in Table 1 respectively; The total DNA was used as a template, and the above-mentioned 4 sets of primers were used for PCR amplification. The PCR amplification reaction system is shown in Table 2.

[0074] Table 1 PCR amplificat...

Embodiment 3

[0087] Embodiment 3 detects the method for oak tree powdery mildew pathogen Erysiphe alphitoides

[0088] 1. Detection method

[0089] (1) High-throughput sequencing

[0090] The leaves with typical powdery mildew lesions randomly found in the oak tree garden where the disease occurred were used as samples to be tested. The lesion area in the oak leaves was cut, and the lesion material was cut and fully ground with liquid nitrogen. The total DNA was extracted using Dingguo Fungal DNA Extraction Kit, according to its operating instructions, the total DNA after extraction is stored at -20°C; according to the Illumina library construction process, the total DNA is constructed into a paired-end high-throughput sequencing library with a fragment size of 500bp; use The Illumina Hiseq2500 sequencer performed high-throughput sequencing on the constructed DNA library, and the sequencing read length was 125 bp at both ends.

[0091] (2) Assembly of microbial genome sequences

[0092]...

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Abstract

The invention discloses an oak powdery mildew pathogen Erysiphe alphitoides Ribosomal RNA genes and their applications. Oak powdery mildew pathogen Erysiphe alphitoides The nucleotide sequence of the ribosomal RNA gene is shown in SEQ ID NO.1, utilizes this sequence to be able to the oak tree powdery mildew pathogen Erysiphe alphitoides For detection, the sequence can also be applied to the study of fungal species classification. In addition, the present invention also provides a group of pathogenic bacteria for detecting oak powdery mildew Erysiphe alphitoides The primers have strong specificity, and based on the primers, a specific detection method for the pathogenic bacteria of oak powdery mildew was established. Erysiphe alphitoides The detection method and detection kit, in the detection of oak powdery mildew pathogen Erysiphe alphitoides has a good application prospect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and more specifically relates to an oak powdery mildew pathogen Erysiphealphitoides ribosomal RNA gene and its application. Background technique [0002] In the existing fungal research methods, ribosomal DNA (ribosome DNA, rDNA) sequences are often sequenced and compared for the identification of fungi. Ribosomes have important functions in cells, and many genes encoded by rDNA are closely related to the reaction process of protein synthesis and play a decisive role in protein biosynthesis. The rDNA sequence is divided into a transcribed region and a non-transcribed region. The transcribed region consists of genes encoding ribosomal 5.8S, 18S, and 28S protein structures and two intergenic transcribed spacers (Internal Tanscribed Spacer, ITS) ITS1 and ITS2. form a transcription unit. [0003] The rDNA sequences encoding 5.8S, 18S, and 28S in rDNA are relatively conserved and can be used to analyze ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/31C12N15/11C12Q1/6895C12Q1/6869C12Q1/04
CPCC07K14/37C12Q1/6869C12Q1/6895C12Q2531/113C12Q2535/122C12Q2537/165
Inventor 刘吉平王狗旦孙勋勋
Owner SOUTH CHINA AGRI UNIV
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