Mulberry powdery mildew pathogenic bacteria Phyllactinia mori ribosome RNA sequence and application thereof

A DNA sequence and ribosome technology, which can be used in DNA/RNA fragmentation, recombinant DNA technology, determination/inspection of microorganisms, etc.

Active Publication Date: 2017-06-06
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it is difficult to study the classification of fungal species based on the rDNA sequences of 5.8S, 18S, and 28S, and the species of pathogenic fungi cannot be determined. Therefore, the full-length sequence of rRNA is required for research

Method used

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  • Mulberry powdery mildew pathogenic bacteria Phyllactinia mori ribosome RNA sequence and application thereof
  • Mulberry powdery mildew pathogenic bacteria Phyllactinia mori ribosome RNA sequence and application thereof
  • Mulberry powdery mildew pathogenic bacteria Phyllactinia mori ribosome RNA sequence and application thereof

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Embodiment 1

[0043] The leaves with typical powdery mildew lesions were randomly found in the mulberry field where the disease occurred, collected, and the lesions in the mulberry leaves were cut. The cut lesions were fully ground with liquid nitrogen, and the total RNA of the mulberry diseased leaves was extracted; The extracted total RNA was stored at -80°C. The total RNA was reverse-transcribed into a cDNA library; the designed primer sequences and universal primer sequences ITS1, ITS4, and ITS5 are shown in SEQ.ID.NO2~SEQ.ID.NO8, respectively, see Table 1. The cDNA library was used as a template for PCR amplification; the PCR reaction system is shown in Table 2.

[0044] Table 1 PCR verification primer sequence list

[0045]

[0046] Table 2 PCR reaction system (20μL)

[0047]

[0048] ITS1 and ITS4 primer set PCR program: pre-denaturation at 94°C for 5 minutes; 94°C for 30s, 55°C for 30s, 72°C for 1min, 30 cycles; 72°C for 10min.

[0049] ITS4 and ITS5 primer set PCR program:...

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Abstract

The invention discloses a full-length cDNA sequence of mulberry powdery mildew pathogenic bacteria Phyllactinia mori ribosome RNA and application thereof. The full-length cDNA sequence of the mulberry powdery mildew pathogenic bacteria Phyllactinia mori ribosome RNA is shown in SEQ.ID.NO 1, and two pairs of primer sequences contained in the full-length cDNA sequence are shown as SEQ.ID.NO 2 to 5. According to the full-length cDNA sequence of the mulberry powdery mildew pathogenic bacteria Phyllactinia mori ribosome RNA, the cDNA sequence of the Phyllactinia mori ribosome RNA is applied to detecting Phyllactinia mori bacteria, and qualitative and quantitative results can be obtained at the same time. The results show that fungus with the highest relative abundance in study on fungus on mulberry powdery mildew leaves is Phyllactinia mori pathogenic bacteria. In addition, the cDNA sequence of the Phyllactinia mori ribosome RNA can be applied to study on fungus species classification.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a ribosomal RNA sequence of a mulberry powdery mildew pathogen Phyllactinia mori and an application thereof. 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 transcribed spacers (Internal Tanscribed Spacer, ITS) ITS1 and ITS2 between the genes. form a transcription unit. [0003] The rDNA sequences encoding 5.8S, 18S, and 28S in rDNA are relatively conservative, and can be use...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
CPCC12N15/11C12Q1/6895C12Q2600/178
Inventor 刘吉平刘希
Owner SOUTH CHINA AGRI UNIV
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