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Chitin synthase and gene thereof, and application of the chitin synthase

A chitin synthase and amino acid technology, applied in the field of chitin synthase in Oomycota, can solve problems such as economic losses

Active Publication Date: 2016-08-24
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] There are many important plant pathogenic oomycetes in Oomycota, which can infect a variety of host plants. For example, Phytophthora capsici can infect peppers and tomatoes in Solanaceae, green beans and kidney beans in leguminous plants, and cucurbits. most plants in the family, causing serious economic losses

Method used

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  • Chitin synthase and gene thereof, and application of the chitin synthase
  • Chitin synthase and gene thereof, and application of the chitin synthase
  • Chitin synthase and gene thereof, and application of the chitin synthase

Examples

Experimental program
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Effect test

Embodiment 1

[0051] 1. Cloning of chitin synthase PSCHS1 gene

[0052] 1.1 Extraction of Phytophthora soybean total RNA

[0053] Phytophthora sojae strain PS6 was cultured in the dark at 25°C for 6 days on solid medium V8, and the bacteria cake with a diameter of 5 mm was picked from the edge of the colony and inoculated on a PDA plate covered with cellophane, and the bacteria were scraped after 6 days of dark culture at 25°C. For silk, take about 30 mg of mycelium, put it into a 2ml centrifuge tube without RNase, and add two steel balls with a diameter of 5mm, freeze it in liquid nitrogen and grind it to powder with a ball mill. Phytophthora sojae RNA was extracted using the SV Total RNA extraction kit from Promega, and the specific method steps refer to the instructions for plant tissue RNA extraction in the kit.

[0054] 1.2 Synthesis of first-strand cDNA by reverse transcription of Phytophthora sojae

[0055] First-strand cDNA was synthesized using Takara’s RT reagent Kit with gDNA...

Embodiment 2

[0127] 1. Cloning of the chitin synthase PCCHS gene of Phytophthora capsici

[0128] Operation is with embodiment 1.

[0129] The difference is:

[0130] 1) In this embodiment, Phytophthora capsici strain LT1534 was selected, and the culture time was 4 days.

[0131] 2) The primers designed to clone the chitin synthase PCCHS gene of Phytophthora capsici are PCCHS-F (SEQ ID No. 19): ATGAGCGACCTTCCACCT; PCCHS-R (SEQ ID No. 20): TTACGCGACCGAGGAGAT. Sequencing results show that the PCCHS gene consists of 2745 deoxynucleotides (see SEQ ID No. 21), which encodes 914 amino acids (see SEQ ID No. 22 for the sequence).

[0132] 2. Analysis of the expression pattern of PCCHS gene in different developmental stages of Phytophthora capsici

[0133] Operation is with embodiment 1.

[0134] The difference is:

[0135] Part of the deoxynucleotide fragments in Phytophthora capsici 40S ribosomal protein S3A gene (WS21), Ubc gene and β-tublin gene were used as internal references in the fluo...

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Abstract

The invention relates to a chitin synthase, particularly a chitin synthase in Oomycota. The amino acid sequence of the chitin synthase has more than 75% (preferably more than 80%, more preferably more than 90% and most preferably more than 95%) of similarity to the amino acid sequence disclosed as SEQ ID No.4, and has the amino acid sequence with the same function as the amino acid sequence disclosed as SEQ ID No.4. The activity level of the chitin synthase can regulate the yields of the active sporangia and active zoospore of oomycetes, thereby influencing the pathogenicity of the oomycetes or the disease development degree of the host.

Description

technical field [0001] The present invention relates to chitin synthase, in particular to chitin synthase in Oomycota. Background technique [0002] There are many important plant pathogenic oomycetes in Oomycota, which can infect a variety of host plants. For example, Phytophthora capsici can infect peppers and tomatoes in Solanaceae, green beans and kidney beans in leguminous plants, and cucurbits. most plants in the family, causing serious economic losses. Among them, pepper blight is a devastating oomycete disease in pepper planting and production, which occurs in most parts of my country. Pepper blight has the characteristics of multiple infection routes, short course of disease, and fast spreading speed. When the onset conditions are suitable, short-term It can cause a large number of wilting and death of peppers, or even extinction. Soybean root rot caused by Phytophthora sojae infection is one of the important diseases that seriously endanger soybean production, cau...

Claims

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Application Information

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IPC IPC(8): C12N9/10C12N15/54C12N15/11C12Q1/68A01P3/00
CPCC12N9/1051C12Q1/6895C12Q2600/136C12Q2600/158C12Q2600/166C12Y204/01016
Inventor 刘西莉张灿王为镇牟文君蔡萌刁永朝刘利刘鹏飞李健强
Owner CHINA AGRI UNIV
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