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Polygalacturonase PCIPG21 from phytophora capsic, and coding gene and application thereof

A technology of encoding and amino acids, which is applied in the fields of application, genetic engineering, and plant genetic improvement, and can solve problems such as agricultural production losses

Inactive Publication Date: 2012-08-29
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Worldwide, phytopathogenic oomycetes can easily cause a variety of plant diseases, causing serious losses to agricultural production

Method used

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  • Polygalacturonase PCIPG21 from phytophora capsic, and coding gene and application thereof
  • Polygalacturonase PCIPG21 from phytophora capsic, and coding gene and application thereof
  • Polygalacturonase PCIPG21 from phytophora capsic, and coding gene and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Embodiment 1, the preparation of polygalacturonase

[0050] 1. Expression of polygalacturonase PCIPG21, PCIPG2 and PCIPG5

[0051] 1.1 Preparation of the coding genes of polygalacturonase PCIPG21, PCIPG2 and PCIPG5

[0052] The inventors of the present application discovered polygalacturonase PCIPG21 gene, PCIPG2 gene and PCIPG5 gene from P.capsici SD33 (J.Phytopathol 157:585-591, 2009) (Shandong Agricultural University) . Wherein, the nucleotide sequence of the polygalacturonase PCIPG21 gene is shown in sequence 1 in the sequence listing, its coding sequence is the 1-1173rd position of sequence 1, and it encodes the protein of sequence 2. Positions 1-57 of Sequence 1 encode a signal peptide consisting of 19 amino acid residues. The nucleotide sequence of the PCIPG2 gene is shown as sequence 3 in the sequence listing, and its coding sequence is the 106th-1191th position of the sequence 3. Positions 106-162 of Sequence 3 encode a signal peptide consisting of 19 amin...

Embodiment 2

[0100] Embodiment 2, polygalacturonase is to the degradation of capsicum leaf cell wall

[0101] The polygalacturonases PCIPG21, PCIPG2 and PCIPG5 purified in Example 1 were inoculated with pepper leaves by acupuncture. The specific method is as follows: take pepper seedlings of Zhongjiao No. 6 at the 4-6 leaf stage, and inoculate pepper leaves by acupuncture method. The inoculation concentrations of PCIPG21, PCIPG2 and PCIPG5 are all 700ug / mL. At the same time, PCIPG21, PCIPG2 and PCIPG5 inoculated and inactivated with sterile water were used as controls. During the period of 1-7 days after protein inoculation of pepper leaves, samples were taken every 24 hours, and the degradation of cell wall was observed by transmission electron microscope. Among them, the passivated PCIPG21, PCIPG2 and PCIPG5 were obtained according to the following method: the purified polygalacturonase PCIPG21, PCIPG2 and PCIPG5 in Example 1 were boiled in hot water for 5 minutes and then used for acti...

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Abstract

The invention discloses polygalacturonase PCIPG21 from phytophora capsic, and a coding gene and application thereof. The polygalacturonase PCIPG21 is the following protein shown in a), b) or c): a) the protein comprises the amino acid sequence shown as the 20th to the 390th site of the sequence 2 in the sequence table; b) the protein comprises the amino acid sequence shown as the sequence 2 in the sequence table; and c) the protein is derived from a) and formed by substituting and / or deleting and / or adding one or more amino acid residues in the amino sequence shown as the sequence 2 in the sequence table or the 20th to the 390th site of the sequence 2 in the sequence table, and has the polygalacturonase activity. The polygalacturonase PCIPG21 has high enzyme activity, wherein the specificactivity of the polygalacturonase is 63 + / - 0.2 U / mg protein; and the specific activity of pectinase is 43 + / - 0.2 U / mg.

Description

technical field [0001] The invention relates to a polygalacturonase, its encoding gene and application, in particular to the polygalacturonase from Phytophthora capsici and its encoding gene and application. Background technique [0002] During the interaction between plant pathogenic oomycetes and hosts, they secrete a variety of important cell wall degrading enzymes or pathogenic enzymes, among which polygalacturonase (PG: EC 3.2.1.15) is a kind of enzyme secreted by various plant pathogenic oomycetes. Important pathogenic enzymes, this type of enzyme destroys the host defense system by degrading the pectin and mesogelatin of the host plant cell wall, thereby enhancing the affinity of the pathogen to the host, so this type of enzyme is an important pathogenic factor. Studies have shown that polygalacturonase is a cell wall-binding protease that can catalyze the cleavage of α-(1,4)-polygalacturonic acid in pectin molecules, degrade pectin in the cell wall, and lead to softe...

Claims

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Application Information

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IPC IPC(8): C12N9/26C12N15/56C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10C12N15/11C12P19/14
Inventor 张修国
Owner SHANDONG AGRICULTURAL UNIVERSITY
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