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Pectate lyase PCPEL16 from phytophthora capsici, and coding gene and application thereof

A pectin lyase, coding technology, applied in lyase, application, genetic engineering and other directions, can solve problems such as agricultural production loss

Inactive Publication Date: 2012-09-19
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Worldwide, phytopathogenic oomycetes can easily cause a variety of plant diseases, causing serious losses to agricultural production

Method used

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  • Pectate lyase PCPEL16 from phytophthora capsici, and coding gene and application thereof
  • Pectate lyase PCPEL16 from phytophthora capsici, and coding gene and application thereof
  • Pectate lyase PCPEL16 from phytophthora capsici, and coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1, the preparation of pectin lyase

[0054] 1. Expression of pectin lyases PCPEL16 and PCPEL1

[0055] 1.1 Preparation of genes encoding pectin lyases PCPEL16 and PCPEL1

[0056] The inventors of the present application discovered pectin lyase PCPEL16 and PCPEL1 genes from P. capsici SD33 (J. Phytopathol 157:585-591, 2009) (Shandong Agricultural University). Wherein, the nucleotide sequence of the pectin lyase PCPEL16 gene is shown in sequence 1 in the sequence table, and its coding sequence is the 1-1326th position of sequence 1, which encodes the protein of sequence 2; the 1-66th position of sequence 1 encodes Signal peptide consisting of 22 amino acid residues. The nucleotide sequence of the PCPEL1 gene is shown as sequence 3 in the sequence listing, and its coding sequence is the 1-1233rd position of the sequence 3; the 1-63rd position of the sequence 3 encodes a signal peptide consisting of 21 amino acid residues.

[0057] According to the pectin lya...

Embodiment 2

[0099] Embodiment 2, pectin lyase is to the degradation of capsicum leaf cell wall

[0100] The pectin lyases PCPEL16 and PCPEL1 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, and the inoculation concentration of PCPEL16 and PCPEL1 are both 700ug / mL. At the same time, PCPEL16 and PCPEL1 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. Wherein, the passivated PCPEL16 and PCPEL1 were obtained according to the following method: the purified pectin lyases PCPEL16 and PCPEL1 in Example 1 were boiled in hot water for 5 minutes and then used for activity detection.

[0101] The pectin lyase PCPEL16 de...

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Abstract

The invention discloses pectate lyase PCPEL16 from phytophthora capsici, and a coding gene and an application thereof. The pectate lyase PCPEL16 provided by the invention is protein of a) or b) or c), wherein a) protein consisting of amino acid sequences shown by 23-441 sites of a sequence 2 in a sequence table; b) protein consisting of amino acid sequences shown by the sequence 2 in the sequencetable; and c) protein which is obtained by carrying out substitution and / or deletion and / or addition of one or more amino acid residue on the amino acid sequences shown by the sequence 2 in the sequence table or the amino acid sequences shown by 23-441 sites of the sequence 2, has the activity of the pectate lyase and is derived from a). The pectate lyase PCPEL16 disclosed by the invention has higher enzymatic activity, and the specific activity reaches 28+ / -0.2U / mg protein, so that the pectate lyase PCPEL16 can be used for food processing, such as production of juice and wine.

Description

technical field [0001] The invention relates to a pectin lyase, its encoding gene and application, in particular to the pectin lyase from Phytophthora capsici and its encoding gene and application. Background technique [0002] Pepper Phytophthora Blight is a devastating disease that occurs worldwide. The pathogen causing the disease is Phytophthora capsici Leonian, which belongs to Oomycetes, Peronosporales, Pythiaceae, and Phytophthora. Pathogenic bacteria usually overwinter in diseased plant residues and soil with mycelium, chlamydospores and oospores, and can also overwinter on other host plants and become the source of infection in the second year. When the conditions are suitable, the pathogen after overwintering will spread to various parts of the host along with irrigation and air flow, causing disease and re-infection. In the onset season, the general rate of diseased plants is about 20%, and the serious ones reach more than 80%. In recent years, with the continu...

Claims

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Application Information

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IPC IPC(8): C12N9/88C12N15/60C12N15/63C12N1/19C12N1/21C12N1/13C12N1/15C12N5/10A23L2/04C12R1/645
Inventor 张修国
Owner SHANDONG AGRICULTURAL UNIVERSITY
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