Rubber tree disease-resistant related protein GLPs and coding gene and application thereof

A technology for encoding genes and related proteins, which is applied in the application field of disease resistance correlation of rubber tree germin protein gene, and can solve the problems that the molecular mechanism of disease resistance has not been reported.

Inactive Publication Date: 2017-02-22
ENVIRONMENT & PLANT PROTECTION INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the gene of the rubber tree germin-like protein GLPs a...

Method used

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  • Rubber tree disease-resistant related protein GLPs and coding gene and application thereof
  • Rubber tree disease-resistant related protein GLPs and coding gene and application thereof
  • Rubber tree disease-resistant related protein GLPs and coding gene and application thereof

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Experimental program
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Effect test

Embodiment 1

[0029] Embodiment 1. Acquisition of rubber tree germin protein (GLPs) and its coding gene

[0030] Analyzing the nucleotide sequences of the germin-like proteins (GLPs) of Arabidopsis thaliana, poplar and Jatropha curcas that have been registered in NCBI, and by searching the EST sequence database established by us after rubber tree leaves were infected by Corynesporium multiprime About 200bp of the Hevea germinoid proteins (GLPs) gene EST assembled sequence (contig), design a pair of specific primers to amplify the cDNA fragment, and finally obtain the full-length cDNA sequence including the complete reading frame.

[0031] The specific method is as follows:

[0032] cDNA fragment cloning

[0033] Specific primers were designed as follows:

[0034] F (5' end): 5'-GATGAGTCCTGAGTAACGGTG-3'

[0035] R (3' end): 5'-TGACTGCGTACCAATTCCGTC-3'.

[0036] Respectively using Hevea brasiliensis Reyan 7-33-97 (cultivated by the Rubber Research Institute of the Chinese Academy of Tropic...

Embodiment 2

[0040] Embodiment 2. Hevea germin-like proteins (GLPs) bioinformatics analysis

[0041] The Blastp results of the amino acid sequence of the HbGLP01 gene showed that the sequence was highly similar to the typical β-barrel-shaped conserved structure sequence of the cupin-1 family between the 63rd and 209th amino acid residues ( image 3 ). It is speculated that the gene is a member of the Cupin superprotein family. SignalP 4.0 predicts that its signal peptide exists in the 6-25 amino acid sequence region near the N-terminal of the sequence, and the possible cleavage site is between the 22nd and 23rd amino acid residues ( Figure 4 ), the signal peptide can guide the amino acid sequence into the extracellular matrix. The results of subcellular localization prediction by Wolf Psort also indicated that the protein might be localized in the extracellular matrix. The analysis of the physicochemical properties of the protein shows that the encoded 231 aa protein has a molecular we...

Embodiment 3

[0043] Example 3. Analysis of HbGLP01 gene expression pattern

[0044] Specific expression of HbGLP01 gene in Hevea resistant and susceptible germplasm

[0045] Select the conserved region of the gene to design specific fluorescent expression upstream and downstream primers GX-F: 5'-TAATATTGCCAGCGAAACTTC-3' and GX-R: 5'-CCATTTGCTTCATAATCGATA-3', and use qRT-PCR to analyze the receptors of rubber tree varieties IAN873 and PR107 Differential expression of HbGLP01 gene after infection with Corynemycetes sp. The results showed that the expression of HbGLP01 gene in IAN873 (resistant) and PR107 (susceptible) leaves was significantly up-regulated first and then decreased after infection. The expression level of IAN873 began to increase 6 hours after inoculation, reached the peak at 24 hours and then decreased, while the expression level of PR107 rose slowly at 6-24 hours after inoculation, and was significantly up-regulated and reached the peak at 48 hours, and the peak expression ...

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Abstract

The invention discloses rubber tree disease-resistant related protein GLPs and a coding gene and application thereof. The protein has one of following amino acid residue sequences including an amino acid residue sequence shown as SEQ ID No.1 in a sequence list or is protein derived from SEQ ID No.1 by subjecting the amino acid residue sequence shown as the SEQ ID No.1 in the sequence list to substitution and/or deletion and/or addition of one or multiple amino acid residues and having functions of the rubber tree disease-resistant related protein.

Description

technical field [0001] The invention belongs to the field of molecular biology and relates to a disease-resistance related application of a rubber tree germin protein gene. Background technique [0002] Germin-like proteins (GLPs) are a family of proteins that widely exist in plants. They exist in the extracellular matrix in the form of glycoproteins through ionic bonds. Under certain conditions, it exhibits important biological functions. (Li Ying, Liu Shenkui. Research progress on plant germin-like proteins[J]. China Agricultural Science Bulletin, 2014, 30(30): 246-254.) This gene contains a β-sheet barrel domain and belongs to the cupin superfamily (Druka A, Kudrna D, Kannangara CG, von Wettstein D, Kleinhofs A. Physical and genetic mapping of barley (Hordeum vulgare) germin-like cDNAs [J]. Proc Natl AcadSci USA (2002) 99 (2): 850 ~ 855.). GLPs have a variety of functions, not only related to the normal growth and development of plants, but also participate in the defen...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N5/10A01H5/00
CPCC07K14/415C12N15/8282
Inventor 黄贵修李博勋刘先宝时涛蔡吉苗
Owner ENVIRONMENT & PLANT PROTECTION INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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