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Rubber tree tapping panel dryness associated protein HbMC1 and encoding gene and application thereof

A rubber tree and dead bark technology, applied in application, genetic engineering, plant genetic improvement, etc., can solve the problem of no research report on the metacaspase family gene of rubber tree.

Active Publication Date: 2016-01-20
RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] So far, there is no research report on the metacaspase family genes of rubber tree

Method used

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  • Rubber tree tapping panel dryness associated protein HbMC1 and encoding gene and application thereof
  • Rubber tree tapping panel dryness associated protein HbMC1 and encoding gene and application thereof
  • Rubber tree tapping panel dryness associated protein HbMC1 and encoding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0111] Example 1. Acquisition and sequence analysis of rubber tree dead skin-related protein HbMC1 and its coding gene

[0112] The applicant used RNA-Seq technology to analyze the difference in gene expression between healthy and dead rubber tree bark, and found that an EST was differentially expressed between healthy and dead rubber tree bark, and highly expressed in dead rubber tree bark. Then the applicant used the EST sequence as a probe to perform homologous comparison analysis on Hevea brasiliensis EST (Expressed Sequence Tags), TSA (Transcriptome Shotgun Assembly) and genome databases, and obtained a line from Hevea brasiliensis containing the complete The gene sequence of the coding region, the gene encodes the metacaspase protein, so it is named HbMC1, and the gene encoding the protein is named HbMC1 gene.

[0113] In order to verify the accuracy of the HbMC1 gene sequence, the total RNA of the bark of Hevea brasiliensis Reyan 7-33-97 was extracted using the Universa...

Embodiment 2

[0117] Example 2. Expression pattern analysis of rubber tree dead skin-related gene HbMC1

[0118] (1) HbMC1 gene is highly expressed in dead rubber tree

[0119] The applicant used the bark and latex cDNA of Hevea brasiliensis Reyan 7-33-97 healthy and dead rubber trees as templates to perform real-time fluorescent quantitative PCR. The experiment was repeated three times, and the specific steps of each repeated experiment were as follows:

[0120]Using the reverse-transcribed cDNA of total RNA extracted from the bark and latex of Hevea Reyan 7-33-97 healthy and dead bark respectively as a template, use gene-specific primers (forward primer is 5′-AGGGAGTCTCAGTTCCGGATT-3′ , the reverse primer is 5′-CGGAAGGTAGAAAAGCAGAGCA-3′) for real-time fluorescent quantitative PCR analysis. Real-time fluorescent quantitative PCR was carried out in Bio-RadCFX96 fluorescent quantitative PCR instrument, and a parallel experiment was set for 3 repetitions. reaction system: PremixExTaq TM (...

Embodiment 3

[0126] Example 3. HbMC1 gene can reduce the oxidative stress resistance of transgenic yeast and induce the death of transgenic yeast

[0127] 1. Construction of recombinant yeast

[0128] (1) Construction of yeast expression vector

[0129] A primer pair for amplifying the coding region (23rd to 1126th nucleotides at the 5' end) was designed according to sequence 2 in the sequence listing, and EcoRI and XhoI restriction sites and protective bases were introduced at the 5' ends of the forward and reverse primers, respectively. Forward primer: 5′-G GAATTC ATGCATATGCTGGTGGACTG-3' (the recognition sequence of EcoRI is underlined), reverse primer: 5'-CCG CTCGAG TCACAAAGAAAAAGGTTTTG-3' (recognition sequence of XhoI is underlined). Using the bark cDNA as a template, the target gene was amplified by PCR. The PCR reaction system is: 2 μL of cDNA template, FastPfuBuffer 5μL, 2.5mMdNTPs 2μL, forward and reverse primers (10μmol / L) each 0.5μL, FastPfuDNAPolymerase (2.5U / L) 0.5μL,...

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Abstract

The invention discloses rubber tree tapping panel dryness associated protein HbMC1 and an encoding gene and application thereof. The rubber tree tapping panel dryness associated protein HbMC1 is A1 or A2. The amino acid sequence of A1 is shown in the sequence 1 of a sequence table. A2 is the protein obtained after substitution and / or missing and / or adding of one or more amino acid residues is conducted on the amino acid sequence shown in the sequence 1, and A2 is derived from A1 and has the same function as A1. Experiments prove that overexpression of the HbMC1 encoding gene can promote cell death under oxidative stress and reduce the resistance of biont on oxidative stress. Expression of the HbMC1 encoding gene is closely related to occurrence of rubber tree tapping panel dryness, and the HbMC1 and the encoding gene thereof can serve as an important target for preventing rubber tree tapping panel dryness.

Description

technical field [0001] The invention relates to rubber tree dead skin-related protein HbMC1 and its coding gene in the field of biotechnology and the application of the two in regulating cell death, oxidative stress resistance and rubber tree dead skin generation. Background technique [0002] Hevea brasiliensis Muell.Arg. has become the main source of natural rubber because of its unique advantages such as high rubber content, good quality, long economic life, and easy harvesting. my country is the world's largest consumer of natural rubber, with an annual consumption of 3.7 million tons. Since my country belongs to a non-traditional rubber planting area, the area of ​​rubber planting is limited, the annual rubber production in my country is less than 800,000 tons, and the self-sufficiency rate is less than 22%. In the case of limited land area for rubber planting, in order to ensure the supply capacity of natural rubber, it is necessary to greatly increase the rubber outp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N15/81C12N1/21C12N1/19A01H5/00
CPCC07K14/415C12N15/8261C12N15/8263C12N15/8271
Inventor 刘辉李德军邓治
Owner RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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