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Mango ethylene receptor gene

An ethylene receptor and mango technology, applied in genetic engineering, plant genetic improvement, fruit and vegetable preservation, etc., can solve the problems of unclear ethylene receptor expression and regulation mechanism, delay the peak of ethylene release, reduce the amount of ethylene released, Ease of cloning and effect of genetic engineering manipulations

Active Publication Date: 2017-05-24
AGRI PROD PROCESSING INST GUANGXI ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] At present, although there have been many reports on the ethylene release and ethylene response of mango fruit, the research on the ethylene signal transduction pathway in the process of mango fruit ripening and aging, especially the expression and regulation mechanism of ethylene receptors is still unclear, and no clones have been cloned. Members of the Ethylene Receptor Gene Family in Mango Fruit

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 Extraction of Mango Ethylene Receptor Gene MiETR1

[0048] In the present invention, the extraction of total RNA and the extraction of cDNA-synthesized total RNA refer to the instructions of Bioteke (produced by Beijing Baitaike Biotechnology Co., Ltd.) Universal Plant Total RNA Extraction Kit (spin column type), and are stored in a -80°C refrigerator. The first strand of cDNA was synthesized according to the manual of RevertAidTM First-Strand Synthesis System, and the synthesized cDNA was used for cloning of conserved regions, 3′RACE and ORF cloning. The first strand of 5′RACE cDNA was synthesized with reference to the TransScriptⅡFirst-Strand cDNA Synthesis SuperMix kit, and 0.2 μL 2 pmol / 0.2 μL 5′ETR-specific primer GSP: 5′-GCTGCCATCTTCAAGCCT-3′; The purification of the first-strand synthesis product, the addition of homopolymer tails, and the synthesis of the second-strand cDNA all refer to the kit of 5′RACE System for Rapid Amplification of cDNAEnds, Vers...

Embodiment 2

[0118] Example 2 Verification or expression characteristic analysis of mango ethylene receptor gene MiETR1

[0119] Use the ProtParam tool to analyze the amino acid sequence deduced from MiETR1 nucleotides, the results show that: the total amount of MiETR1 protein molecules is 86385.1, the isoelectric point pI9.09, the total number of negatively charged and positively charged amino acid residues is 75 and 88 respectively, and its molecular formula is C 3891 h 6264 N 1064 o 1080 S 36 , the total atomic number is 12335, the fat coefficient is 106.84, the instability coefficient is 39.39, and the hydrophilicity is 0.065, indicating that it is a stable hydrophobic protein;

[0120] ProtScale analysis of hydrophilicity and hydrophobicity prediction shows that: the maximum hydrophobicity value of MiETR1 is +4.500, the minimum hydrophilicity value is -4.500, and there are 20 maximum hydrophobic peaks with a value above +2.000; 14 hydrophilic peaks with a value of +2.000 Below -2....

Embodiment 3

[0129] Example 3 Effect of 1-methylcyclopropene repressor gene MiETR1 expression on mango aging

[0130] Effect of 1.1-MCP treatment on MiETR1 expression

[0131] 1-MCP is 1-methylcyclopropene (1-methylcyclopropene, 1-MCP) is a new type of ethylene receptor inhibitor, which has the advantages of non-toxic, tasteless, easy to use, low cost, safe and efficient.

[0132] Take 1-MCP to treat freshly picked mango samples, and process them according to conventional methods. The treatment time is 0, 2, 4, 6, 8, 10, 12, 14, and 16 days, and a blank control group is set at the same time. After each time point, the mango samples were taken to extract the MiETR1 gene according to the methods of Examples 1 and 2, and the expression level of the MiETR1 gene was detected. The results showed that the expression level of MiETR1 was significantly reduced after being treated with 1-MCP, and the reduction was most obvious at 4-10 days. The result is as Figure 9 shown.

[0133] 2.1-Effect of...

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Abstract

The invention relates to the field of biotechnology, in particular to a mango ethylene receptor gene, which is named as MiETR1. Base sequences of the gene MiETR1 are as shown in SEQ ID NO:1 in a sequence table; the gene MiETR1 totally has 2570 base sequences including 113 3' untranslated regions, one polyadenylic acid tail and one opening decode regions with 2220 base sequences; an amino acid sequence of the gene MiETR1 is as shown in SEQ ID NO:2 in the sequence table. According to the mango ethylene receptor gene provided by the invention, the mango ethylene receptor gene MiETR1 is obtained through separating and cloning from mango, and an ethylene inhibitor is acted on the ethylene receptor gene MiETR1, so that an expression of the ethylene receptor gene MiETR1 can be remarkably inhibited, the ethylene release rate is reduced, the ethylene release peak is delayed, and the aim of delaying the ripening and senescence of the mango is achieved.

Description

[0001] 【Technical field】 [0002] The invention relates to the field of biotechnology, in particular to a mango ethylene receptor gene. [0003] 【Background technique】 [0004] Mango (Mangnifera indica L.) is a famous tropical fruit with unique flavor, tender meat and high nutritional value. It plays an important role in both production and trade. In recent years, the annual global mango production has reached 27 million tons. Mango is a typical respiratory climacteric fruit. There is an obvious respiration climacteric during postharvest fruit ripening. The respiration intensity is the highest when the fruit reaches full ripeness, and then gradually decreases as the fruit tissue ages. Post-harvest mangoes are very sensitive to ethylene. Before the jump begins, the ethylene concentration in the tissues is extremely low. Before the jump, ethylene increases significantly, causing a respiratory jump (Zhang Jin, et al. "Study on the Quality Characteristics Evaluation of Six Mango ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415A23B7/154
CPCA23B7/154C07K14/415
Inventor 李丽孙健李昌宝何雪梅盛金凤零东宁郑凤锦刘国明李杰民唐雅园饶川艳肖占仕
Owner AGRI PROD PROCESSING INST GUANGXI ACADEMY OF AGRI SCI
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