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Regulation gene separated from lumnitzera littorea and application method of regulation gene

A technology for regulating genes and application methods, which can be used in applications, genetic engineering, plant genetic improvement, etc., and can solve problems such as unclear molecular mechanisms.

Active Publication Date: 2020-05-12
HAINAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, as an endangered species with a narrow distribution of thermophiles, the molecular mechanism by which MADS-box genes respond to high-temperature environments to promote plant thermomorphogenesis in Prunus japonica is still unclear.

Method used

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  • Regulation gene separated from lumnitzera littorea and application method of regulation gene
  • Regulation gene separated from lumnitzera littorea and application method of regulation gene
  • Regulation gene separated from lumnitzera littorea and application method of regulation gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Cloning method and sequence analysis of LlMADS11 gene

[0025] The cloning method and sequence analysis of the above-mentioned LlMADS11 gene include the following steps.

[0026] S1: Extraction and detection of the RNA of Honglan Plum The leaves of Honglan Plum were used as materials, and the RNA was extracted using the polysaccharide polyphenol / complex plant RNA rapid extraction kit of Beijing Aidelai Biotechnology Co., Ltd. (refer to the instruction manual for the specific method).

[0027] Detection of total RNA quality and integrity of Honglan plum: take 5μL RNA solution and dilute it to 100μL, measure the absorbance values ​​at 260, 280 and 230nm with a UV nucleic acid protein detector and calculate the ratio of A260nm / A280nm; at the same time, take 5μL of RNA sample with 1% The integrity of total RNA was detected by agarose gel electrophoresis. When the ratio of A260 / A280 is between 1.8-2.0, the 28S and 18S rRNA bands are clear and the brightness ratio is between...

Embodiment 2

[0044] Application of LlMADS11 gene in regulating plant thermomorphogenesis:

[0045] 1. Acquisition of LlMADS11 gene overexpressing plant lines

[0046] The plasmid containing the target gene L1MADS11 obtained in Example 1 was transferred into Agrobacterium competent GV3101 by freeze-thaw method, and cultured on a plate containing 50 mg / L kanamycin and 50 mg / L rifampicin 2 -3 days, PCR amplification was carried out with primers specific to LlMADS11, and the size of the target fragment was the same. The strains were activated in YEB medium at 28°C, and Arabidopsis Col-0 was transformed by the dipping method. Infest three times and wait until the seeds are ripe for harvest. Under a green fluorescent light, the red seeds were selected as the positive transgenic seeds by using glasses that filter green light. Plant the T1 generation, take the leaves in the T1 generation to extract genomic DNA by CTAB method, carry out PCR amplification with LlMADS11 specific primers, carry out ...

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Abstract

The invention discloses a flowering regulation gene LliMADS11 separated from an endangered mangrove plant lumnitzera littorea and an application method of the flowering regulation gene LliMADS11 in thermomorphogenesis, wherein the nucleotide sequence is shown as SEQ ID NO.1, and the amino acid sequence coded by the LliMADS11 gene is shown as SEQ ID NO.2. The LliMADS11 gene contains a structural domain of an MADS family. A recombinant plasmid is obtained after the LliMADS11 gene is connected with a pBinGlyRed vector, and agrobacterium is transformed by a freeze-thaw method. After the arabidopsis thaliana is transformed by a pollen tube channel method, the rosette leaves of transgenic arabidopsis thaliana is 11 leaves fewer in average than the rosette leaves of wild arabidopsis thaliana, andthe flowering period of the transgenic arabidopsis thaliana is obviously put forward. According to the invention, the flowering induction mechanism of the lumnitzera littorea is preliminarily known from the viewpoint of molecular biology for the first time, so that a foundation is laid for regulating the flowering time through over-expression of the target gene in lumnitzera littorea or expression inhibition of the target gene in lumnitzera littorea, and the application method has important significance for clarifying the danger of lumnitzera littorea from the molecular level.

Description

technical field [0001] The present invention relates to a kind of regulatory gene isolated from red olive plum and an application method thereof, belonging to the technical field of bioengineering. Background technique [0002] According to the prediction results of NOAA model, by 2020, the global surface temperature will be at least 0.5℃ higher than the average temperature from 1986 to 2005, and many parts of the earth will be accompanied by a phenomenon of extreme climate - uneven hot and cold, but the overall global temperature is rising. Under high temperature, plants will appear early flowering, thinning and smaller leaves, and elongation and growth of hypocotyl. These biological phenomena are collectively referred to as thermomorphogenesis. At present, the research on thermomorphogenesis has attracted a lot of attention, but the components of this regulatory pathway still need to be identified, and its potential mechanism of action still needs to be further studied. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82A01H5/02A01H6/20
CPCC07K14/415C12N15/827
Inventor 张静文李佳文杨勇张颖
Owner HAINAN NORMAL UNIV
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