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A kind of llimads11 gene isolated from red olive plum and its application method

An application method and isolated technology that can be used in applications, genetic engineering, plant genetic improvement, etc., and can solve problems such as unclear molecular mechanisms

Active Publication Date: 2022-04-05
HAINAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 thermal morphogenesis in Prunus japonica is still unclear.

Method used

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  • A kind of llimads11 gene isolated from red olive plum and its application method
  • A kind of llimads11 gene isolated from red olive plum and its application method
  • A kind of llimads11 gene isolated from red olive plum and its application method

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 L1MADS11 gene include the following steps.

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

[0027] Detection of the quality and integrity of the total RNA of Red Olive Plum: Take 5 μL RNA solution and dilute it to 100 μL, measure the light absorption values ​​at 260, 280 and 230 nm with an ultraviolet nucleic acid protein detector and calculate the ratio of A260nm / A280nm; at the same time, take 5 μL 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 ...

Embodiment 2

[0041] Application method of LlMADS11 gene in regulating plant thermal morphogenesis:

[0042] 1. Obtaining of LlMADS11 Gene Overexpression Plant Strains

[0043] The plasmid containing the target gene L1MADS11 obtained in Example 1 is transferred to Agrobacterium competent GV3101 by freeze-thaw method, and is cultivated on the plate containing the kanamycin of 50mg / L and the rifampicin of 50mg / L. On -3 days, use LlMADS11-specific primers for PCR amplification. If the target fragments have the same size, activate the strains in YEB medium at 28°C, and transform Arabidopsis Col-0 by the flower dipping method. Infestation three times and wait until the seeds are mature to harvest. Under green fluorescent light, use glasses that filter green light to select red seeds as positive transgenic seeds. Plant the T1 generation, and extract the genomic DNA from the leaves in the T1 generation using the CTAB method, and perform PCR amplification with LlMADS11-specific primers, and perfo...

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Abstract

The invention discloses a LliMADS11 gene isolated from Prunus oleifera and an application method thereof. The gene has the property of promoting flowering of plants. The present invention preliminarily understands the flowering induction mechanism of Prunus vulgaris from the perspective of molecular biology for the first time, and lays the foundation for regulating the flowering time by overexpressing or inhibiting the expression of the target gene in Prunus vulgaris in the future. The endangerment of red olive plum is of great significance.

Description

technical field [0001] The invention relates to a LliMADS11 gene isolated from Prunus oleracea and an application method thereof, belonging to the technical field of bioengineering. Background technique [0002] According to the prediction results of the NOAA model, by 2020, the global surface temperature will be at least 0.5°C higher than the average temperature from 1986 to 2005, and there will be an extreme climate in many regions of the earth-the phenomenon of uneven cold and hot, but the overall global temperature is on the rise. Under high temperature, plants will appear early flowering, leaves become thinner and smaller, and hypocotyls elongate and grow. These biological phenomena are collectively called thermomorphogenesis. At present, the research on thermomorphogenesis has attracted wide attention, but the components of this regulatory pathway have yet to be identified, and its potential mechanism of action still needs further in-depth research. [0003] The enda...

Claims

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Application Information

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Patent Type & Authority Patents(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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