Application of arabidopsis gene SPOC1 in regulating and controlling flowering stages of plants

A technology of Arabidopsis and genes, applied in the field of molecular biology, can solve the problems of key regulatory factors of target genes and unclear molecular regulatory mechanisms

Active Publication Date: 2017-05-24
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the expression level of ADF gene directly affects the regulation of plant growth and development, especially the flowering stage, the target gene, the key regulatory factors involved and the molecular regulation mechanism are not clear.

Method used

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  • Application of arabidopsis gene SPOC1 in regulating and controlling flowering stages of plants
  • Application of arabidopsis gene SPOC1 in regulating and controlling flowering stages of plants
  • Application of arabidopsis gene SPOC1 in regulating and controlling flowering stages of plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Embodiment 1, T-NDA insertion mutant homozygous screening

[0019] 1. Obtain sterile vaccine

[0020] Seeds of wild-type Arabidopsis thaliana and SPOC1 gene T-DNA insertion mutant (SALK_099124C) were purchased from ABRC (Arabidopsis Biological Resource Center). The full-length gene coding (CDS) sequence of the SPOC1 gene is shown in SEQ ID No:1.

[0021] Arabidopsis thaliana seeds were sterilized with 1% NaClO solution (V / V) for 8 minutes, washed with sterile Tween water for 1 minute after shaking for 1 minute, dried and sown on 1 / 2 MS medium. After being placed at 4°C for 3 days, the light culture was placed in a light incubator (16h light / 8h dark, 22°C). After 10 days of light culture, the seedlings were moved into the soil to continue culturing.

[0022] 2. Arabidopsis plant DNA extraction

[0023] (1) Take 2 leaves and place them in a 1.5ml EP tube, add 800μl of DNA extraction solution;

[0024] (2) Water bath at 60°C for 1 hour, and mix by inverting every 10 mi...

Embodiment 2

[0046] Example 2. SPOC1 (AT5G25520) expression analysis in spoc1 mutants

[0047] 1. Extraction of Arabidopsis total RNA

[0048] (1) Take 2 pieces of rosette leaves of Arabidopsis thaliana and put them into a pre-cooled mortar, add liquid nitrogen and quickly grind them into a uniform powder. During the grinding process, ensure that the material is immersed in liquid nitrogen;

[0049] (2) After the liquid nitrogen volatilizes, quickly transfer the material to a pre-cooled 1.5ml RNase-free centrifuge tube, add 1ml TRIZOL solution for every 50-100mg tissue material, mix well, and place it at room temperature for 5min, then 12000rpm, 4℃ Centrifuge for 10 minutes;

[0050] (3) Take 800 μl of the supernatant to a new 1.5ml RNase-free centrifuge tube, add 0.2ml of chloroform, oscillate for 15sec to mix well, leave at room temperature for 5min, centrifuge at 12000rpm, 4℃ for 15min;

[0051] (4) Take 450 μl of supernatant, add an equal volume of isopropanol, mix well, and place a...

Embodiment 3

[0072] Embodiment 3.spoc1 mutants compare with wild type flowering time

[0073] 1. Obtain sterile vaccine

[0074] Arabidopsis thaliana seeds were sterilized with 1% NaClO solution (V / V) for 8 minutes, washed with sterile Tween water for 1 minute after shaking for 1 minute, dried and sown on 1 / 2 MS medium. After being placed at 4°C for 3 days, the light culture was placed in a light incubator (16h light / 8h dark, 22°C). After 10 days of light culture, the seedlings were moved into the soil to continue culturing.

[0075] 2. Statistical analysis of flowering time and number of rosette leaves

[0076] 40 Arabidopsis wild-type plants and 40 spoc1 mutant plants with normal growth were selected, and the days of plant growth and the number of rosette leaves when the first flower of the plants opened were counted.

[0077] Statistical results such as image 3 As shown, the flowering time of the spoc1 mutant was later than that of the wild type, and the number of rosette leaves was...

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Abstract

The invention discloses application of an arabidopsis gene SPOC1 in regulating and controlling flowering stages of plants. The application is implemented as follows: screening out T-DNA of the arabidopsis gene SPOC1, inserting the arabidopsis gene SPOC1 into a mutant homozygote plant; and comparing the flowering time and rosette leaf quantity of the mutant homozygote plant with those of a wild-type plant growing under same conditions so as to discover that the knockout of the SPOC1 gene causes expression changes of flowering relevant genes CO, RGL2, FUL, LFY and AP1 in the arabidopsis plants, and the flowering stage of the mutant homozygote plant is later than that of the wild-type plant. The technical scheme provided by the invention has great significance in regulating and controlling flowering stages of plants.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and specifically relates to the application of the Arabidopsis gene SPOC1 in regulating the flowering period of plants. Background technique [0002] Actin filament depolymerization factor (ADF / cofilin) ​​widely exists in eukaryotic cells and plays a very important role in maintaining cell morphology, cell transport, energy conversion, information transmission, cell division and differentiation. Our previous studies showed that the overexpression of Arabidopsis ADF1 in transgenic plants can cause the disappearance of microfilament bundles in different cell types, while reducing the expression of ADF1 (gene silencing) can promote the formation of microfilament bundles; ADF1 expression The change of ADF1 gene expression will also affect the flowering period and plant growth, and the reduction of ADF1 expression will lead to flowering delay (the wild type blooms in 35 ± 1 days, and grows 1...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29A01H5/00
Inventor 董春海赵宇航于延冲乔龙飞崔宪奎王琨杨靖丽侯晓敏杨洪兵
Owner QINGDAO AGRI UNIV
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