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A kind of peony myb protein and its coding gene and application

A technology of peony and protein, applied in application, genetic engineering, plant genetic improvement, etc., can solve problems such as changing flowering time

Active Publication Date: 2020-11-24
LUOYANG NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For the MYB gene related to anthocyanin regulation that has been reported so far, it has not been found to change the flowering time

Method used

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  • A kind of peony myb protein and its coding gene and application
  • A kind of peony myb protein and its coding gene and application
  • A kind of peony myb protein and its coding gene and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] 1. Extraction of plant total RNA by CTAB method

[0059] (1) Preheat 15 mL of CTAB extract in a 65°C water bath (add 300 μL of β-mercaptoethanol);

[0060] (2) Grinding 2 to 3 g of fresh or -70°C frozen Mudan Erqiao material in liquid nitrogen;

[0061] (3) Transfer the sample to a centrifuge tube with CTAB extract, immediately vortex vigorously for 30-60 seconds, and put it back in a 65°C water bath for a short time (4-5 minutes);

[0062] (4) Add an equal volume of chloroform / isoamyl alcohol (24:1) and vortex, centrifuge at 10,000 rpm for 15 minutes at room temperature to precipitate protein;

[0063] (5) Transfer the supernatant to a new centrifuge tube, repeat the extraction once, and precipitate the protein;

[0064] (6) Transfer the supernatant to a new centrifuge tube, add an equal volume of 4M LiCl to make the final concentration of LiCl 2M, and precipitate at 4°C overnight (<16h) to denature the RNA;

[0065] (7) Centrifuge at 15,000 rpm at 4°C for 1 h to pr...

Embodiment 2

[0086] Example 2 Gene expression vector construction and tobacco genetic transformation

[0087] (1) Design primers with enzyme cleavage sites at the 5' end to amplify the tree peony MYB gene (see Table 1 and Table 2 for the amplification system and procedures), and the primers are as follows (the underlined parts are the Sal I and Pst I enzyme cleavage sites):

[0088] Forward primer MYB-F2: 5'-ACGC GTC GAC ATGGAGGGAATGTTAGGATTGAGAAAAG-3';

[0089] Reverse primer MYB-R2: 5'-ATTGG CTGCAG TTAATTCACCGCTTGCCCTTCAGCACTTAATAG-3';

[0090] (2) Carry out double digestion of the PCR product with Sal I and Pst I and connect it into the plant expression vector pCAMBIA1301 vector, utilize the electric shock method to transfer the expression vector into Agrobacterium Gv3101, and adopt the Agrobacterium-mediated method to transform tobacco, wherein, containing Agrobacterium Gv3101 with empty vector pCAMBIA1301 served as a control. The specific method is as follows:

[0091] ①Rapid P...

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Abstract

The invention belongs to the field of plant gene breeding, and in particular relates to a peony MYB protein, its encoding gene and its application. The amino acid sequence of the peony MYB protein is shown in SEQ ID No.1, the nucleotide sequence of the gene encoding the protein is shown in SEQ ID No.2, and the gene can regulate the synthesis of anthocyanins in tobacco leaves , to make tobacco leaves mottled, which is obviously different from the effects of other anthocyanin-regulated genes that have been reported, and can make tobacco bloom in advance. Thus, this gene has a dual role in the simultaneous regulation of anthocyanins and flowering time, which has not been found in other species. The peony MYB protein and its encoding gene provided by the invention can not only provide important gene resources for peony breeding, but also can be applied to the breeding of other flower plants.

Description

technical field [0001] The invention belongs to the field of plant gene breeding, and in particular relates to a peony MYB protein, its encoding gene and its application. Background technique [0002] Colored plants are one of the key traits that people pay attention to in the breeding process, especially in the field of ornamental plant breeding, a new colored variety can produce huge economic benefits. For a long time, color plant breeding has been dominated by traditional hybrid breeding, which has disadvantages such as long breeding cycle and difficulty in obtaining specific traits. With the development of molecular biology technology, researchers have begun to pay attention to the use of genetic engineering methods to carry out flower breeding research. Its typical feature is that the breeding cycle is greatly shortened, and the use of functional genes can effectively obtain the target traits. The main component of the color of most plants is anthocyanin, and the diffe...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/02A01H6/72
CPCC07K14/415C12N15/827
Inventor 张延召许书贞彭正峰程彦伟杨学义张晓申
Owner LUOYANG NORMAL UNIV
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