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Gene PeMYB4 sequence for regulating and controlling petal color of phalaenopsis equestris and application thereof

A Phalaenopsis petal and gene sequence technology, which can be applied in application, genetic engineering, plant genetic improvement, etc., can solve the problem of anthocyanin accumulation and other issues

Active Publication Date: 2021-03-19
SHANGHAI NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Overexpression of two MYB genes, SlANT1 or SlAN2, leads to accumulation of anthocyanins

Method used

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  • Gene PeMYB4 sequence for regulating and controlling petal color of phalaenopsis equestris and application thereof
  • Gene PeMYB4 sequence for regulating and controlling petal color of phalaenopsis equestris and application thereof
  • Gene PeMYB4 sequence for regulating and controlling petal color of phalaenopsis equestris and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Cloning of embodiment 1 Phalaenopsis PeMYB4 gene

[0036] The total RNA of wild-type Phalaenopsis petals was extracted, the extraction kit was RNAplant (commercially available), and the total RNA was reverse-transcribed into cDNA using a reverse transcription kit (commercially available). Primers were designed according to the transcriptome sequencing results. The primer sequences are shown in SEQ ID NO.3 and SEQ ID NO.4. A 738bp band was amplified from the Phalaenopsis cDNA by RT-PCR. The PCR product was recovered to obtain the PeMYB4 gene, its nucleotide sequence is shown in SEQ ID NO.1, and the amino acid sequence encoded by the nucleotide sequence is shown in SEQ ID NO.2, consisting of 245 amino acid residues, with a molecular weight of 28.35 kilodaltons.

Embodiment 2

[0037] Example 2 Verification of the expression profile of Phalaenopsis phalaenopsis in Xiaolanyu at different developmental stages

[0038] (1) extract as figure 1 As shown, the RNA of Phalaenopsis japonica at different developmental stages was extracted using RNAplant (commercially available), and the total RNA was reverse-transcribed into cDNA using a reverse transcription kit (commercially available).

[0039] (2) Design primers according to transcriptome sequencing data, and the primer sequences are shown in SEQ ID NO.5 and SEQ ID NO.6.

[0040] (3) Using the cDNA obtained by reverse transcription of Phalaenopsis japonica at different developmental stages as a template, the expression profile of PeMYB4 at different developmental stages was verified as follows: figure 2 As shown, the expression of PeMYB4 was significantly induced at different developmental stages. It indicated that PeMYB4 may be involved in the formation of petal color of Phalaenopsis in Xiaolanyu.

Embodiment 3

[0041] Example 3 Verification of the expression profile of PeMYB4 in different tissues of Xiaolanyu Phalaenopsis

[0042] (1) extract as image 3As shown, the RNA of Phalaenopsis xiaolanyu in different tissue parts was extracted using RNAplant (commercially available), and the total RNA was reverse-transcribed into cDNA using a reverse transcription kit (commercially available).

[0043] (2) Design primers according to transcriptome sequencing data, and the primer sequences are shown in SEQ ID NO.5 and SEQ ID NO.6.

[0044] (3) Using the cDNA obtained by reverse transcription of Phalaenopsis orchid in different tissue parts as a template, the expression profile of PeMYB4 in different tissue parts was verified as follows: Figure 4 As shown, the expression of PeMYB4 is different in different tissues. Higher expression in petals, lips and sepals. It indicated that PeMYB4 may be involved in the formation of petal color of Phalaenopsis orchid in Xiaolanyu.

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Abstract

The invention discloses a gene sequence for regulating and controlling the petal color of phalaenopsis equestris. The gene sequence comprises one or a plurality of the following nucleotide sequences:1, a nucleotide sequence shown in SEQ ID NO.1; 2, a nucleotide sequence generated by substitution, deletion or addition derivation of one or a plurality of nucleotides of the nucleotide sequence shownin SEQ ID NO.1; and 3, a nucleotide sequence with homology of at least 80% with that in SEQ ID NO.1. The gene sequence PeMYB4 disclosed by the invention has an important effect of regulating and controlling the flower color development of the phalaenopsis equestris, and in a VIGS strain of the phalaenopsis equestris, the color of the phalaenopsis equestris is obviously lightened, so that an important foundation is provided for the breeding of the phalaenopsis equestris.

Description

technical field [0001] The invention relates to a gene sequence and its application, in particular to a gene sequence for regulating the petal color of Xiaolanyu Phalaenopsis orchid and its application. Background technique [0002] Flower plants have important ornamental and economic value, and flower color, as one of the important ornamental traits, has very important research significance; and flower color is mainly regulated by the MYB transcription factor family. The MYB gene family widely exists in eukaryotic cells and is one of the largest gene families in plants (Dubos et al., 2010). The MYB gene family has a highly conserved DNA-binding domain near the N-terminus of the sequence, which is composed of 1-4 incomplete repeat sequences in series, each repeat sequence contains about 52 amino acid residues, and the second repeat sequence And the third repeating sequence constitutes a helix-turn-helix (HTH) structure, which contains 3 tryptophans and is separated by 18-19...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/83A01H5/02A01H6/62
CPCC07K14/415C12N15/8218C12N15/825
Inventor 明凤王锐徐庆于毛婵娟李威
Owner SHANGHAI NORMAL UNIVERSITY
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