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Peony PsMYB12 transcription factor as well as encoding gene and application thereof

A technology that encodes genes and genes, applied in applications, genetic engineering, plant genetic improvement, etc., can solve problems such as lack of experimental evidence

Active Publication Date: 2018-04-20
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the analysis of the petal transcriptome analysis of the F1 generation of the purple-spotted peony and 'Fengdan' and their hybrids suggests that CHS, DFR, ANS and GST may play an important role in the formation of the purple-spotted peony. ANS and GST regulate the formation of macules, but both lack experimental evidence
No functional identification and research has been carried out on the genes involved in the formation of spots, especially the MYB family genes that regulate the formation of spots have not been reported

Method used

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  • Peony PsMYB12 transcription factor as well as encoding gene and application thereof
  • Peony PsMYB12 transcription factor as well as encoding gene and application thereof
  • Peony PsMYB12 transcription factor as well as encoding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1, the cloning of peony PsMYB12 gene

[0042] 1. Gene cloning

[0043] Gene cloning was carried out using the S2 stage petals of the purple-spotted peony cultivar 'Qinghai Lake Yinbo' as material.

[0044] (1) Plant total RNA extraction

[0045] Refer to the product manual of RNAprep Pure PlantKit from Tiangen Company to extract total RNA from plant materials. The specific method is as follows:

[0046] 1) Take 50-100mg of petals in the S2 stage, quickly and fully grind them into powder in liquid nitrogen, immediately pour them into a 1.5mL EP tube, add 500μL of SL (add 5% mercaptoethanol before use), and immediately vortex vigorously Shake and mix;

[0047] 2) Centrifuge at 12,000rpm for 2min;

[0048] 3) Transfer the supernatant to the filter column CS in the collection tube, centrifuge at 12,000rpm for 2min, carefully pipette the supernatant in the collection tube into a new 1.5mL EP tube without RNase, try to avoid the tip from touching the collection...

Embodiment 2

[0108] Example 2, Comparison of expression levels of PsMYB12 gene in different stages of peony petal development and in different tissues

[0109] The purple-spotted peony variety 'Qinghai Lake Yinbo' was divided into four stages from the colorless period to the full-bloom period ( figure 1 ), samples were taken every two weeks in the first two periods, and once a week in the latter two periods. The petals of 3 different plants were taken each time as 3 independent experimental repetitions. Total RNA was extracted from all samples and reverse-transcribed into cDNA, which was used as a template for fluorescent quantitative PCR.

[0110] 1. RNA extraction

[0111] The RNA extraction method was the same as that in Example 1 above.

[0112] 2. Reverse transcription

[0113] For reverse transcription, refer to the product manual of Tiangen FastQuant RT Kit for reverse transcription, the method is as follows:

[0114] Since the concentration of RNA extracted from multi-stage sa...

Embodiment 3

[0134] Example 3, VIGS silences PsMYB12

[0135] Tobacco rattle virus (TRV) is a virus composed of two strands, TRV1 and TRV2. The VIGS carrier used in the present invention comes from tobacco rattle virus carrier (tobacco rattle virus, TRV), and TRV1 / TRV2 is a gift from Professor Ma Nan of the Ornamental Plant Postharvest and Stress Physiology Laboratory of China Agricultural University (the non-patent literature that has recorded the carrier is: Tian J,Pei H X,Zhang S,Chen J W,Chen W,Yang R Y,Meng Y L,You J,Gao J P,Ma N.2014.TRV-GFP:amodified Tobacco rattle virus vector for efficient and visualizable analysis of gene function.J Exp Bot, 65:311-322.), which has a Kanna selection marker and a 35S promoter, and pTRV2 has multiple cloning sites such as BamH I and Xho I. The modified virus is used for related research. TRV1 mainly plays the role of virus operation in plants, and TRV2 is an expression vector containing multiple cloning sites.

[0136]PsMYB12 was cloned into an ...

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Abstract

The invention discloses a peony PsMYB12 transcription factor as well as an encoding gene and application thereof. The peony PsMYB12 transcription factor is a protein of (a) or (b) which is described as follows: (a) a protein consists of an amino acid sequence shown by a sequence 1 in a sequence table; (b) proteins which are derived from (a), are associated with peony color spot formation and / or anthocyanin synthesis and are formed by replacing and / or deleting and / or adding one or more amino acid residues in the amino acid sequence shown by the sequence 1 in the sequence table. By means of VIGSsilencing and heterologous excess transformation analysis, the results show that the peony PsMYB12 transcription factor can regulate the expression of key enzyme genes in an anthocyanin synthesis route. The peony PsMYB12 transcription factor is speculated to play an important role in the synthesis and accumulation of anthocyanin in peony flower spots and the formation of color spots. The specifically expressed gene is cloned for the first time and the functions of the gene are verified, so that a foundation is laid for the molecular mechanism of the formation of the color spots, and technicalbasis and thought are provided for further analysis of the synthesis and regulation of flavonoids in peony flowers.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the tree peony PsMYB12 transcription factor and its coding gene and application. Background technique [0002] Peony (Paeonia suffruticosa Andrews) is a deciduous subshrub of Paeoniaceae (Paeoniaceae), Paeonia (Paeonia) peony group (Sec. Moutan DC). It occupies a very important position in the history of flower development in China and even in the world. According to the traditional color system, the peony flower color can be roughly divided into 9 major color systems: red, pink, purple, white, yellow, black, green, blue, and double color. The color spot at the base of peony petals is one of its important features. In addition to its great ornamental value, the color spot has become an important basis for the classification and evolution of peony species. It is worth mentioning that the purple-spotted peony (P.rockii) and some Yunnan peony (P.delayvaii) in its wild species, as well...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N15/86A01H5/00A01H6/82
CPCC07K14/415C12N15/825C12N15/86C12N2770/00043
Inventor 舒庆艳朱瑾刘政安王亮生
Owner INST OF BOTANY CHINESE ACAD OF SCI
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