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Anthocyanin synthesis related protein IbMYB113 as well as coding gene and application thereof

A technique for encoding genes and related proteins, which is applied in the field of cloning and application of anthocyanin synthesis-related protein IbMYB113 and its encoding genes. question

Active Publication Date: 2021-11-09
YUNNAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

IbMYB1-2 can regulate the biosynthesis and accumulation of anthocyanins in purple sweet potato flesh. However, due to the polyploidy, genome heterozygosity, and complex genetic background of sweet potato, new anthocyanin biosynthesis regulatory genes in sweet potato have not been discovered, such as Regulatory genes of anthocyanin biosynthesis and accumulation in sweet potato red potato skin are still unclear and need further study

Method used

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  • Anthocyanin synthesis related protein IbMYB113 as well as coding gene and application thereof
  • Anthocyanin synthesis related protein IbMYB113 as well as coding gene and application thereof
  • Anthocyanin synthesis related protein IbMYB113 as well as coding gene and application thereof

Examples

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Embodiment example 1

[0024] Implementation case 1. Cloning of sweet potato anthocyanin synthesis-related protein IbMYB113 and its coding gene

[0025] The cloning steps of IbMYB113 gene are as follows:

[0026] 1. The Trizol reagent produced by Beijing Quanshijin Biotechnology Co., Ltd. was used to extract the total RNA of sweet potato. The specific operation steps are as follows:

[0027] ①Take an appropriate amount of sweet potato red potato skin and quickly grind it into powder with liquid nitrogen in a mortar;

[0028] ②Transfer about 0.1g of the ground sample into a 1.5mL centrifuge tube, add 1.0mL Trizol reagent, vortex for 15sec, and then stand at room temperature for 5min;

[0029] ③Add 0.2mL chloroform, vortex for 15sec, and then let stand at room temperature for 3min;

[0030] ④ Centrifuge at 4°C and 12,000 rpm for 15 minutes, take about 550 μL of the supernatant into another new 1.5 mL centrifuge tube, add 0.5 mL of isopropanol, mix well, and let stand at room temperature for 10 minut...

Embodiment example 2

[0053] Implementation Case 2. Application of Sweet Potato IbMYB113 Protein in Regulating Plant Anthocyanin Synthesis and Accumulation

[0054] 1. Obtaining of whole purple Nicotiana overexpressing IbMYB113 transgene

[0055] 1. Plant expression vector construction

[0056] Produced by Nanjing Novizan Biotechnology Co., Ltd. II One Step CloningKit (C112) constructs the plant expression vector, the specific steps are as follows:

[0057] ① Linearized vector preparation: The vector pHELLSGATE12 was double digested with restriction enzymes XhoI and XbaI, and a linearized vector fragment of about 13798 bp was recovered.

[0058] ② Insertion fragment acquisition: Using the IbMYB113 plasmid DNA obtained in Example 1 as a template, introduce a homologous sequence at both ends of the linearized vector at the 5' end of the forward and reverse amplification primers of the IbMYB113 gene, that is, the forward primer OE-F: 5'- TTGGAGAGGACACGCTCGAGATGGCTAATTCATCATGTGC-3' (underlined is t...

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Abstract

The invention relates to an anthocyanin synthesis related protein IbMYB113 as well as a coding gene and application thereof. The amino acid sequence of the IbMYB113 protein is as shown in SEQ ID NO: 2; the nucleotide sequence of the coding gene is shown as SEQ ID NO: 1. According to the invention, IbMYB113 protein and a coding gene thereof are cloned from red sweet potato peel, the IbMYB113 gene is connected to a plant expression vector, a transgenic tobacco plant over-expressing IbMYB113 is obtained through agrobacterium-mediated genetic transformation, pigment accumulation of the whole plant is shown, and the content of delphinidin and cornflower pigment is detected to be obviously increased by high performance liquid chromatography. The IbMYB113 protein and the coding gene thereof provided by the invention have important theoretical significance and application value for promoting synthesis and accumulation of plant anthocyanin.

Description

technical field [0001] The invention belongs to the field of plant biotechnology, in particular to the cloning and application of anthocyanin synthesis-related protein IbMYB113 and its encoding gene. Background technique [0002] Sweet potato is one of the important food crops, economic crops and energy crops in the world. The color of potato skin mainly includes white, light yellow, yellow, light red, red, purple, dark purple, etc.; the color of potato flesh mainly includes white, yellow, orange, red , purple, deep purple, mixed colors, etc. Red and purple sweet potato varieties mainly contain anthocyanins, which are very popular among consumers because of their beautiful potato appearance and excellent health functions. [0003] Anthocyanin is a water-soluble natural pigment widely present in higher plants and is the most important metabolite in the flavonoid metabolic pathway. At present, more than 635 kinds of anthocyanins have been found in nature. There are mainly 6 ...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/84C12N1/21C12N5/10A01H5/02A01H6/82C12R1/01C12R1/19
CPCC07K14/415C12N15/825Y02A40/146
Inventor 郭华春李茂兴李有涵王琼
Owner YUNNAN AGRICULTURAL UNIVERSITY
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