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Anthurium andraeanum transcription factor AaMYB3 as well as coding gene and application thereof

A technique for encoding genes and transcription factors, which is applied in the field of Anthurium AaMYB3 transcription factor and its encoding genes and applications, and can solve problems such as transcription regulation research that has not been reported yet

Active Publication Date: 2018-02-16
TROPICAL CORP STRAIN RESOURCE INST CHINESE ACAD OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Transcription factors regulating anthocyanin biosynthesis in Anthurium have been identified, however, the transcriptional regulation of anthurium proanthocyanidin biosynthesis has not been reported

Method used

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  • Anthurium andraeanum transcription factor AaMYB3 as well as coding gene and application thereof
  • Anthurium andraeanum transcription factor AaMYB3 as well as coding gene and application thereof
  • Anthurium andraeanum transcription factor AaMYB3 as well as coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1, the cloning of Anthurium AaMYB3 gene

[0033] Using anthurium variety Anthurium 'Tropical' (seedlings purchased from Kunming Anzuhua Gardening Co., Ltd.) spathe as material, using Tiangen polysaccharide polyphenol plant total RNA extraction kit (purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd. company, product catalog number is DP441) method to extract total RNA, use Thermo Scientific reverse transcription kit (purchased from Thermo Fisher Scientific, product catalog number is K1622) to synthesize cDNA, and use this as a template. Design primers for full-length cDNA amplification: F: 5'-ATGGGCAGGAGACCCTGTT-3' (sequence 4 in the sequence listing); R: 5'-CGCCATTACTTCACCCATTC-3' (sequence 5 in the sequence listing). The above template was used for amplification, and the reaction program was 94°C for 3min, 94°C for 30S, 60°C for 30S, 72°C for 1min for 30S, 30 cycles, 72°C. The DNA polymerase used was Takara ExTaq enzyme. Amplification system...

Embodiment 2

[0035] Example 2, Comparison of expression levels of AaMYB3 gene in different stages of Anthurium development and in different tissues

[0036]The development of flowers (including spathe and spadix) of Anthurium variety Anthurium 'Tropical' was divided into 5 stages, stage 1, flowers (including spathe and spadix) were all drawn out from the protective sheath; stage 2, pedicel Elongated but the spadix is ​​tightly wrapped around the inflorescence; stage 3, the spathula is semi-coiled; stage 4, the spathula has just fully unfolded; stage 5, the middle and lower 2 / 3 of the spadix turn white. The leaves were divided into young leaf stage (deep red leaves curled up) and mature stage (green mature leaves). Using the Tiangen Polysaccharide Polyphenol Plant Total RNA Extraction Kit to extract total RNA from the spathe and spadix of the five developmental stages of the Anthurium 'Tropical' (red) flower, as well as the young leaves and mature leaves, respectively, using The Thermo Sci...

Embodiment 3

[0042] Example 3, construction and functional analysis of Anthurium AaMYB3 gene expression vector

[0043] Using the sequenced pMD 18-T-AaMYB3 plasmid as a template, use Extaq enzyme (purchased from Bao Biological Engineering (Dalian) Co., Ltd., product catalog number is RR001A) to amplify (primer sequence used for amplification: amplification primer: F : 5'-ATGGGCAGGAGACCCTGTT-3'; R: 5'-CGCCATTACTTCACCCATTC-3') AaMYB3 fragment containing complete ORF and stop codon (sequence 2 in the sequence listing from the 1st position to the 891st position at the 5' end), through The TA clone is connected to pCXSN (T-vector) (the public can obtain it from the Institute of Tropical Crops Variety Resources, Chinese Academy of Tropical Agricultural Sciences, and the non-patent literature that has recorded pCXSN (T-vector) is: Chen S, Songkumarn P, Liu J, Wang G.A versatile zero background T-Vector system for gene cloning and functional genomics.Plant Physiology, 2009,150:1111–1121), and sequ...

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Abstract

The invention discloses an anthurium andraeanum transcription factor AaMYB3 as well as a coding gene and an application thereof. The anthurium andraeanum transcription factor AaMYB3 is protein shown in the following a) or b): a) protein consisting of an amino acid sequence shown in sequence 1 in a sequence table; and b) protein which is obtained after the amino acid sequence shown in sequence 1 inthe sequence table is subjected to substitution and / or deficiency and / or addition of one or more amino acid residues, is related to synthesis of proanthocyanidins and is derived from a). The transcription factor AaMYB3 is obtained from the anthurium andraeanum and belongs to an MYB transcription factor family for regulating synthesis of proanthocyanidins. A complete coding frame of the gene is obtained through RT-PCR (reverse transcription-polymerase chain reaction), then, an overexpression vector is constructed and allogenic transformation of tobacco is performed for stable expression, and the transcription factor is found to have the capacity of prompting accumulation of proanthocyanidins.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to anthurium AaMYB3 transcription factor and its coding gene and application. Background technique [0002] Proanthocyanidins, also known as condensed tannins, are an important class of secondary metabolites, belonging to flavonoids, and most of their biosynthetic pathways are shared with anthocyanins. Proanthocyanidins are produced by the polymerization of 2,3-trans-(+)-flavan-3-ol (such as catechin) or 2,3-cis-(-)-flavan-3-ol (such as epicatechin) It usually exists in the form of oligomers or polymers, and its physiological functions in plants are similar to those of other flavonoid compounds, which mainly play the role of anti-oxidation and enhancing resistance to biotic and abiotic stress. Similar to anthocyanins, the metabolism of proanthocyanidins in plants is regulated by many factors: first, its metabolism has strict time and tissue-specific characteristics; second, its metabol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/11C12N15/82A01H5/00A01H6/82
CPCC07K14/415C12N15/825
Inventor 李崇晖黄素荣杨光穗尹俊梅
Owner TROPICAL CORP STRAIN RESOURCE INST CHINESE ACAD OF TROPICAL AGRI SCI
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