Tulip flavonoid-3'-hydroxylase TfF3' H protein and its coding gene

A technology of flavonoids and tulips, applied in the molecular field, can solve problems such as the unknown

Inactive Publication Date: 2014-02-19
SHANGHAI JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no literature report related to tulip F3'H protein and its coding gene sequence

Method used

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  • Tulip flavonoid-3'-hydroxylase TfF3' H protein and its coding gene
  • Tulip flavonoid-3'-hydroxylase TfF3' H protein and its coding gene
  • Tulip flavonoid-3'-hydroxylase TfF3' H protein and its coding gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 , the cloning of tulip TfF3'H gene

[0052] 1. Acquisition of plant material

[0053] Healthy, uniform-sized tulip bulbs (Tulipa fosteriana 'Shangnongzaoxia', approved by the Shanghai Crop Variety Approval Committee. No.: Shanghai Nongpin Huahua 2011 No. 004) were planted and managed in the field according to the routine, and the flowers were fully opened. Petal tissue was collected when the petals were fully colored for RNA extraction.

[0054] 2. Extraction of RNA

[0055] Total RNA was extracted using the "RNA prep pure plant total RNA extraction kit" (RNA prep pure Plant Kit: Tiangen Biochemical Technology (Beijing) Co., Ltd.). The integrity of RNA was identified by formaldehyde denaturing gel electrophoresis, and then the purity and concentration of RNA were determined on a spectrophotometer (Thermo Scientific NANODROP1000 Spectrophotometer).

[0056] 3. Full-length cloning of the gene

[0057] According to the amino acid conserved sequence of F3'H ...

Embodiment 2

[0071] Example 2 , Sequence information and homology analysis of tulip TfF3'H gene

[0072] The full-length CDS open reading frame sequence of tulip TfF3'H is 1533bp, and the detailed sequence is shown in the sequence shown in SEQ ID NO.3; the amino acid sequence of tulip TfF3'H is deduced according to the CDS open reading frame sequence, with a total of 510 amino acid residues , with a molecular weight of 55482.2 Daltons and an isoelectric point (pI) of 6.98. See the sequence shown in SEQ ID NO.4 for the detailed sequence.

[0073] The CDS open reading frame sequence of tulip TfF3'H and the amino acid sequence of its encoded protein were stored in the Non-redundant GenBank+EMBL+DDBJ+PDB and Non-redundant GenBank CDS translations+PDB+SwissProt+Superdate+PIR databases using the BLAST program Nucleotide and protein homology searches were carried out, and it was found that it has 82% identity with the lily F3'H gene (GenBank accession number AB699162.1) at the nucleotide level,...

Embodiment 3

[0074] Example 3 , Expression differences of tulip F3'H gene in different developmental stages of flowers and in different tissues of tulip

[0075] 1. Acquisition of materials

[0076] In the 4 different developmental stages of tulip flowers (buds, petals are not colored; buds, petals are beginning to color; flowers are partially open, petals are not fully colored; flowers are fully open, petals are fully colored), the petals are collected in the field, and the leaves are collected at the same time , aerial stems, stamens, pistils, and petals (mixed samples of petals at each coloring stage), the samples were wrapped in aluminum platinum paper and immediately put into liquid nitrogen, and then transferred to -80°C ultra-low temperature refrigerator for storage until use.

[0077] 2. Extraction of RNA

[0078] RNA prep pure Plant Total RNA Extraction Kit (RNA prep pure Plant Kit: Tiangen Biochemical Technology (Beijing) Co., Ltd.) was used to extract the petals of tulip flow...

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Abstract

The invention relates to a tulip flavonoid 3'- hydroxylase TfF3' H protein, and a coding gene and a probe thereof. The protein is (a) or (b) as follows: (a) a protein composed of an amino acid sequence shown as a SEQ ID NO.4; and (b) a protein obtained by substitution, deletion or addition of one or several amino acids on the amino acid sequence shown as the SEQ ID NO.4, having tulip hydrogen flavonoid 3'-hydroxylase activity and derived from (a). The invention also provides a nucleic acid sequence encoding the protein, and the probe to detect the above nucleic acid sequence. The invention employs genetic engineering technology to regulate spatio-temporal expression characteristic of the TfF3' H gene of tulip, so as to provide theoretical basis for color change and color innovation, and has important application value.

Description

technical field [0001] The invention relates to a key enzyme in the synthesis pathway of tulip anthocyanins and its encoding gene, in particular to a tulip flavonoid-3'-hydroxylase TfF3'H protein and its encoding gene, belonging to the field of molecular technology. Background technique [0002] Flower color is an important factor affecting plant pollination, and also determines the commercial value and ornamental value of a flower. Changing flower colors and creating new flower colors can increase the commodity value and ornamental value of flowers. The color of flowers is the result of the accumulation of anthocyanins in petal cells, mainly including flavonoids, carotenoids and betaines. Flavonoids are a large class of anthocyanins that give flowers their full range of color, from yellow to purple. [0003] Flavonoid-3'-hydroxylase (flavonoid3'-hydroxylase, F3'H, once called dihydroflavonol-3'-hydroxylase in the previous literature, is now mostly called flavonoid-3'-hydr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/02C12N15/53
CPCC12N9/0073C12Y114/13021
Inventor 袁媛史益敏唐东芹马晓红
Owner SHANGHAI JIAOTONG UNIV
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