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Tulip chalcone synthase TfCHS protein, and coding gene and probe thereof

A chalcone synthase, tulip technology, applied in genetic engineering, plant genetic improvement, enzymes and other directions, can solve problems such as the absence of tulip CHS protein

Inactive Publication Date: 2013-03-13
SHANGHAI JIAO TONG 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 CHS protein and its coding gene

Method used

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  • Tulip chalcone synthase TfCHS protein, and coding gene and probe thereof
  • Tulip chalcone synthase TfCHS protein, and coding gene and probe thereof
  • Tulip chalcone synthase TfCHS protein, and coding gene and probe thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 , Tulip TfCHS gene cloning

[0044] 1. Acquisition of plant material

[0045] 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. When the petals are fully colored, the petal tissue is collected for RNA extraction;

[0046] 2. Extraction of RNA

[0047] Total RNA was extracted with "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 NANODROP 1000 Spectrophotometer);

[0048] 3. Full-length cloning of genes

[0049] According to the conserved amino acid sequence of the CHS gene, usi...

Embodiment 2

[0060] Example 2 , Sequence information and homology analysis of the TfCHS gene of tulip

[0061] The new tulip TfCHS full-length CDS open reading frame sequence of the present invention is 1179bp, and the detailed sequence is shown in the sequence shown in SEQ ID NO.3. According to the CDS open reading frame sequence, the amino acid sequence of tulip TfCHS is deduced, with a total of 392 amino acid residues. The molecular weight is 42861.3 Daltons, the isoelectric point (pI) is 5.90, and the detailed sequence is shown in SEQ ID NO.4;

[0062] The CDS open reading frame sequence of tulip TfCHS and the amino acid sequence of its encoded protein were analyzed by BLAST program in Non-redundant GenBank+EMBL+DDBJ+PDB and Non-redundant GenBank CDS translations+PDB+SwissProt+Superdate+PIR database Acid and protein homology search, it was found that it has 86% identity with lily CHS gene (GenBank accession number HQ161731.1) at the nucleotide level, such as figure 1and figure 2 S...

Embodiment 3

[0063] Example 3 , Expression difference of tulip TfCHS gene in different developmental stages of flowers and in different tissues of tulip

[0064] 1. Obtaining materials: During the four different development stages of tulip flowers (buds, petals are not colored; buds, petals are beginning to be colored; flowers are partially open, petals are not fully colored; flowers are fully open, petals are fully colored), and other tulips are collected in the field. For the petals, the leaves, aboveground stems, stamens, pistils, and petals (the mixed samples of petals at each coloring stage) were collected at the same time, and the samples were wrapped in aluminum platinum paper and immediately put into liquid nitrogen, and then transferred to -80°C ultra-low temperature Store in the refrigerator for later use;

[0065] 2. RNA extraction: RNA prep pure Plant Total RNA Extraction Kit (RNA prep pure Plant Kit: Tiangen Biochemical Technology (Beijing) Co., Ltd.) was used to extract the...

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Abstract

The invention relates to a tulip chalcone synthase TfCHS 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 chalcone synthase 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 provides theoretical basis for regulation of spatio-temporal expression characteristic of the TfCHS gene of tulip for color change and color innovation by using a genetic engineering technology, and has important application value.

Description

technical field [0001] The invention relates to a key enzyme in the synthetic pathway of tulip anthocyanins, its encoding gene and probe, and in particular to a tulip chalcone synthase TfCHS protein, its encoding gene and its probe. Background technique [0002] The color of flowers is one of the important factors that determine the commodity value and ornamental value of flowers. Changing and innovating 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 color from yellow to purple. The biosynthetic pathway of flavonoids can be divided into three stages, the first stage is from phenylalanine to coumaryl-CoA, which is common to many secondary metabolisms; the second stage is from coumaryl-CoA to dihydroflavonol , which is a key reaction ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/10C12N15/54C12Q1/68C12N15/11
CPCC12N9/1037C12Y203/01074
Inventor 袁媛史益敏唐东芹马晓红
Owner SHANGHAI JIAO TONG UNIV
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