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Application of a Paeonia lactiflora PlTDC gene in changing plant flower colors

A technology of peony and gene, applied in the application field of peony PlTDC gene in changing plant flower color, and can solve problems such as changing plant flower color

Active Publication Date: 2020-10-27
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

And the application of peony PlTDC in changing the flower color of plants has not been reported yet.

Method used

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  • Application of a Paeonia lactiflora PlTDC gene in changing plant flower colors
  • Application of a Paeonia lactiflora PlTDC gene in changing plant flower colors
  • Application of a Paeonia lactiflora PlTDC gene in changing plant flower colors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Expression of Paeoniae officinale PlTDC Gene Overexpression Vector in Tobacco

[0021] Peony PlTDC gene overexpression vector construction: design primers containing restriction sites BamH I and Kpn I for amplifying the PlTDC sequence (upstream primer PlTDC-F: 5'-CGCGGATCCATGGGTAGTCTTGAAC-3', downstream primer PlTDC-R: 5' -GGGTACCCTAACACCCCTTTGAGTAT-3'). PCR product ligated into pEASY TM -T5 Zero CloningVector vector, transformed into Trans-T 1 Competent cells were then screened for positive clones on LB plates supplemented with 0.1% ampicillin, and positive plasmids were extracted. Take the binary expression vector pCAMBIA1301 plasmid and the PlTDC plasmid containing the restriction site for double enzyme digestion with BamH I and Kpn I respectively. The reaction system is: 2.0 μL 0.5×K Buffer, 10 μL pCAMBIA1301 plasmid or PlTDC containing the restriction site Plasmid, 1.0 μL BamH I, 1.0 μL Kpn I, 6.0 μL ddH 2 O; react at 37°C for 1.5h. The double-digest...

Embodiment 2

[0024] Embodiment 2 Tobacco plant petal color correlation index determination

[0025] Determination of the a* value of the petal color parameter of tobacco plants: the a* value of the petal color parameter was measured by a portable imaging spectrophotometer (RM 200QC, American X-Rite Company). The a* value shows that the petal color of transgenic PlTDC tobacco is lighter, and the results are shown in figure 2 .

[0026] Determination of anthocyanin content in tobacco plant petals: Weigh 1.0g of fresh petals, grind the petals into powder with liquid nitrogen, and use 6mL of extracting solution (CH 3 OH:ddH 2 (O:HCl; 70:29.9:0.1; V:V:V) was dissolved, and the anthocyanins were extracted by shaking at 4°C for 24 hours under light-shading conditions, and then centrifuged at a speed of 12,000rpm for 10 minutes with a centrifuge, and the supernatant was drawn and passed Filter through a 0.22 μm microporous membrane, and measure the absorbance at 520 nm with a spectrophotometer...

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Abstract

The invention discloses an application of a Paeonia lactiflora PlTDC gene in changing plant flower colors. The accession number of the Paeonia lactiflora PlTDC gene is KY765554. The constructed PlTDCgene overexpression vector is transformed into tobacco for expression, so that the anthocyanin content in plants, particularly in tobacco petals, is reduced, anthocyanin synthesis related gene expression is inhibited, and a novel light pink tobacco germplasm is created.

Description

technical field [0001] The invention belongs to the technical field of new application of genes, and in particular relates to the application of peony PlTDC gene in changing the flower color of plants. Background technique [0002] Tryptophan decarboxylase (TDC; EC4.1.1.28) is located in the metabolic pathway of tryptophan, which catalyzes the decarboxylation reaction of tryptophan to generate tryptamine. The biochemical characterization of TDC was first started from periwinkle, a soluble cytosolic protein, and the monomer exists as a homodimer with a molecular weight of 54,000. In the study of the TDC gene of periwinkle, it was found that the TDC enzyme showed high substrate specificity for tryptophan, and the Km value was 0.075mmol / L, while the TDC enzyme in snakeroot, rice and Rauwolfia The Km values ​​were 0.72mmol / L, 0.69mmol / L and 2.89mmol / L, respectively. [0003] The TDC gene was first cloned in periwinkle (J04521) in 1993, and then it was also isolated in plants s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82A01H5/02A01H6/82
CPCC07K14/415C12N15/825
Inventor 赵大球陶俊王荣孟家松孙静张克亮
Owner YANGZHOU UNIV
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