Application of pltdc Gene of Paeoniae Alba in Changing Flower Color of Plants

A peony and gene technology, applied in the application field of peony PlTDC gene in changing plant flower color, can solve the problem of changing plant flower color, etc.

Active Publication Date: 2021-03-19
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 pltdc Gene of Paeoniae Alba in Changing Flower Color of Plants
  • Application of pltdc Gene of Paeoniae Alba in Changing Flower Color of Plants
  • Application of pltdc Gene of Paeoniae Alba in Changing Flower Color of Plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Expression of excess expression vectors in Tobacco

[0021] Survey of peony PLTDC gene excess expression vector: design containing enzyme dug point BAMH I and KPN I for amplifying PLTDC sequences (upstream primers PLTDC-F: 5'-cgcggggatgggtcttgaac-3 ", downstream primer PLTDC-R: 5 ' -Gggtacccctaaccccttgagtat-3 '). PCR product is connected to PEASY TM -T5 Zero CloningVector carrier, trans-t 1 Apositive cells, followed by screening of positive clones on LB plates attached 0.1% ampicillin, and extracting yang-positive plasmid. The binary expression vector PCAMBIA1301 plasmid and the enzyme-containing PLTDC plasmid were used to perform bisase digestion with BamH I and KPN I, the reaction system: 2.0 μL 0.5 × k buffer, 10 μL of PCAMBIA1301 plasmid or release point PLTDC Plasmid, 1.0 μL of Bamh I, 1.0 μL of KPN I, 6.0 μL DDH 2 O; 37 ° C reaction 1.5 h. The bisase digestion was analyzed by agarose gel electrophoresis using Takara Minibest Agarose Gel Dna Extra MiniBest Ag...

Embodiment 2

[0024] Example 2 Determination of Correlation Indicators of Tobacco Plant Petal Color

[0025] Tobacco plant petals color parameter a * value measurement: using portable imaging spectroscopic chromogenic instrument (RM 200QC, US Ai Li Company) to measure the petal color parameter a * value, compared to wild type tobacco, the PLTDC gene tobacco petals have a significant lower A * value indicates that the PLTDC gene tobacco flower petals are shallow, figure 2 .

[0026] Determination of glycosides in petals of tobacco plants: 1.0 g of fresh petals, ground petals into powder, use 6 ml extract (CH 3 OH: DDH 2 O: HCl; 70: 29.9: 0.1; V: v: V) dissolved, under light shielding conditions were oscillated at 4 ° C for 24 hours for coloridin extraction, then centrifuge with a centrifuge at 12,000 rpm rotational speed for 10 min, absorb the supernatant, pass through 0.22 μm microporous filter filtration, the absorbance of 520 nm was measured by a spectrophotometer. From image 3 It can be se...

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Abstract

The invention discloses the application of peony P1TDC gene in changing plant flower color, and the accession number of the peony P1TDC gene is KY765554. The invention transforms the constructed PlTDC gene overexpression vector into tobacco for expression, reduces the content of anthocyanins in plants, especially tobacco petals, inhibits the expression of genes related to anthocyanin synthesis, and creates a new light pink tobacco germplasm.

Description

Technical field [0001] The present invention belongs to the field of gene new use techniques, and the application of peony PLTDC gene is changed in a variety of plants. Background technique [0002] Tryptophan Decarboxylase, TDC; EC4.1.1.28) is located in a tryptophan metabolic pathway, catalytic chloride (Tryptophan) for decarboxyphine. Tiyptamine. TDC Biochemical Characteristics The earliest begins in Changchun, is a soluble cytoprotein, and the monomer is present as a two polymer having a molecular weight of 54,000. In the study of the Changchun flower TDC gene, TDC enzymes were found to show high substrate specificity for toner, and the KM value was 0.075mol / L, while TDC enzyme in short snake roots, rice and Radu The KM values ​​were 0.72 mmol / L, 0.69 mmol / L and 2.89 mmol / L, respectively. [0003] The TDC gene was first cloned in 1993 in 1993, followed by seminating in rice (AK069031), AK069039), Shu (KU842377) and other plants. At present, the TDC gene is more applic...

Claims

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Application Information

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Patent Type & Authority Patents(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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