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Method for cultivating blue chrysanthemum by synthesizing related genes through co-transfecting delphinin

A technology of delphinium and gene, applied in the direction of genetic engineering, plant genetic improvement, botany equipment and methods, etc.

Active Publication Date: 2021-08-20
NANJING AGRICULTURAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no relevant report on whether DgAA7GT, DgAA7BG and DgSCPL2 have similar catalytic activities in heterologous plants, thus making the flower color of chrysanthemum blue.

Method used

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  • Method for cultivating blue chrysanthemum by synthesizing related genes through co-transfecting delphinin
  • Method for cultivating blue chrysanthemum by synthesizing related genes through co-transfecting delphinin
  • Method for cultivating blue chrysanthemum by synthesizing related genes through co-transfecting delphinin

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Experimental program
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Effect test

Embodiment 1

[0099] (1) Construction of R4-CamF3'5'HCtA3'5'GT plant expression vector

[0100] 1) Construction of d2F3HPCtA3'5'GT vector

[0101] According to the promoter sequence (Chrysanthemum F3HPromoter, CmF3HP, GenBank: FW570860.1) of the chrysanthemum gene F3H published on the public database (NCBI), and at the 3' end of the promoter sequence, add as shown in SEQ ID NO.1 The 5'UTR sequence of NtADH (tobacco alcohol dehydrogenase gene), and the multiple cloning site (MCS) of the intermediate vector pYL322d2 (GenBank: KY420077.1), the Xho I restriction site is introduced upstream of the sequence, and the Pst I is introduced downstream Restriction site, entrusted to General Biological Company to synthesize, carry out double enzyme digestion on the pYL322d2 (GenBank: KY420077.1) plasmid and the synthesized fragments Xho I and Pst I respectively; 20 μL reaction system: 10× FastDigest Buffer 2.0 μL, pYL322d2 plasmid 1.0 μL, Xho I 1.0 μL, Pst I 1.0 μL, ddH 2 O 15.0 μL; synthetic sequence...

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Abstract

The invention discloses a method for cultivating blue chrysanthemum by co-transforming Consolida ajacis (L.) Schur DgAA7GT, DgAA7BG-GT1 and DgSCPL2 genes. The method comprises the following steps: constructing a co-expression vector containing five genes, namely DgAA7GT, DgAA7BG-GT1 and DgSCPL2, Campanula medium L. Cam F3'5'H and Clitoria ternatea Linn. CtA3'5'GT, and then introducing cut chrysanthemum. Researches prove that the exogenous delphinin synthetic genes DgAA7GT, DgAA7BG-GT1 and DgSCPL2 genes can enable the chrysanthemum to form pure blue flower color under the assistance of Campanula medium L. CamF3'5'H and Clitoria ternatea Linn. CtA3'5'GT genes. The method disclosed by the invention can fill the blank that the color of the chrysanthemum cannot be changed into pure blue color only by using Campanula medium L. CamF3'5'H and Clitoria ternatea Linn. CtA3'5'GT genes in the prior art, provides a novel and practical method for breeding blue chrysanthemum by utilizing a genetic engineering technology, and can effectively promote the breeding process of a chrysanthemum biotechnology.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering technology and transgenic breeding, and relates to a method for cultivating blue chrysanthemums by co-transferring genes related to delphinidin synthesis, and specifically involves including delphinium delphinidin (cyanodelphin), aster The plant expression vector 380N-CamF3'5'H- 7BG-SCPL2-7GT-Ct3'5'GT construction, transformed cells, cultivation and identification methods of cut chrysanthemum transfected with 380N-CamF3'5'H-7BG-SCPL2-7GT-Ct3'5'GT vector and application of transgenic plants. Background technique [0002] Chrysanthemum flowers are rich in color, but only blue is lacking, because chrysanthemum lacks the biosynthetic pathway for delphinidin. The flavonoid 3', 5' hydroxylase (F3'5'H) gene is called "blue gene". The real reason why the flower color of chrysanthemum lacks blue is the lack of F3'5'H gene. Flavonoid 3', 5' hydroxylase (F3'5'H) catalyzes naringenin to produce pent...

Claims

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Application Information

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IPC IPC(8): C12N15/84C12N15/66A01H5/02A01H6/14
CPCC12N15/825C12N15/66C12N9/0073C12N9/1051C12N9/1029C12N9/2445C12Y114/13088C12Y204/01249C12Y302/01021Y02A50/30
Inventor 蒋甲福祝钦泷林娇阳韩笑盈罗宇婷吴慧莹周李杰陈素梅房伟民陈发棣
Owner NANJING AGRICULTURAL UNIVERSITY
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