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Method for cultivating lilies containing delphinin in the petals

A delphinidin and lily technology, applied in biochemical equipment and methods, botany equipment and methods, plant regeneration, etc., can solve problems such as insufficient overexpression

Inactive Publication Date: 2015-02-11
SUNTORY HLDG LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In many cases, simply overexpressing the F3'5'H gene is not sufficient to achieve this goal, and additional genetic manipulations including additional genetic manipulations are required

Method used

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  • Method for cultivating lilies containing delphinin in the petals
  • Method for cultivating lilies containing delphinin in the petals
  • Method for cultivating lilies containing delphinin in the petals

Examples

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Comparison scheme
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Embodiment

[0088] The present invention will be described more specifically by way of examples below, but the scope of the present invention is not limited to these examples.

reference example 1

[0089] [Reference Example 1: Expression of Vinca major F3'5'H gene in lily]

[0090] The plasmid pNAVFH (refer to Non-Patent Document 9, a binary vector for expressing the F3'5'H gene of Vinca vinca in plants) was transformed into the Agrobacterium system EHA101. The recombinant Agrobacterium was used to transform lily variety Marshal (pink flowers). The transformation of lily is carried out by a known method such as the method described in Non-Patent Document 10, but is not limited thereto.

[0091] To investigate whether the transferred F3'5'H gene functions in lilies and whether delphinidin accumulation is formed in the petals, it takes 2 to 3 years after obtaining the transformed lily tissue culture plants. By focusing on the coloring of anthocyanins found in cultured petioles and analyzing the anthocyanins in the petioles, at least it can be quickly judged that the transgene is completely or barely functional. Tissue culture plants of transformed lilies of the obtained ...

reference example 2

[0092] [Reference Example 2: Expression of Cineraria F3'5'H Gene in Lily]

[0093] A DNA fragment of about 1.7 kb obtained by digesting plasmid pSPB2774 containing the F3'5'H cDNA Ci5a18 from Cineraria cineraria (refer to Patent Document 9) with SmaI and XhoI, and a DNA fragment obtained by digesting plasmid pBI221 with HindIII and BamHI A DNA fragment of the cauliflower mosaic virus 35S promoter (hereinafter also referred to as "35S promoter") was ligated to a DNA fragment including a replicon obtained by digesting plasmid pSPB176 (see Patent Document 7) with HindIII and SalI , resulting in plasmid pSPB3472. The DNA fragment obtained by digesting the plasmid with AscI and PacI (including the sequence connected with 35S promoter, Ci5a18 and nopaline synthase terminator) was ligated with the DNA fragment of #493 digested with AscI and PacI to obtain plasmid pSPB3376. It was transformed into lily using Agrobacterium as described in Reference Example 1 to obtain the tissue cultu...

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Abstract

Provided is a method for producing lilies containing a blue pigment (delphinidin) in the petals thereof by introducing a foreign gene into lilies. The method pertaining to the present invention is a method for producing lilies containing delphinidin in the petals thereof, including the following steps: introducing, into a lily, a F3'5'H gene derived from a campanula and comprising a nucleotide sequence encoding a peptide having flavonoid 3'5'-hydroxylase (F3'5'H) activity, such as a nucleotide sequence represented by SEQ ID NO: 1 or SEQ ID NO: 11; while also introducing a F3'H gene fragment derived from a lily and comprising a nucleotide sequence encoding a peptide having flavonoid 3'-hydroxylase activity, such as a nucleotide sequence represented by SEQ ID NO: 3 or SEQ ID NO: 16; and inhibiting the expression of endogenous F3'H expression, which acts on cyanidin synthesis in lily petals, while the F3'5'H gene that has been introduced acts to cause the synthesis of delphinidin.

Description

technical field [0001] The present invention relates to a method for producing lilies with blue pigment (delphinidin) in petals by transferring exogenous genes into lilies. More specifically, the present invention relates to a method for producing a lily having a blue pigment in petals and a lily produced by the method, wherein, in order to synthesize delphinidin-derived flavonoid 3', The endogenous flavonoid 3'-hydroxylase (F3'H) that plays a role in the synthesis of red pigment (anthocyanin) is inhibited by RNAi method when the 5'-hydroxylase (F3'5'H) gene is transferred into lily ) expression, and the fragment of F3'H gene of lily was transferred to synthesize delphinidin in lily petals. Background technique [0002] So far, it has been reported to pass through in carnation (referring to patent document 1), chrysanthemum (referring to patent document 1), rose (referring to patent document 1, patent document 2), Phalaenopsis (referring to patent document 3), Whelan (refer...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/52C12N15/63A01H1/00A01H5/00
CPCA01H5/0272C12N15/825A01H6/56A01H4/008C12N15/8205
Inventor 田中良和中村典子小林仁奥原宏明近藤正刚小池洋介星洋介野水利和
Owner SUNTORY HLDG LTD
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