Method for production of moth orchid having modified flower color

A manufacturing method and a technology for Phalaenopsis are applied in the field of changing the flower color of Phalaenopsis, which can solve the problems of difficulty in changing the flower color of Phalaenopsis to a desired color and the like

Inactive Publication Date: 2010-03-10
ISHIHARA SANGYO KAISHA LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, it is extremely difficult to change the flower color of Phalaenopsis plants to the desired color despite market demand.

Method used

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  • Method for production of moth orchid having modified flower color
  • Method for production of moth orchid having modified flower color
  • Method for production of moth orchid having modified flower color

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1 Gene introduction into petals of Phalaenopsis

[0061] In the examples of the present specification, unless otherwise stated, various genes were introduced into petals of Phalaenopsis plants using the gene introduction method described below, and their functions were evaluated. All genes adopt a DNA structure with a promoter on the 5' side and a terminator on the 3' side, and are introduced in the form of expression in petal cells.

[0062] The flower buds of Phalaenopsis were sterilized with 1% sodium hypochlorite aqueous solution for 5 minutes, washed 3 times with sterilized water, then the flower buds were decomposed, and the lateral sepals, back sepals, and petals were placed on the bed containing NDM salt (Tokuhara and Mii , Plant Cell Reports (1993) 13:7-11) and 0.6% agarose on agar medium. In addition, as a flower bud, a flower bud of about 15 mm in length of a plant of the genus Phalaenopsis genus Phalaenopsis was used.

[0063]The introduced DNA was...

Embodiment 2

[0065] Example 2 Preparation of expression vector (pBS-P35T35) for gene introduction

[0066] pBS-P35T35 has cauliflower mosaic virus 35S promoter (Hohn et al., Curent Topics in Microbiology and Immunology (1982) 96:194-236) and tobacco mosaic virus Ω sequence (Gallie et al., Nucleic AcidsResearch (1987) 15:3257-3273), restriction enzyme SwaI site and cauliflower mosaic virus 35S terminator figure 2 ). A plasmid substantially functionally equivalent to this pBS-P35T35 can be constructed as follows.

[0067] After the oligonucleotide SAS-S (5'-CTAGCTAGCGGCGCGCCTGCAGGATATCATTTAAATCCCGGG-3'; sequence number: 17) and the oligonucleotide SAS-AS (5'-CCCGGGATTTAAATGATATCCTGCAGGCGCGCCGCTAGCTAG-3'; sequence number: 18) were heat-denatured, slowly After returning to room temperature, the product was treated with NheI and ligated to the XbaI-EcoRV site of pBluescriptIISK-(Stratagene) to prepare a plasmid DNA with changed restriction enzyme sites, which was pBS-SAS. Using the genome D...

Embodiment 3

[0069] The separation of CHS gene, CHI gene, F3H gene, F3'H gene, DFR gene and ANS gene of embodiment 3 Phalaenopsis and preparation of DNA for transient expression

[0070] (1) Isolation of the CHS gene (PhCHS3) of Phalaenopsis

[0071] Using RNeasy Plant Mini Kit (QIAGEN), total RNA was extracted from the petals of Phalaenopsis (Dtps.SogoVivien×Dtps.Sogo Yenlin) just before flowering, and using the RNA as a template, SuperscriptII First-Strand Synthesis System (Invitrogen) was used to Prepare cDNA. Next, RT-PCR was performed using the cDNA as a template. The primers used in the PCR reaction were PhCHS3 F1 (5'-AAGCTTGTGAGAGACGACGGA-3'; sequence number: 21) and PhCHS3R1 ( 5'-TGGCCCTAATCCTTCAAATT-3'; SEQ ID NO: 22). In the reaction, the steps of 94° C. for 30 seconds, 55° C. for 30 seconds, and 72° C. for 1 minute were repeated 25 cycles. Using this reaction solution as a template, the reaction product was amplified again under the same conditions. The resulting reaction p...

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Abstract

Disclosed is a method for modifying the flower color of a moth orchid, particularly a method for producing a moth orchid having the modification of its flower color from white line to red line. Specifically disclosed is a method for producing a moth orchid having a modified flower color, which comprises introducing a gene encoding flavanone 3-hydroxylase, a gene encoding flavonoid 3'-hydroxylase,a gene encoding dihydroflavonol 4-reductase and a gene encoding anthocyanidin synthase into a moth orchid, and allowing these genes to express in the moth orchid.

Description

technical field [0001] The invention relates to a method for changing flower color of Phalaenopsis plants by using gene recombination technology. More specifically, it relates to a method for producing a safflower-colored Phalaenopsis plant by introducing a gene encoding an enzyme of an anthocyanin biosynthetic system into a Phalaenopsis plant. Background technique [0002] In ornamental plants, the color of flowers is a particularly important property, and has been produced by cross breeding of flowers of various colors since ancient times. However, in the random breeding method, in order to introduce only a specific property such as flower color into a specific variety, it is necessary to repeat backcrossing for several generations, which requires a lot of labor and time. Furthermore, the cycle of mating and breeding differs depending on the plant, and most orchids such as Phalaenopsis take a long time until flowering, and it is virtually impossible to produce varieties w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H5/00A01H1/00C12N15/09
CPCC12N15/8243C12N9/0071C12N9/0004A01H5/10
Inventor 荒木智史铃木崇纪片山孝一
Owner ISHIHARA SANGYO KAISHA LTD
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