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Method of producing sterile plant, plant obtained by using the same and use thereof

A production method and plant technology, applied in the plant field, can solve the problems of limited RNA interference effect, experimental failure, difficulty in obtaining overlapping petal plants, etc.

Inactive Publication Date: 2006-12-13
JAPAN SCI & TECH CORP +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A technology that inhibits anther dehiscence by producing a chimeric protein containing a transcription factor that promotes the transcription of anther dehiscence-related genes and a functional peptide that converts the transcription factor into a transcriptional repressor has not been seen before
In addition, in the conventional method of breeding by crossbreeding, in order to produce plants with the desired traits, a long period of time and the experience of skilled technicians are required
Therefore, it is very difficult to easily and reliably obtain overlapping petal plant bodies in a short period of time.
[0064] It is also conceivable to suppress the function of the AG gene by the above-mentioned RNAi method to obtain a plant body with overlapping petals. However, in the RNAi method, it is difficult to determine the target site when suppressing gene expression, and there are various problems such as: inevitable experimental failure, difficulty Establishment of constructs, low transfection efficiency due to different cells, limited effect of RNA interference, etc.
Therefore, it is very difficult to obtain overlapping petal plant bodies easily and reliably in a short period of time by the RNAi method

Method used

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  • Method of producing sterile plant, plant obtained by using the same and use thereof
  • Method of producing sterile plant, plant obtained by using the same and use thereof
  • Method of producing sterile plant, plant obtained by using the same and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0370] In the following examples, the polynucleotide encoding the 12-amino acid peptide LLDLDLELRLGFA (SRDX) (SEQ ID NO: 17), one of the transcriptional repression transition peptides, was combined with the APETALA3 gene and linked to the cauliflower flower that functions in plant cells Under the 35S promoter of the leaf virus, a recombinant expression vector was constructed, and introduced into Canina chinensis by using the Agrobacterium method, thereby changing the shape and quality of Canina chinensis.

[0371] (1) Construction of vector pBIG2 for plant shape and quality change

[0372] Plasmid p35S-GFP produced by Clontech (Clontech, USA) was digested with restriction enzymes Hind III and BamHI, and a DNA fragment containing the cauliflower mosaic virus 35S promoter was separated and recovered by agarose gel electrophoresis.

[0373] The vector p-BIG-HYG (Becker, D. 1900 Nucleic acid Research, 18: 203) obtained from Michigan State University was cleaved using restriction e...

Embodiment 2

[0404] In this example, by constructing a recombinant expression vector that incorporates the following polynucleotide, the recombinant expression vector is introduced into Canina chinensis by using the Agrobacterium method, thereby changing the shape and quality of Canina chinensis, the above polynucleotide Below the NACAD1 gene, a polynucleotide encoding 12 amino acid peptide LLDLDLELRLGFA (SRDX) (SEQ ID NO. 17) is combined. One of the transcriptional repression transition peptides between the transcriptional termination regions of the synthetase gene.

[0405]

[0406] Such as figure 2 As shown, p35SG was constructed as a vector for shape modification vector construction according to the following steps (1)-(4).

[0407] (1) Using primers attL1-F (SEQ ID NO: 142), attL1-R (SEQ ID NO: 143), attL2-F (SEQ ID NO: 144) attL2-R (SEQ ID NO: 145), amplify the Invitrogen (Invitrogen Corporation) attL1, attL2 regions on the pENTR vector. The obtained attL1 fragment was digested ...

Embodiment 3

[0450] In this example, by constructing a recombinant expression vector that incorporates the following polynucleotide, the recombinant expression vector is introduced into Canina chinensis by using the Agrobacterium method, thereby changing the shape and quality of Canina chinensis, the above polynucleotide A polynucleotide encoding 12 amino acid peptide LLDLDLELRLGFA (SRDX) (SEQ ID NO. 17) is combined below the C. chinensis MYB26 gene, and this 12 amino acid peptide LLDLDLELRLGFA (SRDX) (SEQ ID NO. 17) is the 35S promoter of cauliflower mosaic virus and One of the transcriptional repression transition peptides between the transcription termination regions of the choline synthase gene.

[0451]

[0452] Such as figure 2 As shown, p35SG was constructed as a vector for shape modification vector construction in the same manner as in Example 2.

[0453]

[0454] Such as image 3 As shown, p35SSRDXG, which is a construction vector into which a polynucleotide encoding a trans...

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Abstract

Transcription of a gene associated with formation of floral organs is suppressed to produce a sterile plant. A plant cell is transfected with a chimeric gene that includes (i) a coding gene of a transcription factor that promotes expression of a gene associated with formation of floral organs, and (ii) a polynucleotide that encodes a functional peptide that converts an arbitrary transcription factor into a transcription repressor, and a chimeric protein in which the transcription factor is fused with the functional peptide is expressed in the plant cell. The expression of the gene associated with formation of floral organs is dominantly suppressed by the chimeric protein, and as a result a male sterile plant is produced that cannot properly form pollen. The chimeric protein also suppresses expression of a gene associated with dehiscence of anther, and as a result a plant is produced in which dehiscence of anther is suppressed. Further, the chimeric protein suppresses expression of target genes of a transcription factor associated with formation of stamen and pistil, and as a result a double flowered plant is produced.

Description

technical field [0001] The present invention relates to a technique for producing a sterile plant body, and more specifically, to a method for producing a male sterile plant, a method for producing a plant body in which anther dehiscence is suppressed, a method for producing a plant body with overlapped petals, and the production method using these The obtained plant body and its utilization. Background technique [0002] When different varieties are mated and hybrids are made, the offspring show better shape (character) than their parents. Call it heterosis. It is now common to cross crops to produce superior varieties through mating that takes advantage of this heterosis. For example, the main varieties of vegetables and grains are almost all improved through such mating. [0003] To reap the benefits of heterosis, mating must occur between mutually distinct breeds. Therefore, in mating varieties, be sure to make it impossible for self-pollination. Here, in plants in ...

Claims

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Application Information

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IPC IPC(8): A01H1/00A01H5/00C12N15/62C12N15/09
Inventor 高木优平津圭一郎光田展隆
Owner JAPAN SCI & TECH CORP
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