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Application of transcription factor gene RrMYB11 in regulating and controlling rose types

A technology of rosette type and transcription factor, applied in the field of plant genetic engineering

Inactive Publication Date: 2013-12-04
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, no MYB transcription factors have been reported in roses

Method used

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  • Application of transcription factor gene RrMYB11 in regulating and controlling rose types
  • Application of transcription factor gene RrMYB11 in regulating and controlling rose types
  • Application of transcription factor gene RrMYB11 in regulating and controlling rose types

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Isolation and cloning of the RrMYB11 gene

[0027]In the early stage of the present invention, the breeding literature of Fenghua rose (also known as "Pingyin No. 1", http: / / tc.cctv.com / 20100412 / 103621.shtml) has been published, see: Lu Chuanrun (ping Yin Rose Research Institute), new varieties of roses-Fenghua Rose and its cultivation techniques, Shandong Forestry Science and Technology, 2007, 5:77; Huazhong Agricultural University College of Horticulture and Forestry, Horticultural Plant Biology Flower Practice Teaching Base from Pingyin County, Pingyin County, Shandong Province Introduced by the research institute) the transcriptome sequencing was performed on flowers at different periods (transcriptome sequencing was completed by Shenzhen Huada Gene Technology Co., Ltd.), and the full-length sequence of the RrMYB11 gene was obtained from the transcriptome sequencing results (see GenBank accession number: FR828544. 1, http: / / www.ncbi.nlm.nih.gov / nuccore / FR...

Embodiment 2

[0038] Embodiment 2: Construction and transformation of RrMYB11 gene overexpression vector

[0039] MYB transcription factors are involved in almost all aspects of plant development and metabolism. In order to better clarify the function of this gene, the applicant overexpressed it in tobacco and verified it from the phenotype of transgenic plants. The specific steps are: at first the positive clone obtained in Example 1 The plasmid was double-digested with BamH I and Sal I, and the target fragment was recovered; at the same time, the genetic transformation vector pCAMBIA2300s carrying the double tobacco mosaic virus promoter 35S (the genetic transformation vector came from Huazhong Agricultural University, Wuhan City, Hubei Province) was digested with the same method. Established and donated by the State Key Laboratory of Crop Genetic Improvement). After the enzyme digestion, use the enzyme-digested fragment containing the RrMYB11 gene and the enzyme-digested pCAMBIA2300s (...

Embodiment 3

[0064] Example 3: RrMYB11 gene transgenic T 0 Phenotype observation and RT-PCR detection of offspring in the field

[0065] After the transgenic tobacco plants were transplanted to the flowering stage, the flower shape of the transgenic tobacco was compared with that of the non-transformed tobacco, and it was found that the flower shape of the RrMYB11-transformed tobacco changed: the corolla of the non-transformed tobacco was five-sided shape, no substantive split petals ( image 3 A); the RrMYB11 gene-transferred tobacco petals are invaginated to form a five-pointed star, and the outline of 5 petals with obvious deep cracks appears, and the tail tip of the petals is heart-shaped ( image 3 B).

[0066] In order to verify whether the change of the flower type of transgenic tobacco is related to the RrMYB11 gene transferred, the present invention has adopted the commonly used RT-PCR method to detect the RrMYB11 gene expression in some transgenic tobacco plants (results see ...

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Abstract

The invention belongs to the technical field of plant genetic engineering, and particularly relates to separation, clone, functional identification and application of a rose DNA fragment. The rose DNA fragment comprises a rose transcription factor gene RrMYB11, wherein the nucleotide sequence of the rose transcription factor gene RrMYB11is shown in a sequence list SEQ ID NO: 1, and the corresponding protein sequence is shown in SEQ ID NO: 2. The gene fragment can change the flower type of a plant, and can be transferred into the plant directly to change the flower type of the transgenic plant.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering. It specifically relates to the separation and cloning, functional verification and application of a rose gene fragment. The gene is related to the development of plant petals. After the complete translation region of the gene is combined with the cauliflower mosaic virus promoter, it is directly transferred into normal plants, and the petal development and flower type of the transgenic plants change. Background technique [0002] Transcription factors, also known as trans-acting factors, refer to DNA-binding proteins that can specifically interact with cis-acting elements in the gene promoter region, and activate or inhibit certain genes through the interaction between them and other related proteins. Transcription of these genes (Zhang Chunyu, Long Yan, Feng Ji, Meng Jinling Plant Gene Regulation at the Transcriptional Level and Its Biological Significance. Heredity. 2007, 29...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H5/00
Inventor 宁国贵邢文包满珠王秀卿包颖
Owner HUAZHONG AGRI UNIV
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