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Application of GhGDEF3 gene of gerbera hybrida in double flower formation

A gene and double petal technology, applied in the application field of gerbera GhGDEF3 gene in the formation of double petal flowers, can solve the complex developmental process of double petal flowers, how GhGDEF3 gene regulates the petal properties of gerbera chrysanthemums, etc., and achieve important application value Effect

Active Publication Date: 2018-10-16
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the research on the flower development model has been relatively in-depth, and many valuable conclusions have been drawn, and some genes with important application prospects have been isolated, the formation of double-petaled flowers is a very complicated developmental process
There is no report on how the GhGDEF3 gene regulates the petal properties of gerbera

Method used

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  • Application of GhGDEF3 gene of gerbera hybrida in double flower formation
  • Application of GhGDEF3 gene of gerbera hybrida in double flower formation
  • Application of GhGDEF3 gene of gerbera hybrida in double flower formation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Phenotype analysis of double-petal 'Fenxiu' gerbera and its semi-double mutant

[0025] During the blooming period, count the petals of semi-double and double flowers, border tongue flowers, transition flowers and disc flowers, and count 10 flowers for each flower type. Measure the length and width of marginal tongue flowers and transition flowers, and observe the development of disk flowers. The length of the petal is the longest length from the flower tip to the base, and the width is measured at the widest point perpendicular to the length of the petal. Each experiment was repeated three times, and 20-30 petals were measured each time.

[0026] The results showed that the overall morphology and vegetative growth of the semi-double mutant had no difference compared with the double-petal 'Fenxiu' gerbera, and the mutant phenotype was manifested in the inflorescence ( figure 1 ). It can be seen from Table 1 that the size of the double-petal type transitiona...

Embodiment 2

[0033] Example 2 Expression characteristics of GhGDEF3 gene in double-petal 'Fenxiu' gerbera and semi-double mutants thereof

[0034] 1. Total RNA extraction

[0035] The double-petal 'Fenxiu' gerbera and its semi-double-petal mutants were used to extract RNA from the marginal tongue flowers, transition flowers, disk flowers and stamens of the inflorescence initial opening, half-opening, full-blooming and decaying stages, respectively, and the RNA extraction and The purification process was extracted according to the Magen RNA plus kit, and the extraction steps were performed according to the instructions. 1 μg of total RNA was used for RNA reverse transcription using the PrimeScript reverse transcription kit (TaKaRa), and the cDNA product obtained by reverse transcription was diluted 10 times and then used for real-time fluorescent quantitative PCR.

[0036] 2. Fluorescent quantitative PCR detection of GhGDEF3 gene

[0037] The gene of Gerbera chrysanthemum GhGDEF3 is numbe...

Embodiment 3

[0045] The virus silencing (VIGS) experiment of embodiment 3GhGDEF3

[0046] 1. Cloning of GhGDEF3 gene

[0047] According to the nucleotide sequence of the target gene GhGDEF3 on NCBI, design and synthesize the following gene full-length cloning primers:

[0048] Upstream primer (SEQ ID NO: 7): 5'-ATGGCGAGAGGGAAGATCCAG-3';

[0049] Downstream primer (SEQ ID NO: 8): 5'-TTAGCCAAAGCAAAGCATATGTGGC-3'.

[0050] Using the cDNA of double-petal 'Fenxiu' gerbera as a template, use ExTaq enzyme for PCR amplification, use TBE buffer to make 1.0% agarose gel, and then perform agarose gel electrophoresis on the PCR product, cut it under ultraviolet light Get an agarose gel containing the target DNA, use Shanghai Sangon's Gel Recovery Kit for gel recovery, and use 40 μL of eluent to elute the recovered product. The recovered product was subjected to A-tailing reaction using rTaq DNA polymerase (TaKaRa), and the reaction condition was 72° C. for 30 min. The ligation vector used pMD19-T (T...

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Abstract

The invention belongs to the technical field of gene engineering, and particularly discloses application of a GhGDEF3 gene of gerbera hybrida in double flower formation. The invention firstly discovers that expression of the GhGDEF3 gene in the gerbera hybrida can control the length of a transition flower; during inhibition of the expression of the gene, the double gerbera hybrida can become semidouble, so that the gene has important influence on the formation of double flowers of the gerbera hybrida; a theoretical guidance is provided for the molecular research on the flower type of the gerbera hybrida and the breeding of the gerbera hybrida; the GhGDEF3 gene has an important application value.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to the application of the gerbera chrysanthemum GhGDEF3 gene in the formation of double flowers. Background technique [0002] Gerbera (Gerbera hybrida) is a perennial evergreen perennial herbaceous flower of the genus Gerbera in the family Asteraceae, also known as Gerbera, and is one of the five largest cut flowers in the world. Gerbera has three flower types: single petal, semi-double petal and double petal, which are divided according to the length and presence or absence of transitional flowers. Among them, those with only marginal tongue flowers and central disk flowers are defined as single petals; semi-double flowers not only have marginal tongue flowers and central disk flowers, but also have a tongue similar to marginal tongue flowers between marginal tongue flowers and central disk flowers The flower structure, that is, the transition flower, its length is le...

Claims

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Application Information

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IPC IPC(8): C12N15/84C12N15/29A01H5/02A01H6/14
CPCC07K14/415C12N15/8218C12N15/827
Inventor 范燕萍罗燕羽余让才李昕悦玉云祎岳跃冲
Owner SOUTH CHINA AGRI UNIV
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