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Genes, proteins, gene expression cassettes, expression vectors, host cells, methods and applications for controlling rice flowering time

A flowering time and gene technology, applied in the field of molecular genetics, can solve problems such as unclear member functions, and achieve the effects of improving ecological adaptability, reducing planting costs, and shortening the growth period

Active Publication Date: 2021-07-13
JIANGHAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the large number of F-box protein genes in rice, the functions of many members are still unclear

Method used

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  • Genes, proteins, gene expression cassettes, expression vectors, host cells, methods and applications for controlling rice flowering time
  • Genes, proteins, gene expression cassettes, expression vectors, host cells, methods and applications for controlling rice flowering time
  • Genes, proteins, gene expression cassettes, expression vectors, host cells, methods and applications for controlling rice flowering time

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Rice OsFBO14 Gene Sequence and Interference Fragment Sequence

[0041] The sequence of the coding region of the rice OsFBO14 (LOC_Os03g27250) gene is shown in SEQ ID NO.1, and the protein encoded by it contains the F-box domain ( figure 1 ), the sequence is shown in SEQ ID NO.2. There is no information about the function of this gene in rice and the specific traits it controls. In order to clarify the function of the gene in rice, the present invention intends to use RNAi method to reduce the expression level of OsFBO14 gene in rice. The present invention selects the region between the 1115th and 1529th nucleotides of the sequence shown in SEQ ID NO.1 as the interference fragment for constructing the RNAi vector (see SEQ ID NO.3 for the sequence).

Embodiment 2

[0042] Example 2 Construction of rice OsFBO14 gene RNAi vector.

[0043] Primers (SEQ ID NO.4 and SEQ ID NO.5) were designed according to the full-length cDNA sequence of OsFBO14, and the OsFBO14 interference fragment with restriction sites was amplified by PCR. Construct the interference fragment and the reverse complementary sequence of the fragment on the expression vector, the specific steps are as follows:

[0044] 1. Digest the OsFBO14 interference fragment with KpnI and BamHI, recover the target band, and digest the expression vector plasmid pDS1301 with KpnI and BamHI (Chu et al., Promoter mutations of an essential gene for pollen development result in disease resistance in rice. Genes Dev, 2006, 20:1250-1255.), connection (the endonucleases used are all purchased from TAKARA company, usage and dosage are according to the product manual of this company; Ligase is the product of Invitrogen company, usage and dosage are according to the company's Product Manual);

[00...

Embodiment 3

[0050] Example 3 Preparation of rice OsFBO14 gene RNAi transgenic plants

[0051] The invention adopts the Agrobacterium-mediated method to transfer the OsFBO14 gene expression vector into rice, and the Agrobacterium strain used is EHA105. The rice seeds were sterilized to induce callus, and then co-cultured with Agrobacterium to infect the callus, and the transformed callus was screened by selective culture, and then the plants were regenerated on the selective medium. The specific process is as follows:

[0052] 1. Induction of rice callus: Take the mature rice seeds of Zhonghua 11 (ZH11) variety (bred and provided by the Institute of Crop Science, Chinese Academy of Agricultural Sciences), manually or mechanically dehull, select plump, smooth and clean seeds without bacterial spots, and put them in In a 100ml sterile beaker, pour 70% alcohol (15ml) into a sterilized 2min; pour off the alcohol, add 100ml 30% sodium hypochlorite (NaClO) solution, soak for 30min; pour off the...

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Abstract

The invention discloses a gene, protein, gene expression box, expression vector, host cell, method and application for controlling flowering time of rice. The invention provides the sequence of the OsFBO14 gene and a method for reducing the expression of the OsFBO14 gene by means of RNAi technology to advance the flowering time of rice. The growth period of rice can be artificially adjusted by increasing or decreasing the expression of OsFBO14 gene according to the climate characteristics of the planting site, so as to improve the ecological adaptability of rice.

Description

technical field [0001] The invention belongs to the field of molecular genetics, and specifically refers to a gene, protein, gene expression box, expression vector, host cell, method and application for controlling flowering time of rice. Background technique [0002] In higher plants, flowering represents the transition process from vegetative growth to reproductive growth, which plays an important role in the whole growth and development stages of plants. The biological trait of when to flower is affected by the plant's own genetic factors and external environmental factors. Under the influence of this dual effect, a series of flowering induction processes have a general rule in higher plants, that is, the plant leaves produce flower-forming substances (or Called florigen), the flower-forming substance is transported from the leaf to the apex growth point through the transport tissue, stimulating the apical meristem to form a flower. [0003] Flowering period is an impor...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/82C12N5/10C12N1/21C07K14/415A01H5/10A01H5/02A01H6/46C12R1/19C12R1/01
CPCC07K14/415C12N15/827
Inventor 李甜甜花梦婷陈利红高利芬周俊飞彭海方治伟李论
Owner JIANGHAN UNIVERSITY
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