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Pleiotropic gene for controlling vegetative growth and development of floral organs of rice and application thereof

A technology for vegetative growth and flower organs, applied in the fields of application, genetic engineering, plant genetic improvement, etc.

Inactive Publication Date: 2012-01-18
FUJIAN AGRI & FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007]Currently, there is no report of a mutant similar to ddf1 in rice

Method used

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  • Pleiotropic gene for controlling vegetative growth and development of floral organs of rice and application thereof
  • Pleiotropic gene for controlling vegetative growth and development of floral organs of rice and application thereof
  • Pleiotropic gene for controlling vegetative growth and development of floral organs of rice and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Embodiment 1: Rice DDF1 map-based cloning

[0056] 1. Rice material:

[0057] Rice ( Oryza sativa L.) Mutant df1 , the original wild-type parent is indica rice. The mutant was obtained by the inventor from the intermediate material of indica rice breeding.

[0058] 2. Analyze and target groups:

[0059] mutant heterozygote ( DDF1ddf1 ) was crossed with the wild-type variety DZ60, F 1 F2 population was obtained by selfing in the first generation, and 3632 individuals were selected at the rice tillering stage df1 The plants with the mutant phenotype were used as the positioning population, and about 1 gram of leaves were taken from each plant to extract the total DNA.

[0060] 3. Mapping using rice microsatellite markers (RM and SSR) and InDel markers DDF1 Gene:

[0061] The total DNA of rice was rapidly extracted by the rice micro-method. Take about 0.3 grams of rice leaves, put them into a 1.5ml centrifuge tube, and quickly freeze them with liquid ni...

Embodiment 2

[0068] Embodiment 2 complementary experiment

[0069] 1. Construction of complementary experimental vectors

[0070] According to the sequence of the indica rice Minghui 86 gene, the pCAMBIA1300 vector was used to construct the following Figure 4 shown DDF1 Candidate Gene Complementation Experiment Vector pCAMBIA1300- DDF1 . The specific process of vector construction is: design a pair of primers with restriction sites in the candidate F-box gene region and promoter region, use high-fidelity enzymes to perform high-fidelity PCR amplification and sequencing, and select clones with completely correct sequences , after enzyme digestion and ligation, construct as Figure 4 The indicated expression vector was then transformed into Agrobacterium. The DNA sequence of the complementary vector is 8869bp, including a 3740bp fragment before the start codon and a 609bp fragment after the stop codon. The primers used to amplify the DNA sequence of the complementary experiment are ...

Embodiment 3

[0078] Example 3 Inhibition of rice endogenous DDF1 The expression of the new germplasm of ideal plant type and fertility

[0079] First extract the total RNA of wild-type rice leaves, then reverse transcribe it into cDNA, and use the cDNA as a template to PCR amplify DDF1 cDNA sequence; the PCR product was recovered and purified and connected to the pGEMT-easy vector; after sequencing confirmed that it was completely correct, at the same time use Bam H1 and Sac 1 Digest the pGEMT-easy vector and pTCK303 vector with double enzymes, recover the digested products respectively, and connect them with T4 DNA ligase. DDF1 The cDNA was reversely connected to the downstream of the Ubiquitin promoter of the pTCK303 vector and constructed as Figure 6 The indicated expression vector was then transformed into Agrobacterium. amplify DDF1 The primer sequences used in the cDNA are (the underlines are restriction sites respectively Sac 1 and Bam H1): Upstream primer: 5'-C GA...

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Abstract

The invention belongs to the technical field of plant gene engineering, and discloses a pleiotropic gene DDF1 (dwarf and delayed flowering 1) for controlling vegetative growth and development of floral organs of rice and application thereof, particularly mapping, cloning, function verification and application of a pleiotropic gene for controlling vegetative growth and development of floral organsof rice. The nucleotide sequence of the gene DDF1 is disclosed as SEQ ID NO.1 in the sequence table, and the amino acid sequence coded by the gene DDF1 is disclosed as SEQ ID NO.2 in the sequence table. The DDF1 determines the vegetative growth and development of floral organs of rice, especially the development of stamen. Gene engineering technology can be utilized to purposefully regulate the expression of the gene in rice, thereby regulating the plant type and fertility of rice, and creating new species of rice seeds. Therefore, the invention has wide application prospects in genetic improvement of rice.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering. Specifically, it relates to a pleiotropic gene for vegetative growth and floral organ development in rice cloned using map-based cloning technology DDF1 ( Dwarf and Deformed Flower 1 ), and the function of the gene was verified by transgene complementation experiment. At the same time, it also involves the use of this gene to regulate rice vegetative growth and reproductive development, that is, through the use of genetic engineering methods to regulate DDF1 Gene expression, thereby controlling the plant type and fertility of rice, and creating new germplasm for rice breeding. Therefore, the present invention has good application prospect to genetically improved rice. Background technique [0002] Plant growth and development is an extremely complex process. It is generally believed that growth is the increase in plant volume, while development is the process of changing the str...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82C12N15/84A01H5/00
Inventor 段远霖吴为人李生平陈志伟周元昌官华忠兰涛郑蕾蕾
Owner FUJIAN AGRI & FORESTRY UNIV
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