Transcription factor gene Osspl10 of Oryza sativa and application of transcription factor gene Osspl10

A rice transcription factor and gene technology, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problems of rice SPL gene lack of systematicness, lack of understanding of the impact of yield-related traits, etc., to achieve large application potential, panicle size Reduce and promote the effect of breeding

Inactive Publication Date: 2019-12-03
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, there is a lack of systematic research on the rice SPL gene, and there is no report on the function and impact of the Osspl10 gene in rice. rice The function of the transcription factor gene Osspl10 and its impact on yield-related traits remain poorly understood

Method used

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  • Transcription factor gene Osspl10 of Oryza sativa and application of transcription factor gene Osspl10
  • Transcription factor gene Osspl10 of Oryza sativa and application of transcription factor gene Osspl10
  • Transcription factor gene Osspl10 of Oryza sativa and application of transcription factor gene Osspl10

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Embodiment 1

[0036] A kind of rice transcription factor gene Osspl10 of the present invention, described rice transcription factor gene Osspl10 has following nucleotide sequence:

[0037] (1) The nucleotide sequence of the rice transcription factor gene Osspl10 is as shown in SEQ ID NO: 1, or

[0038] (2) The nucleotide sequence is a nucleotide sequence similar to the DNA sequence shown in SEQ ID NO: 1 with a degree of similarity ≥ 95%, or

[0039] (3) The function of the nucleotide sequence is equivalent to a subfragment of the DNA sequence shown in SEQ ID NO: 1, or the DNA sequence shown in SEQ ID NO: 1 is replaced, deleted or increased by one or more nucleosides Nucleotide sequences that encode the same amino acid sequence.

[0040] The present invention relates to a DNA fragment of Osspl10 gene of rice. The DNA fragment of this gene is shown in the sequence table as SEQ ID NO:1. The Osspl10 gene can encode a transcription factor of the SPL family, and its amino acid sequence is show...

Embodiment 2

[0051] Target Design of Osspl10 Gene

[0052] Such as Figure 1 to Figure 3 As shown, using the TP309 genome and public database sequencing species Nipponbare as the reference sequence, according to the characteristics of the specific recognition sequence of the CRISPR / Cas9 system, a perl language program was written to screen suitable targets, and the qualified results were manually screened The site with the best specificity and located in the exon sequence at the front of the gene was selected as the target for subsequent experiments.

Embodiment 3

[0054] Construction of CRISPR / Cas9 expression vector

[0055] Add the cohesive end sequence and protective bases after digestion to the target sequence and complementary sequence, as the primer sequence sgRNA-F / sgRNA-R and artificially synthesize it. For the sgRNA-F / sgRNA-R sequence, refer to the sequence listing, the sgRNA-F sequence is shown in SEQ ID NO:3, and the sgRNA-R sequence is shown in SEQ ID NO:4. Take the synthesized primers sgRNA-F / sgRNA-R and dilute each 4 μM to 25 μL, and perform annealing reaction in the following procedure: 95°C for 3 minutes; 95°C to 25°C for slow cooling; -1°C for 30 seconds; 16°C for 5 minutes.

[0056] The annealed primer double-stranded DNA fragment is further phosphorylated. The phosphorylation reaction system was as follows: 2 μL of 10×T4 nucleotide kinase buffer; 2 μL of T4 nucleotide kinase; 5 μL of ATP (10 mMol); 11 μL of annealed DNA fragment. The phosphorylation reaction procedure is as follows: 60 min at 37°C; 10 min at 70°C.

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Abstract

The invention belongs to the technical field of plant genetic engineering, and particularly relates to a transcription factor gene Osspl10 of oryza sativa and an application of the transcription factor gene Osspl10. The invention discloses a nucleotide sequence of the gene Osspl10 of oryza sativa and a nucleotide sequence of an SPL family transcription factor coded with the gene Ospl10. A knockoutmutant oryza sativa plant with gene Osspl10 frameshift mutation and loss mutation shows notable reduction of stem length, tiller number and panicle type size. These results indicate that the gene Osspl10 can notably influence yield and correlated character of the oryza sativa, and can have important value in the field of oryza sativa character improvement.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and in particular relates to a rice transcription factor gene Osspl10 and an application thereof. Background technique [0002] Rice (Oryza sativa) is an important economic crop that provides staple food for more than half of the world's population. As the world's largest rice producer, my country's rice production accounts for two-fifths of the world's total rice production. About 30% of the total domestic grain output is rice, and rice production occupies an important position in the national economy. [0003] In order to meet the growing population's demand for food, the improvement of rice traits has always been a hot research topic in the field of botany. With the continuous advancement of high-throughput sequencing and molecular biology technologies, major research breakthroughs in rice functional genomics are continuously providing new theoretical basis for trait improvem...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82C07K14/415A01H5/00A01H6/46C12N1/21
CPCC07K14/415C12N15/8213C12N15/8205C12N15/8261
Inventor 田大成孙晓光张小辉
Owner NANJING UNIV
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