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Application of Rice Auxin Responsive Genes

An auxin and gene technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc.

Active Publication Date: 2021-06-04
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Auxin is an important factor that regulates plant growth and development. There is no report on the regulation of rice seed vigor by auxin-responsive genes, and there is no report on the application of using auxin-responsive genes to screen and cultivate rice varieties with high vigor

Method used

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  • Application of Rice Auxin Responsive Genes
  • Application of Rice Auxin Responsive Genes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Gene Cloning

[0033] The sequence of the OsSAUR33 gene was cloned by PCR using the japonica rice variety Nipponbare cDNA as a template. The sequence of the upstream primer is shown in SEQ ID NO.3, and the sequence of the downstream primer is shown in SEQ ID NO.4. The nucleotide sequence and amino acid sequence of the rice OsSAUR33 gene are obtained, the nucleotide sequence is shown in SEQ ID NO.1, and the amino acid sequence is shown in SEQ ID NO.2.

Embodiment 2

[0034] Example 2: Mutant construction

[0035] login to website http: / / www.genome.arizona.edu / crispr / CRISPRsearch.html , screening targets. The target sequence is shown in SEQ ID NO.5 and SEQ ID NO.6, and the primers are designed based on the target sequence. The primer structure is shown in SEQ ID NO.7 / SEQ ID NO.8, SEQ ID NO.9 / SEQ ID NO.10 shown. Using pCBC-MT1T2 as a template, PCR amplification with four primers was carried out, and the PCR product was purified and recovered to obtain MT1T2-PCR vector. The pHUE411 vector was digested with BsaI, and the pHUE411+MT1T2-PCR vector was obtained by homologous recombination.

[0036] The obtained vector containing pHUE411+MT1T2-PCR was transformed into Agrobacterium; the Agrobacterium with the transformed plasmid was transformed into the wild-type japonica rice variety Nipponbare; the PCR amplification product was sequenced and compared with the wild type to screen homozygous mutants. The primer sequence is shown in SEQ ID NO....

Embodiment 3

[0037] Example 3: Phenotype Analysis of Gene Mutants

[0038] Seed germination experiments were performed using the constructed OsSAUR33CRISPR / Cas9 mutants ossaur33-1, ossaur33-2, and ossaur33-3 seeds, as well as wild-type Nipponbare (WT) rice varieties. The specific method is as follows: each time repeatedly select 50 healthy and plump seeds, sterilize the surface with 0.1% mercuric chloride solution for 5 minutes, rinse with distilled water for 3 times, dry the surface of the seeds, and place them on a petri dish with two layers of filter paper (diameter 9cm), add 10mL of distilled water, place at 25°C under light / dark conditions for 12h and culture for 7d, and finally count the seedling rate. The experiment was repeated 3 times. The results showed that compared with the control seeds, the mutant seeds germinated slower and the seedling growth was significantly weaker ( figure 1 ). It can be seen that the gene plays an important role in improving the germination speed of ...

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Abstract

The invention discloses the application of the rice auxin response gene. The nucleotide sequence of the gene OsSAUR33 is shown in SEQ ID NO.1, and the amino acid sequence encoding the corresponding protein is shown in SEQ ID NO.2 in the sequence table. The present invention reports for the first time in rice that the OsSAUR33 gene can regulate rice seed vigor, and tests show that mutation of the gene affects seed germination and seedling formation. It is proved that the OsSAUR33 gene of the present invention regulates rice seed vigor, and the use of the gene is helpful for screening and cultivating rice varieties with high seed vigor.

Description

technical field [0001] The invention belongs to the field of seed biotechnology and relates to the application of rice auxin response genes. Background technique [0002] Rice (Oryza sativa L.) is one of the most important food crops in the world. In recent years, with the development of the economy and the shortage of rural labor force, direct-seeding rice production has become more and more common, and the cultivation of high-vigorance rice varieties is of great significance. Breeding rice varieties with high vigor can improve the germination ability of field seeds and promote field seedling formation, which is of great significance to ensure the yield of direct-seeding rice. Auxin is an important factor that regulates plant growth and development. There is no report on the research on the regulation of rice seed vigor by auxin-responsive genes, and there is no report on the application of using auxin-responsive genes to screen and cultivate rice varieties with high vigor...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/6851C12N15/29A01H5/10A01H6/46
CPCC07K14/415C12N15/8261C12Q1/6851C12Q1/6895C12Q2600/158C12Q2600/166C12Q2531/113C12Q2563/107C12Q2545/101
Inventor 王州飞赵佳何永奇
Owner SOUTH CHINA AGRI UNIV
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