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Application of Rice Auxin Glycosyltransferase Gene

A technology of glycosyltransferase and auxin, applied in the direction of transferase, application, genetic engineering, etc., can solve the problems of uneven growth of seedlings, affecting crop yield, and unreported application

Active Publication Date: 2020-11-10
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Direct-directed rice production requires high seed sowing quality. Poor seed quality will lead to slow seed germination, low seedling rate in the field, uneven growth of seedlings, and even seriously affect crop yield.
Auxin is an important factor that regulates plant growth and development. There are few reports on the regulation of rice seed germination and seedling growth by auxin glycosyltransferase genes. The application of auxin glycosyltransferase genes to screen and cultivate rice varieties with high vigor Not reported

Method used

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  • Application of Rice Auxin Glycosyltransferase Gene
  • Application of Rice Auxin Glycosyltransferase Gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Gene Cloning

[0033] The sequence of the OsIAGLU gene was cloned by PCR using the japonica rice variety Nipponbare cDNA as a template. The sequence of the upstream primer of PCR 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 OsIAGLU gene are obtained, the nucleotide sequence is shown in SEQ ID NO.1 in the sequence table, 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, SEQ ID NO.6, and the primers are designed based on the target sequence, and the primer sequences are shown in SEQ ID NO.7 / SEQ ID NO.8, SEQ ID NO.9 / SEQ ID NO.10 . 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 pHUE411+MT1T2-PCR vector was transformed into Agrobacterium; the Agrobacterium with the transformation plasmid was transformed into the wild-type japonica rice variety Nipponbare; the PCR amplification product was sequenced, compared with the wild type, and the homozygous mutant was screened. The upstream primers used As shown in SEQ ID ...

Embodiment 3

[0037] Example 3: Phenotype Analysis of Gene Mutants

[0038] Seeds of the successful OsIAGLU CRISPR / Cas9 mutants osiaglu-1, osiaglu-2, and osiaglu-3 constructed in Example 2, and wild-type Nipponbare (WT) rice varieties were used for seed germination experiments. 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 15°C under the conditions of light / darkness 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 ...

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Abstract

The invention discloses application of an oryza sativa L. growth auxin glycosyl transferase gene. The gene OsIAGLU has a nucleotide sequence of SEQ ID NO.1 shown in the description, and a corresponding protein encoded by using the gene has an amino acid sequence of SEQ ID NO.2 shown in the description. By adopting the application, the research that the oryza sativa L. seed activity can be regulated and controlled by the OsIAGLU gene in oryza sativa L. is reported for a first time, and experiments show that by mutating the gene, seed germination and seedling establishing capabilities under normal conditions can be affected. Results show that the OsIAGLU gene is capable of regulating and controlling the oryza sativa L. seed activity, and by adopting the gene, varieties of high oryza sativa L. seed activity can be screened and bred, and production of direct sowing oryza sativa L. can be facilitated.

Description

technical field [0001] The invention belongs to the field of seed biotechnology and relates to the application of rice auxin glycosyltransferase gene. Background technique [0002] Rice (Oryza sativa L.) is one of the food crops with the longest cultivation history in my country. In recent years, direct-seeding rice production has become more common as the economy has developed and rural labor has become increasingly scarce. Direct-directed rice production requires high seed sowing quality. Poor seed quality will lead to slow seed germination, low seedling rate in the field, uneven growth of seedlings, and even seriously affect crop yield. Auxin is an important factor that regulates plant growth and development. There are few reports on the regulation of rice seed germination and seedling growth by the auxin glycosyltransferase gene. The application of the auxin glycosyltransferase gene to screen and cultivate rice varieties with high vitality Not reported. Therefore, the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/54C12N9/10C12N15/82A01H5/10A01H6/46
CPCC12N9/1048C12N15/8267
Inventor 王州飞何永奇赵佳
Owner SOUTH CHINA AGRI UNIV
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