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Application of rice auxin glycosyltransferase gene in cultivation of flooding-resistant rice varieties

A technology of glycosyltransferase and auxin, applied in application, genetic engineering, plant genetic improvement and other directions, can solve the problems of reducing seed germination rate, affecting crop yield, poor seedling formation in the field, etc.

Active Publication Date: 2021-03-16
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the production of direct-seeding rice, due to uneven land preparation or flooding after direct-seeding, the germination rate of seeds will be reduced, the seedlings in the field will be poor, and the growth of seedlings will be uneven, which will even seriously affect the crop yield.
Auxin is an important factor that regulates plant growth and development. There is no report on the regulation of rice flood tolerance by auxin glycosyltransferase gene, and there is no report on the application of using auxin glycosyltransferase gene to screen and cultivate flood-tolerant rice varieties. to report

Method used

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  • Application of rice auxin glycosyltransferase gene in cultivation of flooding-resistant rice varieties
  • Application of rice auxin glycosyltransferase gene in cultivation of flooding-resistant rice varieties

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Gene Cloning

[0046] The sequence of the OsIAGLU gene was cloned by PCR using the japonica rice variety Nipponbare cDNA as a template. The upstream primer sequence of PCR is shown in SEQ ID NO.3 in the sequence table, and the downstream primer sequence is shown in SEQ ID NO.4 in the sequence table. 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

[0047] Example 2: Analysis of OsIAGLU gene expression at different times of seed flooding

[0048]Using the japonica rice variety Nipponbare, select 15 healthy and full seeds each time, disinfect the surface with 0.1% mercuric chloride solution for 5 minutes, rinse with distilled water for 3 times, dry the surface of the seeds, put them in a test tube, add 8 cm deep distilled water, They were cultured at 25°C under light / dark conditions for 12 hours each, and samples were taken after flooding for 0, 12, 24, 36, 48, 60, 72, and 96 hours. The seeds were frozen in liquid nitrogen and quickly ground into powder, and the samples were stored at -80°C. The experiment was repeated 3 times.

[0049] Use the TransZol Plant kit (Transgen, www.transgen.com) kit to extract the RNA of each sample; II Reverse Transcriptase system (Vazyme Biotech Co., Ltd) kit was reverse-transcribed to form cDNA, which was used as a template; analyzed with fluorescent quantitative PCR, and fluorescent qu...

Embodiment 3

[0050] Example 3: Mutant construction

[0051] login to website http: / / www.genome.arizona.edu / crispr / CRISPRsearch.html , screening targets. The target sequence is shown in the sequence list SEQ ID NO.5, SEQ ID NO.6, and primers are designed based on the target sequence, and the primer structure is shown in the sequence list 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 MT1T2-PCR gel recovery product was constructed on the pHUE411 vector by homologous recombination to obtain the pHUE411+MT1T2-PCR vector.

[0052] The obtained plasmid containing the pHUE411+MT1T2-PCR vector 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 sequ...

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Abstract

The invention discloses an application of a rice auxin glycosyl transferase gene in cultivation of flooding-resistant rice varieties. The nucleotide sequence of the gene OsIAGLU is shown as SEQ ID NO.1, and the encoded amino acid sequence of the corresponding protein is shown as SEQ ID NO.2 in a sequence table. According to the invention, the fact that the OsIAGLU gene can regulate and control coleoptile growth under the flooding condition in rice is reported for the first time, tests show that mutation of the gene reduces the coleoptile growth capacity of the seed under the flooding condition, and overexpression of the gene improves the coleoptile growth capacity of the seed under the flooding condition. It is proved that the OsIAGLU gene regulates and controls the coleoptile growth of the rice seed under flooding, and the gene is beneficial to screening and cultivating of flooding-resistant rice varieties and the production of direct seeding rice.

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. In the production of direct-seeding rice, due to uneven land preparation or flooding after direct-seeding, the seed germination rate will be reduced, the seedlings in the field will be poor, and the growth of seedlings will be uneven, which will even seriously affect the crop yield. Auxin is an important factor that regulates plant growth and development. There is no report on the regulation of rice flood tolerance by auxin glycosyltransferase gene, and there is no report on the application of using auxin glycosyltransferase gene to screen and cultivate...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/54C12N15/82A01H5/10A01H6/46C12Q1/6895
CPCC12N9/1048C12N15/8271C12Q1/6895C12Q2600/158
Inventor 何永奇王州飞赵佳孙珊黄成威
Owner SOUTH CHINA AGRI UNIV
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