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Functions and applications of rice lipase gene Os07g0586800 and encoded protein thereof

A protein-encoding and rice technology, applied in the field of plant genetic engineering, can solve the problems of easy loss of resistance, difficulty in use, and variation of varieties, and achieve the effect of efficient fixed-point editing and improving resistance

Active Publication Date: 2019-12-31
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are currently 42 rice bacterial blight resistance genes / loci reported (http: / / www.shigen.nig.ac.jp / rice / oryzabase / gene / list), but most of the reported Blight genes have a narrow resistance spectrum or are difficult to use. Only genes such as Xa3, Xa4, Xa21 and Xa23 have been widely used in production.
[0003] Because Xanthomonas oryzae pv. oryzae is easy to mutate, the co-evolution of rice and Xanthomonas oryzae pv.

Method used

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  • Functions and applications of rice lipase gene Os07g0586800 and encoded protein thereof
  • Functions and applications of rice lipase gene Os07g0586800 and encoded protein thereof
  • Functions and applications of rice lipase gene Os07g0586800 and encoded protein thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1 Fixed-point editing method of rice lipase gene Os07g0586800 based on Fn-CRISPR / Cpf1 system

[0062] 1. Sequence analysis and target sequence screening of rice lipase gene Os07g0586800

[0063] The amino acid sequence of the protein encoded by the rice lipase gene Os07g0586800 is shown in SEQ ID NO.1, and the nucleotide sequence is shown in SEQ ID NO.2. Sequence analysis shows that the gene includes 1 exon in total, located in SEQ ID NO. No. 66-827 of the NO.2 sequence.

[0064] According to the sequence analysis, the sequence on the first exon of the rice lipase gene Os07g0586800 was used as the Os07g0586800-T1 target sequence of the rice lipase gene Os07g0586800 fixed-point editing method based on CRISPR / Cpf1 technology.

[0065] After a large number of screenings, the present invention determines the positive-sense strand targeting the first exon of the rice Os07g0586800 gene using Fn-CRISPR / Cpf1 technology, using the 495th to 517th positions of Os07g058680...

Embodiment 2

[0078] Example 2 Application of fixed-point editing method based on Fn-CRISPR / Cpf1 technology in rice varieties

[0079] The recombinant Agrobacterium EH105-Fn-Os07g0586800-1 was used to infect the callus induced by mature embryos of the rice variety Nipponbare, and the obtained rice transformed plants were respectively named NIP-Fn-Os07g0586800-1; the specific method is as follows:

[0080]1. Inoculate the recombinant Agrobacterium EH105-Fn-Os07g0586800-1 obtained in Example 1 in YEB liquid medium (containing 50 μg / ml kanamycin and 20 μg / ml rifampicin), shake at 28°C and 200rpm Grow to OD 600 0.6-0.8; centrifuge at 5000rpm, 4°C for 5min, and resuspend the precipitated bacteria in AAM liquid medium (acetosyringone concentration is 200μM / L, pH 5.2) to OD 600 It is 0.6-0.8.

[0081] 2. Remove the glumes from the mature seeds of the rice variety Nipponbare, soak them in 75% ethanol for 1 min, then sterilize them in NaClO solution (mixed with water 1:2, add 1 drop of Tween 20) f...

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Abstract

The invention relates to the technical field of plant genetic engineering, and particularly relates to functions and applications of rice lipase gene Os07g0586800 and an encoded protein thereof. The invention finds that the rice lipase gene Os07g0586800 and the encoded protein participate in the regulation and control of the immune response of the rice to bacterial blight, and can obviously improve the resistance of the rice to the bacterial blight by destroying the biological functions of the encoded protein of the Os07g0586800 gene. The invention realizes high-efficiency fixed-point editingof the Os07g0586800 gene by using the CRISPR / Cpf1 technology, and the length of a lesion is obviously shortened after the fixed-point edited rice is inoculated with the bacterial blight pathogenic bacteria IV and GD 1358. The new function of the Os07g0586800 gene provided by the invention provides a new method for plant breeding and disease resistance, and has very important application value in agricultural production.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, in particular to the application of rice lipase gene Os07g0586800 and its encoded protein in regulating plant disease resistance. Background technique [0002] Rice bacterial blight caused by Xanthomonas oryzae pv.oryzae is an important bacterial disease that restricts rice production. It is a serious hazard to the rice planting industry, generally causing a 20% reduction in rice production- About 30%, seriously up to 50%. Breeding and planting disease-resistant varieties using disease-resistant genes is the most economical and effective measure to control rice bacterial blight. There are currently 42 rice bacterial blight resistance genes / loci reported (http: / / www.shigen.nig.ac.jp / rice / oryzabase / gene / list), but most of the reported Blight genes have a narrow resistance spectrum or are difficult to use. Only genes such as Xa3, Xa4, Xa21 and Xa23 have been widely used in produc...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N15/113C12N9/22C12N15/55A01H5/00A01H6/46
CPCC12N9/20C12N9/22C12N15/8205C12N15/8218C12N15/8281C12Y301/01003
Inventor 周永力李全林
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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