Wx mutant protein based on gene editing technology and application of gene of Wx mutant protein in plant breeding

Abstract

The invention discloses a rice Wx mutant protein, a mutant gene and an application of the mutant protein and the mutant gene. The invention further discloses a breeding method for creating quality-improved rice which is transparent in appearance and properly reduced in amylose content by utilizing gene editing. According to the invention, a gene editing technology is used for carrying out single base editing on the Wx gene for the first time, and a new material without T-DNA, transparent in appearance, properly reduced in amylose content, improved in quality and stable in inheritance can be obtained in a T2 generation by screening offspring. Compared with breeding such as chemical mutagenesis and hybridization transformation, a gene editing directional improved molecular breeding technology has the advantages of quickness, accuracy, high efficiency and the like, gene function marker genotype selection is combined, or other mature rice varieties are directly used as background materialsfor conversion to obtain corresponding materials, so that the breeding efficiency can be greatly improved, and the breeding process is greatly accelerated.

Description

technical field [0001] The invention belongs to the fields of crop genetics and breeding, innovation of new resources for crop quality improvement, and breeding of good-tasting crops, and specifically relates to the application of Wx mutant protein and its gene in plant breeding based on gene editing technology. Background technique [0002] Rice (Oryza sativa L.) is an important food crop in China and even in the world, and plays a decisive role in ensuring food security. [0003] Rice quality refers to the basic characteristics that rice must possess in commercial circulation, including four main aspects: milling quality, appearance quality, cooking and eating quality, and nutritional quality, among which appearance quality and cooking and eating quality are particularly important. Seed endosperm is the main edible part of rice. From the analysis of chemical composition, the main component of endosperm is starch, which generally accounts for 90% of the dry weight of endosp...

AI Technical Summary

Problems solved by technology

For quality traits, backcrossing is a commonly used method. Generally, 4-6 generations of backcrossing are required, and at least 3-5 generations are required for homozygous selfing. To improve a single trait and play a role in production, at least 6-8 years, the breeding cycle is long, it is difficult to meet the urgent needs of the market and the people for high-quality soft fragrant rice varieties

If we want to tap new soft rice genetic resources, we mainly rely on chemical mutagenesis at present. As we all know, the frequency of chemical mutations is low, and the initial investment is large. It may take 2-3 years to screen and obtain a stable material. At the same time, chemical mutagenesis may cause wild There are multiple gene mutations in the type material, and it needs to be improved through hybridization to eliminate bad gene mutations when it is applied to production

[0007] 2. Most soft rice japonica rice varieties have poor appearance quality

For single-base mutations, enzyme-cut target markers can be designed, but the process is relatively cumbersome. Allele-specific PCR is developed, and genotypes with high and low amylose content can be distinguished by two PCRs

[0013] At present, if scientific and technical personnel want to obtain new materials or new genes of soft rice with AC reduced to 8-12% and the appearance remains transparent, they need to undergo chemical or radiation mutagenesis, which requires a lot of work and the effect is not ideal.

Simultaneously, adopt conventional chemical mutagenesis and conventional transgenic breeding, its breeding life will be at least 6-8 years, and the breeding cycle is long, be difficult to satisfy the urgent demand of market and common people to high-quality soft fragrant rice variety

[0014] At the same time, there is no relevant report on the use of gene editing technology to perform site-directed mutations on the Wx gene of rice varieties to create new soft rice alleles

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