In rice production, the prevention and control of gramineous weeds is a difficult point, especially mimic gramineous weeds such as barnyardgrass, stephanotis, and weedy rice are difficult to control, which seriously affects the yield and quality of rice. Although pendimethalin, a dinitroaniline herbicide, is ideal for controlling gramineous weeds, it is easy to cause rice phytotoxicity. The emerging plant single-base editing technology provides a brand-new breeding idea and way to solve the problem of weed control.

Recently, the Crop Pest Functional Genomics Research Innovation Team of the Institute of Plant Protection of the Chinese Academy of Agricultural Sciences published an online research paper entitled "Developing a novel artificial rice germplasm for dinitroaniline herbicide resistance bybase editing of OsTubA2" in the "Plant Biotechnology Journal". Base editing technology to create new rice germplasms resistant to dinitroaniline herbicides pendimethalin and trifluralin, and found that the target site improvement program is also applicable to other rice varieties and other major crops resistant to such herbicides Rapid improvement of traits.

The researchers used the previously developed adenine base editor rBE14 tool (F Yan et al, Mol Plant, 2018) to perform site-directed nucleotide editing on the rice α-tubulin gene OsTubA2, and obtained the 268th amino acid residue. The base Met is mutated into a Thr mutant.

The mutant can grow normally on a high-dose drug-loaded medium and produce fertile offspring. Its tolerance to the target herbicide pendimethalin reaches 3 times the recommended dose, and it also has a good tolerance to trifluralin However, wild-type rice cannot survive on the same or even lower dose of drug-loaded medium, indicating that the Met268Thr mutant has higher resistance to the target herbicide. Further studies have shown that the mutation can be stably inherited in the offspring and will not affect rice yield and other agronomic traits. Bioinformatics analysis revealed that the Met268 locus of the OsTubA2 gene is highly conserved in more than 4000 rice germplasm in nature and major crops such as corn, wheat and sorghum.

Therefore, the scheme used in this study is also suitable for precise and rapid molecular breeding of other rice varieties and other crops that are resistant to dinitroaniline herbicides. This research is an example of the application of single-base editing technology developed in recent years to the creation of herbicide-resistant rice breeding materials and rapid improvement of traits. It accelerates the process of traditional crop breeding and saves a lot of manpower and material resources. The development of herbicide resistance traits and the mining and application of other important genes are useful for reference.

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