Expression cassette and vector for recovering male fertility of rice OsCYP704B2 mutant as well as detection method and application thereof

Abstract

The invention relates to the field of plant molecular biology, in particular to an expression cassette and a vector for recovering male fertility of rice OsCYP704B2 mutant as well as a detection method and application of the expression cassette and the vector. The invention provides a plant anther promoter as shown in SEQ ID NO.1 and the rice OsCYP704B2 gene expression cassette which is driven bythe promoter and is subjected to sequence optimization. The anther promoter provided by the invention can drive genes to be efficiently expressed in plant anther; according to the rice OsCYP704B2 geneexpression cassette provided by the invention, high-level expression of the OsCYP704B2 gene in anther under the driving of the anther promoter can be realized; the rice male sterility caused by OsCYP704B2 function deletion is completely recovered, and the rice male sterility promoter has high genetic stability and can be used for recovering rice male fertility or preparing a rice male genic malesterility maintainer line and a sterile line.

Description

technical field [0001] The invention relates to the field of plant molecular biology, in particular to an expression cassette for restoring the male fertility of rice OsCYP704B2 mutants, a detection method and application thereof. Background technique [0002] Rice is an important food crop, and hybrid rice has played an important role in increasing food production. Hybrid rice is the selection of two rice parents with different genetic backgrounds and complementary traits to produce the first-generation hybrids with heterosis for rice production. Hybrid rice has obvious heterosis phenomenon, which can increase yield by 30% compared with conventional rice. Hybrid rice can be divided into three-line hybrid rice and two-line hybrid rice. Three-line hybrid rice (three-line hybrid rice) means that the production of this hybrid rice requires three rice lines: rice cytoplasmic male sterile line, rice cytoplasmic male sterile line and rice cytoplasmic male sterile restoration Ti...

AI Technical Summary

Problems solved by technology

The three-line breeding method has problems such as disease susceptibility, poor quality, and low breeding efficiency, and due to the constraints of the restoration relationship, 95% of rice resources cannot be used for hybrid breeding

Finally, the larger the transformed fragment, the lower the transformation efficiency

Therefore, Li et al. used the rice original genome sequence up to 3.8kb (excluding the flanking vector sequence) as a restoration gene fragment, which has a higher possibility of genetic instability in engineering production applications, and also increases vector construction and Difficulty of transformation; also difficult to identify the genotype of the mutation site

In summary, the recovery DNA fragments and vectors developed by Li et al. are not suitable for engineered seed breeding techniques, and there is an urgent need to develop recovery DNA fragments and vectors with higher genetic stability, easier construction and transformation, and convenient detection of mutant genotypes

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