Mutation site of opaque2 gene in Zheng 58/opaque2 near-isogenic line and its application

Abstract

The present invention discloses the opaque2 gene mutation site in Zheng 58 / opaque2 near-isogenic line and its high-efficiency molecular marker and application. The sequencing analysis of this application found that 10 bases were missing at 713bp after the ATG of the o2 gene in Zheng 58 / o2, and ORF prediction O2 protein translation is prematurely terminated. The intragenic co-dominant molecular marker o2-indel-1 was developed for the mutation deletion site, and the marker was used for backcross breeding. The results showed that o2-indel-1 was completely linked to the o2 mutant phenotype, and the misselection rate was 0. Therefore, the analysis of the mutation site of opaque2 gene is helpful for the development of high-efficiency molecular markers and improves the selection efficiency of high-quality protein maize breeding.

Description

technical field [0001] The invention belongs to the field of agricultural biological breeding, and in particular relates to an opaque2 gene mutation site in a Zheng 58 / opaque2 near-isogenic line and its high-efficiency molecular marker and application. Background technique [0002] Corn (Zea mays L.) is the world's largest food crop, and it is also an important feed and economic dual-purpose crop. The protein content of corn kernels is generally about 10%. From a nutritional point of view, the protein quality of ordinary corn is relatively poor. The content of lysine in common corn is only about 0.23%, while the feed lysine content usually required by livestock and poultry is 0.6%-0.8%. At present, the insufficient part needs to be supplemented by adding more synthetic lysine, bean cake and fish meal, etc. Supplement, which greatly increases the cost of feed. [0003] In 1964, Mertz et al. first discovered that the endosperm lysine content of maize opaque2 mutant was 70% h...

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Problems solved by technology

Chen et al. (2014) found that CA339 is a mutant in which a mutation in the promoter region leads to no expression of the o2 gene. CA339 was used as the o2 donor material, Liao 2345 was used as the acceptor material, and the molecular marker phi057 in the CA339o2 gene was used for backcrossing. During the process, it was found that the grain phenotype and gliadin expression of Liao 2345 / o2-1 returned to normal phenotype, and it was found that the CA339o2 gene CDS was introduced, but the CA339 mutant promoter sequence was not introduced into Liao 2345, resulting in the The o2 gene is normally expressed and functions under the drive of the Liao 2345 promoter

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