Opaque2 gene mutation site in Zheng 58/opaque2 near-isogenic line and application thereof

A technology of near-isogenic lines and loci, applied in the field of agricultural biological breeding

Active Publication Date: 2018-09-21
HENAN ACAD OF AGRI SCI
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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

Method used

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  • Opaque2 gene mutation site in Zheng 58/opaque2 near-isogenic line and application thereof
  • Opaque2 gene mutation site in Zheng 58/opaque2 near-isogenic line and application thereof
  • Opaque2 gene mutation site in Zheng 58/opaque2 near-isogenic line and application thereof

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Embodiment Construction

[0031] 1 Materials and methods

[0032] 1.1 Test material

[0033] The Zheng 58 / o2 near-isogenic line and the control material Zheng 58 were from the Institute of Food Crops, Henan Academy of Agricultural Sciences. Zheng 58 / o2 near-isogenic line is the self-bred offspring of Ye 478 / o2 and Zheng 58 backcrossed for six generations.

[0034] 1.2 Detection of Amino Acids in Corn Grain

[0035] 1. Sample preparation: Take 3 ears for each sample, and take the middle 1 / 3 of the kernels, mix them thoroughly, and grind them through a 60-mesh sieve. Take 2g-10g of the sieved sample, put it into a flat weighing bottle (a small petri dish can be used), put the bottle cap on the edge of the bottle, place it in an oven at 101-105°C for 2-4 hours, take it out and cover it. Cool in a desiccator for 0.5 hour and weigh. After weighing, reheat for 1 hour, cool for 0.5 hour, and weigh. Repeat until the weighing is within 2 mg before and after, and calculate the water content of the sample. ...

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Abstract

The invention discloses an opaque2 gene mutation site in a Zheng 58/opaque2 near-isogenic line and a high-performance molecular marker and application thereof. By virtue of sequencing analysis, ten basic groups are omitted at 713bp site after ATG of o2 gene in Zheng 58/o2, and ORF predicts that O2 protein translation is terminated in advance. A co-dominance molecule marker o2-indel-1 in the gene is developed for the mutation omission site, the marker is used for backcross transformation, the result shows that o2-indel-1 and o2 mutant phenotype are completely linked, and an error rate is 0. Therefore, the parsing of the opaque2 gene mutation site facilitates the development of the high-performance molecular marker, and can improve the high-quality protein corn breeding selection efficiency.

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...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/11
CPCC07K14/415C12Q1/6895C12Q2600/156
Inventor 韩小花铁双贵许蒙蒙岳润清郭书磊夏彦陈娜娜刘璐池海锋傅晓雷
Owner HENAN ACAD OF AGRI SCI
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