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OsNramp5 mutant as well as screening method and application thereof

A technology of mutants and expression vectors, applied in the field of OsNramp5 mutants and their screening, can solve problems such as intellectual property infringement, gene editing product transgenic safety management obstacles, etc., to improve efficiency and accuracy, reduce cadmium content, and accelerate mutation rate Effect

Active Publication Date: 2022-02-08
湖南省核农学与航天育种研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The industrialization of gene editing products has the policy obstacle of "transgenic safety management", and there may be a risk of infringement of the original intellectual property rights of gene editing technology from abroad

Method used

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  • OsNramp5 mutant as well as screening method and application thereof
  • OsNramp5 mutant as well as screening method and application thereof
  • OsNramp5 mutant as well as screening method and application thereof

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Effect test

Embodiment 1

[0059] In this embodiment, according to the agronomic traits of the breeding target, the wild-type rice OsNramp5 gene was found in the Gene bank database, and the Oligo6.0 software was used to design probes for this gene, wherein the nucleotide sequence of the wild-type rice OsNramp5 gene included: The sequence shown in SEQ ID NO:3. Commissioned Epson Biotechnology Co., Ltd. to synthesize biotin-labeled probes.

[0060] Probes are designed according to the wild-type rice OsNramp5 gene, the probes include Os07t0257200-1 to Os07t0257200-20, and the sequences of the probes include sequences shown in SEQ ID NO.4 to SEQ ID NO.23.

[0061] Os07t0257200-1

[0062] GTCACTACCACCATTCTCTTCTTCGTCTACTTCCAGCTAGCCTGAGCTCTAGCTTAGCTCTAGCTTAGCCTGAAGAAGCTAAGAGAGGAAGCAACAATGGAGATTGAGAGAGAGAGCAGT (SEQ ID NO. 4)

[0063] Os07t0257200-2

[0064] GAGAGAGGGAGCATCAGCTGGAGAGCTAGTGCGGCACATGATCAAGATGCCAAGAAGCTCGACGCAGATGATCAGCTGCTAATGAAGGTCAGTTGCTACCTTAATTATCACAAAGTTGTC (SEQ ID NO. 5)

[0065] Os07t02...

Embodiment 2

[0103] In this embodiment, the screening of OsNramp5 mutants includes the following:

[0104] 1. Carry out radiation mutagenesis treatment to plants, including as follows:

[0105] 1) 2000 dry rice seeds of model XF1822, using heavy ions 12 C +6 Radiation mutagenesis treatment with a dose of 120Gy and a dose rate of 1.5Gy / Min was planted to obtain M1 generation seeds.

[0106] 2) Plant the seedling-raised plants of the M1 generation in individual plants, strictly self-fertilize the M1 generation, harvest the seeds of each ear of each individual plant according to the ear harvesting method, and obtain the M2 generation seeds.

[0107] 3) Plant the plants of the seed seedlings of the M2 generation in single plants, plant 14000 plants, 100 plants in each group, and number each group; take each sample group as a unit, when the seedlings grow to two leaves and one heart, the Among them, each individual plant uses a hole puncher with a diameter of 6 mm to take an equal amount of ...

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Abstract

The invention relates to the field of plant biotechnology breeding, in particular to an OsNramp5 mutant as well as a screening method and application thereof. The invention discloses an OsNramp5 mutant, and a nucleotide sequence for coding the OsNramp5 mutant comprises a sequence as shown in SEQ ID NO. 1 or SEQ ID NO. 2. The mutation rate of the plant genome is increased by adopting radiation mutagenesis, the mutation of the OsNramp5 allele is detected from the radiation mutagenesis population by applying a high-throughput molecular screening technology in the seedling stage of the plant M2 generation after mutagenesis, and the screening efficiency and accuracy of the OsNramp5 mutant are improved. The OsNramp5 mutant is screened out, and compared with a wild plant, the cadmium content in seeds of the plant containing the OsNramp5 mutant is reduced by 90% or above.

Description

technical field [0001] The invention relates to the field of plant breeding, in particular to the OsNramp5 mutant and its screening method and application. Background technique [0002] Crop radiation mutation breeding plays an important role in ensuring world food security and increasing nutrient supply. According to the latest statistics of the United Nations FAO / IAEA Mutant Variety Database, by the end of 2019, more than 60 countries had bred 3,299 plant mutant varieties on 214 plant species and passed commercial registration. The number of new varieties bred indirectly is 1,050, accounting for nearly one-third of the number of mutant varieties bred in the world. As a traditional biological breeding technology, nuclear radiation mutation breeding technology has produced huge social and economic effects. Under long-term natural environmental conditions, the organism itself interacts with the external environment. In order to adapt to the change of the environment, the gen...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/10A01H6/20A01H6/46A01H6/82A01H6/54A01H6/74
CPCC07K14/415C12N15/8271
Inventor 杨震彭选明张莉张渊海张勇张逸妍柏连阳余应弘
Owner 湖南省核农学与航天育种研究所
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