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Application of a coding gene that increases plant iron content and increases tolerance to iron deficiency stress

A technology that encodes genes, iron content, applied in the field of bioengineering, can solve problems such as undiscovered

Active Publication Date: 2022-03-18
HEFEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] mMDH2 (AT3G15020) expresses mitochondrial malate dehydrogenase in Arabidopsis thaliana, and its currently widely reported function is to catalyze the interconversion of malate and oxaloacetate in the tricarboxylic acid cycle, which has superoxide in plant cells Different types of enzymosomal malate dehydrogenase and chloroplast malate dehydrogenase have different functions, but so far, no mMDH2 Involved in the regulation of plant iron content

Method used

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  • Application of a coding gene that increases plant iron content and increases tolerance to iron deficiency stress
  • Application of a coding gene that increases plant iron content and increases tolerance to iron deficiency stress
  • Application of a coding gene that increases plant iron content and increases tolerance to iron deficiency stress

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Experimental program
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Effect test

Embodiment 1

[0025] mmdh2 Plant acquisition and its response to iron deficiency

[0026] Compared with the use of iron-free MS medium to create stress conditions to screen the Arabidopsis seed mutant library, two mutants sensitive to iron deficiency were obtained, and the screening and identification found that the mutants were mMDH2 loss-of-function mutants, named mmdh2 . Using qRT-PCR, the mmdh2 The mutants were identified at the transcript level, and it was found that in the two mutants mMDH2 The expression levels of the genes were significantly lower than those of the wild type. Because the seed is a T-DNA insertion mutation, specific primers are used for PCR amplification and sequencing, and the sequencing results are compared in the NCBI database Blast, and the information of the T-DNA insertion site is obtained through comprehensive analysis, among which mmdh2- 1. mmdh2-2 mutant of The T-DNA insertion sites are respectively in the gene mMDH2 (AT3G15020) 168bp and 222bp...

Embodiment 2

[0029] Breeding iron-deficiency-tolerant Arabidopsis

[0030] 1,mMDH2 Gene Overexpression Transgenic Lines mMDH2-OE11, OE13 the acquisition

[0031] To further verify the function of this gene in the regulation of iron deficiency stress in plants, we constructed mMDH2 Gene overexpression vector ( 35S::mMDH2 ). Firstly, the target fragment is amplified. The wild-type Arabidopsis was normally cultured on MS medium for two weeks, the total RNA of the plant was extracted, and cDNA was synthesized by reverse transcription using a reverse transcription kit. Using the synthesized cDNA as a template, PCR was performed to amplify a sufficient amount of target product. Using the PCR product as a template, the second amplification was carried out to introduce double restriction sites XhoI and EcoRI. The PCR product and vector pBI121 were digested and recovered. Then, the recovered and purified target DNA fragment and the carrier were ligated overnight with T4 DNA ligase to form ...

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Abstract

The invention relates to the application of a coding gene for increasing the iron content of plants and increasing the tolerance to iron deficiency stress, and belongs to the technical field of bioengineering. The coding gene sequence for increasing iron content in plants and increasing tolerance to iron deficiency stress is shown in SEQ ID No: 1 in the sequence table. In the present invention, the coding gene for increasing the iron content of the plant and increasing the tolerance to iron deficiency stress is transferred into the plant, so that the coding gene for the iron content in the plant is overexpressed, so that the plant shows tolerance to iron deficiency and has Higher iron content than common plants. The coding gene for increasing the iron content of plants and increasing the tolerance to iron deficiency stress provided by the invention can provide gene resources and technical support for breeding of crops resistant to iron deficiency stress.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and specifically relates to a gene encoding for increasing iron content in plants, and at the same time, relates to using the gene to increase iron content in plants and tolerance to an iron-deficiency environment. Background technique [0002] Iron is a metallic element and one of the most commonly used metals in life. For plants, iron is also one of the essential micronutrients, directly involved in many physiological and biochemical processes. Iron in plants is the prosthetic group of various enzymes such as cytochrome oxidase, peroxidase, etc., and plays an important role in respiratory electron transfer; at the same time, iron is an essential element for the synthesis of chlorophyll. Therefore, iron deficiency can lead to yellowing and premature senescence of plant leaves, and in severe cases, it will cause a decline in crop yield and quality. Although the content of iron in most so...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82C12N15/53A01H5/00A01H6/20
CPCC12N15/8243C12N15/8271C12N9/0006C12Y101/01037
Inventor 曹树青吴席江力朱香豫陶曼芝王婷婷贾亚峰宋慧樊婷婷
Owner HEFEI UNIV OF TECH