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Application of atcipk23 gene in improving plant tolerance to low iron

A transgenic plant, plant technology, applied in applications, plant peptides, plant products, etc., can solve problems such as poor iron absorption capacity

Active Publication Date: 2019-05-28
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to mechanism I, the roots of plants convert Fe 3+ reduced to Fe 2+ is the rate-limiting step for iron uptake, so iron uptake is poorer in mechanism I plants compared to mechanism II plants

Method used

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  • Application of atcipk23 gene in improving plant tolerance to low iron
  • Application of atcipk23 gene in improving plant tolerance to low iron
  • Application of atcipk23 gene in improving plant tolerance to low iron

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1, AtCIPK23 Gene Knockout Effects on Plant Phenotypes

[0056] Disinfect the seeds of AtCIPK23 gene mutants lks1-2, lks1-3 and wild-type Arabidopsis col-0 (WT, purchased from Arabidopsis Biological Resource Center (ABRC stocks, product catalog number is CS60000), and sow them on iron-supplying agarose for cultivation. base (Fe 3+ -EDTA at a concentration of 50 μM), vernalization in the dark at 4°C for 3 days, moved to an artificial climate chamber for 5 days, and then transferred the seedlings to low-iron medium (Fe 3+ -EDTA at a concentration of 0.5 μM) and iron-sufficient medium (Fe 3+ -EDTA at a concentration of 50 μM) for 5 days and treated with iron deficiency. There were 120 seeds for each strain, the experiment was repeated three times, and the results were averaged.

[0057] 1) Phenotype

[0058] Observe the phenotype of the plants grown in low-iron or iron-sufficient medium for 5 days, harvest the aboveground parts of the plants, and measure the abo...

Embodiment 2

[0069] Embodiment 2, AtCIPK23 gene overexpression improves plant resistance to low iron stress

[0070] 1. Construction of transgenic Arabidopsis thaliana

[0071] 1. Construction of AtCIPK23 overexpression recombinant vector

[0072] According to the coding region sequence of AtCIPK23, the 5' end primer was designed: 5'-CGTAGTTGGAATAGGTTC ATGGCTTCTC GAACAACGCC -3' (coding region underlined), 3' end primer: 5'-CCAGTATGGAGTTGGGTTC TTATGTCGA CTGTTTTGC -3' (the coding region is underlined).

[0073] Cut about 0.2g of wild-type Arabidopsis seedlings and grind them in liquid nitrogen; then add 800 μL of freshly prepared extraction buffer (0.1M pH8.0 Tris-HCl; 50mM EDTA; 0.5M NaCl; 1% SDS; 1% β-mercaptoethanol), shake vigorously to suspend it; incubate in a water bath at 65°C for 30 minutes, mix by inverting every 5 minutes; then add 250 μL of pre-cooled 5M potassium acetate, mix by inverting immediately, and place on ice for 5 minutes; add Equal amount of phenol / chlorofor...

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Abstract

The invention discloses an application of an AtCIPK23 gene in improvement of a plant capacity of tolerance to iron deficiency. The invention provides an application of any one biological material of the following 1)-4) in regulation of the plant capacity of tolerance to iron deficiency: 1) an AtCIPK23 protein; 2) an AtCIPK23 protein encoding gene; and 3) a recombinant vector containing the AtCIPK23 protein encoding gene. An experiment proves that the AtCIPK23 gene is subjected to surface expression, and transgenic arabidopsis is obtained; under the force of iron deficiency, the iron content in leaves and grains is higher than that of the wild type, the ferric reductase (FCR) activity is higher than that of the wild type, the iron absorption capacity is higher than that of the wild type, and the AtCIPK23 gene is proved to improve the plant tolerance to iron deficiency.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of AtCIPK23 gene in improving the ability of plants to tolerate low iron. Background technique [0002] Iron (Fe) is an essential mineral for humans and is of great significance to human health. my country is a country where iron deficiency and iron deficiency anemia occur more seriously in the world (Hu Rongrong, 2004). Therefore, starting from the food that humans depend on for survival, improving the iron absorption capacity of plants will provide an effective way to improve human iron nutrition. Iron is an essential trace element for plant growth and participates in various metabolic processes such as plant photosynthesis and respiration. Although the content of iron in soil is high, iron in soil often exists in the form of oxides, which cannot be absorbed and utilized by plants (Guerinot and Yi, 1994). Therefore, the content of available iron in the soil is lo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/20
CPCC07K14/415C12N15/8271
Inventor 张文浩田秋英张欣欣
Owner INST OF BOTANY CHINESE ACAD OF SCI
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