Method adopting OsRMC protein to cultivate transgenic plant with improved iron absorption capability

A technology of transgenic plants and absorption capacity, applied in botany equipment and methods, biochemical equipment and methods, applications, etc., can solve the problems of inability to absorb and utilize plants, and achieve the effect of increasing plant yield

Inactive Publication Date: 2013-05-01
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the content of iron in the soil is high, the iron in the soil often exists in

Method used

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  • Method adopting OsRMC protein to cultivate transgenic plant with improved iron absorption capability
  • Method adopting OsRMC protein to cultivate transgenic plant with improved iron absorption capability
  • Method adopting OsRMC protein to cultivate transgenic plant with improved iron absorption capability

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Embodiment 1, construction of recombinant plasmid

[0048] 1. Construction of OsRMC gene overexpression vector

[0049] 1. Extract the total RNA of the rice variety "Zhonghua 10" and reverse transcribe it into cDNA.

[0050] 2. Using the cDNA in step 1 as a template, perform PCR amplification with a primer pair composed of F1 and R1 to obtain a PCR amplification product.

[0051] F1: 5'-CGC GGATCC ATGGCGCGGTGCACTTTG-3';

[0052] R1: 5'-CGG GGTACC CTACTCACGCAGCACCACC-3'.

[0053] 3. Digest the PCR amplified product in step 2 with restriction endonucleases BamHI and KpnI, and recover the digested product.

[0054] 4. Digest the vector pUN1301 with restriction endonucleases BamHI and KpnI to recover a vector backbone of about 13876bp.

[0055] 5. Digest the product of step 3 and the vector skeleton of step 4 to obtain the OsRMC gene overexpression vector (also known as recombinant plasmid pUN::OsRMC). According to the sequencing results, the structure of the OsR...

Embodiment 2

[0067] Embodiment 2, OsRMC gene overexpression rice or the acquisition of OsRMC gene suppressed expression rice

[0068] 1. Obtaining of OsRMC gene overexpressed rice

[0069] The OsRMC gene overexpression vector constructed in Example 1 was introduced into the rice variety "Zhonghua 10" through the method of rice genetic transformation mediated by Agrobacterium strain EHA105 (Xu et al., 2005, Plant Cell Rep. Mature embryo callus, after co-cultivation, hygromycin resistance screening, differentiation, rooting, seedling hardening, transplanting, T 0 Substitute plants.

[0070] Will T 0 T 1 Generation seeds, cultivate T 1 generation seed to get T 1 Substitute plants. Will T 0 Generation plants and T 1 The generation plants were carried out GUS staining respectively, and the method was as follows: Take 2-3 mm of the blade tip of rice seedlings grown for two weeks and use GUS staining solution (solvent is pH7. / L EDTA, 0.5mmol / L potassium ferricyanide, and 1mg / mL X-Gluc) ...

Embodiment 3

[0089] Embodiment 3, the tolerance effect of transgenic plants to low iron stress

[0090] Randomly select the T of an OsRMC gene overexpression line (OE3 line) 5 For the 1-week-old seedlings of generation plants, T 5 For the 1-week-old seedlings of the generation plants, the 1-week-old seedlings of the rice variety "Zhonghua 10" (WT) were taken, and the T 5 For the 1-week-old seedlings of the generation plant, take the T of the empty vector plant B 5 The 1-week-old seedlings of generation plants were identified as follows:

[0091] Experimental group: plants were continuously cultured in nutrient solution B for 20 days;

[0092] Control group: the plants were continuously cultured in nutrient solution A for 20 days.

[0093] After the grouping process is completed, take pictures, measure the phenotypic parameters of the plants (dry weight of the aboveground part, dry weight of the underground part and plant height), and detect the chlorophyll content of the leaves of the ...

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Abstract

The invention discloses a method adopting OsRMC protein to cultivate a transgenic plant with improved iron absorption capability, which includes the step that the transgenic plant with the iron-deficiency tolerance and/or iron absorption and/or zinc absorption higher than that of a target plant can be obtained through transferring an encoding gene of the OsRMC protein to a target plant, wherein the OsRMC protein is (a) or (b): (a), the protein consists of an amino acid sequence represented in sequence 1 in a sequence table; and (b), the protein derived from the sequence 1, obtained through performing replacement and/or deletion and/or addition of one or more amino acid residues for the amino acid sequence in the sequence 1, and related to iron-deficiency tolerance and/or iron absorption and/or zinc absorption of the plant. The method is of great value on cultivating plants with high iron-deficiency tolerance, iron absorption and zinc absorption, as well as improving the output of plants.

Description

technical field [0001] The invention relates to a method for cultivating transgenic plants with increased iron absorption capacity using OsRMC protein. Background technique [0002] Iron (Fe) is an essential trace element for plants and plays an important role in plant life activities, such as photosynthesis, respiration, and nitrogen metabolism. Although the content of iron in soil is high, iron in soil often exists in the form of oxide or hydrated oxide, which cannot be absorbed and utilized by plants. Therefore, the content of available iron in the soil is not high, and the production of crops is often affected by iron deficiency. [0003] To adapt to iron-deficient environments, plants have developed a series of strategies to increase the availability of iron in the soil, thereby increasing iron uptake. According to the different types of plants, they can be divided into Mechanism I plants and Mechanism II plants. Mechanism I plants include dicotyledonous plants and n...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N15/84A01H5/00C12N5/10C12N1/21
Inventor 张文浩杨安
Owner INST OF BOTANY CHINESE ACAD OF SCI
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