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Application of nitrate transporter and coding gene thereof in increasing number of root nodules of hairy roots of crops

A technology for transgenic plants and hairy roots, which is applied in the field of genetic engineering to achieve the effects of improving the nitrogen-fixing capacity of nodules, increasing the number of nodules of hairy roots, and improving soybean yield

Pending Publication Date: 2022-04-08
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no research and report on improving the number of root nodules of legumes and crops

Method used

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  • Application of nitrate transporter and coding gene thereof in increasing number of root nodules of hairy roots of crops

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1, gene

[0022] This example involves soybean GmNRT1.5c The gene, the nucleotide sequence of its cDNA reading frame is shown in SEQ ID NO:1. A sequence corresponding to this gene that is completely complementary to SEQ ID NO: 1, nucleotides that are substituted, deleted or increased by one or more nucleotides on the basis of the sequence shown in SEQ ID NO: 1, and have the same function sequence. Considering the degeneracy of codons, without changing the amino acid sequence, appropriately modifying the base sequence of the above cDNA reading frame is also a manifestation of the technical solution of the present invention.

Embodiment 2

[0023] Embodiment 2, protein

[0024] This example relates to the protein corresponding to the nucleotide sequence of Example 1. without affecting soybean GmNRT1.5c On the premise of protein structure and activity, soybean GmNRT1.5c Various substitutions, deletions or additions of one or more amino acids, or terminal modifications to the amino acid sequence of the protein are also a manifestation of the technical solution of the present invention.

Embodiment 3

[0025] Embodiment 3, the acquisition of gene sequence

[0026] soybean GmNRT1.5c The cDNA reading frame of the gene can be obtained by the following method: using the soybean variety Williams 82 in the laboratory as a material, extract its RNA, reverse transcribe it into cDNA, design primers based on the existing Williams 82 genome sequence to clone soybean GmNRT1.5c Gene CDS, the primers include the upstream primer shown in SEQ ID NO:2 and the downstream primer shown in SEQ ID NO:3. Use high-fidelity DNA enzymes for PCR amplification. The PCR reaction system is: cDNA template: 1 μL; front primer (F): 0.5 μL; back primer (R): 0.5 μL; buffer: 15 μL; ddH 2 O: 19 μL; dNTP: 5 μL Total reaction system 30 μL. Reaction conditions: 94 °C: 2 min, 98 °C: 20 s, 56 °C: 30 s, 68 °C: 2 min, 68 °C: 10 min, 16 °C: ∞; where, 98 °C: 20 s, 56 °C: 30 s, 68 °C: 2 min, 38 cycles in three steps. 30 μL of PCR amplification products were analyzed by 1% agarose gel electrophoresis. The band of a...

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PUM

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Abstract

The invention discloses a nitrate transporter and an application of a coding gene of the nitrate transporter in increasing the number of root nodules of hairy roots of crops, a nucleotide sequence shown in SEQ ID NO: 1 or a nucleotide sequence with the same function as SEQ ID NO: 1 in a plant body is over-expressed, and a transgenic plant with the number of root nodules on hairy roots increased compared with that of wild plants is obtained. The method has important scientific research and practical significance for improving the nodulation and nitrogen fixation capacity of the soybeans and increasing the yield of the soybeans.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to the application of a nitrate transporter and its coding gene in increasing the number of hairy root nodules of crops, thereby improving the ability of nodulation and nitrogen fixation of crops. Background technique [0002] Plants mainly obtain the nitrogen source they need from nature in three ways: the absorbed NO 3 - Restore to organic nitrogen, directly absorb ammonium or organic nitrogen in the soil, or fix N in the air through nitrogen-fixing bacteria 2 and converted into a form usable by plants. [0003] For most non-legumes, NO 3 - It is the main form of nitrogen utilization. In order to adapt to the change of nitrogen in the soil, plants have evolved two transport systems, the low-affinity transport system (LATS) and the high-affinity transport system (HATS). NRT1.5 is a low-affinity bi-transporter localized in the plasma membrane and expressed in peric...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82A01H5/06A01H6/54
Inventor 王幼宁王小迪刘超李霞
Owner HUAZHONG AGRI UNIV
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