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microRNA for promoting plant nodulation and nitrogen fixation and application thereof

A plant and nitrogen fixation technology, applied in the field of small RNA, can solve problems that have not been seen yet

Inactive Publication Date: 2014-04-02
中国科学院遗传与发育生物学研究所农业资源研究中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there are no reports about the regulation of miR167 on legume root nodule development

Method used

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  • microRNA for promoting plant nodulation and nitrogen fixation and application thereof
  • microRNA for promoting plant nodulation and nitrogen fixation and application thereof
  • microRNA for promoting plant nodulation and nitrogen fixation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1, Analysis of the expression pattern of gma-miR167c

[0022] 1. Real-time fluorescent quantitative PCR of gma-miR167c: The expression characteristics of gma-miR167c in roots at different stages of root nodule development after rhizobia infection were analyzed by real-time fluorescent quantitative PCR technology;

[0023] (1) Material acquisition: The material used in the experiment is Wilimas82 (Williams 82, hereinafter referred to as W82), and the material is processed according to the following process: Soybean seeds are sterilized with 70% sprinkler for 30 seconds, sown in vermiculite pearls soaked in low-nitrogen nutrient solution Rock (3:1) mixed matrix, cultured in the culture room, 16h light / 8h dark, light intensity 7000LUX, temperature 26°C, relative humidity 70%. Seven days after sowing, each plant was inoculated with 30ml of bradyrhizobium USDA110 bacterial solution (OD600:0.08), and the roots were taken at 0, 1, 3, 5, and 10 days after inoculation; ...

Embodiment 2

[0041] Example 2, the method for cultivating soybeans with high nitrogen fixation capacity based on the function of gma-miR167c

[0042] 1. Extraction of total RNA from functional leaf tissue of Wilimas82 soybean

[0043] The mortar has been treated at 180°C for 8 hours or burned to eliminate RNase contamination; reagents such as chloroform, isopropanol, and ethanol should be freshly opened and uncontaminated; other equipment such as pipette tips, centrifuge tubes and reagents such as ultrapure water , NaAc, were all treated with 1‰ DEPC water overnight, then sterilized at 121°C for 30 minutes, and the pipette tip and centrifuge tube were dried at 65°C for later use; Trizol method was used to extract soybean total RNA:

[0044] (1) Take 50mg of material (leaf) and grind it with liquid nitrogen, add 1ml of redzol reagent, after fully homogenizing, inhale the homogenate into a 1.5ml centrifuge tube, and place it at room temperature for 5 minutes;

[0045] (2) Add 200μl chlorofo...

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Abstract

The invention discloses microRNA for promoting plant nodulation and nitrogen fixation and an application thereof. A nucleotide sequence of the microRNA is shown as SEQ ID NO:1; a precursor sequence of the microRNA is shown as SEQ ID NO:3; coding genes of the precursor sequence of the microRNA are shown as SEQ ID NO:4. The application method comprises the following steps: constructing an overexpression recombinant vector containing the genes shown as SEQ ID NO:3, constructing a transformant by utilizing the recombinant expression vector, infecting a target plant by utilizing the transformant, and screening to obtain a transgenic plant which has higher nitrogen fixation capacity compared with a normal plant. The microRNA has high significance for improving the soybean yield, and novel gene resources are provided for cultivating novel soybean types with high nitrogen fixation capacity.

Description

technical field [0001] The invention relates to a small RNA related to root nodule development in plants, in particular to microRNA167c derived from soybean and related to root nodule development and its application in cultivating new soybean varieties with high nitrogen fixation capacity. Background technique [0002] Soybean is a very important crop with special economic value. The nodules on the roots of soybeans can convert nitrogen in the air into ammonia nitrogen that plants can absorb and use, thus providing the necessary nitrogen nutrients for the growth and development of soybeans. The symbiotic nitrogen fixation mediated by root nodules not only affects the normal growth and yield of soybean, but also helps to save energy and reduce environmental chemical pollution. At present, improving nitrogen fixation efficiency has become one of the important ways to increase soybean yield and ensure sustainable agricultural development. [0003] MicroRNA (miRNA) is a class ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/11C12N15/82A01H5/00
Inventor 李霞王幼宁陈亮李科学
Owner 中国科学院遗传与发育生物学研究所农业资源研究中心
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