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A method for cultivating transgenic plants that increases the number of root nodules and promotes the development of root nodules

A technology of transgenic plants and cultivation methods, applied in the fields of botanical equipment and methods, chemical instruments and methods, biochemical equipment and methods, etc., can solve problems such as unreported research, and achieve the goal of increasing the number of nodules, promoting the maturation of nodules, and significantly increasing the number of nodules. The effect of practical value

Active Publication Date: 2021-08-31
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the research on the regulation of GmSGF14H and GmSGF14I in soybean nodule number and nodule development has not been reported.

Method used

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  • A method for cultivating transgenic plants that increases the number of root nodules and promotes the development of root nodules
  • A method for cultivating transgenic plants that increases the number of root nodules and promotes the development of root nodules
  • A method for cultivating transgenic plants that increases the number of root nodules and promotes the development of root nodules

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1. Nomenclature and sequence of genes.

[0042] The invention relates to coding genes of soybean 14-3-3 family proteins GmSGF14H and GmSGF14I and applications thereof. The overexpression vector constructed by the present invention can specifically overexpress the GmSGF14H and GmSGF14I Gene. The genes are located on soybean chromosomes 4 and 6 respectively, and are named as follows, GmSGF14H (gene number: Glyma.04G099900), having the coding sequence shown in SEQ ID NO: 1; GmSGF14I (Gene number: Glyma.06G101500), having the coding sequence shown in SEQ ID NO:2.

Embodiment 2

[0043] Example 2. Expression analysis of GmSGF14H and GmSGF14I genes in response to rhizobia infection.

[0044] 1) Material acquisition: The material used in the experiment was Williams 82 (W82 for short); the material was carried out according to the following process: soybean seeds were sterilized with 70% alcohol for 30 sec, and sowed in the vermiculite substrate soaked in low-nitrogen nutrient solution. They were cultured in a culture room with 16 h light / 8 h dark, light intensity 7000 LUX, temperature 26 °C, and relative humidity 70%. 15 days after sowing, inoculate with rhizobia, each plant was inoculated with 30 mL of bradyrhizobium USDA110 bacterial solution (OD600=0.08), and the soybean roots were collected on the 28th day after inoculation.

[0045] 2) Isolation of mRNA: Trizol method was used to extract soybean total RNA. ①Put the tissue into a grinder and grind it with liquid nitrogen for 3 times, then add 0.1-0.2 g of the ground tissue into a 1 mL centrifuge tube...

Embodiment 3

[0054] Example 3. Cultivation of transgenic soybeans with high ability of nodulation and nitrogen fixation.

[0055] 1) Extraction of total RNA from root tissue of Wilimas82 soybean.

[0056] The mortar has been treated at 180 ℃ 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 treated with 1 ‰ DEPC water overnight, and then sterilized at 121 ℃ for 30 minutes by high temperature and humidity.

[0057] (1) Take 100 mg of soybean root material and grind it with liquid nitrogen, add 1 mL of TRI pure reagent, and homogenize thoroughly, then suck the homogenate into a 1.5 mL centrifuge tube, and place it at room temperature for 5 min.

[0058] (2) Add 200 μl chloroform, vortex to mix, let stand for 5 minutes, 4 ℃, 12000 r / min, centrifuge for 10 minutes.

[0059] (3) Take th...

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Abstract

The invention discloses a transgenic plant breeding method for increasing the number of root nodules and promoting the development of root nodules, overexpressing the gene shown in SEQ ID NO: 1 or SEQ ID NO: 2 in the sequence list in the plant or its homologous / equivalent Genes to obtain transgenic plants with increased number of nodules and / or enhanced nodule development compared to normal plants. Using the method of the invention to construct transgenic plants can significantly increase the number of root nodules and promote the maturity of root nodules, and has great practical value in theoretical research and agricultural and forestry applications.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to genes related to plant nodulation and nitrogen fixation and applications thereof. Background technique [0002] Soybean plays an important role in the national economy. Its unique symbiotic nitrogen fixation ability can convert nitrogen in the air into ammonia nitrogen that can be absorbed and utilized by plants, which not only provides the necessary nitrogen nutrition for its own growth and development, but also helps reduce Environmental pollution caused by excessive fertilization. Making full use of the symbiotic nitrogen fixation ability mediated by soybean root nodules, mining and improving the efficiency of soybean symbiotic nitrogen fixation has become one of the important ways for the sustainable development of agriculture. [0003] Root nodules are places for symbiotic nitrogen fixation, and the number of root nodules, especially the number of root nodules w...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/82C07K14/415A01H5/00A01H6/54
CPCC07K14/415C12N15/8261
Inventor 纪洪涛李霞何晴晴
Owner HUAZHONG AGRI UNIV
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