Application of disease-resistant gene Glyma.04G016500.1 of soybean

A technology of disease resistance gene and soybean mosaic virus is applied in the field of genetic engineering to achieve the effects of improving disease resistance, simple synthesis method and high synthesis efficiency

Active Publication Date: 2020-10-27
ZHEJIANG NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This expansion suggests that the E3 ligase may play a role...

Method used

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  • Application of disease-resistant gene Glyma.04G016500.1 of soybean
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  • Application of disease-resistant gene Glyma.04G016500.1 of soybean

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1, Glyma.04G016500.1 disease resistance gene sequence synthesis

[0067] 1. Extraction of total RNA, using the Trizol method to extract total RNA:

[0068] (1) Get the leaves of soybean seedling plants, cut the leaves of the plants into pieces, remove the petioles and thicker veins, put them into a mortar pre-cooled in liquid nitrogen, add liquid nitrogen to the mortar and grind to powder, and The powder was transferred to a 1.5 mL centrifuge tube cooled in liquid nitrogen.

[0069] (2) Add 1mL Trizol to a 1.5mL centrifuge tube, and vortex the centrifuge tube. After shaking for a while, open the cap and deflate the centrifuge tube to prevent the air pressure in the centrifuge tube from increasing and causing the liquid to splash, thereby obtaining a grinding liquid .

[0070] (3) Put the obtained grinding solution into a new centrifuge tube, and put the centrifuge tube filled with the grinding solution on a centrifuge for centrifugation. The temperature of the...

Embodiment 2

[0107] Example 2, construction of BPMV-Glyma.04G016500.1 vector:

[0108] 1. Enzyme digestion system of vector BPMV-RNA1

[0109]

[0110] Place the centrifuge tube in a 37°C water bath for enzyme digestion for 3 hours, and then perform purification, that is, add 3 volumes of Buffer PCR-A to the PCR, enzyme digestion, enzyme labeling, or sequencing reaction solution (if Buffer PCR-A is insufficient 100 μL, add to 100 μL); after mixing, transfer to a DNA preparation tube, place the DNA preparation tube in a 2 mL centrifuge tube, centrifuge at 12,000 rpm for 1 min, and discard the filtrate. Put the preparation tube back into the 2mL centrifuge tube, add 0.7mL Buffer W2, centrifuge at 12000rpm for 1min, and discard the filtrate. Place the preparation tube in a clean 1.5mL centrifuge tube, add 20 μL ddH to the center of the DNA preparation membrane 2 O, stand at room temperature for 1 min. Centrifuge at 12000rpm for 1min to elute DNA.

[0111] 2. Enzyme digestion system for...

Embodiment 3

[0135] Embodiment 3, gene bombardment creates soybean silent plant

[0136] 1. Preparation of microprojectiles

[0137] (1) Weigh 3 mg of gold powder (0.6 μm in diameter) and put it into a 1.5 mL centrifuge tube;

[0138] (2) Add 50 μL of absolute ethanol to the centrifuge tube, and vortex for 2-3 minutes;

[0139] (3) Place the vortexed centrifuge tube in an ice bath for 5 minutes and allow the gold powder to settle, centrifuge at 10,000 rpm for 30 seconds, and discard the supernatant;

[0140] (4) Repeat (2), (3) steps 2 times;

[0141] (5) Add 50 μL sterile water to the centrifuge tube, vortex for 2-3 minutes, and fully resuspend;

[0142] (6) Place the centrifuge tube in an ice bath for 5 minutes, centrifuge at 10,000 rpm for 30 seconds, and discard the supernatant;

[0143] (7) Repeat (5), (6) steps 2 times;

[0144] (8) Add 50 μL of sterile water to the centrifuge tube, vortex for 1 min, and store at -20°C.

[0145] 2. DNA embedding

[0146] (1) Put the microproje...

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PUM

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Abstract

The invention relates to the technical field of gene engineering, and discloses an application of a disease-resistant gene Glyma.04G016500.1 of soybean. The gene is used for improving the disease resistance of the soybean. A sequence of cDNA nucleotide of the disease-resistant gene Glyma.04G016500.1 of the soybean is as shown in SEQ ID NO: 1. The invention also provides a method for preparing a silencing plant GmSAUL1 at the same time, and the gene Glyma.04G016500.1 of the soybean is silenced.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to soybean Glyma.04G016500.1 disease-resistant gene, its encoded protein GmSAUL1 and its synthesis method. Background technique [0002] Soybean is a very important crop that can provide protein for humans. However, soybeans are very vulnerable to biotic and abiotic stresses during the growth process, such as soybean spot fungus, soybean spot fungus, soybean root rot, soybean downy mildew, and soybean mosaic virus. These diseases will affect the yield of soybeans, so Research on improving the disease resistance of soybean is very necessary. [0003] E3 ligases are a large family. There are less than 700 E3 coding genes in the human genome. Compared with the human genome, higher plants have greatly expanded the E3 gene family. In the Arabidopsis genome, for example, there are more than 1400 E3-encoding genes in Arabidopsis. There are more than 1300 E3 coding genes in ...

Claims

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Application Information

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IPC IPC(8): C12N15/52C12N15/82A01H5/00A01H6/54
CPCC12N9/93C12N15/8218C12N15/8283C12N15/8281C12Y603/02019Y02A40/146
Inventor 刘建中李俊梅钟晨丽
Owner ZHEJIANG NORMAL UNIVERSITY
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