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MicroRNA (micro ribonucleic acid) related to soybean phytophthora resistance

A technology against Phytophthora root rot and Phytophthora, applied in the field of microRNA, can solve the problem of loss of resistance of disease-resistant varieties

Inactive Publication Date: 2015-06-24
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, with the growth of the planting years of this variety, new physiological races continue to appear, and the original disease-resistant varieties will gradually lose their resistance.

Method used

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  • MicroRNA (micro ribonucleic acid) related to soybean phytophthora resistance
  • MicroRNA (micro ribonucleic acid) related to soybean phytophthora resistance
  • MicroRNA (micro ribonucleic acid) related to soybean phytophthora resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1 Prediction and screening of disease-resistant gma-miR482c-3p

[0017] (1) Screening genes related to Phytophthora root rot resistance in soybean

[0018] The expression profiling of Suinong No. 10 soybean variety infected with Phytophthora root rot physiological race No. 1 was carried out. Five samples of SCKL, SML, STL0.5, STL2 and STL4 infecting the same physiological race 1 of Phytophthora root rot were used, and total RNA was extracted as sequencing samples with the RNeasy Mini Kit of Qiagen Company. Expression profile sequencing was completed by BGI. Among them, SCKL is a sample without any treatment, SML is a sample not inoculated with Phytophthora root rot, and STL0.5, STL2, and STL4 are samples taken at 0.5h, 2h, and 4h after inoculation with Phytophthora root rot. Inoculation location and method such as Figure 4 As shown, in the expression profile sequencing data of the five samples of SCKL, SML, STL0.5, STL2, and STL4, a total of 33683 spliced ​​...

Embodiment 2

[0025] Embodiment 2 Phytophthora root rot resistance inspection

[0026] According to prediction, the target gene of gma-miR482c-3p is Glyma08g42971, which is differentially expressed during soybean infection with Phytophthora, and is related to both disease resistance and immunity in SoyBase. In this example, for gma-miR482c-3p, the gma-miR482c-3p in Suinong No. 10 soybean variety was tested for resistance to Phytophthora root rot by using the designed specific reverse transcription primer and specific PCR primer pair.

[0027] First of all, the total RNA in the sample should be extracted, and the total RNA of each sample should be reverse transcribed twice, one is to reverse transcribe gma-miR482c-3p according to the specific reverse transcription primer, and the other is to reverse transcribe gma-miR482c-3p according to the general RNA Reverse transcription primers reverse transcribe all RNA. Next, quantitative PCR was performed on the two reverse transcription products us...

Embodiment 3

[0060] Example 3 Cluster analysis of miRNA-target gene relationship

[0061] (1) Use the clustalx software to perform a pairwise comparison of the miRNAs and genes screened and predicted in Example 1, and present the pairwise comparison results using mega to obtain a guide tree. And the miRNAs and target genes were classified according to the distance between the two genes presented in the guide tree.

[0062] (2) Due to the short miRNA sequence and high family similarity, the miRNAs were divided into family MIRNA forms, and then the miRNA families and target genes were linked.

[0063] (3) Classify and organize the relationship between miRNA and target gene in the connection, as shown in Table 4, it is found that among the predicted miRNA-target gene relationships, most of the miRNAs have a tendency to regulate their target genes, according to With this tendency (Table 3), the miRNA-target gene relationship can be divided into 8 categories a1b1, a1b2, a1b3, a21b3, a22b1, a22...

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Abstract

The invention discloses a microRNA (micro ribonucleic acid) related to soybean phytophthora resistance and belongs to the field of molecular biology of soybeans. 223 phytophthora-resistant genes and 256 miRNAs are obtained by means of screening known relevant disease-resistant immune genes. A phytophthora-resistance-related miRNA-Target predicting system is established by means of expression profile sequencing, rigorous bioinformatics analysis and real-time quantitative PCR (polymerase chain reaction) detection and can predict functions of the miRNAs, so that screening and identifying efficiency for soybean phytophthora resisting genes and the miRNAs is improved greatly. A MIRNA-target relation classification method is provided, that a Plant-pathogen interaction path plays a key role in phytophthora resisting is discovered, and that a gma-miR482c-3p serves as a main ingredient of the Plant-pathogen interaction path and is closely related to the soybean phytophthora resistance is verified.

Description

technical field [0001] The invention relates to a microRNA related to soybean phytophthora resistance and belongs to the field of soybean molecular biology. Background technique [0002] Soybean is an important crop, rich in protein and fat and active ingredients beneficial to the human body such as soap II, isoflavones, phospholipids, etc. It is one of the main sources of vegetable protein and oil in the world. At present, my country's soybean planting area, total output and per unit yield rank fourth in the world, and its production level lags behind the world's soybean production. In 2012, China's soybean planting area was 7.18 million hectares, with a total output of 12.8 million tons, while imported soybeans reached a record 57.5 million tons, accounting for four-fifths of my country's soybean market. There are many reasons why my country's soybean production lags behind, the supply is seriously insufficient, and the domestic soybean industry is severely impacted by im...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/31C12N15/10A01N57/16A01P3/00
Inventor 朱荣胜陈庆山辛大伟刘春燕胡振帮蒋洪蔚齐照明韩雪王志豪
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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