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Phytophthora root rot resistance gene and application

A technology for Phytophthora sojae and root rot, applied in the directions of application, genetic engineering, plant genetic improvement, etc., can solve the problems of no gene of Phytophthora sojae root rot found, no soybean, and low efficiency of variety screening.

Active Publication Date: 2016-05-25
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, although the researchers have sequenced the entire soybean genome, they have not yet found a gene in the soybean genome that is capable of resisting Phytophthora sojae root rot
Without the key gene of soybean Phytophthora root rot resistance, it is very unfavorable for the selection of disease-resistant soybean varieties in the agricultural breeding process, and the efficiency of variety selection is very low

Method used

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  • Phytophthora root rot resistance gene and application
  • Phytophthora root rot resistance gene and application
  • Phytophthora root rot resistance gene and application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] 1 Reagents used in the experiment

[0036] 1) Reagents for extracting soybean RNA

[0037] TRIzolReagent was purchased from Invitrogen Company; M-MLV reverse transcriptase, RNase-freeDNaseI, RNasin, Taq enzyme, dNTP, Oligo (dT), and DTT were purchased from TaKaRa Company; DEPC was purchased from Sigma Company; agarose (Agarose) for electrophoresis The reagents were imported from Spain; QuantiFastSYBRGreenPCRKit was purchased from QIAGEN.

[0038] 2 Genomic RNA extraction method

[0039] Preparation:

[0040] (1) DEPC (0.1%) water preparation: 1mLDEPC was added to 1000mL deionized water, and magnetically stirred for 24h.

[0041] (2) Put the mortar, tweezers, beaker, measuring cylinder, glass rod, etc. into the oven, and dry heat sterilization at 180°C for more than 6 hours.

[0042] (3) Put the centrifuge tube, pipette tip, etc. into the reagent bottle that has been sterilized by dry heat, fully soak it with the prepared DEPC (0.1%) water, seal the bottle mouth with...

Embodiment 2

[0080] Real-time quantitative analysis was performed on the control and treated RNA extracted from the different tissues of the resistant variety Suinong 10 and the susceptible variety Hefeng 25 at different times. The expression levels of all changed (results Such as Figure 1-3 shown). Among the resistant varieties: the reaction time of the stem is the longest, and the reaction of the leaf is the most sensitive; among the susceptible varieties: the reaction of the stem is the most sensitive. In the middle leaves of the disease-resistant variety Suinong 10, the expression peak was reached at 6 hours, and the expression level was 12 times, while in the susceptible variety Hefeng 25, there was only a slight up-regulation at 12 hours, and the expression level was not high, about 3 times, Such as figure 1 ; In the resistant variety Suinong 10, the roots and stems responded slowly, reaching the peak at 12 hours, and the expression levels reached 14 times and 18 times respect...

Embodiment 3

[0082] Real-time quantitative analysis was performed on the control and treated RNA extracted from the different tissues of the resistant variety Jiusan H05-23 and the susceptible variety Baojiao 06-6046 at different times, and it was found that the GmDRR gene in the resistant variety soybean GmDRR gene in each tissue from 0h to 48h expression levels have changed. In the disease-resistant variety Jiusan H05-23, the response of leaves is the most sensitive, reaching the peak expression at 6 hours, and the expression level is 8 times. The reaction of roots and stems is slower than that of leaves, reaching the peak at 12 hours, and the expression level is 11 times and 7 times, the change trend was consistent with the change trend of the gene in Suinong 10 after being inoculated with Phytophthora soybean No. The amount is 14 times and 4 times respectively ( Figure 4 with Figure 5 ), the expression level in the roots was relatively low, which was slightly different from the...

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Abstract

The invention discloses a phytophthora root rot resistance gene and application, and belongs to the technical field of soybean genetic breeding. The phytophthora root rot resistance gene is the GmDRR gene of soybeans. The nucleotide sequence of the gene is shown in SEQ ID NO.1. The gene can be used for genetic breeding of plants, especially soybeans. Meanwhile, the invention further provides an application method of the gene, and particularly provides a method for identifying or screening varieties resistant to phytophthora root rot through the gene. By detecting the expression amount of the gene, the resistance to phytophthora root rot can be predicated, and the selection efficiency of phytophthora root rot resistance is greatly improved.

Description

technical field [0001] The invention relates to a soybean Phytophthora root rot resistance gene and its application, belonging to the technical field of soybean genetic breeding. Background technique [0002] Soybean Phytophthora root rot is caused by Phytophthorasojae (Phytophthorasojae), one of the world-wide diseases that seriously endanger soybean production. Its damage area is very large and it is a very serious soil-borne disease. So far, its damage momentum in some major soybean producing areas is unstoppable. Therefore, the mining of new Phytophthora root rot resistance genes is very important. Finding soybean inland genes for soybean Phytophthora root rot resistance, and comparing the differences of resistant soybean resources at the gene level, has important theoretical and practical implications for understanding the mechanism of soybean resistance to Phytophthora root rot at the molecular level and for cultivating disease-resistant varieties. significance. [...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12Q1/68
Inventor 陈庆山辛大伟朱荣胜胡振帮蒋洪蔚齐照明韩雪杨蕊
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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