Soybean mosaic virus resistant gene GmNN1 and application of functional markers of soybean mosaic virus resistant gene GmNN1
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A leaf virus and soybean technology, applied in the application field of soybean anti-mosaic virus gene GmNN1 and its functional markers, can solve the problem that there is no soybean anti-mosaic virus functional markers
Active Publication Date: 2015-12-16
HEBEI AGRICULTURAL UNIV.
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Embodiment 1
[0058] Example 1: Identification of Mosaic Virus Resistance in Different Soybean Varieties
[0059] Select neat and uniform seeds of soybean varieties and sow them in the insect-proof net room. After the first pair of true leaves unfold, select seedlings with consistent growth and inoculation with soybean mosaic virus SMV3, and use plants that have not been inoculated with the virus as controls. Subsequently, the disease incidence of the varieties was observed at 1, 2, 3, and 4 weeks after inoculation, and the mosaic virus resistance of the varieties was identified. The identification results showed that Jidou 12 and Wuxing 2 were resistant varieties, and Ji NF58 and Nannong 1138-2 were susceptible varieties.
Embodiment 2
[0060] Embodiment 2: Soybean leaf total RNA is extracted
[0061] Total RNA was extracted from the leaves of soybean varieties Jidou 12, Wuxing 2, Ji NF58 and Nannong 1138-2, and the operation steps were referred to the operation guide of the TRNzolTotalRNAReagent kit.
Embodiment 3
[0062] Embodiment 3: synthetic cDNA method
[0063] Ⅰ. cDNA synthesis for gene cloning
[0064] The cDNA synthesis was carried out according to the operation guide of PrimeScriptTM1stStrandcDNASynthesisKit, and the reaction procedure was as follows:
[0065] Prepare the following reaction solution in a 0.2mL centrifuge tube:
[0066]
[0067] Gently pipette the pipette tip to mix, incubate at 65°C for 5min, and place on ice to cool.
[0068] Continue to add the following reactants to the above tube:
[0069]
[0070] Mix gently, store at 42°C for 60 minutes, 90°C for 5 minutes, and store at 4°C. After the reaction, 5 μL was taken for 2% agarose gel electrophoresis detection.
[0071] Ⅱ. cDNA Synthesis for Fluorescence Quantitative PCR
[0072] Follow the instructions of PrimeScriptRTreagentKitWithgDNAEraser, the reaction procedure is as follows:
[0073] A. Removal of Genomic DNA
[0074]
[0075] Incubate at 42°C for 2 minutes and store at 4°C.
[0076] B. Sy...
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Abstract
The invention provides a soybean mosaic virus resistant gene GmNN1 and a susceptible allelic gene thereof. Nucleotide sequences of the soybean mosaic virus resistant gene GmNN1 and the allelic gene are respectively shown as SEQ ID No.1 and SEQ ID No.2. The soybean mosaic virus resistant gene GmNN1 and the allelic gene have the advantages that difference of expression of the GmNN1 among different soybean resistant and susceptible varieties is analyzed, participation of the GmNN1 to SMV (soybean mosaic virus) resistant reaction is clarified, difference between GmNN1 gene sequences of the different soybean SMV resistant and susceptible varieties is studied, functional markers GmNN1-AC/GmNN1-GT are developed according to the difference between the sequences, association between the markers and the disease resistance of soybean is verified by the aid of SMV resistant and susceptible RIL (recombinant inbred line) populations, and the different SMV resistant and susceptible soybean varieties can be differentiated by the aid of the markers; functional genes and selection markers can be provided for SMV resistant soybean breeding by study results. The invention further provides a method for identifying the functional markers of the soybean mosaic virus resistant gene GmNN1 and application of the soybean mosaic virus resistant gene GmNN1 to breeding selection.
Description
technical field [0001] The invention belongs to the field of plant molecular biology, and in particular relates to the application of a soybean anti-mosaic virus gene GmNN1 and a functional marker thereof. Background technique [0002] Soybean is an important grain and oil crop in my country. It contains a variety of physiologically active substances and has a wide range of industrial uses. It occupies an irreplaceable important position in the national economy. Soybean is also the main source of plant protein in the human food structure, and it is an important health food and high-quality protein feed in the market today. In recent years, the world's demand for soybeans has been increasing year by year, while two-thirds of my country's soybean consumption depends on imports. Therefore, improving soybean yield has become an important task and research direction of soybean breeding in my country and even in the world. [0003] Soybean mosaic virus (SMV) is a global soybean ...
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