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A wheat gene for improving plant scab resistance and its application

A technology for head blight and plants, applied in the field of biogenetic engineering, can solve the problems of less research and achieve the effects of overcoming the difficulty of distant hybridization, improving the efficiency of variety breeding, and shortening the breeding time

Active Publication Date: 2020-12-18
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The degree of degradation of the cell wall by degrading enzymes is affected by the composition and relative content of the cell wall, and plays an important role in the host-pathogen interaction, but there are still relatively few studies on this aspect in cereal crops

Method used

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  • A wheat gene for improving plant scab resistance and its application
  • A wheat gene for improving plant scab resistance and its application
  • A wheat gene for improving plant scab resistance and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 TaXAT Gene acquisition and analysis

[0038] According to the obtained EST sequence of the target gene, use Primer Premier 5.0 to design primers for 5' and 3' RACE nested amplification: 5'-Primer (its sequence is shown in SEQ ID NO.3) and 3'-Primer ( Its sequence is shown in SEQ ID NO.4);

[0039] Using the ear cDNA of Sumai No. 3 inoculated with Gibberella and treated for 8 hours as a template, the 3’ and 5’ RACE amplifications were performed using the cDNA Amplification kit:

[0040] PCR reaction system: 2×SeqAmp Buffer 25.0 μl, SeqAmp DNA Polymerase 1.0 μl, cDNA template 2.5 μl, 10×Universal Primer A Mix 5 μl, 5’ or 3’-Primer (10 μM) 1 μl, ddH 2 O 15.5 μl.

[0041]PCR reaction program: 94°C for 3 minutes; 94°C for 30 seconds, 55°C for 30 seconds, 72°C for 2 minutes, 30 cycles; 72°C for 5 minutes;

[0042] The PCR products were subjected to gel electrophoresis on 1% agarose gel, and the electrophoresis results were as follows: figure 1 as shown, fig...

Embodiment 2

[0045] Example 2 Construction of Plant Expression Vectors

[0046] Using the spike cDNA of Sumai 3 as a template, the primers TaXATFXba (shown in SEQ ID NO.5) and TaXATRSac (shown in SEQ ID NO.6) were used to amplify TaXAT complete open reading frame, and introduced restriction enzyme sites Xba I and Sac I, recovery of target fragments.

[0047] PCR reaction system: 10×Ex buffer 5 µl, MgCl2 (25mM) 4 µl, dNTP (2.5mM) 1.6 µl, TaXATFXba primer (10uM) 1 µl, TaXATRSac primer (10uM) 1 µl, ExTaq 0.25 µl, ddH2O 13.7 µl.

[0048] PCR reaction program: 94°C for 3 minutes; 94°C for 15 seconds, 55°C for 30 seconds, 72°C for 2 minutes, 30 cycles; 72°C for 5 minutes;

[0049] restriction endonuclease Xba I and Sac I digested pCAMBIA2301-CaMV35S and the target gene fragment respectively, digested at 37°C for 15 minutes, and recovered again.

[0050] Enzyme digestion system: target fragment 200ng or vector 1000ng, 10× Buffer 2μl, Xba I 1μl, Sac I 1μl,ddH 2 O supplemented to 20...

Embodiment 3

[0061] Example 3 Arabidopsis genetic transformation and gene function verification

[0062] The experiment was divided into transgenic group and control wild group:

[0063] Transgenome: Genetic transformation of Arabidopsis thaliana using flower dip method (see Clough SJ, Bent AF(1998) Floral dip: a simplified method for Agrobacterium-mediatedtransformation of Arabidopsis thaliana. Plant J 16: 735–743.), will implement The plant expression vector pCAMBIA2301-CaMV35S- obtained in Example 2 TaXAT Transferred into the Arabidopsis plant, after being screened by MS medium containing kanamycin (final concentration 50mg / L), the Arabidopsis plant that grew true leaves and was green was transplanted on the soil substrate, placed in Normal culture in the incubator for 6-8 weeks. The culture conditions are: relative humidity 80%, constant temperature 20-24 ℃, light intensity 80-200umol / M 2 / S, and the photoperiod was 8 h in the dark and 16 h in the light.

[0064] Control wild grou...

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Abstract

The invention discloses a wheat gene capable of improving the resistance to gibberellic disease of plant and applications of the wheat gene. The amino acid sequence of the wheat resistant TaXAT gene is shown as SEQ ID NO.2, and the gene belongs to glycosyl transferase family 61. The gene is subjected to functional verification by transforming arabidopsis thaliana, the result shows that the gene can improve the resistance to gibberellic disease, and through overexpression of the gene, the capacity in resisting gibberellic disease of the plant is obviously enhanced.

Description

technical field [0001] The invention belongs to the technical field of biogenetic engineering, in particular to the cloning of a wheat xylan-arabinosyltransferase gene TaXAT and its application in improving plant scab resistance. Background technique [0002] Fusarium head blight (Fusarium Head Blight) is caused by Fusarium graminearum, one of the important diseases that harm wheat, barley, oats, corn, rice, rye and other cereal crops, seriously affecting the yield and quality of my country's grain production. Fusarium scab occurs in large areas in the middle and lower reaches of the Yangtze River and Northeast my country, and the annual output will be reduced by 20-40% during the pandemic. In recent years, with changes in farming patterns and climate change, head blight has spread to other major producing areas in my country. In addition, the mycotoxins carried by diseased wheat grains affect the quality of seeds, endanger the health of humans and animals, and become one o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/54C12N9/10C12N15/82A01H5/00A01H6/20
CPCC12N9/1077C12N15/8282
Inventor 吴磊丁彬彬张旭姜朋张瑜马鸿翔
Owner JIANGSU ACAD OF AGRI SCI
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