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Application of gene BEC1019 in improvement of full rot resistance of wheat

A BEC1019, wheat total eclipse technology, applied in the field of agricultural biology, can solve problems such as inability to effectively control a large number of diseases

Active Publication Date: 2019-12-31
ZHOUKOU NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, due to the rapid emergence of toxic strains and fungicide-resistant strains, traditional breeding has been unable to effectively control a large number of crop diseases, and breeding disease-resistant varieties has become the main way to improve plant disease resistance.

Method used

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  • Application of gene BEC1019 in improvement of full rot resistance of wheat
  • Application of gene BEC1019 in improvement of full rot resistance of wheat
  • Application of gene BEC1019 in improvement of full rot resistance of wheat

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Clone BEC1019 Gene Homologous Sequence

[0031] The roots and stem bases of wheat var. graminearum infested for 7 days were taken, ground thoroughly with liquid nitrogen, and the DNA of the pathogen was extracted in a pre-cooled mortar, and primers were designed to amplify BEC1019 of the wheat var. Source gene, PCR reaction system (50 μL): 2×Easy pfu Mix 25 μL, fungal DNA template 2 μL, primers 1 μL, ddH2O 21 μL, PCR reaction conditions: pre-denaturation at 95°C for 5 minutes; denaturation at 95°C for 30 seconds, annealing at 58°C for 30 seconds, 72°C Extend at ℃ for 30s, 35 cycles; extend at 72℃ for another 5min, store at 4℃ for later use.

[0032] The primer sequences are as follows:

[0033] BEC1019-F: 5'-ATGCAGTCTGTATTGCTTTT-3',

[0034] BEC1019-R: 5'-CTAGACACAATGAACCTCGC-3'.

[0035] After the fragments were detected by electrophoresis and purified, 4 μL of PCR products were taken respectively, added to 1 μL of pEASY-Blunt cloning vector, and reacted a...

Embodiment 2

[0036] Example 2 Obtaining BEC1019 Overexpression and Silenced Transgenic Plants

[0037] Using Gateway technology, first construct the full-length and 264bp fragments (368-632bp) of the BEC1019 gene into the intermediate vector pDONR207, and the primer sequences are as follows:

[0038] F: 5'-GGGGACAAGTTTGTACAAAAAAGCAGGCTTCGTGATGACCCGGACAAAA-3'

[0039] R: 5'-GGGGACCACTTTGTACAAGAAAGCTGGGTAGGGCATCTTGGTAACCA-3'.

[0040] The PCR reaction system is (5 μL): 3.5 μL of PCR recovered product, 1 μL of pDONR207 plasmid, BPClonase TM II Enzyme Mix (Invitrogen) 0.5 μL. After mixing evenly, react in a low-temperature constant temperature tank at 25°C for 1 to 3 hours; then transfer it into competent cells E.coli DH5α, apply it on LB medium containing gentamicin after transformation, grow overnight at 37°C, and pick The single clones were cultured in liquid LB medium containing gentamicin, and the positive clones were sequenced after extracting the plasmids to obtain recombinant plas...

Embodiment 3

[0045] Example 3 Analysis of the expression pattern of the BEC1019 gene in the wheat var.

[0046] Seven time points, including 0, 2, 3, 4, 5, 6, and 7 days after the infection of the root system and stem base of Zhoumai 26 by the wheat var. cDNA. The 18SrRNA of wheat var. graminearum was used as an internal reference gene to analyze the expression of BEC1019 gene. Quantitative PCR, three simultaneous amplification and detection of each template in each quantitative experiment, and three biological repetitions, the primer sequences are as follows:

[0047] Ggt-18S rRNA-F: 5'-CGAACTCGGTCGTTTAGAGG-3';

[0048] Ggt-18S rRNA-R: 5'-GGTATGTTCACAGGGGTTGG-3';

[0049] BEC1019-F: 5'-TCATGTGGACATCGTCGGTC-3',

[0050] BEC1019-R: 5'-CACGCTGATGTCAAACGCAT-3'.

[0051] The expression analysis was carried out at different time points in the roots of wheat roots infected by Triticum graminearum var. figure 1 As shown, the results of qRT-PCR indicated that the peak was reached on the 4th ...

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Abstract

The invention belongs to the technical field of agricultural biology and particularly relates to application of a gene BEC1019 in improvement of full rot resistance of wheat. A method for improving the full rot resistance of wheat comprises a step of silencing the gene BEC1019 in the wheat. The gene BEC1019 is an important effector gene in full rot of the wheat, the gene can be used for enhancingresistance of the wheat to multiple pathogenic bacteria, and new wheat species with durable and broad spectrum resistance can be bred.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology, and in particular relates to the application of gene BEC1019 in improving the resistance to take-all disease of wheat. Background technique [0002] Wheat is one of the most productive and important crops in the world in the 21st century. Wheat production is crucial to global food security, but its yield and quality are greatly affected by various pests and diseases. As a measure to curb the rising incidence of plant fungal diseases, planting disease-resistant varieties and applying various fungicides have become the main methods adopted by farmers, and the latter is very easy to pollute the environment. In addition, due to the rapid emergence of toxic strains and fungicide-resistant strains, traditional breeding has been unable to effectively control a large number of crop diseases, and breeding disease-resistant varieties has become the main way to improve plant disease resistance. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31C12N15/82A01H5/00A01H6/46
CPCC07K14/37C12N15/8218C12N15/8282
Inventor 张怡李成伟于德水李晓丽张菊徐克东雷杰
Owner ZHOUKOU NORMAL UNIV
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