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Application of puccinia striiformis (PSTG_17694) gene in prevention and treatment of puccinia striiformis and cultivation method of anti-puccinia-striiformis wheat

A technology for wheat stripe rust and wheat stripe rust, which is applied in the field of genetic engineering and crop molecular breeding, can solve the problems of small number of key pathogenic genes, difficulty in genetics research of wheat-stripe rust, slow progress, etc., and achieve growth inhibition Effects of Reproduction, Significant Wheat Stripe Rust Resistance

Active Publication Date: 2018-09-21
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Due to its biotrophic and obligate parasitic characteristics, genetic transformation of wheat stripe rust is difficult; and wheat itself is also difficult to carry out genetic transformation, so the genetic research of wheat-stripe rust interaction is very difficult and the progress is slow
At present, the number of infection or pathogenic key genes identified in wheat stripe rust is relatively small, and no genes have been effectively used in wheat disease resistance breeding

Method used

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  • Application of puccinia striiformis (PSTG_17694) gene in prevention and treatment of puccinia striiformis and cultivation method of anti-puccinia-striiformis wheat
  • Application of puccinia striiformis (PSTG_17694) gene in prevention and treatment of puccinia striiformis and cultivation method of anti-puccinia-striiformis wheat
  • Application of puccinia striiformis (PSTG_17694) gene in prevention and treatment of puccinia striiformis and cultivation method of anti-puccinia-striiformis wheat

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] (1) PCR amplification to obtain the PSTG_17694 gene fragment

[0040] Taking the complete genome sequence of wheat stripe rust physiological race PST-78 as a reference (GenBank accession number AJIL00000000.1 or BROAD download link ftp: / / ftp.broadinstitute.org / pub / annotation / fungi / puccinia / genomes / puccinia_striifo rmis_pst- 78 / ), using the coding region sequence of hypothetical protein gene PSTG_17694 (GenBank: KNE88861.1) as a template to design PCR primers.

[0041] Forward primer sequence CQM17694-F2, as shown in Seq ID No.3:

[0042] 5`-caagcggccgcTCATTCGACCAAAGAAAGGC-3`;

[0043] Reverse primer sequence CQM17694-R2, as shown in SeqID No.4:

[0044] 5`-tcaggcgcgccGATCACCTCGTTTCCATGCT-3`.

[0045] The length of the amplified segment is 490bp. Using the sequence alignment of the amplified segments to compare the whole genome data of wheat stripe rust PST-78 ( https: / / genome.jgi.doe.gov / Pucst_PST78_1 / Pucst_PST78_1.home.html ) and the NCBI database wheat stripe ru...

Embodiment 2

[0058] Embodiment 2 Anti-stripe rust resistance identification of wheat stripe rust

[0059] Propagation of fresh spores of wheat stripe rust: plant the susceptible variety Huixianhong in the greenhouse, inject the spore aqueous solution with a syringe during the one-leaf-one-heart stage, then spray water with a watering can and cover with a plastic film to keep it moist. Inoculation can be repeated 2-3 times to ensure full onset, with an interval of 1 week between each time. Preparation of the spore aqueous solution: Take the dried and frozen (-80°C) spores, suspend them with an appropriate amount of tap water until they are light orange, place them on a shaker at room temperature and shake them for 30 minutes (180rpm), and then inoculate them by injection. Sporadic disease can be seen about 20 days after inoculation, and a large number of leaves can be seen after about 30 days. A large number of fresh spores can be collected for inoculation identification of transgenic mater...

Embodiment 3

[0064] Example 3 Identification of PSTG_17694 Gene Expression in Stripe Rust-resistant Wheat

[0065] Twelve days after inoculation of wheat seedling leaves, the surface of wild-type CB037 leaves was covered with a large number of spore piles. About 100 mg of leaves of CB037 and transgenic materials were taken at different time points after inoculation, RNA was extracted by Trizol method, and cDNA was prepared for real-time fluorescent quantitative PCR. The real-time fluorescent quantitative PCR primer sequences are CQM_17694-F3 and CQM_17694-R3, and the specific primer sequences are shown in Seq ID No.7 and Seq ID No.8 in the sequence table. Taking the α-tubulin gene of wheat stripe rust as an internal reference (Huang Xueling et al., Journal of Agricultural Biotechnology, 2012, 20(2): 181-187), the primer TUBA-F / R sequence is shown in the sequence table Seq ID No.9, SeqID Shown in No.10.

[0066]The PCR system is: 5 μl 2×SYBR GreenMaster, 1 μl each of the forward and rever...

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Abstract

The invention provides application of a puccinia striiformis (PSTG_17694) gene in prevention and treatment of puccinia striiformis. The sequence of the PSTG_17694 gene is shown in Seq ID No.1; the PSTG_17694 gene can effectively regulate and control growth and propagation of puccinia striiformis and is used as a molecular target of transcriptional regulation and control or used as a gene target for inhibiting a protein function, and the PSTG_17694 gene is silenced to achieve the effect of inhibiting the growth and propagation of puccinia striiformis. According to a cultivation method for anti-puccinia-striiformis wheat by using the gene, the obtained wheat has obvious puccinia striiformis resistance.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering and crop molecular breeding, and in particular relates to the application of wheat stripe rust PSTG_17694 gene in the prevention and treatment of stripe rust and the cultivation method of stripe rust resistant wheat. Background technique [0002] Wheat is the second largest grain crop in my country, with an average annual sowing area of ​​about 360 million mu. The stability and improvement of wheat production are related to the national economy and people's livelihood. However, for many years, fungal diseases have posed a serious threat to wheat production, such as rust, powdery mildew, scab, sheath blight and other diseases, seriously affecting the yield and quality of wheat. Among them, wheat stripe rust, commonly known as "jaundice", is caused by obligate parasitic stripe rust (Puccinia striiformis Westend.f.sp.tritici). In my country, the incidence of wheat stripe rust is rampant...

Claims

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Application Information

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IPC IPC(8): C12N15/31C12N15/82
CPCC07K14/37C12N15/8218C12N15/8282
Inventor 吴佳洁黄德华张会飞刘强倪飞付道林
Owner SHANDONG AGRICULTURAL UNIVERSITY
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