Rice sheath blight resistance gene SBR11 as well as molecular marker and application thereof

A technology of molecular markers and sheath blight, which is applied in the fields of application, genetic engineering, and plant genetic improvement, can solve problems such as poor results, and achieve the effect of speeding up the breeding process and improving resistance

Active Publication Date: 2021-05-25
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For a long time, the control of sheath blight has mainly relied on chemical agents, but the effect has been poor

Method used

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  • Rice sheath blight resistance gene SBR11 as well as molecular marker and application thereof
  • Rice sheath blight resistance gene SBR11 as well as molecular marker and application thereof
  • Rice sheath blight resistance gene SBR11 as well as molecular marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1, verification of candidate genes

[0036] In this practice, a near-isogenic line constructed by LE and TQ was used, and a candidate gene SBR11 for resistance to sheath blight was detected on chromosome 11 of LE. In order to verify the function of the gene, the present invention constructs SBR11 overexpression (SBR11OE) and RNAi vector (SBR11RNAi), and transfers the SBR11-OE vector into the susceptible near-isogenic line NIL-SBR11 TQ (NIL), the SBR11RNAi vector was transferred into LE. The field resistance identification results showed that compared with the control, the resistance of the SBR11OE transgenic line to sheath blight was significantly increased ( figure 1 ), the resistance of SBR11RNAi transgenic lines to sheath blight was significantly reduced ( figure 2 ). The full-length gene of SBR11 was cloned from LE, and transformed into the susceptible near-isogenic line NIL, and a blank control (#8) was set at the same time. The inoculation identifi...

Embodiment 2

[0037] Embodiment 2, SBR11 functional analysis

[0038] The resistant allele of SBR11 (SBR11) came from LE, and the indica variety TQ carried the susceptible allele (sbr11) at this locus. The sequencing results showed ( Figure 4), SBR11 not only had 11 SNP differences in the upstream 2kb of ATG between the resistant and sensitive allele donor parents, but also had 5 mutations in the coding region, of which the 190th, 926th and 1492nd positions were sense mutations. The induced expression level of SBR11 after the infection of sheath blight was significantly higher than that of sbr11( Figure 5 ). In order to verify the gene function of SBR11, the present invention uses the overexpression promoter Ubiquitin to drive the coding regions of the two alleles SBR11 and sbr11 respectively, and transfers them into the susceptible near-isogenic line NIL. Transgenic lines were inoculated in vitro with sheath blight in greenhouse ( Figure 6 ) and field inoculation identification ( ...

Embodiment 3

[0042] Example 3, the development of molecular markers

[0043] The present invention develops dCAPS markers for gene selection for the base variation of SNP190 and SNP926, and the specific information of these two dCAPS markers is as follows:

[0044] SNP190: Located at the 4851244th base of rice chromosome 11, that is, the 190th base of the SBR11 gene, the molecular marker polymorphism is T / A, and the sequence of 176 bp before and after the SNP site is shown in SEQ ID No.3 , the 154th bp base is the SNP site 190; the base of the SBR11 disease-resistant allele at this site is T, which will not be recognized by the restriction endonuclease PvuII, and the expanded fragment size is 176 bp. The base of the susceptible allele sbr11 at this site is A, which will be recognized by the restriction endonuclease PvuII, and the 21bp fragment will be cut off, leaving only a 155bp fragment.

[0045] SNP926: Located at the 4851980th base of rice chromosome 11, that is, the 926th base of th...

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Abstract

The invention discloses a rice sheath blight resistance gene SBR11 as well as a molecular marker and application thereof, and belongs to the technical field of crop molecular biology. Genetic transformation confirms that the SBR11 gene positively regulates the resistance of rice to sheath blight, allele function and haplotype analysis confirms that gene function variation is caused by SNP (Single Nucleotide Polymorphism) at 190th and 926th sites of a gene coding region, and further, the invention also develops two molecular markers for detecting the genotypes of the two sites, the molecular markers comprise the SNP site 190 and the SNP site 926th, and the SNP site 190 and the SNP site 926th can be used for detecting the genotypes of the two sites. The molecular marker can be used for detecting and selecting SBR11 alleles, more resources are provided for rice disease-resistant molecular design and breeding, and the breeding process is accelerated.

Description

technical field [0001] The invention belongs to the technical field of crop molecular biology and relates to a rice sheath blight resistance gene SBR11 and its molecular marker and application. Background technique [0002] Rice (Oryza sativa L.) is the most important food crop in my country. Rice sheath blight caused by strong saprophytic fungus Rhizoctonia solani Kühn is one of the three major diseases of rice in my country. Due to its wide occurrence area and difficult control, sheath blight is the first among all rice diseases in terms of yield loss. For a long time, the control of sheath blight has mainly relied on chemical agents, but the effect has been poor. Promoting and breeding disease-resistant varieties is the most cost-effective measure to control diseases. The resistance of rice to sheath blight is a quantitative trait resistance, which is controlled by multiple genes or QTL (Quantitative traitlocus), and cloning genes related to sheath blight resistance ha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12Q1/6895C12Q1/6858C12N15/11
CPCC07K14/415C12Q1/6895C12Q1/6858C12Q2600/13C12Q2600/156C12Q2531/113C12Q2521/301C12Q2521/313C12Q2565/125
Inventor 王雨左示敏薛芗陈宗祥冯志明胡珂鸣潘学彪
Owner YANGZHOU UNIV
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