Development and Application of Functional Markers for Rice Blast Resistance Gene 
            <i>Bsr-d1</i> in Rice

Wang, Jun; Zhao, Jieyu; Yang, Xu; Fan, Fangjun; Jun, Zhu; Li, Wenqi; Wang, Fangquan; Fei, Yun-Yan; WeiGong, Zhong; Yang, Jie

doi:10.3724/sp.j.1006.2018.01612

Cited by 3 publications

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“…It has been reported that there is a single nucleotide change among the promoter of the Bsr-d1 (-618bp, A to G), which causes reduced gene expression of the gene through the binding of the repressive MYB transcription factor, thereby inhibiting H 2 O 2 degradation and enhancing disease resistance . In this study, we detected the blast resistance allele of Bsr-d1 gene in a total of 256 japonica rice accessions collected from Jilin province in Northeast China using its functional marker (Wang et al 2018), and found that none of the accessions had the blast resistance allele of Bsr-d1 gene (Fig. 1B).…”

Section: Discussionmentioning

confidence: 89%

“…Genomic DNA was extracted from 10-day-old rice seedlings of 256 japonica rice accessions in Jilin province of Northeast China (Table S3) by CTAB method, which were provided by Rice Research Institute of Jilin Academy of Agricultural Sciences. Molecular marker assisted PCR was carried out using blast resistance marker of Bsr-d1-F/3Bsr-d1-R and blast susceptible marker Bsr-d1-F/3bsr-d1-R (Wang et al 2018), the PCR products were analyzed by 2.5% agarose gel electrophoresis, 25-µL mixture was applied.…”

Section: Allelic Analysis Of Blast Resistance Functional Allele Of Bs...mentioning

confidence: 99%

“…Natural allele of Bsr-d1 gene in Digu causes a single nucleotide change (A to G) in the promoter region of -618 bp, which causes reduced expression of the gene through the binding of the restrictive MYB transcription factor, consequently, inhibits H 2 O 2 degradation and increases the resistance to rice blast. According to a review of 3,000 sequenced rice genomes, 10% of rice had this allele ; Wang et al (2018) developed a functional marker for Bsr-d1 gene and screened the distribution of Bsr-d1 gene in 324 rice varieties, the results showed the Bsr-d1 gene was not detected in 271 japonica rice varieties, which indicated that the Bsr-d1 gene was mainly distributed in indica rice varieties.…”

Section: Introductionmentioning

confidence: 99%

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CRISPR/Cas9-mediated knockout of Bsr-d1 enhances the blast resistance of rice in Northeast China

Zhang,

Lin,

et al. 2024

Plant Cell Rep

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Rice blast is a globaldisease, which has a signi cant negative impact on rice yield and quality. Due to the complexity and changeability of the physiological races of rice blast, controlling the rice blast is challenging in agricultural production. Bsr-d1, a negative transcription factor confers broad-spectrum resistance to rice blast which was identi ed from the indica rice cultivar Digu, however, the biological function in japonica rice varieties is still unclear. In this study, we analyzed the blast resistance allele of OsBsr-d1 in a total of 256 japonica rice varieties of Jilin province in Northeast China and found that it does not exist in these varieties. Therefore, we developed Bsr-d1 knockout mutants by CRISPR/Cas9 system using a japonica rice variety Jigeng88 (JG88) as a recipient variety. Compared with the wild-type JG88, the homozygous Bsr-d1 mutant lines, KO#1 and KO#2, showed enhanced leaf blast resistance at seedling stage to several Magnaporthe oryzae (M. oryzae) races collected from Jilin Province in Northeast China. Physiological and biochemical indicators revealed that the homozygous mutant lines produced more hydrogen peroxide compared to JG88 when infected by M. oryzae. Comparative RNA-seq revealed that the DEGs were mainly involved in the synthesis of amide compounds, zinc nger proteins and transmembrane transporters, etc. In summary, our results indicate that the Bsr-d1 knockout mutants developed through gene editing technology can enhance the broad-spectrum resistance of rice in Northeast China to rice blast. This study not only provides a theoretical basis for disease resistance breeding of Bsr-d1 gene in Northeast China, but also provides new germplasm resources for disease resistance rice breeding. Key MessageBlast resistance allele of OsBsr-d1 doesn't exist in most Japonica rice varieties of Jilin province in China. Bsr-d1 knockout mutants developed by CRISPR/Cas9 enhance the broad-spectrum resistance to rice blast in Northeast China.

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Section: Discussionmentioning

confidence: 89%

Section: Allelic Analysis Of Blast Resistance Functional Allele Of Bs...mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

CRISPR/Cas9-mediated knockout of Bsr-d1 enhances the blast resistance of rice in Northeast China

Zhang,

Lin,

et al. 2024

Plant Cell Rep

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“…Gene-specific markers developed from DNA polymorphisms among the alleles are particularly useful for MAS breeding and screening large amounts of germplasm for allelic diversity (Brar and Khush 1997;Kumari et al 2013;Nawaz et al 2017;Shomura et al 2008;Xu et al 2005). Such markers have been developed for genes including Pit, Pi-2, Pi-9, and Pig-m (Hayashi et al 2010;Tian et al 2016;Wang et al 2019). In the case of the Pik locus, which displays high similarity among Pi-1, Pik-h, Pi-k, Pik-m, Pik-s, Pik-p, and wild rice clusters from the same region (Hua et al 2012;Zhai et al 2011), it is difficult to differentiate these genes from each other by simple sequence repeat (SSR) markers.…”

Section: Introductionmentioning

confidence: 99%

Two novel gene-specific markers at pik locus facilitate the application of rice blast resistant alleles in breeding

Tian

Chen

Lin

et al. 2019

Preprint

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Background: Rice blast disease, caused by Magnaporthe oryzae, is a major constraint for rice production in the world. Introgression of blast-durable resistance genes into high-yielding rice cultivars has been considered an agricultural priority in an effort to control the disease. The blast resistance Pik locus, located on chromosome 11, contains at least six important resistance genes, but these genes have not been widely employed in resistance breeding since existing markers hardly satisfy current breeding needs owing to their limited scope of application.Results: In the present study, two PCR-based markers, Pikp-Del and Pi1-In, were developed to target the specific InDel (insertion/deletion) of the Pik-p and Pi-1 genes, respectively. The two markers precisely distinguished Pik-p, Pi-1, and the K-type alleles at the Pik locus, which is a necessary element for functional genes from rice varieties. Conclusions:Two gene-specific markers of Pi-kp and Pi1 identified that only several old varieties contain the two genes, nearly half these varieties yet carry the K-type alleles. Therefore, these identified varieties can be new gene sources for developing blast resistant rice. The two newly developed markers should be highly useful for using Pi-kp, Pi1 and other resistance genes at the Pik locus in marker-assisted selection (MAS) breeding programs.

show abstract

CRISPR/Cas9-mediated knockout of Bsr-d1 enhances blast resistance of rice in Northeast China

Zhang,

Lin,

et al. 2024

Preprint

View full text Add to dashboard Cite

Rice blast is a globaldisease, which has a significant negative impact on rice yield and quality. Due to the complexity and changeability of the physiological races of rice blast, controlling the rice blast is challenging in agricultural production. Bsr-d1, a negative transcription factor confers broad-spectrum resistance to rice blast which was identified from the indica rice cultivar Digu, however, the biological function in japonica rice varieties is still unclear. In this study, we analyzed the blast resistance allele of OsBsr-d1 in a total of 256 japonica rice varieties of Jilin province in Northeast China and found that it does not exist in these varieties. Therefore, we developed Bsr-d1 knockout mutants by CRISPR/Cas9 system using a japonica rice variety Jigeng88 (JG88) as a recipient variety. Compared with the wild-type JG88, the homozygous Bsr-d1 mutant lines, KO#1 and KO#2, showed enhanced leaf blast resistance at seedling stage to several Magnaporthe oryzae (M. oryzae) races collected from Jilin Province in Northeast China. Physiological and biochemical indicators revealed that the homozygous mutant lines produced more hydrogen peroxide compared to JG88 when infected by M. oryzae. Comparative RNA-seq revealed that the DEGs were mainly involved in the synthesis of amide compounds, zinc finger proteins and transmembrane transporters, etc. In summary, our results indicate that the Bsr-d1 knockout mutants developed through gene editing technology can enhance the broad-spectrum resistance of rice in Northeast China to rice blast. This study not only provides a theoretical basis for disease resistance breeding of Bsr-d1 gene in Northeast China, but also provides new germplasm resources for disease resistance rice breeding.

show abstract

Development and Application of Functional Markers for Rice Blast Resistance Gene Bsr-d1 in Rice

Cited by 3 publications

References 0 publications

CRISPR/Cas9-mediated knockout of Bsr-d1 enhances the blast resistance of rice in Northeast China

CRISPR/Cas9-mediated knockout of Bsr-d1 enhances the blast resistance of rice in Northeast China

Two novel gene-specific markers at pik locus facilitate the application of rice blast resistant alleles in breeding

CRISPR/Cas9-mediated knockout of Bsr-d1 enhances blast resistance of rice in Northeast China

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