Resynthesis of &lt;i&gt;Brassica juncea&lt;/i&gt; for resistance to &lt;i&gt;Plasmodiophora brassicae&lt;/i&gt; pathotype 3

Hasan, Muhammad Jakir; Rahman, Habibur

doi:10.1270/jsbbs.18010

Cited by 15 publications

(8 citation statements)

References 35 publications

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“…Hagimori et al [ 54 ] carried out somatic hybridization between broccoli and radish and obtained hybrid offspring with 36 chromosomes, which could be used as bridge germplasm for subsequent CR broccoli cultivation. Hasan and Rahman [ 55 ] obtained the B. juncea which was resistant to race 3 of P. brassicae using the method of distant hybridization. Liu et al [ 56 ] identified three B. rapa with high resistance to Pb4 from 50 materials, which were then used as donor parents to cross with rapeseed, and obtained CR progenies.…”

Section: Discussionmentioning

confidence: 99%

Introgression of clubroot resistant gene into Brassica oleracea L. from Brassica rapa based on homoeologous exchange

Zhu

Yang

Zhang

et al. 2022

Horticulture Research

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Clubroot is a soil-borne disease in cabbage (Brassica oleracea L.) caused by Plasmodiophora brassicae, which poses a great threat to cabbage production. However, clubroot resistance (CR) genes in Brassica rapa could be introduced into the cabbage via breeding to make it clubroot resistant. In this study, CR genes from B. rapa were introduced into the cabbage genome and the mechanism of gene introgression was explored. Two methods were used to create CR materials: 1) The fertility of CR Ogura CMS cabbage germplasms containing CRa was restored by using an Ogura CMS restorer. After cytoplasmic replacement and microspore culture, CRa-positive microspore individuals were obtained. 2) Distant hybridization was performed between cabbage and B. rapa, which contained three CR genes (CRa, CRb and Pb8.1). Finally, BC2 individuals containing all three CR genes were obtained. Inoculation results showed that both CRa-positive microspore individuals and BC2 individuals containing three CR genes were resistant to race 4 of P. brassicae. Sequencing results from CRa-positive microspore individuals with specific molecular markers and genome-wide association study (GWAS) showed penetration at the homologous position of the cabbage genome by a 3.42 Mb CRa containing fragment from B. rapa; indicating homoeologous exchange (HE) as the theoretical basis for the introgression of CR resistance. The successful introduction of CR into the cabbage genome in the present study can provide useful clues for creating introgression lines within other species of interest.

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Section: Discussionmentioning

confidence: 99%

Introgression of clubroot resistant gene into Brassica oleracea L. from Brassica rapa based on homoeologous exchange

Zhu

Yang

Zhang

et al. 2022

Horticulture Research

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“…Due to the evolutionary history of the Brassica genus, valuable traits of interest already identified in diploid Brassica genomes have the potential to be exploited in amphidiploid Brassica species through modern genetic editing techniques, or via interspecific crossing/resynthesis. This approach was used to great effect when introducing clubroot disease resistance from B. rapa into B. juncea material, wherein natural resistance is not evident ( Hasan and Rahman, 2018 ) and to introduce turnip yellows virus resistance from Brassica oleracea and B. rapa into B. napus ( Greer et al, 2021 ). Potential therefore exists for the generation of enhanced TuMV-resistant B. juncea cultivars using resistances already identified on the “A” genome of other Brassica species via resynthesis.…”

Section: Discussionmentioning

confidence: 99%

Characterization and Mapping of retr04, retr05 and retr06 Broad-Spectrum Resistances to Turnip Mosaic Virus in Brassica juncea, and the Development of Robust Methods for Utilizing Recalcitrant Genotyping Data

et al. 2022

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Turnip mosaic virus (TuMV) induces disease in susceptible hosts, notably impacting cultivation of important crop species of the Brassica genus. Few effective plant viral disease management strategies exist with the majority of current approaches aiming to mitigate the virus indirectly through control of aphid vector species. Multiple sources of genetic resistance to TuMV have been identified previously, although the majority are strain-specific and have not been exploited commercially. Here, two Brassica juncea lines (TWBJ14 and TWBJ20) with resistance against important TuMV isolates (UK 1, vVIR24, CDN 1, and GBR 6) representing the most prevalent pathotypes of TuMV (1, 3, 4, and 4, respectively) and known to overcome other sources of resistance, have been identified and characterized. Genetic inheritance of both resistances was determined to be based on a recessive two-gene model. Using both single nucleotide polymorphism (SNP) array and genotyping by sequencing (GBS) methods, quantitative trait loci (QTL) analyses were performed using first backcross (BC1) genetic mapping populations segregating for TuMV resistance. Pairs of statistically significant TuMV resistance-associated QTLs with additive interactive effects were identified on chromosomes A03 and A06 for both TWBJ14 and TWBJ20 material. Complementation testing between these B. juncea lines indicated that one resistance-linked locus was shared. Following established resistance gene nomenclature for recessive TuMV resistance genes, these new resistance-associated loci have been termed retr04 (chromosome A06, TWBJ14, and TWBJ20), retr05 (A03, TWBJ14), and retr06 (A03, TWBJ20). Genotyping by sequencing data investigated in parallel to robust SNP array data was highly suboptimal, with informative data not established for key BC1 parental samples. This necessitated careful consideration and the development of new methods for processing compromised data. Using reductive screening of potential markers according to allelic variation and the recombination observed across BC1 samples genotyped, compromised GBS data was rendered functional with near-equivalent QTL outputs to the SNP array data. The reductive screening strategy employed here offers an alternative to methods relying upon imputation or artificial correction of genotypic data and may prove effective for similar biparental QTL mapping studies.

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“…rapa ) ( Yu et al , 2021 ; Yang et al , 2022 ). Most of the resistance loci have been mapped to chromosomes A01, A02, A03, A06, and A08 of the A genome, with only a few being mapped to the B or C genome ( Matsumoto et al , 1998 ; Suwabe et al , 2003 , 2006 ; Hirai et al , 2004 ; Piao et al , 2004 ; Sakamoto et al , 2008 ; Werner et al , 2008 ; Ueno et al , 2012 ; Chen et al , 2013 ; Hatakeyama et al , 2013 ; Chu et al , 2014 ; Li et al , 2016 ; Yu et al , 2016 , 2017 , 2021 ; Huang et al , 2017 , 2019 ; Hasan and Rahman, 2018 ; Hirani et al , 2018 ; Pang et al , 2018 ; Chang et al , 2019 ; Laila et al , 2019 ; Fredua-Agyeman et al , 2020 ; Karim et al , 2020 ; Dakouri et al , 2021 ; Rahaman et al , 2022 ). To date, only two clubroot-resistance genes, Crr1a and CRa , have been isolated, and they both encode toll-interleukin-1 receptor, nucleotide binding site, and leucine-rich repeat (TIR-NBS-LRR, TNL) proteins ( Ueno et al , 2012 ; Hatakeyama et al , 2013 ).…”

Section: Introductionmentioning

confidence: 99%

A CRISPR/Cas9-based vector system enables the fast breeding of selection-marker-free canola with Rcr1-rendered clubroot resistance

Hu,

Zhang,

2023

Journal of Experimental Botany

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Breeding for disease resistance in major crops is of crucial importance for global food security and sustainability. However, common biotechnologies such as traditional transgenesis or genome editing do not provide an ideal solution, whereas transgenic crops free of selection markers such as cisgenic/intragenic crops might be suitable. In this study, after cloning and functional verification of the Rcr1 gene for resistance to clubroot (Plasmodiophora brassicae), we confirmed that the genes Rcr1, Rcr2, Rcr4, and CRa from Brassica rapa crops and the resistance gene from B. napus oilseed rape cv. ‘Mendel’ on chromosome A03 were identical in their coding regions. We also determined that Rcr1 has a wide distribution in Brassica breeding materials and renders potent resistance against multiple representative clubroot strains in Canada. We then modified a CRISPR/Cas9-based cisgenic vector system and found that it enabled the fast breeding of selection-marker-free transgenic crops with add-on traits, with selection-marker-free canola (B. napus) germplasms with Rcr1-rendered stable resistance to clubroot disease being successfully developed within 2 years. In the B. napus background, the intragenic vector system was able to remove unwanted residue sequences from the final product with high editing efficiency, and off-target mutations were not detected. Our study demonstrates the potential of applying this breeding strategy to other crops that can be transformed by Agrobacterium. Following the streamlined working procedure, intragenic germplasms can be developed within two generations, which could significantly reduce the breeding time and labor compared to traditional introgression whilst still achieving comparable or even better breeding results.

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Resynthesis of <i>Brassica juncea</i> for resistance to <i>Plasmodiophora brassicae</i> pathotype 3

Cited by 15 publications

References 35 publications

Introgression of clubroot resistant gene into Brassica oleracea L. from Brassica rapa based on homoeologous exchange

Introgression of clubroot resistant gene into Brassica oleracea L. from Brassica rapa based on homoeologous exchange

Characterization and Mapping of retr04, retr05 and retr06 Broad-Spectrum Resistances to Turnip Mosaic Virus in Brassica juncea, and the Development of Robust Methods for Utilizing Recalcitrant Genotyping Data

A CRISPR/Cas9-based vector system enables the fast breeding of selection-marker-free canola with Rcr1-rendered clubroot resistance

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