Identification and Fine-Mapping of Clubroot (Plasmodiophora brassicae) Resistant QTL in Brassica rapa

European fodder turnips (Brassica rapa ssp. rapifera) were identified as sources of clubroot resistance (CR) and have been widely used in Brassica resistance breeding. An F2 population derived from a cross between a resistant turnip and a susceptible Chinese cabbage was used to determine the inheritance and locating the resistance Quantitative Trait Loci (QTLs). The parents showed to be very resistant/susceptible to the field isolates (pathotype 4) of clubroot from Henan in China. After inoculation, 27 very resistant or susceptible individuals were selected to construct bulks, respectively. Next-generation-sequencing-based Bulk Segregant Analysis Sequencing (BSA-Seq) was used and located resistance QTL on chromosome A03 (3.3–7.5 Mb) and A08 (0.01–6.5 Mb), named Bcr1 and Bcr2, respectively. Furthermore, an F3 population including 180 families derived from F2 individuals was phenotyped and used to verify and narrow candidate regions. Ten and seven Kompetitive AlleleSpecific PCR (KASP) markers narrowed the target regions to 4.3–4.78 Mb (A03) and 0.02–0.79 Mb (A08), respectively. The phenotypic variation explained (PVE) of the two QTLs were 33.3% and 13.3% respectively. The two candidate regions contained 99 and 109 genes. In the A03 candidate region, there were three candidate R genes, namely Bra006630, Bra006631 and Bra006632. In the A08 candidate region, there were two candidate R genes, namely Bra030815 and Bra030846.

[1]  Swati Megha,et al.  Clubroot in Brassica: Recent advances in genomics, breeding and disease management. , 2021, Genome.

[2]  Shahida Begum,et al.  QTL-seq analysis of powdery mildew resistance in a Korean cucumber inbred line , 2020, Theoretical and Applied Genetics.

[3]  B. Gossen,et al.  Two Clubroot-Resistance Genes, Rcr3 and Rcr9wa, Mapped in Brassica rapa Using Bulk Segregant RNA Sequencing , 2020, International journal of molecular sciences.

[4]  Zhen Wu,et al.  Identification of heat-tolerance QTLs and high-temperature stress-responsive genes through conventional QTL mapping, QTL-seq and RNA-seq in tomato , 2019, BMC Plant Biology.

[5]  B. Gossen,et al.  Fine mapping of a clubroot resistance gene from turnip using SNP markers identified from bulked segregant RNA-Seq , 2019, Molecular Breeding.

[6]  W. Zhai,et al.  Two QTLs controlling Clubroot resistance identified from Bulked Segregant Sequencing in Pakchoi (Brassica campestris ssp. chinensis Makino) , 2019, Scientific Reports.

[7]  S. Isobe,et al.  Mapping of a novel clubroot resistance QTL using ddRAD-seq in Chinese cabbage (Brassica rapa L.) , 2019, BMC Plant Biology.

[8]  Xiaonan Li,et al.  Identification and Mapping of the Clubroot Resistance Gene CRd in Chinese Cabbage (Brassica rapa ssp. pekinensis) , 2018, Front. Plant Sci..

[9]  R. Komakhin,et al.  The New Clubroot Resistance Locus Is Located on Chromosome A05 in Chinese Cabbage (Brassica rapa L.) , 2018, Russian Journal of Genetics.

[10]  B. Gossen,et al.  Genotyping-by-sequencing reveals three QTL for clubroot resistance to six pathotypes of Plasmodiophora brassicae in Brassica rapa , 2017, Scientific Reports.

[11]  K. Hatakeyama,et al.  The tandem repeated organization of NB-LRR genes in the clubroot-resistant CRb locus in Brassica rapa L. , 2017, Molecular Genetics and Genomics.

[12]  R. Sun,et al.  Mapping of clubroot (Plasmodiophora brassicae) resistance in canola (Brassica napus) , 2016 .

[13]  R. Sun,et al.  Resistance to Plasmodiophora brassicae in Brassica rapa and Brassica juncea genotypes From China. , 2015, Plant disease.

[14]  B. Gossen,et al.  The Role of Primary and Secondary Infection in Host Response to Plasmodiophora brassicae. , 2014, Phytopathology.

[15]  A. Chai,et al.  Research status of clubroot (Plasmodiophora brassicae) on cruciferous crops in China , 2014 .

[16]  K. Falk,et al.  Sources of resistance to Plasmodiophora brassicae (clubroot) pathotypes virulent on canola , 2014 .

[17]  Z. Piao,et al.  Identification of Novel QTLs for Isolate-Specific Partial Resistance to Plasmodiophora brassicae in Brassica rapa , 2013, PloS one.

[18]  S. Strelkov,et al.  Assessment of gene expression profiles in primary and secondary zoospores of Plasmodiophora brassicae by dot blot and real-time PCR. , 2013, Microbiological research.

[19]  R. Terauchi,et al.  QTL-seq: rapid mapping of quantitative trait loci in rice by whole genome resequencing of DNA from two bulked populations. , 2013, The Plant journal : for cell and molecular biology.

[20]  K. Hatakeyama,et al.  Fine mapping of the clubroot resistance gene CRb and development of a useful selectable marker in Brassica rapa , 2013, Breeding science.

[21]  H. Fukuoka,et al.  Identification and Characterization of Crr1a, a Gene for Resistance to Clubroot Disease (Plasmodiophora brassicae Woronin) in Brassica rapa L. , 2013, PloS one.

[22]  H. Matsumura,et al.  Molecular characterization of the CRa gene conferring clubroot resistance in Brassica rapa , 2012, Plant Molecular Biology.

[23]  A. Fleming,et al.  Gall formation in clubroot-infected Arabidopsis results from an increase in existing meristematic activities of the host but is not essential for the completion of the pathogen life cycle. , 2012, The Plant journal : for cell and molecular biology.

[24]  Hiroki Ueno,et al.  Accumulation of Three Clubroot Resistance Genes through Marker-assisted Selection in Chinese Cabbage (Brassica rapa ssp. pekinensis). , 2012 .

[25]  J. Poulain,et al.  The genome of the mesopolyploid crop species Brassica rapa , 2011, Nature Genetics.

[26]  M. DePristo,et al.  The Genome Analysis Toolkit: a MapReduce framework for analyzing next-generation DNA sequencing data. , 2010, Genome research.

[27]  H. Hakonarson,et al.  ANNOVAR: functional annotation of genetic variants from high-throughput sequencing data , 2010, Nucleic acids research.

[28]  Gonçalo R. Abecasis,et al.  The Sequence Alignment/Map format and SAMtools , 2009, Bioinform..

[29]  Richard Durbin,et al.  Sequence analysis Fast and accurate short read alignment with Burrows – Wheeler transform , 2009 .

[30]  G. Dixon,et al.  The Occurrence and Economic Impact of Plasmodiophora brassicae and Clubroot Disease , 2009, Journal of Plant Growth Regulation.

[31]  K. Sakamoto,et al.  Mapping of isolate-specific QTLs for clubroot resistance in Chinese cabbage (Brassica rapa L. ssp. pekinensis) , 2008, Theoretical and Applied Genetics.

[32]  N. Kubo,et al.  Fine mapping of the clubroot resistance gene, Crr3, in Brassica rapa , 2006, Theoretical and Applied Genetics.

[33]  H. Fukuoka,et al.  Simple Sequence Repeat-Based Comparative Genomics Between Brassica rapa and Arabidopsis thaliana: The Genetic Origin of Clubroot Resistance , 2006, Genetics.

[34]  Y. Lim,et al.  SCAR and CAPS mapping of CRb, a gene conferring resistance to Plasmodiophora brassicae in Chinese cabbage (Brassica rapa ssp. pekinensis) , 2004, Theoretical and Applied Genetics.

[35]  H. Fukuoka,et al.  Identification of two loci for resistance to clubroot (Plasmodiophora brassicae Woronin) in Brassica rapa L. , 2003, Theoretical and Applied Genetics.

[36]  M. Sacristán,et al.  Disease response of resynthesized Brassica napus L. lines carrying different combinations of resistance to Plasmodiophora brassicae Wor. , 1996 .

[37]  J. Doyle,et al.  Isolation of plant DNA from fresh tissue , 1990 .

[38]  G. Dixon,et al.  Study of physiologic specialization in Plasmodiophora brassicae: Proposals for attempted rationalization through an international approach , 1975 .

[39]  P. Williams A system for the determination of races of Plasmodiophora brassicae that infect cabbage and Rutabaga , 1966 .