Identification and molecular tagging of two Arabidopsis resistance genes to Botrytis cinerea

Abstract

To map Arabidopsis resistance genes to Botrytis cinerea, Arabidopsis Col-0 ecotype resistant to B. cinerea BC18 isolate and Arabidopsis Ler ecotype susceptible to B. cinerea BC18 isolate were crossed. According to the resistant responses of the F₁, BC₁ and F₂ populations to B. cinerea, we identified two genes, named BC1 and BC2, responsible for the resistance of Arabidopsis Ler ecotype to B. cinerea. Through the method of map-based cloning, BC1 was linked to DNA markers CCR1 and DHS1 on the fourth chromosome of Arabidopsis with genetic distances of 1.2 cM and 1.6 cM for CCR1 and DHS1, respectively, and BC2 was linked to DNA markers CA72/NGA151 and NGA106 on the fifth chromosome with genetic distances of 1.4 cM and 2.4 cM for CA72/NGA151 and NGA106, respectively. Our results are beneficial for chromosome walking so that we can obtain the whole gene sequences, which will facilitate the understanding of their roles and manners of resistance to B. cinerea.

Keywords

Arabidopsis thaliana / Botrytis cinerea / resistant genes / linkage analysis / gene mapping

References

[1] AbuQamar S, Chen X, Dhawan R, Bluhm B, Salmeron J, Lam S, Dietrich R A, Mengiste T (2006). Expression profiling and mutant analysis reveals complex regulatory networks involved in Arabidopsis response to Botrytis infection. Plant J, 48(1): 28-44

[2] Bell C J, Ecker J R (1994). Assignment of 30 microsatellite loci to the linkage map of Arabidopsis. Genomics, 19(1): 137-144

[3] Bent A F, Kunkel B N, Dahlbeck D, Brown K L, Schmidt R, Giraudat J, Leung J, Staskawicz B J (1994). RPS2 of Arabidopsis thaliana: a leucine-rich repeat class of plant disease resistance genes. Science, 265(5180): 1856-1860

[4] Bisgrove S R, Simonich M T, Smith N M, Sattler A, Innes R W (1994). A disease resistance gene in Arabidopsis with specificity for two different pathogen avirulence genes. Plant Cell, 6(7): 927-933

[5] Borhan M H, Holub E B, Beynon J L, Rozwadowski K, Rimmer S R (2004). The Arabidopsis TIR-NB-LRR gene RAC1 confers resistance to Albugo candida (white rust) and is dependent on EDS1 but not PAD4. Mol Plant Microbe Interact, 17(7): 711-719

[6] Cao H, Glazebrook J, Clarke J D, Volko S, Dong X (1997). The Arabidopsis NPR1 gene that controls systemic acquired resistance encodes a novel protein containing ankyrin repeats. Cell, 88(1): 57-63

[7] Dhawan R, Luo H, Foerster A M, Abuqamar S, Du H N, Briggs S D, Scheid O M, Mengiste T (2009). HISTONE MONOUBIQUITINATION1 interacts with a subunit of the mediator complex and regulates defense against necrotrophic fungal pathogens in Arabidopsis. Plant Cell, 21(3): 1000-1019

[8] Ferrari S, Plotnikova J M, De Lorenzo G, Ausubel F M (2003). Arabidopsis local resistance to Botrytis cinerea involves salicylic acid and camalexin and requires EDS4 and PAD2, but not SID2, EDS5 or PAD4. Plant J, 35(2): 193-205

[9] Gassmann W, Hinsch M E, Staskawicz B J (1999). The ArabidopsisRPS4 bacterial-resistance gene is a member of the TIR-NBS-LRR family of disease-resistance genes. Plant J, 20(3): 265-277

[10] Konieczny A, Ausubel F M (1993). A procedure for mapping Arabidopsis mutations using co-dominant ecotype-specific PCR-based markers. Plant J, 4(2): 403-410

[11] Lander E S, Green P, Abrahamson J, Barlow A, Daly M J, Lincoln S E, Newberg L A (1987). MAPMAKER: an interactive computer package for constructing primary genetic linkage maps of experimental and natural populations. Genomics, 1(2): 174-181

[12] Lukowitz W, Gillmor C S, Scheible W R (2000). Positional cloning in Arabidopsis. Why it feels good to have a genome initiative working for you. Plant Physiol, 123(3): 795-805

[13] Mengiste T, Chen X, Salmeron J, Dietrich R (2003). The BOTRYTIS SUSCEPTIBLE1 gene encodes an R2R3MYB transcription factor protein that is required for biotic and abiotic stress responses in Arabidopsis. Plant Cell, 15(11): 2551-2565

[14] Michelmore R W, Paran I, Kesseli R V (1991). Identification of markers linked to disease-resistance genes by bulked segregant analysis: a rapid method to detect markers in specific genomic regions by using segregating populations. Proc Natl Acad Sci USA, 88(21): 9828-9832

[15] Parker J E, Coleman M J, Szabò V, Frost L N, Schmidt R, van der Biezen E A, Moores T, Dean C, Daniels M J, Jones J D (1997). The Arabidopsis downy mildew resistance gene RPP5 shares similarity to the toll and interleukin-1 receptors with N and L6. Plant Cell, 9(6): 879-894

[16] Stewart C N Jr, Via L E (1993). A rapid CTAB DNA isolation technique useful for RAPD fingerprinting and other PCR applications. Biotechniques, 14(5): 748-750

[17] The Arabidopsis Genome Initiative(2000). Analysis of the genome sequence of the flowering plant Arabidopsis thaliana. Nature, 408(6814): 796-815

[18] Veronese P, Chen X, Bluhm B, Salmeron J, Dietrich R, Mengiste T (2004). The BOS loci of Arabidopsis are required for resistance to Botrytis cinerea infection. Plant J, 40(4): 558-574

[19] Veronese P, Nakagami H, Bluhm B, Abuqamar S, Chen X, Salmeron J, Dietrich R A, Hirt H, Mengiste T (2006). The membrane-anchored BOTRYTIS-INDUCED KINASE1 plays distinct roles in Arabidopsis resistance to necrotrophic and biotrophic pathogens. Plant Cell, 18(1): 257-273

[20] Wilson I W, Schiff C L, Hughes D E, Somerville S C (2001). Quantitative trait loci analysis of powdery mildew disease resistance in the Arabidopsis thaliana accession kashmir-1. Genetics, 158(3): 1301-1309

[21] Xing J H, Ban H W, Weng Q Y, Dong J G (2010). Resistance of different Arabidopsis ecotypes to Botrytis cinerea. Journal of Agricultural University of Hebei, 6: 26-30 (in Chinese)

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Higher Education Press and Springer-Verlag Berlin Heidelberg