Gradus

VOL 3, NO 1 (2016): SPRING (APRIL)

 

EXPERIMENTS FOR DEVELOPING MARKERS FOR TOBAMOVIRUS-RESISTANCE IN PEPPER (CAPSICUM SPP.) USING NBS PROFILING APPROACH


Virág Mihálka, C. Gerard van der Linden and István Nagy

Abstract

For generating molecular markers for Tobamovirus-resistance alleles in pepper, we tested the application of NBS-profiling procedure, which specifically produces markers in or near resistance genes. The NBS-profiling method was applied to controlled crosses, in which the donor plants with L1, L3 and L4 resistance alleles were crossed with sensitive C. annuum ‘Albaregia’. Individuals of F2 progenies containing the three different resistance alleles, were phenotypically characterized and used for further analysis. DNA of resistant and susceptible F2 plants was digested and ligated with adapter. In the PCR reaction one adapter primer and one degenerate NBS specific primer were used. The PCR products were analyzed on 6% polyacrylamid gels. Partially co-segregating bands with the resistant phenotype L3 were found.


Keywords

Keywords: Pepper (Capsicum annuum L.), Tobamovirus-resistance, NBS profiling, molecular marker,


References

[1] Brugmans B., Wouters D., van Os H., Hutten R., van der Linden G., Visser R.G., van Eck H.J., van der VossenE.A.G. (2008): Genetic mapping and transcription analyses of resistance gene loci in potato using NBS profiling.Theor Appl Genet, 117: 1379–1388.
[2] Calenge F., van der Linden C.G., van de Weg E., Schouten H.J., van Arkel G., Denancé C., Durel C.E. (2005):Resistance gene analogues identified through the NBS profiling method map close to major genes and QTL fordisease resistance in apple. Theor Appl Genet, 110: 660–668.
[3] Fulton T.M., Chunwongse J., Tanksley S.D. (1995): Microprep protocol for extraction of DNA from tomato andother herbaceous plants. Plant Mol Biol Rep 13 (3): 207-209.
[4] Hayes A.J., Saghai Maroof M.A. (2000) Targeted resistance gene mapping in soybean using modified AFLPs.Theor Appl Genet 100:1279–1283
[5] Hulbert S.H., Webb C.A., Smith S.M., Sun Q (2001): Resistance gene complexes: Evolution and utilization.Annual Rev. Phytopathol., 39:285–312.
[6] Jupe F., Pritchard L., Etherington G.J., MacKenzie K., Cock P.J.A., Wright F., Sharma S.K., Bolser D., BryanG.J., Jones J., Hein I. (2012): Identification and localisation of the NB-LRR gene family within the potatogenome. BMC Genomics, 13, 75.
[7] Kim et al. (2014): Genome sequence of the hot pepper provides insights into the evolution of pungency inCapsicum species. Nature Genetics 46 (3): 270–278
[8] Mantovani P., van der Linden G., Maccaferri M., Sanguineti M.C., Tuberosa R. (2006): Nucleotide-binding site(NBS) profiling of genetic diversity in durum wheat. Genome, 49:1473–1480.
[9] Qin et al. (2014): Whole-genome sequencing of cultivated and wild peppers provides insights into Capsicumdomestication and specialization. Proc Natl Acad Sci U S A 111: 5135–5140.
[10] Salamon P. - Sági Zs. (2004): A paprikát (Capsicum spp.) fertőző tobamovírusok patotípusai és a tobamovirusrezisztencia allélek közötti összefüggések. 9. Tiszántúli Növényvédelmi Fórum, 2004. október 20-24. 256-263.
[11] Salamon Pál (2006): Növények és vírusok kapcsolatai a paprika (Capsicum) -Tobamovírus patoszisztémákban.Doktori értekezés: http://phd.lib.uni-corvinus.hu/343/1/salamon_pal.pdf
[12] Tomita R., Sekine K.T., Mizumoto H., Saks-Moto M., Murai J., Kiba A., Hikichi Y., Suzuki K., Kobayashi K.(2011): Genetic basis for the hierarchical interaction between Tobamo-virus spp. and L resistance gene allelesfrom different pepper species. Molecular Plant-Microbe Interactions, 24 (1): 108–117.
[13] van der Linden C.G., Wouters D.C.A.E., Mihalka V., Kochieva E.Z., Smulders M.J.M., Vosman B. (2004):Efficient targeting of plant disease resistance loci using NBS profiling. Theor Appl Genet 109 (2): 384-393
[14] Vossen J.H., Dezhsetan S., Esselink D., Arens M., Sanz M.J., Verweij W., Verzaux E., van der Linden C.G.(2013): Novel applications of motif-directed profiling to identify disease resistance genes in plants. PlantMethods 9: 37



Copyright (c) 2019 Gradus