A versatile PCR marker for pungency in Capsicum spp.
Pungency in Capsicum spp. is an important quality trait for pepper breeding. The perception of pungency in pepper is due to the presence of a group of compounds named capsaicinoids, only found within the Capsicum genus. How pungency is controlled at genetic and molecular levels has not been completely elucidated. The use of molecular markers to assess pungency trait is required for molecular breeding, despite the difficulty of development of universal markers for this trait. In this work, a DNA sequence possibly related to pungency with a high similarity to Pun1 locus was studied, and sequence analysis of this homolog revealed a 15 bp deletion in non-pungent pepper accessions. An allele-specific pair of primers was designed and specific fragments of 479 bp from non-pungent and 494 bp from pungent accessions were obtained. Polymorphism of this marker, named MAP1, was tested in a wide range of accessions, belonging to several Capsicum species, including pungent and non-pungent accessions of C. annuum L., and pungent accessions of C. chinense, C. baccatum, C. frutescens, C. pubescens, C. galapagoense, C. eximium, C. tovarii, C. cardenasii, and C. chacoense. All these Capsicum accessions were correctly discriminated. The marker suitability to assess pungency in domesticated and wild Capsicum species was demonstrated, and therefore it will be very useful in marker assisted selection (MAS). Moreover, MAP1 was located in a saturated pepper linkage map and its possible relationship with the Pun1 locus has been discussed. Among the available markers for this complex quality trait, the marker developed in this study is the most universal so far.
KeywordsCapsicum spp. Marker assisted selection Molecular marker Pungency
This research was financially supported by “Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria” (INIA)—Spanish Ministry of Science and Innovation (MICINN), grant RTA2008-00095-00-00 and grant A16 for Aragon Government. MJ. R.-M. was supported for a INIA Phd fellowship. We are grateful to M. R. Navarro and E. Fuertes (CITA, Spain) for technical support and M. Carravedo (CITA, Spain) for providing most of the Capscicum seeds.
- Bosland PW, Baral JB (2007) ‘Bhut Jolokia’—The world’s hottest known chile pepper is a putative naturally occurring interspecific hybrid. HortScience 42(2):222–224Google Scholar
- Deshpande RB (1935) Studies in Indian chillies: 4 inheritance of pungency in Capsicum annuum L. Indian J Agric Sci 5:513–516Google Scholar
- Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 19:11–15Google Scholar
- Drummond AJ, Ashton B, Buxton S, Cheung M, Cooper A, Duran C, Field M, Heled J, Kearse M, Markowitz S, Moir R, Stones-Havas S, Sturrock S, Thierer T, Wilson A (2010) Geneious v5.3. Available at http://www.geneious.com
- Estrada B, Pomar F, Díaz J, Merino F, Bernal MA (1998) Effects of mineral fertilizer supplementation on fruits development and pungency in “Padrón” peppers. J Hortic Sci Biotechnol 73:493–497Google Scholar
- Garcés-Claver A, Gil Ortega R, Arnedo-Andrés MS (2007b) Characterization of Capsicum genotypes for pungency trait, and pungency-related DNA sequences. In: Niemirowicz-Szczytt K (ed) Progress in research on Capsicum and eggplant. Warsaw University of Life Sciences Press, Warsaw, pp 181–186Google Scholar
- Harvell K, Bosland PB (1997) The environment and cultural practices produces a significant effect on pungency of chile. HortScience 32:1292Google Scholar
- Paran I, Akler T, Jones G (2010) QTLs for capsaicinoids content in Capsicum. In: Prohens J, Rodriguez-Burruezo A (eds) Advances in genetics and breeding of Capsicum and eggplant. Universidad Politécnica de Valencia, Valencia, pp 273–278Google Scholar
- Park SW, An SJ, Yang HB, Kwon JK, Kang BC (2009) Optimization of high resolution melting analysis and discovery of single nucleotide polymorphism in Capsicum. Hort Environ Biotechnol 50(1):31–39Google Scholar
- Rodríguez-Maza MJ, Garcés-Claver A, Arnedo-Andrés MS (2010) Allelic variation in a putative gene related to pungency in pepper (Capsicum spp.). In: Rallo L et al (eds) Book of abstracts, vol 2. ISHS, Lisboa, p 251Google Scholar
- Rozen S, Skaletsky H (2000) Primer3 on the web for general users and for biologist programmers. In: Krawetz S, Misener S (eds) Bioinformatics methods and protocols: methods in molecular biology. Humana Press, Totowa, pp 365–386Google Scholar
- Ruiz-Lau N, Medina-Lara F, Minero-Garcia Y, Torres-Tapia LW, Peraza-Sanchez SR, Martinez-Estevez M (2011) Capsaicinoids are absent in habanero pepper vegetative organs (Capsicum chinense Jacq.). HortScience 45(2):323–326Google Scholar
- Scoville WL (1912) Note on Capsicum. J Am Pharm Assoc 1:453Google Scholar
- Stewart C, Mazourek M, Stellari GM, O’Connell M, Jahn MM (2007) Genetic control of pungency in C. chinense via the Pun1 locus. Plant J 58:979–991Google Scholar
- Sugita T, Kinoshita T, Kawano T, Yuji K, Yamaguchi K, Nagata R, Shimizu A, Chen L, Kawasaki S, Todoroki A (2005) Rapid construction of a linkage map using high-efficiency genome scanning/AFLP and RAPD, based on an intraspecific, doubled-haploid population of Capsicum annuum. Breed Sci 55:287–295CrossRefGoogle Scholar
- Suzuki T, Fujiwake H, Iwai K (1980) Intracellular localization of capsaicin and its analogues, capsaicinoid, in Capsicum fruit 1. Microscopic investigation of the structure of the placenta of Capsicum annuum var. annuum cv. Karayatsubusa. Plant Cell Physiol 21:839Google Scholar
- Truong HTH, Kim KT, Kim S, Kim HR, Cho MC, Woo JG (2009) Development of gene-based markers for the Pun1 pungency gene in pepper (Capsicum spp.) for marker-assisted selection. Hort Environ Biotechnol 50(4):358–365Google Scholar