Assessment of seed quality and sex determination in hop seedlings using molecular markers

Marília Pereira Machado, Andreza Cerioni Belniaki, André Felipe Bernert, Erik Nunes Gomes, João Carlos Bespalhok Filho, Maristela Panobianco, Cícero Deschamps


Brazil is the world's third largest beer consumer and currently imports all of its hops for the brewing industry. Such a fact justifies the selection of hop genotypes adapted for cultivation locally, which requires high quality seeds and efficient sex determination of the seedlings. The objectives of this study were to develop a methodology to assess hop seed quality and to efficiently determine hop seedling sex through the use of male-specific molecular markers. Freshly harvested hop seeds were germinated with and without pre-chilling (3-5 ° C) for 3, 6 and 12 weeks and then germinated at 20 or 25 ° C in the presence or absence of light, evaluating germination percentage and germination speed index. F1 progenies were obtained from after seed germination in a greenhouse and seedlings sex was determined using male-specific molecular markers. The best conditions for physiological quality assessment of hop seeds used in the present study were pre-chilling for 12 weeks, followed by germination at 25 ° C, and normal seedling counts at 7 and 15 days. The progeny submitted to molecular marker sexing was composed of 61.3% female plants. The established methodologies presented here can be considered efficient and may contribute to expedite hops breeding programs.


Breeding programs; Dioecious plants; Humulus lupulus

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ALEKSANDROV, O. S.; DIVASHUK, M. G.; KARLOV, G. I. Development of a Sex Specific Molecular Marker for Japanese Hop Humulus Japonicus Siebold & Zucc. Russian Journal of Genetics, Springer Nature Switzerland, v.47, p.1016–1020, Aug. 2011. Available from: Accessed Mar. 22, 2019.

BENTSINK, L.; KOORNNEEF, M. Seed dormancy and germination. The Arabidopsis book. v.6, p.1-18, Dec. 2008. Available from: Accessed Jan. 12, 2019.

BRASIL. Ministério da Agricultura, Pecuária e Abastecimento. Regras para Análise de Sementes. Brasília, 2009, 395 p. Available from:

DANILOVA, T. V.; KARLOV, G. I. Application of inter simple sequence repeat (ISSR) polymorphism for detection of sex-specific molecular markers in hop (Humulus lupulus L.). Euphytica, v.151, p.15-21, Oct. 2006. Available from: Accessed Apr. 12, 2019.

DOYLE, J. J.; DOYLE, J. L. Isolation of plant DNA from fresh tissue. Focus, v.12, p.13-15, 1990. Available from: Accessed Feb. 17, 2019.

DURAISAMY, G. S.; MISHRA, A. K.; KOCABEK, T.; MATOUŠEK, J. Identification and characterization of promoters and cis-regulatory elements of genes involved in secondary metabolites production in hop (Humulus lupulus L). Computational Biology and Chemistry, v.64, p.346-352, Feb. 2019Available from: Accessed Feb. 2019.

GOMES, E. M.; MACHADO, M. P.; MIOLA, J.; DESCHAMPS, C. Leaf area and intermittent misting on hop plants propagation by stem cuttings. Revista Colombiana de Ciencias Horticolas, v.12, p.508-513, May. 2018. Available from:

HE, H.; VIDIGAL, D. S.; SNOEK, L. B.; SCHABEL, S.; NIJVEEN, H.; HILHORST, H.; BENTSINK, L. Interaction between parental environment and genotype affects plant and seed performance in Arabidopsis. Journal of Experimental Botany, v.65, p.6603-6615, Dec. 2014. Available from:

INTERNATIONAL SEED TESTING ASSOCIATION (ISTA) International Rules for Seed Testing. ISTA. Bassersdorf, SW, 2015.

INTERNATIONAL HOP GROWERS CONVENTION (IHCG). Economic committee, 2018. Available from: Accessed October 1, 2018.

KAISER, D. K.; MALAVASI, M. M.; MALAVASI, U. C.; DRANSKI, J. A. L.; FREITAS, L.C.N.; KOSMAN, C.R.; ANDRIOLI, K.K. Physiological maturity of seeds and colorimetry of the fruits of Allophylus edulis [(A.St.-Hil., A. Juss. & Cambess.) Hieron. Ex Niederl.]. Journal of Seed Science, Abrates Londrina, v.38, p.92-100, Jun. 2016. Available from: Accessed Mai. 2, 2019.

KARABÍN, M.; HUDCOVÁ, T.; JELÍNEK, L.; DOSTÁLEK, P. Biologically active compounds from hops and prospects for their use. Comprehensive Reviews in Food Science and Food Safety, Wiley Online Library, v.15, p.542-567, Mar. 2016. Available from: Accessed Mai. 8, 2019.

KELLER, K. R. Seed germination in hop, Humulus lupulus L. Agronomy Journal, Wiley Online Library, v.45, p.146-150, Mar. 1953. Available from: Accessed Mai. 2, 2019.

LIBERATORE, C. M.; MATTION, G.; RODOLFI, M.; GANINO, A.; FABBRI, A.; CHIANCONEA, B. Chemical and physical pre-treatments to improve in vitro seed germination of Humulus lupulus L., cv. Columbus. Scientia Horticulturae, Elsevier, v.235, p.86-94, May. 2018. Available from: Accessed Apr. 12, 2019.

MACHADO, M. P.; GOMES, E. N.; FRANCISCO, F.; BERNERT, A. F.; BESPALHOK-FILHO, J. C.; DESCHAMPS, C. Micropropagation and Establishment of Humulus lupulus L. Plantlets Under Field Conditions at Southern Brazil. Journal of Agricultural Science, The Canadian Center of Science and Education, v.10, p.275-281, Jun. 2018. Available from: Accessed Jan. 12, 2019.

MAGUIRE, J. D. Speed of germination - aid in selection and evolution for seedling emergence and vigor. Crop Science, Wiley Online Library, v.2, p.176-177, Mar. 1962. Available from: Accessed Apr. 26, 2019.

MAHARAJAN, T.; CEASAR, S. A.; KRISHNA, T. P. A.; RAMAKRISHNAN, M.; DURAIPANDIYAN, V.; ABDULLA, I. S. Utilization of molecular markers for improving the phosphorus efficiency in crop plants. Plant Breeding, Wiley Online Library, v.137, p.10-26, Oct. 2018. Available from: Accessed Apr. 7, 2019.

NANCE, M. R.; SETZER, W. N. Volatile components of aroma hops (Humulus lupulus L.) commonly used in beer brewing. Journal of Brewing and Distilling, Academic Journals, v.2, p.16-22, Apr. 2011. Available from: Accessed Apr. 22, 2019.

NEWTON, R. J.; HAY, F. R.; ELLIS, R. H. Ecophysiology of seed dormancy and the control of germination in early spring flowering Galanthus nivalis and Narcissus pseudonarcissus (Amaryllidaceae). Botanical Journal of the Linnean Society, Oxford academic, v.177, p. 246-262, Feb. 2015. Available from: Accessed Apr. 7, 2019.

PATZAK, J.; VEJL, P.; SKUPINOVA, S.; NEWADBA, V. Identification of sex in F1 progenies of hop (Humulus lupulus L.). Rostlinna Vyroba, CAAS Agriculture journals, v.48, p.318-321, 2002. Available from: Accessed January 20, 2019.

SAKIYAMA, N. S.; RAMOS, H. C. C.; CAIXETA, E. T. P. Plant breeding with marker-assisted selection in Brazil. Crop Breeding and Applied Biotecnology, Viçosa, v.14, p. 54-60, Mar. 2014. Available from: Accessed January 10, 2019.

SEEFELDER, S.; EHRMAIER, H.; SCHWEIZER, G.; SEIGNER, E. Male and female genetic linkage map of hops, Humulus lupulus. Plant Breeding, Wiley Online Library, v.119, p.249-245, Jun. 2008. Available from: Accessed March 10, 2019.

SIMONS, A. M.; JOHNSTON, M. O. Environmental and genetics sources of diversification in the timing of seed germination: implications for the evolution of bet hedging. Evolution, v.60, p.2280-2292, 2006. Available from: Accessed March 25, 2019.

SINGH, S.; DEY, S. S.; BHATIA, R.; BATLEY, J.; KUMAR, R. Molecular breeding for resistance to black rot [Xanthomonas campestris pv. campestris (Pammel) Dowson] in Brassicas: recent advances. Euphytica, Springer Nature Switzerland, v.214, p.1-17, Oct. 2018. Available from: Accessed March 4, 2019.

SUCIU, T.; SALONTAI, A.; MUNTEAN, L.; FELECAN, V.; VAIDA, L. Recherches concernant la germination des semences de houblon. Notulae Botanicae Horti Agrobotanici, v.9, p.79-84, 1977.

VASCONCELOS, Y. Inovações cervejeiras. Pesquisa FAPESP, v.251, p.18-25, 2017.

VENDEMIATTI, A. Beer Marketing Report, USDA. 2020 . Available from: Accessed Jun. 1, 2020.

XU, Y.; CROUCH, J. H. Marker-assisted selection in plant breeding: from publication to practice. Crop Science, Wiley Online Library, v.48, p.391-407, Mar. 2008. Available from: Accessed Apr. 14, 2019.


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