Summary
Cannabis sativa L. is a dioecious species with sexual dimorphism occurring in a late stage of plant development. Sex is determined by heteromorphic chromosomes (X and Y): male is the heterogametic sex (XY) and female is the homogametic one (XX). The sexual phenotype of Cannabis often shows some flexibility leading to the differentiation of hermaphrodite flowers or bisexual inflorescences (monoecious phenotype). Sex is considered an important trait for hemp genetic improvement; therefore, the study of the mechanism of sexual differentiation is of paramount interest in hemp research. A morphological and molecular study of Cannabis sativa sexual differentiation has been carried out in the Italian dioecious cultivar Fibranova.
Microscopic analysis of male and female apices revealed that their reproductive commitment may occur as soon as the leaves of the fourth node emerge; the genetic expression of male and female apices at this stage has been compared by cDNA-AFLP. A rapid method for the early sex discrimination has been developed, based on the PCR amplification of a male-specific SCAR marker directly from a tissue fragment.
Five of the several cDNA-AFLP polymorphic fragments identified have been confirmed to be differentially expressed in male and female apices at the fourth node. Cloning and sequencing revealed that they belong to nine different mRNAs that were all induced in the female apices at this stage. Four out of them showed a high degree of similarity with known sequences: a putative permease, a SMT3-like protein, a putative kinesin and a RAC-GTP binding protein.
Similar content being viewed by others
References
Altschul, S.F., W. Gish, W. Miller, E.W. Myers & D.J. Lipman, 1990. Basic local alignment search tool. J Mol Biol 215: 403–410.
Bachem, C.W.B., R.S. van der Hoeven, S.M. de Bruijn, D. Vreugdenhil, M. Zabeau & R.G.F. Visser, 1996. Visualization of differential gene expression using a novel method of RNA finger-printing based on AFLP: Analysis of gene expression during potato tuber development. Plant J 9: 745–753.
Bachem, C.W.B., R.J.F.J. Oomen & R.G.F. Visser, 1998. Transcript imaging with cDNA-AFLP: A step-by-step protocol. Plant Mol Biol Rep 16: 157–173.
Baskin, T.I., 2000. The cytoskeleton. In: B. Buchanan, W. Gruissem & R. Jones, (Eds.), Biochemisty and Molecular Biology of Plants, pp. 219–221. American Society of Plant Physiologists, Rockville, Maryland.
Boecke, J.D., 1989. Transposable elements in Saccharomyces cerevisiae. In: D.E. Berg & M.M. Howe (Ed.), Mobile DNA, pp. 335–374. American Society of Microbiology, Washington DC.
Bredemann, G., 1938. Züchtung des Hanfes auf Fasergehaltes. Die Ergebnisse des Jahres 1937. Faserforschung 4: 239–258.
Charlesworth, D. & D.S. Guttman, 1999. The evolution of dioecy and plant sex chromosome systems. In: C.C. Ainsworth (Ed.), Sex Determination in Flowering Plants, pp. 25–33. Bios Scientific Publishers, Oxford, Washington DC.
Corpet, F., 1988. Multiple sequence alignment with hierarchical clustering. Nucl Acids Res 16(22): 10881–10890.
Donnison, I.S. & S.G. Grant, 1999. Male sex-specific DNA in Silene latifolia and other dioecious plant species. In: C.C. Ainsworth (Ed.), Sex Determination in Flowering Plants, pp. 73–88. Bios Scientific Publishers, Oxford–Washington DC.
Faeti, V., G. Mandolino & P. Ranalli, 1996. Genetic diversity of Cannabis sativa germplasm based on RAPD markers. Plant Breeding 115: 367–370.
Forapani, S., A. Carboni, C. Paoletti, V.M.C. Moliterni, P. Ranalli & G. Mandolino, 2001. Comparison of hemp varieties using random amplified polymorphic DNA markers. Crop Sci 41: 1682–1688.
Galoch, E., 1980. The hormonal control of sex differentiation in dioecious plants of hemp (Cannabis sativa). Acta Physiol Plant vol II (n.1): 31–39.
Grant, S., A. Houben, B. Vyskot, J. Siroky, W.H. Pan, J. Macas & H. Saedler, 1994. Genetics of sex determination in flowering plants. Dev Genet 15: 214–230.
Harley, C.B., 1997. Hybridization of Oligo(dT) to RNA on nitrocellulose. Gene Anal Tech 4: 17–22.
Hartings, H., 1999. High resolution fingerprinting of transcribed genes by means of a modified cDNA-AFLP method. Maydica 44: 179–186.
Klimyuk, V.I., B.J. Carrolm, C.M. Thomas & J.D. Jones, 1993. Alkali treatment for rapid preparation of plant material for reliable PCR analysis. Plant J 3: 493–494.
Lacombe, J.-P., 1980. Discrimination des sexes en fonction de caractères végétatifs précoces chez le Chanvre dioïque (Cannabis sativa L.). Physiol Vég 18: 419–430.
Mandolino, G., A. Carboni, S. Forapani & P. Ranalli, 1998. DNA markers associated with sex phenotype in hemp (Cannabis sativa L.). In: Proc Bast Fibrous Plants Today and Tomorrow, St Petersburg, September 28–30, pp. 197–201.
Mandolino, G., A. Carboni, S. Forapani, V. Faeti & P. Ranalli, 1999. Identification of DNA markers linked to the male sex in dioecious hemp (Cannabis sativa L.). Theor Appl Genet 98: 86–92.
Mandolino, G., A. Carboni, M. Bagatta, V.M.C. Moliterni & P. Ranalli, 2002. Occurrence and frequency of putatively Y chromosome linked DNA markers in Cannabis sativa L. Euphytica 126: 211–216.
Mandolino, G. & P. Ranalli, 2002. The applications of molecular markers in genetics and breeding of hemp. J Ind Hemp 7: 7–24.
Mediavilla, V., M. Jonquera, I. Schmid-Slembrouck & A. Soldati, 1998. A decimal code for growth stages of hemp (Cannabis sativa L.). J Ind Hemp Ass 5: 65–74.
Mohan Ram, H.Y. & R. Nath, 1964. The morphology and embryology of Cannabis sativa L. Phytomorphology 14: 414–429.
Mohan Ram, H.Y. & R. Sett, 1982a. Modification of growth and sex expression in Cannabis sativa by aminoethoxyvinylglycine and etephon. Z Planzenphysiol Bd 105: 165–172.
Mohan Ram, H.Y. & R. Sett, 1982b. Induction of fertile male flowers in genetically female Cannabis sativa plants by silver nitrate and silver thiosulphate anionic complex. Theor Appl Genet 62: 369–375.
Peil, A., H. Flachowsky, E. Schumann & W.E. Weber, 2003. Sex-linked AFLP markers indicate a pseudoautosomal region in hemp (Cannabis sativa L.). Theor Appl Genet 107: 102–109.
Sakamoto, K., Y. Akiyama, K. Fukui, H. Kamada & S. Satoh, 1998. Characterization, genome size and morphology of sex chromosomes in hemp (Cannabis sativa L.). Cytologia 63: 459– 464.
Sakamoto, K., N. Ohmido, K. Fukui, H. Kamada & S. Satoh, 2000. Site-specific accumulation of a LINE-like retrotrasposon in a sex chromosome of the dioecious plant Cannabis sativa. Plant Mol Biol 44: 723–732.
Sambrook, J., E.F. Fritisch & T. Maniatis, 1989. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press.
Scutt, C.P., S.E. Robertson, M.E. Willis, K. Kamisugi, Y. Li, M.R. Shenton, R.H. Smith, H. Martin & P.M. Gilmartin, 1999. Molecular approaches to the study of sex determination in dioecious Silene latifolia. In: C.C. Ainsworth (Ed.), Sex Determination in Flowering Plants, pp. 51–72. Bios Scientific Publishers, Oxford, Washington DC.
Valster, A.H., P.K. Hepler & J. Chernoff, 2000. Plant GTP-ases: The Rhos in bloom. Trends Cell Biol 10: 141–146.
Tao, L., A.Y. Cheung & H. Wu, 2002. Plant Rac-Like GTPases are activated by auxin and mediate auxin-responsive gene expression. Plant Cell 14: 2745–2760.
The Angiosperm Phylogeny Group, 2003. An update of the Angiosperm Phylogeny Group classification for the orders and families of flowering plants: APGII. Bot J Linn Soc 141: 399–436.
Westgaard, M., 1958. The mechanism of sex determination in dioecious plants. Adv Genet 9: 217–281.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Moliterni, V.M.C., Cattivelli, L., Ranalli, P. et al. The sexual differentiation of Cannabis sativa L.: A morphological and molecular study. Euphytica 140, 95–106 (2004). https://doi.org/10.1007/s10681-004-4758-7
Issue Date:
DOI: https://doi.org/10.1007/s10681-004-4758-7