Abstract
Salvia is an important genus from the Lamiaceae with approximately 1,000 species. This genus is distributed globally and cultivated for ornamental, culinary, and medicinal uses. We report the construction of the first fingerprinting array for Salvia species enriched with polymorphic and divergent DNA sequences and demonstrate the potential of this array for fingerprinting several economically important members of this genus. In order to generate the Salvia subtracted diversity array (SDA) a suppression subtractive hybridization (SSH) was performed between a pool of Salvia species and a pool of angiosperms and non-angiosperms to selectively isolate Salvia-specific sequences. A total of 285-subtracted genomic DNA (gDNA) fragments were amplified and arrayed. DNA fingerprints were obtained for fifteen Salvia genotypes including three that were not part of the original subtraction pool. Hierarchical cluster analysis indicated that the Salvia-specific SDA was capable of differentiating S. officinalis and S. miltiorrhiza from their closely related species and was also able to reveal genetic relationships consistent with geographical origins. In addition, this approach was capable of isolating highly polymorphic sequences from chloroplast and nuclear DNA without preliminary sequence information. Therefore, SDA is a powerful technique for fingerprinting non-model plants and for identifying new polymorphic loci that may be developed as potential molecular markers.
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Walker, J. B., Sytsma, K. J., Treutlein, J., & Wink, M. (2004). Salvia (Lamiaceae) is not monophyletic: Implications for the systematics, radiation, and ecological specializations of Salvia and tribe Mentheae. American Journal of Botany, 91, 1115–1125.
Bruna, S., Giovannini, A., Benedetti, L. D., Principato, M. C., & Ruffoni, B. (2006). Molecular analysis of Salvia Spp. through RAPD markers. ISHS Acta Horticulturae, 723, 157–160.
Topcu, G. (2006). Bioactive triterpenoids from Salvia species. Journal of Natural Products, 69, 482–487.
Echeverrigaray, S., & Agostini, G. (2006). Genetic relationships between commercial cultivars and Brazilian accessions of Salvia officinalis L. based on RAPD markers. Revista Brasileira de Plantas Medicinais, 8, 13–17.
Longaray-Delamare, A. P., Moschen-Pistorello, I. T., Artico, L., Atti-Serafini, L., & Echeverrigaray, S. (2007). Antibacterial activity of the essential oils of Salvia officinalis L. and Salvia triloba L, cultivated in south Brazil. Food Chemistry, 100, 603–608.
Cai, Z., Lee, F. S. C., Wang, X. R., & Yu, W. J. (2002). A capsule review of recent studies on the application of mass spectrometry in the analysis of Chinese medicinal herbs. Journal of Mass Spectrometry, 37, 1013–1024.
Reales, A., Rivera, D., Palazon, J. A., & Obon, C. (2004). Numerical taxonomy study of Salvia sect. Salvia (Labiatae). Botanical Journal of the Linnean Society, 145, 353–371.
Zhong, G. X., Li, P., Zeng, L. J., Guan, J., Li, D. Q., & Li, S. P. (2009). Chemical characteristics of Salvia miltiorrhiza (Danshen) collected from different locations in China. Journal of Agriculture and Food Chemistry, 57, 6879–6887.
Kiran, U., Khan, S., Mirza, K. J., Ram, M., & Abdin, M. Z. (2010). SCAR markers: A potential tool for authentication of herbal drugs. Fitoterapia, 81, 969–976.
Joshi, K., Chavan, P., Warude, D., & Patwardhan, B. (2004). Molecular markers in herbal drug technology. Current Science India, 87, 159–165.
Canter, P. H., Thomas, H., & Ernst, E. (2005). Bringing medicinal plants into cultivation: Opportunities and challenges for biotechnology. Trends in Biotechnology, 23, 180–185.
Böszörményi, A., Héthelyi, E., Farkas, A., Horváth, G., Papp, N., Lemberkovics, E., et al. (2009). Chemical and genetic relationships among sage (Salvia officinalis L.) cultivars and Judean sage (Salvia judaica Boiss.). Journal of Agriculture and Food Chemistry, 57, 4663–4667.
Skoula, M., El Hilali, I., & Makris, A. M. (1999). Evaluation of the genetic diversity of Salvia fruticosa Mill. clones using RAPD markers and comparison with the essential oil profiles. Biochemical Systematics and Ecology, 27, 559–568.
Braglia, L., Casabianca, V., De Benedetti, L., Pecchioni, N., Mercuri, A., Cervelli, C., et al. (2011). Amplified fragment length polymorphism markers for DNA fingerprinting in the genus Salvia. Plant Biosystems, 145, 274–277.
Jaccoud, D., Peng, K., Feinstein, D., & Kilian, A. (2001). Diversity arrays: A solid state technology for sequence information independent genotyping. Nucleic Acids Research, 29, E25.
Karaca, M., Ince, A. G., Ay, S. T., Turgut, K., & Onus, A. N. (2008). PCR-RFLP and DAMD-PCR genotyping for Salvia species. Journal of the Science of Food and Agriculture, 88, 2508–2516.
Xu, H., Wang, Z. T., Cheng, K. T., Wu, T., Gu, L. H., & Hu, Z. B. (2009). Comparison of rDNA ITS sequences and tanshinones between Salvia miltiorrhiza populations and Salvia species. Botanical Studies, 50, 127–135.
Han, J. P., Liu, C., Li, M. H., Shi, L. C., Song, J. Y., Yao, H., et al. (2010). Relationship between DNA barcoding and chemical classification of Salvia medicinal herbs. Chinese Herbal Medicines, 2, 16–29.
Takano, A., & Okada, H. (2010). Phylogenetic relationships among subgenera, species, and varieties of Japanese Salvia L. (Lamiaceae). Journal of Plant Research, 124, 245–252.
Kingsley, M. T., Straub, T. M., Call, D. R., Daly, D. S., Wunschel, S. C., & Chandler, D. P. (2002). Fingerprinting closely related Xanthomonas pathovars with random nonamer oligonucleotide microarrays. Applied and Environment Microbiology, 68, 6361–6370.
Li, T. -X., Wang, J. -K., Bai, Y. -F., & Lu, Z. -H. (2006). Diversity suppression-subtractive hybridization array for profiling genomic DNA polymorphisms. Journal of Integrative Plant Biology, 48, 460–467.
Jayasinghe, R., Kong, S., Coram, T. E., Kaganovitch, J., Xue, C. C., Li, C. G., et al. (2007). Construction and validation of a prototype microarray for efficient and high-throughput genotyping of angiosperms. Plant Biotechnology Journal, 5, 282–289.
Jayasinghe, R., Hai Niu, L., Coram, T. E., Kong, S., Kaganovitch, J., Xue, C. C. L., et al. (2009). Effectiveness of an innovative prototype subtracted diversity array (SDA) for fingerprinting plant species of medicinal importance. Planta Medica, 75, 1180–1185.
Mantri, N., Olarte, A., Li, C. G., Xue, C., & Pang, E. C. K. (2012). Fingerprinting the Asterid species using subtracted diversity array reveals novel species-specific sequences. PLoS ONE, 7, e34873.
Duncan, B. D., & Isaac, G. (1994). Ferns and allied plants of Victoria, Tasmania and South Australia. Melbourne University Press in association with Monash University, Carlton.
Li, C. G., Sheng, S. J., Pang, E. C., Marriot, P., May, B., Zhou, S. F., et al. (2009). Cultivar variations of Australian-grown Danshen (Salvia miltiorrhiza): Bioactive markers and root yields. Chemistry & Biodiversity, 6, 170–181.
Doyle, J. J., & Doyle, J. L. (1987). A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin, 19, 1–15.
Bremer, B., Bremer, Kr., Chase, M., Fay, M., Reveal, J., Soltis, D., et al. (2009). An update of the Angiosperm Phylogeny Group classification for the orders and families of flowering plants: APG III. Botanical Journal of the Linnean Society, 161, 105–121.
Coram, T. E., & Pang, E. C. K. (2005). Isolation and analysis of candidate ascochyta blight defence genes in chickpea. Part I. Generation and analysis of an expressed sequence tag (EST) library. Physiological and Molecular Plant Pathology, 66, 192–200.
National Center of Biotechnology Information (www.ncbi.nlm.nih.gov).
Gadgil, C., Rink, A., Beattie, C., & Hu, W. S. (2002). A mathematical model for suppression subtractive hybridization. Comparative and Functional Genomics, 3, 405–422.
Lezar, S., Myburg, A. A., Berger, D. K., Wingfield, M. J., & Wingfield, B. D. (2004). Development and assessment of microarray-based DNA fingerprinting in Eucalyptus grandis. Theoretical and Applied Genetics, 109, 1329–1336.
Wittenberg, A. H. J., van der Lee, T., Cayla, C., Kilian, A., Visser, R. G. F., & Schouten, H. J. (2005). Validation of the high-throughput marker technology DArT using the model plant Arabidopsis thaliana. Molecular Genetics and Genomics, 274, 30–39.
Claßen-Bockhoff, R. (2007). Floral construction and pollination biology in the Lamiaceae. Annals of Botany, 100, 359–360.
Pound, L. M., Wallwork, M. A. B., Potts, B. M., & Sedgley, M. (2002). Self-incompatibility in Eucalyptus globulus ssp. globulus (Myrtaceae). Australian Journal of Botany, 50, 365–372.
Dudai, N., Lewinsohn, E., Larkov, O., Katzir, I., Ravid, U., Chaimovitsh, D., et al. (1999). Dynamics of yield components and essential oil production in a commercial hybrid sage (Salvia officinalis x Salvia fruticosa cv. Newe Ya’ar no. 4). Journal of Agriculture and Food Chemistry, 47, 4341–4345.
Cao, J., Wei, Y. J., Qi, L. W., Li, P., Qian, Z. M., Luo, H. W., et al. (2008). Determination of fifteen bioactive components in Radix et Rhizoma Salviae Miltiorrhizae by high-performance liquid chromatography with ultraviolet and mass spectrometric detection. Biomedical Chromatography, 22, 164–172.
Li, M. H. (2008). Investigation of Danshen and related medicinal plants in China. Journal of Ethnopharmacology, 120, 419–426.
Walker, J. B., & Sytsma, K. J. (2007). Staminal evolution in the genus Salvia (Lamiaceae): Molecular phylogenetic evidence for multiple origins of the staminal lever. Annals of Botany, 100, 375–391.
Lee, H. L., Jansen, R. K., Chumley, T. W., & Kim, K. J. (2007). Gene relocations within chloroplast genomes of Jasminum and Menodora (Oleaceae) are due to multiple, overlapping inversions. Molecular Biology and Evolution, 24, 1161–1180.
Fazekas, A. J., Kesanakurti, P. R., Burgess, K. S., Percy, D. M., Graham, S. W., Barrett, S. C. H., et al. (2009). Are plant species inherently harder to discriminate than animal species using DNA barcoding markers? Molecular Ecology Resources, 9, 130–139.
Chase, M. W., Salamin, N., Wilkinson, M., Dunwell, J. M., Kesanakurthi, R. P., Haidar, N., et al. (2005). Land plants and DNA barcodes: Short-term and long-term goals. Philosophical Transactions of the Royal Society of London, Series B: Biological Sciences, 360, 1889–1895.
Alvarez, I., Costa, A., & Feliner, G. N. (2008). Selecting single-copy nuclear genes for plant phylogenetics: A preliminary analysis for the Senecioneae (Asteraceae). Journal of Molecular Evolution, 66, 276–291.
Pang, E. (2011). Using frontier technologies for the quality assurance of medicinal herbs (pp. 1–21). RIRDC publications. RMIT University, Melbourne.
Bertea, C. M., Luciano, P., Bossi, S., Leoni, F., Baiocchi, C., Medana, C., et al. (2006). PCR and PCR-RFLP of the 5S-rRNA-NTS region and salvinorin A analyses for the rapid and unequivocal determination of Salvia divinorum. Phytochemistry, 67, 371–378.
De Mattia, F., Bruni, I., Galimberti, A., Cattaneo, F., Casiraghi, M., & Labra, M. (2011). A comparative study of different DNA barcoding markers for the identification of some members of Lamiaceae. Food Research International, 44, 693–702.
Deng, K. J., Zhang, Y., Xiong, B. Q., Peng, J. H., Zhang, T., Zhao, X. N., et al. (2009). Identification, characterization and utilization of simple sequence repeat markers derived from Salvia miltiorrhiza expressed sequence tags. Yao Xue Xue Bao [Acta Pharmaceutica Sinica], 44, 1165–1172.
Acknowledgments
The authors gratefully acknowledge the support from the Rural Industries Research and Development Corporation, RMIT University and the Australian Postgraduate Scholarship awarded to Alexandra Olarte. We acknowledge the technical support from A/Prof. Reg Lehmann from MediHerb Australia and Claudia Salazar for their assistance with the graphics in this study.
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Olarte, A., Mantri, N., Nugent, G. et al. A gDNA Microarray for Genotyping Salvia Species. Mol Biotechnol 54, 770–783 (2013). https://doi.org/10.1007/s12033-012-9625-5
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DOI: https://doi.org/10.1007/s12033-012-9625-5