Tropane alkaloids and terpenes synthase genes of Datura stramonium (Solanaceae)

Abstract

Background Plants have evolved physical–chemical defense to prevent/diminish damage by their enemies. Chemical defense involves the synthesis’ pathways of specialized toxic, repellent, or anti-nutritive metabolites to herbivores. Molecular evolutionary studies have revealed the origin of new genes, acquisition and functional diversification along time in different plant lineages. Methods Using bioinformatic tools we analyze gene divergence of tropane alkaloids (TAs) and terpene synthases (TPSs) in Datura stramonium and other species of Solanaceae; compared gene and amino acids sequence of TAs and TPSs on genomes, cDNA and proteins sequences of Viridiplantae. We analyzed two recently assembled genomes of D. stramonium (Ticumán and Teotihuacán), transcriptomes of Datura metel and genomes of other Solanaceae. Hence, we analyzed variation of TAs and TPSs to infer genes involved in plant defense and plant responses before stress. We analyzed protein modeling and molecular docking to predict interactions between H6H and ligand; we translated the sequences (Teo19488, Tic8550 and Tic8549) obtained from the two genomes of D. stramonium by using Swiss-Model and Ramachandran plot and MolProbity structure validation of Teo19488 protein model. Results For TAs, we detected an expansion event in the tropinone reductase II (TRII) and the ratio synonymous/non-synonymous substitutions indicate positive selection. In contrast, a contraction event and negative selection was detected in tropinone reductase I (TRI). In Hy-oscyamine 6 b-hydroxylase (H6H), enzyme involved in the production of tropane alkaloids atropine and scopolamine, the synonymous/non-synonymous substitution ratio in its dominion indicates positive selection. For terpenes (TPS), we found 18 DsTPS in D. stramomiun and seven in D. metel; evolutionary analyses detected positive selection in TPS10.1 and TPS10.2 of D. stramonium and D. metel. Comparison of copies of TPSs in D. stramonium detected variation among them in the binding site. Duplication events and differentiation of TAs and TPSs of D. stramonium, as compared to other Solanaceae, suggest their possible involvement on adaptive evolution of defense to herbivores. Protein modeling and docking show that the three protein structures obtained of DsH6H from Teo19488, Tic-8550 and Tic8549 maintain the same interactions and the union site of 2OG-FeII_Oxy with the Hy-o ligand as in 6TTM of D. metel. Conclusion Our results indicate differences in the number of gene copies involved in the synthesis of tropane alkaloids, between the genomes of D. stramonium from two Mexican populations. More copies of genes related to the synthesis of tropane alkaloids (TRI, TRII, H6H, PMT) are found in D. stramonium as compared to Viridiplantae. Likewise, for terpene synthases (TPS), TPS-10 is duplicated in D. stramonium and D. metel. Further studies should be directed to experimentally assess gain (overexpression) or loss (silencing) of function of duplicated genes.

Gene family evolution and natural selection signatures in Datura spp. (Solanaceae)

“…Moreover, several terpenoids are volatilized to attract predators and parasitoids of herbivores (Singh and Sharma, 2015;Lackus et al, 2018;Boncan et al, 2020). Since the four Datura species are attacked by the specialist herbivore L. trilineata daturaphila (Kogan and Goeden, 1970;Castillo et al, 2014;De-la-Cruz et al, 2020b;Goldberg et al, 2020), we speculate that the positive selection (D. pruinosa and D. wrightii), expansion (only in D. wrightii) and physicochemical divergence (only in D. stramonium) detected for terpenoids could be related how these species cope with their natural enemies such as herbivores, pathogens and viruses (Pichersky and Raguso, 2018;Karunanithi and Zerbe, 2019;Boncan et al, 2020;De-la-Cruz et al, 2020a,b;Goldberg et al, 2020Goldberg et al, , 2021Zhou and Pichersky, 2020;Velázquez-Márquez et al, 2021). Thus, positive selection of terpenoid proteins in almost all Datura species could suggest species-specific evolutionary dynamics related to the different biotic and abiotic pressures faced by each Datura species in their natural environments (Castillo et al, 2014;Kariñho-Betancourt et al, 2015;Pichersky and Raguso, 2018;Karunanithi and Zerbe, 2019;De-la-Cruz et al, 2020b;Goldberg et al, 2020Goldberg et al, , 2021De-la-Cruz et al, 2021).…”

“…Within the Solanaceae family, the species of the genus Datura produce high concentrations of tropane alkaloids (Jirschitzka et al, 2012;Kohnen-Johannsen and Kayser, 2019;Céspedes-Méndez et al, 2021), metabolites long used as medicines (Roddick, 1991;Geeta and Gharaibeh, 2007;Benítez et al, 2018;Sahu et al, 2022). Ecological evidence indicates that some of these chemical compounds, as well as some terpenoids, play central roles as defenses against herbivores and possibly against pathogens, viruses, and fungi in Datura (Shonle and Bergelson, 2000;Hare and Elle, 2001;Hare and Smith, 2005;Castillo et al, 2014;Kariñho-Betancourt et al, 2015;Miranda-Pérez et al, 2016;De-la-Cruz et al, 2020a, 2021Goldberg et al, 2020;Céspedes-Méndez et al, 2021;Velázquez-Márquez et al, 2021). Furthermore, there is evidence that these species produce different concentrations of total tropane alkaloids (Doncheva et al, 2006;Kariñho-Betancourt et al, 2015).…”

Modifying Anthocyanins Biosynthesis in Tomato Hairy Roots: A Test Bed for Plant Resistance to Ionizing Radiation and Antioxidant Properties in Space

“…Among upregulated DEGS, Solyc01g105880.4.1 and Solyc09g092470.2.1 (Log 2 Fc = 3.46 and 5.69, respectively; tomato monoterpenoid synthases 2 TPS4 and TPS14 ), are involved in the biosynthesis of monoterpenes and sesquiterpenes, respectively. TPS4 has been found to be upregulated in tomato cultivars resistant to Fusarium oxysporum ( Velázquez-Márquez et al, 2021 ). Solyc11g017240.2.1 (Log 2 Fc = 3.78; tomato chorismate mutase 2 SlCM2 ), catalyzes the first step of the shikimate pathway from phenylalanine to the volatile compounds responsible for tomato fruit aroma and quality and defense from biotic and abiotic stress response ( Tzin et al, 2015 ).…”

A comparative investigation of bioactive compounds present in Datura stramonium L. leaves extract in various solvents (ethanol, aqua, and ether) using TLC, IR, and GC-MS analytical techniques

“…This plants consist of sixty-four various types of tropane alkaloids (Srivastava & Srivastava, 2020). Using bioinformatic tools and applications it was analyzed gene divergence of tropane alkaloids (TAs), terpene synthases (TPSs) and other members of Solanaceae Family (Velazquez-Marquez, de-la-Cruz, Tapia-Lopez, & Nunez-Farfan, 2021).…”