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Solanum tuberosum L. phytoene synthase genes (StPSY1, StPSY2, StPSY3) participate in the potato plant’s response to cold stress
- Authors: Kulakova A.V.1, Shchennikova A.V.1, Kochieva E.Z.1
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Affiliations:
- Institute of Bioengineering, Federal Research Center “Fundamentals of Biotechnology” of the Russian Academy of Sciences
- Issue: Vol 516, No 1 (2024)
- Pages: 3-9
- Section: Articles
- URL: https://journal-vniispk.ru/2686-7389/article/view/263908
- DOI: https://doi.org/10.31857/S2686738924030019
- EDN: https://elibrary.ru/VUVRYO
- ID: 263908
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Abstract
The structure and phylogeny of the genes StPSY1, StPSY2 and StPSY3, encoding phytoene synthases from Solanum tuberosum L., were characterized. The expression of these genes in potato seedlings was studied in response to exposure to cold stress in the dark phase of the diurnal cycle as an imitation of night cooling. It was found that all three genes are activated when the temperature decreases, and the greatest response is observed for StPSY1. The response of the StPSY3 gene to cold stress and photoperiod has been demonstrated for the first time. A search was carried out for cis-regulatory elements in the promoter region and 5´-UTR of the StPSY genes and it was shown that the regulation of all three genes is associated with the response to light. The high level of low-temperature activation of the StPSY1 gene may be associated with the presence of cis-elements associated with sensitivity to cold and ABA.
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About the authors
A. V. Kulakova
Institute of Bioengineering, Federal Research Center “Fundamentals of Biotechnology” of the Russian Academy of Sciences
Author for correspondence.
Email: kulakova_97@mail.ru
Russian Federation, Moscow
A. V. Shchennikova
Institute of Bioengineering, Federal Research Center “Fundamentals of Biotechnology” of the Russian Academy of Sciences
Email: kulakova_97@mail.ru
Russian Federation, Moscow
E. Z. Kochieva
Institute of Bioengineering, Federal Research Center “Fundamentals of Biotechnology” of the Russian Academy of Sciences
Email: kulakova_97@mail.ru
Russian Federation, Moscow
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Fig. 1. a – Comparative alignment of amino acid sequences of StPSY1, StPSY2 and StPSY3. The solid line underlines the phytoene synthase domain of PLN02632, the dashed line – the beginning of the domain in StPSY3 (NCBI_CDD, https://www.ncbi.nlm.nih.gov/). Putative cleavage sites of the transit peptide are in the boxes. Amino acids variable between the three proteins are highlighted in blue, and conserved in two of the three proteins are highlighted in red. b – Phylogenetic dendrogram constructed by comparing 13 amino acid sequences using the maximum likelihood method (Maximum Likelihood, JTT model, bootstrap 1000) in the MEGA7 program (https://www.megasoftware.net/). The length of branches is measured by the number of substitutions per site, and the percentage of trees in which related taxa are grouped together is indicated at the base of branches. The sequences of phytoene synthases PSY1, PSY2, and PSY3 homologues from Solanaceae species S. tuberosum (St), S. lycopersicum (Sl), Capsicum annuum (Ca), and a model species of Dicotyledons, A. thaliana (At), were used for the analysis. NCBI (https://www.ncbi.nlm.nih.gov/) or SolGenomics (https://www.solgenomics.net/) accession numbers are given next to the names of the analyzed proteins.
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Fig. 2. Expression level of the StPSY1 (NCBI Gene ID 102593756), StPSY2 (102589336), and StPSY3 (102603193) genes in the leaf tissue of control (1) and experimental (2) seedlings [0 h 23°C (‘0’); 6 h dark phase, 23°C/3°C (‘6’); 4 h light phase 23°C (‘12’)] of the Lady Claire potato variety. Day and night periods are indicated under the graphs by white and gray boxes, respectively. Primers: PSY1 (5´-catgctcgatggtgctttgtc-3´ and 5´-gacttcctcaagtccatacgca-3´); PSY2 (5´-aactgagctctgctagtagatg-3´ and 5´-gcactagagatcttgcataagca-3´); PSY3 (5´-gcctagtttagccattcaatagac-3´ and 5´-gcctagagttgatcgaacgattc-3´). Data were normalized to two reference genes: ef11 (LOC102600998; 5´-attggaaacggatatgctcca-3´ and 5´-tccttacctgaacgcctgtca-3´) and SEC3A (LOC102599118; 5´-gcttgcacacgccatatcaat-3´ and 5´-tggattttaccaccttccgca-3´). Statistical processing (one-way ANOVA; “multiple comparisons, corrected with Bonferroni test”) was performed using GraphPad Prism v. 8 (GraphPad Software Inc., USA) based on two biological and three technical replicates (p < 0.05 for significant differences); in the table (on the right), p-values for insignificant differences are highlighted in red.
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