Investigating the expression of TYDC2 gene in root, leaf, and stem tissues and its relationship with callus formation rate of these tissues in poppy medicinal plant (Papaver somniferum L.)

Subject Areas : Medicinal Plants

Nazila Bagheri ¹ , Alireza Tari-nejad ² , mohammad majidi ³ , کریم حسن پور ⁴

1 - Faculty of Agriculture, Shahid Madani University of Tabriz
2 - Department of Biotechnology, Shahid Madani University of Azerbaijan, Tabriz, Iran
3 - عضو هیات علمی
4 - University of Tabriz

Received: 2024-09-04 Accepted : 2024-12-06 Published : 2024-12-06

Keywords: auxin hormone, callus formation, gene expression, sequencing ,

Abstract :

Poppy with the scientific name Papaver somniferum L. belongs to the Papaveraceae family. More than 40 types of alkaloids are known in poppy, the most important of which are morphine, codeine, thebaine, noscapine and papaverine. The purpose of this research; Identification of the expression level of TYDC2 gene in three tissues of the stem, root and leaf of the poppy medicinal plant is through NGS and its relationship with the callus formation rate of these tissues is investigated. In this research, the sequence of this gene in three tissues of stem, root and leaf of poppy plant was selected from Gene section of NCBI website and downloaded by SRAtoolkit tool and their quality was evaluated by fastqc tool and heatmap diagram was created with Rstudio software. and other gene expression comparison charts were drawn. then, Morashik - Skoog culture medium was prepared with 2,4-D hormone at a concentration of 2 mg/L, and the mentioned tissues were placed in this medium and compared with the gene expression level. Finally, this gene was evaluated with gene ontology by gprofiler. The results of this survey showed; The expression level of production gene in stem, root and leaf tissues of poppy medicinal plant is different and the expression level in leaf tissue is higher than other tested tissues. The lowest level of expression was also observed in the root tissue and also the results of callus formation of these three plant tissues with 2,4-D hormone (2 mg/liter) in vitro showed that the level of callus formation in leaf tissue is higher than other tissues. and the lowest amount of callus formation was determined in the root tissue, which indicates a direct relationship between gene expression and the amount of callus formation in tissues.

 

References:

Alqethami, A., & Aldhebiani, A. Y. (2021). Medicinal plants used in Jeddah, Saudi Arabia: phytochemical screening. Saudi Journal of Biological Sciences, 28(1), 805-812
. Arumuganathan, K., & Earle, E. (1991). Nuclear DNA content of some important plant species. Plant molecular biology reporter, 9, 208-218
. Bagheri, N., Maleki Zanjani, B., & Ammarlou, A. (2020). A callus study of different explants of the medicinal Myrtle plant (Myrtus communis). Journal of Medicinal Plants Biotechnology, 6(1), 93-101
. Birnie, R., Bryce, S. D., Roome, C., Dussupt, V., Droop, A., Lang, S. H., . . . Stower, M. J. (2008). Gene expression profiling of human prostate cancer stem cells reveals a pro-inflammatory phenotype and the importance of extracellular matrix interactions. Genome biology, 9, 1-13
. Cánovas, R., Moffat, A., & Turpin, A. (2016). CSAM: compressed SAM format. Bioinformatics, 32(24), 3709-3716
. Consortium, G. O. (2008). The gene ontology project in 2008. Nucleic acids research, 36(suppl_1), D440-D444
. D'Antuono, L. F., Moretti, A., & Lovato, A. F. (2002). Seed yield, yield components, oil content and essential oil content and composition of Nigella sativa L. and Nigella damascena L. Industrial crops and products, 15(1), 59-69
. Diaz-Bárcena, A., & Giraldo, P. (2023). Exploring the research evolution of Papaver somniferum and Cannabis sativa: A bibliometric comparative analysis. Industrial Crops and Products, 203, 117143
. Eckert, R. L., Crish, J. F., Efimova, T., Dashti, S. R., Deucher, A., Bone, F., . . . Balasubramanian, S. (2004). Regulation of involucrin gene expression. Journal of Investigative Dermatology, 123(1), 13-22
. Ghasemi, S., Kumleh, H. H., & Kordrostami, M. (2019). Changes in the expression of some genes involved in the biosynthesis of secondary metabolites in Cuminum cyminum L. under UV stress. Protoplasma, 256, 279-290
. Hori, K., Yamada, Y., Purwanto, R., Minakuchi, Y., Toyoda, A., Hirakawa, H., & Sato, F. (2018). Mining of the uncharacterized cytochrome P450 genes involved in alkaloid biosynthesis in California poppy using a draft genome sequence. Plant and Cell Physiology, 59(2), 222-233
. Kanei, M., Horiguchi, G., & Tsukaya, H. (2012). Stable establishment of cotyledon identity during embryogenesis in Arabidopsis by ANGUSTIFOLIA3 and HANABA TARANU. Development, 139(13), 2436-2446
. Lee, D.-K., Geisler, M., & Springer, P. S. (2009). LATERAL ORGAN FUSION1 and LATERAL ORGAN FUSION2 function in lateral organ separation and axillary meristem formation in Arabidopsis
. Lee, M.-L. T., Kuo, F. C., Whitmore, G., & Sklar, J. (2000). Importance of replication in microarray gene expression studies: statistical methods and evidence from repetitive cDNA hybridizations. Proceedings of the National Academy of Sciences, 97(18), 9834-9839
. Marioni, J. C., Mason, C. E., Mane, S. M., Stephens, M., & Gilad, Y. (2008). RNA-seq: an assessment of technical reproducibility and comparison with gene expression arrays. Genome research, 18(9), 1509-1517
. Miranda, M., Vega-Gálvez, A., Quispe-Fuentes, I., Rodríguez, M. J., Maureira, H., & Martínez, E. A. (2012). Nutritional aspects of six quinoa (Chenopodium quinoa Willd.) ecotypes from three geographical areas of Chile. Chilean journal of agricultural research, 72(2), 175
. Romero, I. G., Ruvinsky, I., & Gilad, Y. (2012). Comparative studies of gene expression and the evolution of gene regulation. Nature Reviews Genetics, 13(7), 505-516
. Sakai, H., Honma, T., Aoyama, T., Sato, S., Kato, T., Tabata, S., & Oka, A. (2001). ARR1, a transcription factor for genes immediately responsive to cytokinins. Science, 294(5546), 1519-1521
. Santelia, D., Vincenzetti, V., Azzarello, E., Bovet, L., Fukao, Y., Düchtig, P., . . . Geisler, M. (2005). MDR-like ABC transporter AtPGP4 is involved in auxin-mediated lateral root and root hair development. FEBS letters, 579(24), 5399-5406
. Shinozaki, K., Yamaguchi-Shinozaki, K., & Seki, M. (2003). Regulatory network of gene expression in the drought and cold stress responses. Current opinion in plant biology, 6(5), 410-417
. Yoshikawa, T., & Furuya, T. (1983). Regeneration and in vitro flowering of plants derived from callus cultures of opium poppy (Papaver somniferum). Experientia, 39, 1031-1033