Genome-wide identification and expression profiling of duplicated flavonoid 3'-hydroxylase gene family in Carthamus tinctorius L.

  • Nguyen Q. V. HOANG Jilin Agricultural University, College of Life Sciences, Engineering Research Center of the Chinese Ministry of Education for Bioreactor and Pharmaceutical Development, Changchun, 130118 (CN)
  • Kong JIE Jilin Agricultural University, College of Life Sciences, Engineering Research Center of the Chinese Ministry of Education for Bioreactor and Pharmaceutical Development, Changchun, 130118 (CN)
  • Naveed AHMAD Jilin Agricultural University, College of Life Sciences, Engineering Research Center of the Chinese Ministry of Education for Bioreactor and Pharmaceutical Development, Changchun, 130118; Institute of crop germplasm resources, Shandong Academy of Agricultural Sciences; Shandong Provincial Key Laboratory of Crop Genetic Improvement, Ecology and Physiology, Jinan 250100 (CN)
  • Ma XINTONG Jilin Agricultural University, College of Life Sciences, Engineering Research Center of the Chinese Ministry of Education for Bioreactor and Pharmaceutical Development, Changchun, 130118 (CN)
  • Zhang XINYUE Jilin Agricultural University, College of Life Sciences, Engineering Research Center of the Chinese Ministry of Education for Bioreactor and Pharmaceutical Development, Changchun, 130118 (CN)
  • Wu YUHANG Jilin Agricultural University, College of Life Sciences, Engineering Research Center of the Chinese Ministry of Education for Bioreactor and Pharmaceutical Development, Changchun, 130118 (CN)
  • Wang NAN Jilin Agricultural University, College of Life Sciences, Engineering Research Center of the Chinese Ministry of Education for Bioreactor and Pharmaceutical Development, Changchun, 130118 (CN)
  • Yao NA Jilin Agricultural University, College of Life Sciences, Engineering Research Center of the Chinese Ministry of Education for Bioreactor and Pharmaceutical Development, Changchun, 130118 (CN)
  • Liu XIUMING Jilin Agricultural University, College of Life Sciences, Engineering Research Center of the Chinese Ministry of Education for Bioreactor and Pharmaceutical Development, Changchun, 130118; Wenzhou University, Institute of Life Sciences, Wenzhou, Zhejiang, 325035; Collaborative Innovation Center of Wenzhou, Wenzhou, Zhejiang (CN)
  • Jin LIBO Wenzhou University, Institute of Life Sciences, Wenzhou, Zhejiang, 325035; Collaborative Innovation Center of Wenzhou, Wenzhou, Zhejiang (CN)
Keywords: differential expression, flavonoid 3'-hydroxylase (F3'H), prokaryotic expression, safflower, transient expression

Abstract

Flavonoid 3′-hydroxylase (F3’H) enzyme is essential in determining the flavonoids B-ring hydroxylation pattern. It is mainly implicated in the biosynthetic pathway of cyaniding-based anthocyanins, flavonols, and flavan-3-ols. However, the evolution and regulatory mechanism of these important flavonoid hydroxylases have not been systematically investigated in safflower (Carthamus tinctorius L.). In this study, we identified 22 duplicatedCtF3'H-encoding genes from safflower through genome-wide prediction and conservation analysis. Phylogenetic analysis revealed the pattern of conservation and divergence of CtF3'Hs encoding proteins and their homologs from different plant species. The distribution of conserved protein motifs and cis-regulatory units suggested several structural components that could be crucial in deciphering the final function of CtF3'H proteins. Furthermore, the results of RNA-seq and qRT-PCR assay in different flowering tissues suggested differential expression level of CtF3’H genes during flower development. Based on the unique homology of CtF3’H5 with flavonoid 3’ hydroxylases from other plant species, further validation of CtF3’H5 was carried out. The transient expression of CtF3’H5 in onion epidermal cells implied that the subcellular localization of the fusion construct containing CtF3’H5 and GFP was predominantly detected in the plasma membrane. Similarly, the prokaryotic expression and western blot hybridization of CtF3’H5 demonstrated the detection of a stable 50.3kD target protein. However, more efforts are needed to further extend the functional validation of CtF3’H5 in safflower. This study provides a fundamental gateway for future functional studies and understanding the genetic evolution of F3'Hs in plants.

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Published
2021-11-17
How to Cite
HOANG, N. Q. V., JIE, K., AHMAD, N., XINTONG, M., XINYUE, Z., YUHANG, W., NAN, W., NA, Y., XIUMING, L., & LIBO, J. (2021). Genome-wide identification and expression profiling of duplicated flavonoid 3’-hydroxylase gene family in Carthamus tinctorius L. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 49(4), 12509. https://doi.org/10.15835/nbha49412509
Section
Research Articles
CITATION
DOI: 10.15835/nbha49412509