Comprehensive and evolutionary analysis of Spodoptera litura-inducible Cytochrome P450 monooxygenase gene family in Glycine max elucidate their role in defense

Manisha Yadav; Ruby Panwar; Anjana Rustagi; Amrita Chakraborty; Amit Roy; Indrakant K Singh; Archana Singh

doi:10.3389/fpls.2023.1221526

Comprehensive and evolutionary analysis of Spodoptera litura-inducible Cytochrome P450 monooxygenase gene family in Glycine max elucidate their role in defense

Front Plant Sci. 2023 Nov 2:14:1221526. doi: 10.3389/fpls.2023.1221526. eCollection 2023.

Authors

Manisha Yadav^{1

2}, Ruby Panwar^{1

3}, Anjana Rustagi³, Amrita Chakraborty⁴, Amit Roy⁵, Indrakant K Singh⁶, Archana Singh^{1

2

3

7}

Affiliations

¹ Department of Botany, Hansraj College, University of Delhi, Delhi, India.
² J C Bose Center for Plant Genomics, Hansraj College, University of Delhi, Delhi, India.
³ Department of Botany, Gargi College, University of Delhi, Delhi, India.
⁴ EVA 4.0 Unit, Faculty of Forestry and Wood Sciences, Czech University of Life Sciences Prague, Prague, Czechia.
⁵ Forest Molecular Entomology Lab, EXTEMIT-K, EVA 4.0, Faculty of Forestry and Wood Sciences, Czech University of Life Sciences Prague, Prague, Czechia.
⁶ Molecular Biology Research Lab, Department of Zoology, Deshbandhu College, University of Delhi, New Delhi, India.
⁷ Department of Plant Molecular Biology, University of Delhi, New Delhi, India.

Abstract

Plants being sessile organisms and lacking both circulating phagocytic cells and somatic adaptive immune response, have thrived on various defense mechanisms to fend off insect pests and invasion of pathogens. CYP450s are the versatile enzymes, which thwart plants against insect pests by ubiquitous biosynthesis of phytohormones, antioxidants, and secondary metabolites, utilizing them as feeding deterrents and direct toxins. Therefore, a comprehensive analysis of biotic stress-responsive CYPs from Glycine max was performed to ascertain their function against S. litura-infestation. Phylogenetic analysis and evolutionary studies on conserved domains and motifs disclosed the evolutionary correspondence of these GmCYPs with already characterized members of the CYP450 superfamily and close relatedness to Medicago truncatula. These GmCYPs were mapped on 13 chromosomes; they possess 1-8 exons; they have evolved due to duplication and are localized in endoplasmic reticulumn. Further, identification of methyl-jasmonate, salicylic acid, defense responsive and flavonoid biosynthesis regulating cis-acting elements, their interaction with biotic stress regulating proteins and their differential expression in diverse types of tissues, and during herbivory, depicted their responsiveness to biotic stress. Three-dimensional homology modelling of GmCYPs, docking with heme cofactor required for their catalytic activity and enzyme-substrate interactions were performed to understand the functional mechanism of their action. Moreover, to gain insight into their involvement in plant defense, gene expression analysis was evaluated, which revealed differential expression of 11 GmCYPs upon S. litura-infestation, 12 GmCYPs on wounding while foliar spray of ethylene, methyl-jasmonate and salicylic acid differentially regulated 11 GmCYPs, 6 GmCYPs, and 10 GmCYPs respectively. Our study comprehensively analysed the underlying mechanism of GmCYPs function during S. litura-infestation, which can be further utilized for functional characterization to develop new strategies for enhancing soybean resistance to insect pests.

Keywords: Glycine max; Spodoptera litura; evolutionary analysis; gene expression analysis; herbivory; plant defense.

Grants and funding

Financial support was provided to AS by the Science and Engineering Research Board (SERB), Department of Science and Technology, New Delhi, India (ECR/2017/002478; SPG/2021/002969). MY is grateful to CSIR, MHRD for the JRF fellowship. The authors acknowledge Prof. (Dr.) Rama, Principal, Hansraj College, for providing the facilities and her constant support for research work. AmR, AC, financed by EVA 4.0,” No. CZ.02.1.01/0.0/0.0/16 019/0000803 and “EXTEMIT – K,” No. CZ.02.1.01/0.0/0.0/15_003/0000433 financed by OP RDE. AmR and AC are also supported by “Excellent team grants (2023-24)” from FLD, CZU.