Gibberellin 2-Oxidases in Potato (Solanum tuberosum L.): Cloning, Characterization, In Silico Analysis and Molecular Docking.

Abstract

Gibberellins (GAs; tetracyclic di-terpenoid carboxylic acids) are endogenous plant growth regulators responsible for stimulating plant growth and development from seed germination to plant maturity. In potato (Solanum tuberosum L.), GA levels are known to be crucial in the complex process of tuberization. Gibberellin 2-oxidases (GA2oxs) inactivate bioactive GAs during stolon swelling and early stages of tuberization as evident from the predominant expression of a member of this gene family namely GA2ox1. We isolated and characterized a 1105-bp cDNA clone encoding a 340-aa GA2ox1 form, designated St-GA2ox1, using total RNA from growing tuber of a potato (Solanum tuberosum L.) cultivar, Kufri Chipsona-1 (KC-1) based on RT-PCR approach. A total of 26 GA2ox sequences were also retrieved from potato genome database and analysed. Multiple sequence alignment revealed sequence relatedness between the GA2oxs. Crucial protein motifs were identified. Phylogenetic analysis revealed the evolutionary relationships between the GA2oxs. Three-dimensional structure of St-GA2ox1 was predicted by using AlphaFold tool, validated by the predicted local-distance difference test and Ramachandran Plot. Structural analysis and molecular docking were carried out to identify domains, binding sites and affinity for the ligand. The STRING database and hydropathy analysis revealed the presence of a putative interaction site for other enzymes. Expression Atlas database and semi-quantitative RT-PCR revealed the expression patterns of various GA2ox forms in different potato organs. This comprehensive report would be useful in providing new insights into possible underlying mechanisms involved in tuber development, and could facilitate the targeted alteration of genes responsible to combat the stress and enhance tuber production.