Silver nanoparticles and Chlorella treatments induced glucosinolates and kaempferol key biosynthetic genes in Eruca sativa

Abstract

Background: Microalgae and nanoparticles are currently considered promising tools for numerous agricultural and biotechnological applications. The green microalga Chlorella sp. MF1 and its biosynthesized silver nanoparticles (AgNPs) were used in this study as biofortification agents to enhance glucosinolate and kaempferol levels in Eruca sativa. UV–visible spectroscopy, XRD, FTIR and TEM were comprehensively used for characterizing Chlorella-based AgNPs. Results: The biosynthesized AgNPs were found to be spherical in shape, with size ranging from 1.45 to 5.08 nm. According to FTIR measurements, silver ions were reduced to AgNPs by functional groups such as amide, hydroxyl and carboxylate. Different experimental treatments were conducted, including either soaking seeds of E. sativa or foliar spray with various concentrations of Chlorella suspension (1, 2, 3 and 4 g L−1) and AgNPs (5, 10, 20 and 40 mg L−1). Expression levels of five key genes in the biosynthetic pathway of glucosinolates (MAM1, SUR1, MYB34 and MYB51) and kaempferol (CHS) were assessed using qRT-PCR. The results indicated an upregulation in the gene expression levels in all treatments compared to control, recording the highest level at 40 mg L−1 AgNPs and 4 g L−1Chlorella suspension. In addition, high glucosinolates and kaempferol content was detected in plants whose leaves were sprayed with AgNPs and Chlorella suspension (40 mg L−1 and 4 g L−1) based on HPLC analysis. Sequence analysis of amplified CHS fragments from E. sativa plants treated with AgNPs (40 mg L−1) showed high sequence similarity to A. thaliana CHS gene. However, there were several CHS regions with sequence polymorphism (SNPs and Indels) in foliar sprayed plants. Conclusions: Results of this study evidenced that the application of AgNPs and Chlorella suspension increased glucosinolates and kaempferol content in E. sativa through upregulation of key genes in their biosynthetic pathway.