Molecular biology is one of the most common tools for researchers to explore the molecular mechanisms of biological phenomena. We usually start from the phenotype, then potentially-related genes are located by detecting the change of gene or protein networks, followed by hypothesis verification through overexpression or knockdown of the target genes. Finally, a research result can be reached.
For example, in the research article A γ-glutamylcysteine ligase AcGCL alleviates cadmium-inhibited fructooligosaccharides metabolism by modulating glutathione level in Allium cepa L., the researcher found that fructooligosaccharides (FOS) are important carbohydrates in plants, while cadmium (Cd) toxicity limits growth and development in several plant species. But, whether FOS metabolism is affected by Cd and the molecular mechanisms of tolerance of the effects of Cd toxicity in plants remain enigmatic. The researchers created hypotheses and studied the FOS metabolism under Cd stress in an onion (Allium cepa L.). Results showed that Cd stress can inhibit FOS accumulation in onion, followed by the upregulation of a putative onion γ-glutamylcysteine ligase gene AcGCL. Then, the researchers amplified and cloned AcGCL ORF into the pTF101s vector, revealing that the heterologous expression of the AcGCL protein in Escherichia coli has GCL activity. Furthermore, they overexpressed AcGCL in young onion roots and found that overexpressing AcGCL through increasing glutathione (GSH), phytochelatin (PC), and any other genes’ accumulation, ameliorated Cd phytotoxicity on onion FOS metabolism. Finally, they were able to conclude that the function of AcGCL as a γ-glutamylcysteine ligase can alleviate Cd inhibited FOS metabolism by modulating GSH levels in onions.
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Reference DOI : 10.1016/j.jhazmat.2021.126255