Competitive-IgY- Enzyme Linked Immuno Sorbent Assay (CIgY-ELISA) to detect the cytokinins in Gerbera jamesonii plantlets

ABSTRACT

A competitive hyper-immune yolk Immunoglobulin Y - Enzyme Linked Immune Sorbent Assay (CIgY-ELISA), was developed as an alternative method to detect zeatin and 2ip in plantlets of gerbera. The endogenous level of hormones in the plantlets in vitro of gerbera with one or six weeks after replication was determined with competitive IgY-ELISA set to detect between 1 and 100 pmoles of plant hormone for each 1.0 g tissue. The plantlets of six weeks presented sprouts and root, while the plantlets of one week presented only sprouts. The CIgY-ELISA was set with high independent variables values of sensitivity/specificity of 96/89 percent for zeatin and 94/78 percent for 2ip, with high values of reproducibility (up to 90 percent) for both the cytokinins. Zeatin content varied from 2.2 to 2.8 pmoles.g-1 and from 2.7 to 3.3 pmoles.g-1 on the plantlet with one and six weeks, respectively. The 2ip content did not vary and was detected near the detection limit in all the assays. It was concluded that the observed capabilities of CIgY-ELISA were putative and the competitive assay was a highly robust and stable method, which could be used for the studies on plant physiology for endogenous cytokinins.