Antibody Microarray Expression Profiling Of Maize Roots Treated With Cadmium And Nickel

Abstract

Here, we report the effectiveness of antibody microarray expression profiling (AbMEP) procedure to determine similarities and differences between two maize cultivars (Premia and Blitz) exposed to various concentration of cadmium (Cd), nickel (Ni), as well as simultaneous exposure to both metals (Cd + Ni) for 0, 12 and 24 h. After protein extraction from control (untreated) and heavy metals (HM)-treated root pairs and their fluorescent labelling, the protein extracts were used for the AbMEP procedure and western blot analysis. The results from the microarray were analysed using an internally normalised ratio. Using this highly parallel AbMEP-approach, the timing, dynamics and regulation of the expression of 101 specific genes in untreated and HM-treated roots of maize cultivars were determined. The microarray results revealed 23, 15 and 10 up-regulated/down-regulated proteins in Cd, Ni and Cd + Ni treated cv. Premia roots, respectively. In contrast, the microarray results revealed that 15, 11 and 7 proteins were up-regulated/down-regulated in Cd, Ni and Cd + Ni treated cv. Blitz roots, respectively. The data obtained from the AbMEP array experiment were validated by western blot analyses with more than 89% (the Pearson’s correlation coeficient Rr = 0.78) correlation between the two methods.

Histone Deacetylase Is Required for GA-Induced Programmed Cell Death in Maize Aleurone Layers

“…To investigate how the transcription of these genes changes during phytohormone-induced PCD, two caspase-like protein genes, metacaspase type II (gene identifier 100285605) and putative metacaspase (gene identifier 100191541) (Ahmad et al, 2012), and three PCD inhibition genes, hexokinase (gene identifier 100192075), fdad1 (gene identifier 541686), and ABP9 (GenBank accession no. GU237073) (Zhang et al, 2011;Repka et al, 2015), were selected for analysis in this study. Quantitative real-time PCR analysis revealed that the expression of two metacaspase genes was increased at the late stage of GA-induced PCD, and the expression of three PCD inhibition genes increased initially, which then decreased at the stage of GA-induced PCD (Supplemental Fig.…”