Microarray analysis to identify the differentially expressed genes induced by NROB

Compared to wild type plants, overexpression of NROB results leaf early senescence, biomass and seeds yield decreased more than 70%. Microarray analysis the differential expression genes induced by NROB Young leaves of DAG 30 were harvested from WT and 35S: NROB #2 plants and frozen immediately in liquid nitrogen. Total RNA was extracted using the RNeasy plant mini kit (Qiagen) and was checked for a RIN number to inspect RNA integration by an Agilent Bioanalyzer 2100. Qualified and purified total RNA was amplified and labeled by Low Input Quick Amp Labeling Kit, Two-Color (Agilent technologies, US). Regulated genes were identified with a stringent significance threshold, namely a mean >2.0-fold change (transformants relative to WT control samples) and based on at least two replicates. Microarray analysis result was verified by real-time PCR