SP80-3280 growth and maturation field 1

A field experiment was conducted at the Agricultural Sciences Center of the Federal University of Sao Carlos in Araras (22°21'25''S and 47°23'3''W) in the state of Sao Paulo, Brazil. Trial plots of SP-3280 consisted of four rows of 10 m long and spaced 1.35m apart. The field experiment was initiated in April 2012 and extended up until May 2013, representing the conditions under which one-year-and-a-half sugarcane crops are cultivated. Aiming to carry out observations throughout growth and development, tissue samples of the +1 leaves (L1) and upper (I1), immature (I5) and mature (I9) internodes were collected from two plots (two technical replicates) after 4, 8, 11 and 13 months of planting. RNA was extracted for four biological replicates, two from each plot, using the TriZol method, treated with DNase I and purified. A pool of samples from leaves and a pool of internodes was used as a 'reference sample' for hybridization experiments on a customized 4x44K oligoarray (Agilent Technologies) for sugarcane (CaneRegNet). The oligoarrays were read using the GenePix 4000B scanner device (Molecular Devices) and the fluorescence data was processed by Feature Extraction software 9.5.3 (Agilent Technologies).