Jee 420

Shoot apices were collected at different growth stages. Fresh samples were fixed for 24 h in 4% paraformaldehyde (Sigma, St. Louis, USA) and 2% glutaraldehyde (Sigma) in 0.1 M PBS (pH 7.2), and subsequently washed with PBS buffer (0.1 M, pH 7.2) and dehydrated through a graded alcohol series of 70, 85, 95, and 100% of ethanol, each for 30 min. To prepare them for SEM (JSM–6360LV, Hitachi, Tokyo, Japan) observation, the samples were then washed with 100% ethanol once, post-fixed in 1% (w/v) osmium tetroxide (Alfa Aesar, Massachusetts, USA) for 2 h, dehydrated in a freeze drier (JFD–310, Hitachi, Tokyo, Japan), and sputter-coated with gold palladium in 6 different 30-s bursts (JEE–420, Hitachi, Tokyo, Japan). The samples were analysed with a scanning electron microscope (S-3000N; Hitachi, Japan).

Transgenic Arabidopsis plants for scanning electron microscopy (SEM) were collected and fixed using formalin–acetic acid–alcohol for 2 days and then stored in 70% alcohol. Specimens used for SEM were dehydrated in an alcohol series running up to 100%, treated with isoamylacetate, critical point dried using CO₂, and sputter–coated with gold palladium in six 30 s bursts (JEE–420, Hitachi, Tokyo, Japan). SEM was performed with a JSM–6360LVscanning electron microscope at 15 kV (Hitachi, Tokyo, Japan), and SEM photographs were processed with Adobe Photoshop 8.0 software.

Rice seeds were used for scanning electron microscopy (SEM) analysis. Samples were fixed overnight with 3% glutaraldehyde-sodium phosphate buffer (0.1M) at room temperature and rinsed three times with 0.1M sodium phosphate buffer. The samples were dehydrated through an ethanol series and infiltrated with an isoamyl acetate series. Seeds were then sputter-coated with gold/palladium in six different 30 s pulses (Hitachi JEE-420) and analyzed by scanning electron microscope (Hitachi S-3000N).