Semi-automated analysis of dot blots using ImageJ/Fiji

Commercially available dot blots provide a set of specific antibodies spotted on membranes in a given pattern. If the target analyte is present in the solution that the membrane is incubated with, the detection reaction will result in a chemiluminescence signal which is recorded by film or scanner. In order to know which analytes were detected, the analysis consists of measuring the intensity of the recorded signal on each spot. Manually measuring the entire array (typically ~200 spots) is unreliable and tedious. Fully automatic registration of the blot membrane to the template pattern often fails since there might be only very few positive spots on the membrane. This article presents an ImageJ/Fiji macro that requires minimal user input to perform a robust iterative registration of an adjustable template mask representing the spot pattern to the recorded blot. Once the template mask is matched to the dot blot, the spot intensity of each dot is measured and reported in a results table.

Calcium and potassium contents in nutrient solution on Phoma leaf spot intensity in coffee seedlings

ABSTRACT Coffee is one of the main export commodities of Brazilian agribusiness. Phoma leaf spot [Phoma tarda (Stewart) Boerema & Bollen] is one of the most important coffee fungal diseases in Brazil. Therefore, this study aimed to evaluate the intensity of Phoma leaf spot in coffee seedlings supplied with different rates of Ca+2 and K+. The study was conducted under controlled conditions in a growth chamber, at the Department of Phytopathology - UFLA, from February 2010 to December 2011. The assay was repeated twice under the same conditions. The nutrient solutions consisted of five concentrations of K+ (3, 4, 5, 6, 7 mmol L-1) and Ca+2 (2, 4, 6, 8 and 10 mmol L-1). The experiment was arranged in a randomized block design, with 25 treatments and three replicates, with two plants per plot. The areas under incidence progress curve (AUIPC) and severity (AUSPC) were calculated. At the lowest rate of Ca2+ (2 mmol L-1) and highest K+ (6 and 7 mmol L-1), approximately, the AUIPC was the smallest. For the AUSPC, the lowest rates of Ca+2 and K+ resulted in the lowest severities. Supply of Ca+2 and K+ in nutrient solution reduced AUIPC and AUSPC of Phoma leaf spot, and these nutrients can be recommended for the management of the disease.