Biochemistry
RuBisCo activity was measured by non-radioactive microplate-based assay,
which determines the product (3-phosphoglycerate; 3-PGA) in an enzymatic
cycle between glycerol-3-phosphate dehydrogenase and
glycerol-3-phosphate oxidase (Sulpice et al., 2007). The activity of
triosephosphate isomerase was measured according to the method of
Benítez-Cardoza et al. (2001) by monitoring NADH consumption at 340 nm.
ATP concentration was measured by using ATP Assay System Bioluminescence
Detection Kit (FF2000) as described by Promega, Wisconsin, USA. ATP
synthase was measured according to the method of Cross and Kohlbrenner
(1978). Phosphoenolpyruvate carboxylase activities were monitored as
absorbance changes at 340 nm. The AGPase activity was assayed according
to Nakamura et al. (1989) in a reaction mixture (5 mM ADPG, 50 mM MgCl2,
100 mM Hepes–NaOH). Production of NADPH was monitored at 340 nm. Total
amylase activity was determined as described by Bhatia and Singh (2002).
Soluble proteins were determined according to (Lowry et al., 1951) with
bovine serum albumin as a standard.
MDA was extracted in 2 mL of 80% (v/v) ethanol and measured using a
thiobarbituric acid-MDA assay (Hodges et al., 1999).
H2O2 determination, was done by using
Xylenol Orange reagent (Avramova et al, 2015a).