A mitochondrial glycerol-3-phosphate (G-3-P) shuttle that channels cytosolic reducing equivalent to mitochondria for respiration through oxidoreduction of G-3-P has been extensively studied in yeast and animal systems. Here, we report evidence for the operation of such a shuttle in Arabidopsis thaliana. We studied Arabidopsis mutants defective in a cytosolic G-3-P dehydrogenase, GPDHc1, which, based on models described for other systems, functions as the cytosolic component of a G-3-P shuttle. We found that the gpdhc1 T-DNA insertional mutants exhibited increased NADH/NAD þ ratios compared with wild-type plants under standard growth conditions, as well as impaired adjustment of NADH/NAD þ ratios under stress simulated by abscisic acid treatment. The altered redox state of the NAD(H) pool was correlated with shifts in the profiles of metabolites concerning intracellular redox exchange. The impairment in maintaining cellular redox homeostasis was manifest by a higher steady state level of reactive oxygen species under standard growth conditions and by a significantly augmented hydrogen peroxide production under stress. Loss of GPDHc1 affected mitochondrial respiration, particularly through a diminished capacity of the alternative oxidase respiration pathway. We propose a model that outlines potential involvements of a mitochondrial G-3-P shuttle in plant cells for redox homeostasis.

Proanthocyanidin biosynthesis in the seed coat of yellow-seeded, canola quality Brassica napus YN01-429 is constrained at the committed step catalyzed by dihydroflavonol 4-reductase Akhov, Leonid; Ashe, Paula; Tan, Yifang; Datla, Raju; Selvaraj, Gopalan Proanthocyanidin biosynthesis in the seed coat of yellow-seeded, canola quality Brassica napus YN01-429 is constrained at the committed step catalyzed by dihydroflavonol 4-reductase 1 Leonid Akhov, Paula Ashe, Yifang Tan, Raju Datla, and Gopalan Selvaraj Abstract: The yellow seed characteristic in Brassica napus L. is desirable because of its association with higher oil content and better quality of oil-extracted meal. YN01-429 is a yellow-seeded canola-quality germplasm developed in Canada arising from several years of research. Seed-coat pigmentation is due to oxidized proanthocyanidins (PA; condensed tannins) derived from phenylpropanoids and malonyl CoA. We found PA accumulation to be most robust in young seed coats (20 d post anthesis; dpa) of a related black-seeded line N89-53 and only very little PA in YN01-429, which also contained much less extractable phenolics. The flavonol content, however, did not show as great a difference between these two lines. Furthermore, sinapine, a product of the general phenylpropanoid metabolism, was present at comparable levels in the embryos of both lines. Dihydroflavonol reductase (DFR) activity that commits phenolics to PA synthesis was lower in YN01-429 seed coats. The results of Southern blot and in silico analyses were indicative of two copies of the DFR gene in B. napus. Both copies were functional in YN01-429, ruling out homeoallelic repression or silencing, but together they showed very low expression levels (17-fold fewer transcripts) relative to DFR activity in N89-53 seed coats. These results collectively suggest that YN01-429 differs in regulatory circuits that impact the PA synthesis branch much more than the flavonol synthesis branch in the seed coats and such circuits do not impinge upon general phenylpropanoid metabolism in the embryos.Key words: condensed tannin, dihydroflavonol reductase, flavonoids, oilseed rape, rapeseed, transparent testa. Résumé :Le jaune caractéristique des grains du Brassica napus L. est désirable, compte tenu de son association avec une haute teneur en huile et une meilleure qualité de la farine extraite de l'huile. Le YN01-429 constitue un germplasme à graine jaune de qualité canola résultant de plusieurs années de recherches. La pigmentation des téguments est attribuéeà des proanthocyanidines oxydées (PA; tannins condensés) dérivées de phénylpropanoïdes et du CoA du malonyl. Les auteurs ont trouvé une plus forte accumulation de PA chez les jeunes téguments (20 jours après l'anthèse; dpa) d'une lignée apparentéeà grains noirs N89-53 et très peu de PA chez la lignée YN01-429 qui contient à la fois beaucoup moins de phénols extractibles. Cependant, la teneur en flavonols ne montre pas autant de différences entre ces deux lignées. De plus, la sinapine, un produit du métabolisme géné...