An established Arabidopsis thaliana var. Landsberg erecta cell suspension culture accumulates chlorophyll and exhibits a stay-green phenotype in response to high external sucrose concentrations.
An established cell suspension culture of Arabidopsis thaliana var. Landsberg erecta was grown in liquid media containing 0-15%(w/v) sucrose. Exponential growth rates of about 0.40d-1were maintained between 1.5-6%(w/v) sucrose, which decreased to about 0.30d-1between 6 and 15%(w/v) sucrose. Despite the presence of external sucrose, cells maintained a stay-green phenotype at 0-15% (w/v) sucrose. Sucrose stimulated transcript levels of genes involved in the chlorophyll biosynthetic pathway (ChlH, ChlI2, DVR). Although most of the genes associated with photosystem II and photosystem I reaction centers and light harvesting complexes as well as genes associated with the cytochrome b6f and the ATP synthase complexes were downregulated or remained unaffected by high sucrose, immunoblotting indicated that protein levels of PsaA, Lhcb2 and Rubisco per gram fresh weight changed minimallyon a Chl basis as a function of external sucrose concentration. The green cell culture was photosynthetically competent based on light-dependent, CO2-saturated rates of O2evolution as well as Fv/Fm and P700 oxidation. Similar to Arabidopsis WT seedlings, the suspension cells etiolated in the dark and but remained green in the light. However, the exponential growth rate of the cell suspension cultures in the dark (0.45±0.07d-1) was comparable to that in the light (0.42±0.02d-1). High external sucrose levels induced feedback inhibition of photosynthesis as indicated by the increase in excitation pressure measured as a function of external sucrose concentration. Regardless, the cell suspension culture still maintained a stay-green phenotype in the light at sucrose concentrations from 0 to 15%(w/v) due, in part, to a stimulation of photoprotection through nonphotochemical quenching. The stay-green, sugar-insensitive phenotype of the cell suspension contrasted with the sugar-dependent, non-green phenotype of Arabidopsis Landsberg erecta WT seedlings grown at comparable external sucrose concentrations. It appears that the commonly used Arabidopsis thaliana var. Landsberg erecta cell suspension culture has undergone significant genetic change since its original generation in 1993. We suggest that this genetic alteration has inhibited the sucrose sensing/signaling pathway coupled with a stimulation of chlorophyll an accumulation in the light with minimal effects on the composition and function of its photosynthetic apparatus. Therefore, caution must be exercised in the interpretation of physiological and biochemical data obtained from experimental use of this culture in any comparison with wild-type Arabidopsis seedlings.
Department of Biology and the Biotron Centre for Experimental Climate Change Research, University of Western Ontario, London, ON, N6A 5B7 Canada,. Electronic address: avemccar@gmail.com.
Avery
A
McCarthy
Department of Biology and the Biotron Centre for Experimental Climate Change Research, University of Western Ontario, London, ON, N6A 5B7 Canada,. Electronic address: michellechung86@gmail.com.
Michelle
M
Chung
Department of Biology and the Biotron Centre for Experimental Climate Change Research, University of Western Ontario, London, ON, N6A 5B7 Canada,. Electronic address: aivanov@uwo.ca.
Alexander G
AG
Ivanov
Department of Biology and the Biotron Centre for Experimental Climate Change Research, University of Western Ontario, London, ON, N6A 5B7 Canada,. Electronic address: mkrol1410@gmail.com.
Marianna
M
Krol
Department of Biology and the Biotron Centre for Experimental Climate Change Research, University of Western Ontario, London, ON, N6A 5B7 Canada,. Electronic address: inmanm@gmail.com.
Michael
M
Inman
Department of Biology and the Biotron Centre for Experimental Climate Change Research, University of Western Ontario, London, ON, N6A 5B7 Canada,. Electronic address: dmaxwell@uwo.ca.
Denis P
DP
Maxwell
Department of Biology and the Biotron Centre for Experimental Climate Change Research, University of Western Ontario, London, ON, N6A 5B7 Canada,. Electronic address: nhuner@uwo.ca.
Norman P A
NPA
Hüner
D017360:Arabidopsis / Q000235:genetics / Q000502:physiology* / Q000528:radiation effects / Q000648:ultrastructure; D002478:Cells, Cultured; D002734:Chlorophyll / Q000378:metabolism; D008027:Light; D009154:Mutation; D010084:Oxidation-Reduction; D010641:Phenotype; D010788:Photosynthesis / Q000528:radiation effects; D045331:Photosystem I Protein Complex / Q000378:metabolism; D045332:Photosystem II Protein Complex / Q000378:metabolism; D012273:Ribulose-Bisphosphate Carboxylase / Q000378:metabolism; D013395:Sucrose / Q000378:metabolism*
D045331:Photosystem I Protein Complex; D045332:Photosystem II Protein Complex; D002734:Chlorophyll; D013395:Sucrose; D012273:Ribulose-Bisphosphate Carboxylase
Arabidopsis; Cell suspension; Inhibition of photosynthesis; Photosynthetic capacity; Sugar insensitive; Sugar signaling