Optimizing environmental conditions for mass application of mechano-dwarfing stimuli to Arabidopsis

Obtaining uniform mechano-dwarfing of Arabidopsis thaliana (L.) Heynh. seedlings within dense plantings is problematic. Alternative forms of mechano-stimulation were applied to seedlings in effort to obtain uniform growth reduction compared with undisturbed controls in both greenhouse and controlled growth environments. Arabidopsis grown under low photosynthetic photon flux (PPF) artificial light grew upright with limited leaf expansion, which enhanced mechano-responsiveness compared to that of rosette-growing plants under filtered sunlight or high PPF artificial light. Hypocotyls of seedlings grown at PPFs > 60 micromoles m-2 s-1 elongated less and had 6% less sensitivity to mechanical stress than seedlings grown at PPFs < 60 micromoles m-2 s-1. Fluorescent lamps alone (F) or fluorescent plus incandescent (F+I) lamps were compared for seedling responses to mechanical stress. Under F lighting, hypocotyl elongation was reduced 25% to 40% by twice-daily brush or plate treatments, and brushed seedlings exhibited more growth reduction than did plate treatments. Seedlings grown under F+I lamps exhibited similar stress-induced growth reduction compared to seedlings grown under F only, but stressed F+I seedlings lodged to a greater extent due to excessive hypocotyl elongation. Temperature-response studies using standardized F-only lighting indicated increased hypocotyl elongation but decreased leaf expansion, and decreased mechano-responsivity to brushing over the temperature range from 20 to 28 degrees C. Daylength studies indicated similar degrees of mechano-inhibition of hypocotyl elongation over the daylength range of 12, 16, 20, and 24 hours, whereas fresh weight of stressed seedling shoots declined compared to controls. A combination of environmental growth parameters that give repeatable, visual mechanical dwarfing of Arabidopsis include low-PPF fluorescent lighting from 55 to 60 micromoles m-2 s-1, ambient temperatures from 22 to 25 degrees C, and twice-daily brush treatments.