- This is a somewhat polished protocol as found in a publication.
- This is not an example for a typical lab protocol or method.
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# Plant Material and Growth Conditions
*Talinum triangulare* plants were grown in Miracle-Gro Potting Mix (Miracle-Gro) in “Short-One” treepots, 1.6 l (Stuewe and Sons). The experiment was initiated with 28-d-old plants in a controlled environment chamber (Environmental Growth Chambers) maintained under 12 h light (30°C, 37% relative humidity)/12 h dark (22°C) cycles. Photon flux density at leaf level was 425 mmol m<sup>-2</sup> s<sup>-1</sup>. Irrigation was withheld on day 1 and recommenced on day 14. Leaves were harvested when plants were well-watered as well as after 4, 9, and 12 days of water deprivation and watered for two days following the drought period.
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*Talinum triangulare* plants were grown in Miracle-Gro Potting Mix (Miracle-Gro) in “Short-One” treepots, 1.6 l (Stuewe and Sons). The experiment was initiated with 28-d-old plants in a controlled environment chamber (Environmental Growth Chambers) maintained under 12 h light (30°C, 37% relative humidity)/12 h dark (22°C) cycles. Photon flux density at leaf level was 425 mmol m<sup>-2</sup> s<sup>-1</sup>.
# Treatment and harvest
Irrigation was withheld on day 1 and recommenced on day 14. Leaves were harvested when plants were well-watered as well as after 4, 9, and 12 days of water deprivation and watered for two days following the drought period. The topmost mature unshaded leaves (of approximately 3–4.5 cm length) of *T. triangulare* were harvested in the middle of the light or the middle of the dark period and immediately frozen in liquid nitrogen.
