Supplemental Materials
Supplemental Figure S1 Pictures of 9-day-old wild-type rice
seedlings and two biuret hydrolase overexpressing lines. From
left to right: wild type and overexpressing lines B3-9-1 and B2-3-3.
Upper: seedlings grown in the control culture solution. Lower: seedlings
grown in the culture solution supplemented with 0.3 mmol
L-1 biuret. Bars show 10 cm.
Supplemental Figure S2 Allantoin concentration in 9-day-old
rice shoots measured by colorimetric and HPLC-UV. Plants were grown
hydroponically under the three biuret treatments. Control: plants did
not receive biuret; NB: plants were grown without biuret for three days
after sowing, and with 0.3 mmol L-1 biuret
supplemented in the culture solution from the fourth day; BN: plants
were grown with 0.3 mmol L-1 biuret for 6 days after
sowing and transferred to new culture solution without biuret on the
seventh day. Fresh shoots of 9-day-old seedlings were ground under
liquid N2 and extracted with 10-fold volume of distilled
water. After centrifugation, the supernatant was used for allantoin
determination. Gray boxes indicate mean allantoin concentration
determined colorimetrically and black boxes indicate that determined by
the HPLC method. Symbols indicate each sample. The statistical
significance of the differences between the methods was determined
through paired t-test (n = 2). ns: not significant.
Supplemental Figure S3 Relative expression levels of (a)OsALN and (b) OsUO in rice seedlings. Rice plants were
hydroponically grown with or without 0.3 mmol L-1biuret supplied in the culture solution. Measurements were made using
plants grown in an independent trial, as shown in Figure 5. The relative
expression levels were normalized to those of Ubiquitin andActin1 and expressed on a log2 scale. Gray and black squares
indicate control and biuret-treated plants, respectively; data represent
the mean ± SD (n = 4). Asterisks indicate statistically significant
differences between treatments at the time point (Welch’s t-test).
*p < 0.05; **p < 0.01; ***p< 0.001.
Supplemental Data S1 Peak intensities in Metabolome analysis.