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Fig. 6. zip-1 and rdr6-11 affect the amplitude of
SPL3 expression, but not its temporal pattern. (A)
Northern analysis of SPL3 and miR156 RNA in SD-grown plants
at 14 dap (6 leaves) and 21 dap (12 leaves). zip-1 and
rdr6-11 produce a modest increase in SPL3 mRNA without
affecting the level of miR156. hst-6 reduces the accumulation of
miR156 and produces a dramatic increase in SPL3 mRNA at 21
dap. U6 was used as a loading control for miR156 and 25S
rRNA was the loading control for SPL3. (B) Northern analysis
of SPL3 mRNA in SD-grown wild-type and mutant plants of different
ages. Hybridization intensity was determined for three blots using NIH image,
averaged and normalized to the intensity in wild-type plants at 14 dap.
(C) GUS activity in the 3rd and 4th leaf of primary transgenic plants
transformed with 35S::GUS-SPL3 (G-SPL3) or
35S::GUS-SPL3m (G-SPL3m). More than 30 plants of each
genotype were analyzed. The difference between the expression of these
constructs in wild type and zip-1 is not statistically significant.
(D) Northern blot of SPL4, SPL9 and SPL15 mRNA in
flowers of wild-type, sgs3-11 and rdr6-11 plants. (E)
RT-PCR analysis of the SPL3 antisense transcript (SPL3
AS).
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