Extended Data Fig. 3: Genetic complementation of pinna1.
From: A molecular framework underlying the compound leaf pattern of Medicago truncatula
a, 3-week-old plants of pinna1-2 and two 35S::PINNA1-GFP/pinna1-2 transgenic lines with partially rescued (#2-3) and completely rescued (#2-1) phenotypes. This experiment was repeated independently three times with similar results. Scare bar, 2 cm. b,c, Normal RT-PCR (b) and RT-qPCR (c) analysis of PINNA1 expression level in young leaves of seven T0 independent lines (35S::PINNA1-GFP) compared to pinna1-1 and pinna1-2. The normal RT-PCR experiment was repeated independently two times with similar results. The RT-qPCR experiment was repeated three times using independent biological replicates with similar results, and one representative result is shown (mean ± SD, n = 3). P, 35S::PINNA1-GFP plasmid. M, molecular weight markers. PINNA1 gene structure, the positions of Tnt1 insertions in pinna1-1 and pinna1-2, and the RT-qPCR primer-pair are shown in the top panel. d, A 7-week-old 35S::PINNA1-GFP/pinna1-2 transgenic plant (#2-12) with a completely rescued phenotype. This experiment was repeated independently three times with similar results. Scare bar, 5 cm. e, Western blot analysis of T1 progeny from the 35S:PINNA1-GFP/pinna1-2 transgenic plant (#2-12) using an anti-GFP antibody. A DR5::GFP transgenic plant used as positive control. 3 to 10, independent T1 plants. This experiment was repeated independently three times with similar results. f, A 7-week-old pinna1-2/pPINNA1:GFP-PINNA1 transgenic line (#G1) with a completely rescued phenotype. This experiment was repeated independently three times with similar results. Scare bar, 5 cm. g, Normal RT-PCR analysis of PINNA1 expression level in shoot buds of two independent pinna1-2/pPINNA1::GFP-PINNA1 transgenic lines. P, pPINNA1:GFP-PINNA1 plasmid. This experiment was repeated independently three times with similar results.
