Figure 3
Phenotypes of plants treated with pTRV1 and pTRV2-EScaAG1, pTRV2-EScaAG2, or pTRV2-EScaAG1/2 and expression analysis of the VIGS treated plants. (A) Wild type phenotype of an E. californica flower treated with pTRV2-E. (B) Phenotype of an EScaAG1 VIGS treated plant showing full homeotic conversions of stamens into petals. (C) Phenotype of an EScaAG2 VIGS treated plant showing full homeotic conversions of stamens and carpels into petal-like structures. (D) Phenotype of a flower silenced for EScaAG1/2 showing homeotic conversions of stamens into petals. (E) A mock treated plant with disassembled floral organs. (F) Disassembled flower of a plant treated with pTRV2-EScaAG1/2 showing homeotic conversions of stamens into petals. The same phenotype was also achieved with plants silenced for EScaAG1 or EScaAG2 individually. (G) Transverse hand section of a flower from a mock treated plant. (H) Transverse hand section of a flower from a plant silenced for EScaAG1/2 showing homeotic conversions of stamens into petals. (I) Disassembled flower of a plant silenced for EScaAG1 showing partial homeotic conversions of only the outer whorl stamens. (J) Disassembled flower of a plant treated with pTRV2-EScaAG2 showing that the inner whorl of stamens is converted into petal-like structures. (K) Disassembled flower of a plant treated with pTRV2-EScaAG1/2 exhibiting homeotic conversions of the innermost and outermost stamens whorls into petals while the middle whorls show mild deviation from wild type stamens while the center whorl stamens remain more stamen-like. (L) Transverse section of a flower of an untreated plant. (M) Transverse section of a flower from an EScaAG1 VIGS treated plant showing homeotic conversion of stamens into petals, petal-stamen mosaic structures, malformed stamens and a gynoecium lacking tissue differentiation, ovules, and placenta. (N) Real-Time PCR analysis of the first bud of individual E. californica plants treated with VIGS and untreated (U). Plants were treated with pTRV2-EScaAG1 are abbreviated as VIGS AG1, plants treated with pTRV2-EScaAG2 as VIGS AG2. Numbers below indicated individual plants and the relative expression level of EScaAG1 and EScaAG2 in untreated plants was set to 1. Abbreviations: cw, carpel wall; ov, ovule; p, petal; pl, placenta; se, sepal; pst, petaloid stamens; st, stamen.