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. 2016 Apr 4;211(3):940–951. doi: 10.1111/nph.13938

Figure 2.

Figure 2

Constitutive expression of PHYSCODILLO proteins in Physcomitrella patens leads to arrested protoplast regeneration. (a) Numbers of stable transgenic lines generated as a result of transforming P. patens protoplasts with constructs encoding combinations of proteins (GFP or PHYSCODILLOs) driven from the constitutive maize UBIQUITIN promoter (pUBI) or the inducible heat‐shock promoter (pHSP). Using the constitutive promoter (pUBI) to drive PHYSCODILLO‐GFP fusion proteins results in no recovery of stable transgenic lines. pUBI driving green fluorescent protein (GFP) alone, or pHSP driving PHYSCODILLO‐GFP fusions leads to normal transgenic line generation. (b) Transient transformation of P. patens protoplasts with pUBI::PHYSCODILLO1‐GFP leads to the formation of small, arrested GFP‐expressing colonies that show no further cell growth (zoomed image) compared with untransformed protoplast regenerating colonies (asterisks). (c) A control pUBI::GFP fusion protein is not lethal, and mature regenerated transformants are obtained.