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Fig. 5 | BMC Plant Biology

Fig. 5

From: Microtubule organization changes severely after mannitol and n-butanol treatments inducing microspore embryogenesis in bread wheat

Fig. 5

Mannitol stress recovery (MAN-R4) relates to MT regeneration in uni-nucleated microspores and bi-nucleate structures. A1-A6 Late uni-nucleate microspore. Microspore with a large vacuole (V) and nucleus (N) located close to the exine. A1 Randomly orientated thin and long CMT. A2 Long EMT arranged in thin bundles forming a net-like structure wrapped around the N. A3 Long, single, and thin EMT in the cytoplasmic layer underlying the exine. A4-A6 3D model of the subcellular volume of a similar structure. MT recovery observed in the endoplasm, where long and thin EMT bundles assembling a more complex structure causing association of the nucleus with the sporoderm. Superior visualisation axis model (A4). Volumetric ortho-slicer view from the forward (A5) and opposite site of the microspore model (A6). B1-B3 Bi-nucleate pollen-like structure with the generative-like nucleus (GLN) and vegetative-like nucleus (VLN) and small vacuoles (V) fragmented by cytoplasmic strands. B1 Short fragmented CMT. B2-B3 Short EMT in the cytoplasm and in the proximity of the VLN and thick bundles of EMT surrounding the GLN. C1-C3 Bi-nucleate embryogenic structure with similar nuclei (N) and a large vacuole (V). C1 Both, disoriented and longitudinally oriented CMT. C2 Subcortical MT seem to irradiate towards nuclei. C2-C3 Both, short and long EMT thin bundles forming a basket-like structure surrounding both nuclei. Cortex F – CMT; z-series projected as a maximum intensity projection from cortex optical sections collected from the front side (F) of microspore. Middle – optical section at the middle of the z-series. Middle+Cortex B – Endoplasmic and cortical MT; z-series projected as a maximum intensity projection from middle and cortical optical sections collected from the back side (B) of microspore. Inserts on the upper-right corner in the first panels show a corresponding image in the differential interference contrast (DIC). The dashed line indicates the axis between the operculum (O) and the opposite pole. Prj – z-series projected as a maximum intensity from the front side (Prj F) or from the back side (Prj B) of microspore. Green fluorescence (Alexa 488) shows α-tubulin. Red fluorescence of nuclei is caused by PI staining. Red autofluorescence of exine. Scales =20 μm

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