Arabidopsis Apoplast TET8 Positively Correlates to Leaf Senescence and tet3tet8 Double Mutants are Delayed in Leaf Senescence.

Extracellular vesicles (EVs) are membrane-bound exosomes secreted into the apoplast. Two distinct populations of EVs have been described in Arabidopsis: PEN1-associated and TET8-associated. We previously noted early leaf senescence in the pen1 single and pen1pen3 double mutant. Both PEN1 and PEN3 are abundant in EV proteomes suggesting EVs might regulate leaf senescence in soil-grown plants. We observed that TET8 is more abundant in the apoplast of early senescing pen1 and pen1pen3 mutant rosettes and in older WT rosettes. The increase in apoplast TET8 in the pen1 mutant did not correspond to increased TET8 mRNA levels. In addition, apoplast TET8 was more abundant in the early leaf senescence myb59 mutant, meaning the increase in apoplast TET8 protein during leaf senescence is not dependent on pen1 or pen3 . Genetic analysis showed a significant delay in leaf senescence in tet3tet8 double mutants after six weeks of growth suggesting that these two tetraspanin paralogs operate additively and are positive regulators of leaf senescence. This is opposite of the effect of pen1 and pen1pen3 mutants that show early senescence and suggest PEN1 to be a negative regulator of leaf senescence. Our work provides initial support that PEN1-associated EVs and TET8-associated EVs may have opposite effects on soil-grown plants undergoing age-related leaf senescence.

Subcellular positioning during cell division and cell plate formation in maize.

Introduction: During proliferative plant cell division, the new cell wall, called the cell plate, is first built in the middle of the cell and then expands outward to complete cytokinesis. This dynamic process requires coordinated movement and arrangement of the cytoskeleton and organelles. Methods: Here we use live-cell markers to track the dynamic reorganization of microtubules, nuclei, endoplasmic reticulum, and endomembrane compartments during division and the formation of the cell plate in maize leaf epidermal cells. Results: The microtubule plus-end localized protein END BINDING1 (EB1) highlighted increasing microtubule dynamicity during mitosis to support rapid changes in microtubule structures. The localization of the cell-plate specific syntaxin KNOLLE, several RAB-GTPases, as well as two plasma membrane localized proteins was assessed after treatment with the cytokinesis-specific callose-deposition inhibitor Endosidin7 (ES7) and the microtubule-disrupting herbicide chlorpropham (CIPC). While ES7 caused cell plate defects in Arabidopsis thaliana, it did not alter callose accumulation, or disrupt cell plate formation in maize. In contrast, CIPC treatment of maize epidermal cells occasionally produced irregular cell plates that split or fragmented, but did not otherwise disrupt the accumulation of cell-plate localized proteins. Discussion: Together, these markers provide a robust suite of tools to examine subcellular trafficking and organellar organization during mitosis and cell plate formation in maize.