OsSEC24, a functional SEC24-like protein in rice, improves tolerance to iron deficiency and high pH by enhancing H(+) secretion mediated by PM-H(+)-ATPase.

Iron is abundant in the soil, but its low solubility in neutral or alkaline soils limits its uptake. Plants can rely on rhizosphere acidification to increase iron solubility. OsSEC27p was previously found to be a highly up-regulated gene in iron-deficient rice roots. Here, pH-dependent complementation assays using yeast mutants sec24Δ/SEC24 and sec27Δ/SEC27 showed that OsSEC27 could functionally complement SEC24 but not SEC27 in yeast; thus, it was renamed as OsSEC24. We found that OsSEC24-transgenic tobacco plants increased the length and number of roots under iron deficiency at pH 8.0. To explore how OsSEC24 confers tolerance to iron deficiency, we utilized transgenic tobacco, rice and rice protoplasts. H(+) flux measurements using Non-invasive Micro-test Technology (NMT) indicated that the transgenic OsSEC24 tobacco and rice enhanced H(+) efflux under iron deficiency. Conversely, the application of plasma membrane PM-H(+)-ATPase inhibitor vanadate elucidated that H(+) secretion increased by OsSEC24 was mediated by PM-H(+)-ATPase. OsPMA2 was used as a representative of iron deficiency-responsive PM-H(+)-ATPases in rice root via RT-PCR analysis. In transgenic rice protoplasts OsPMA2 was packaged into OsSEC24 vesicles after export from the ER through confocal-microscopy observation. Together, OsSEC24 vesicles, along with PM-H(+)-ATPases stimulate roots formation under iron deficiency by enhancing rhizosphere acidification.

Mitochondrial dynamics and its responds to proteasome defection during Picea wilsonii pollen tube development.

Tip growth of pollen tubes is essential for higher plant sexual reproduction and has been proposed to be highly regulated by the ubiquitin/proteasome pathway (UPP). The dynamics of mitochondria and the functions of the UPP on mitochondrial dynamics during pollen tube development are still poorly understood. In the present study, using real-time laser scanning and transmission electron microscope, it was revealed that mitochondria in Picea wilsonii, are either ellipsoid or filamentous with various lengths. Time-lapse images indicated that the two forms of mitochondria interconvert frequently through opposite process of fusion and fission. Examination of mitochondrial morphology during four key stages of in vitro pollen tube development revealed a link between mitochondrial remodeling and the process of pollen tube elongation. We also report that MG132, a specific proteasome inhibitor, not only strongly disturbed the mitochondrial remodeling but also significantly reduced mitochondrial membrane potential during pollen tube development. This finding provides new insight into the function of the proteasome in tip growth of pollen tubes.

[Ultracytochemical studies of aloin in Aloe arborescens leaves].

Lead acetate precipitation method was used for ultracytochemical localization of aloin. The processes of aloin production, transport and storage were studied by transmission electron microscope. Results showed that aloin was produced in the plastids of the assimilating tissue. The aloin was transported through the plastid membrane to the surrounding endoplasmic reticulum and enveloped in the vesicles by the endoplasmic reticulum elements, the vesicles approached and later fused with the plasmalemma. Some vesicles of the plastid membrane directly fused with the plasmalemma. The vesicles released their contents into the apoplast through exocytosis, and finally reached the vascular bundle sheath by apoplastic translocation. Aloin was transported to the internal tangential wall of vascular bundle sheath cell through endoplasmic reticulum vesicles, and reached the cytoplasm of aloin cell by means of plasmodesmata. Finally, aloin was stored in the vacuoles of aloin cell.

[Effects of MG132 , an inhibitor of proteasome , on the pollen germination and tube growth of Pecea wilsonii].

The ubiquitin/proteasome system is regarded as a major pathway of proteolysis in eukaryotic cells, in which the proteasome acts as primary protease for its function of degrading substrate proteins to short peptides. In the present paper, cytological, statistical studies and Fourier transform infrared (FTIR) analysis on the effects of MG132, an inhibitor of proteasome, on the pollen germination and tube growth of Pecea wilsonii were carried out in an artificial experimental system. It is showed that MG132 significantly reduced the germination rate and tube growth. Furthermore, MG132 treatment lead to vacuolization occurred both in tube cytoplasm and generative cell. While DMSO and non-proteasome inhibitor E-64 do not have similar effects. FTIR analysis revealed that MG132 treatment markedly reduced the contents of wall-bound proteins and pectin at the apex of tube. Those findings provided evidence that by inhibiting the activity of proteasme, MG132 strongly affects pollen germination and tube growth of P. wilsonii, and that UPP plays an important role in organization and maintaining polarized growth of pollen tube. Inhibition of UPP will induce apoptosis of pollen tube.