Evaluation of vacuum infiltration vitrification and cryo-mesh cryopreservation techniques with Arabidopsis thaliana shoot tips.

BACKGROUND: Novel cryo-techniques are continuously being developed that may better improve cryogenic survival in plants, with the aim of reducing exposure times to otherwise toxic cryoprotective agents whilst maximising regeneration rates. OBJECTIVE: This study used cryo-mesh and vacuum infiltration vitrification with two vitrification solutions (PVS2 and PVS3) to develop an optimised cryopreservation protocol for Arabidopsis thaliana. MATERIALS AND METHODS: Shoot tips from 10-day old seedlings of wild type A. thaliana were cryopreserved using either vacuum infiltration vitrification or the cryo-mesh technique. Shoot tips were treated for up to 60 min in increments of 10 min with PVS2 and PVS3, and for an additional 180 and 300 min incubation for cryo-mesh prior to exposure to liquid nitrogen. RESULTS: Both methods resulted in very high regeneration rates, but which decreased after longer exposure to the vitrification solutions. The highest regeneration rate for vacuum-infiltration vitrification was attained after only 30 min incubation in PVS2 (92.5%) and 50 min incubation in PVS3 (93.6%). In the case of cryo-mesh the highest regeneration was observed after 180 min incubation in either PVS2 (100%) or PVS3 (92.2%). CONCLUSION: Vacuum-infiltration vitrification is more effective than cryo-mesh by reducing exposure times to cryoprotective solutions whilst achieving very high regeneration rates of shoot tips of A. thaliana. https://doi.org/10.54680/fr22610110712.

Unraveling the role of mitochondria during oxidative stress in plants.

The sedentary habit of plants means that they must stand and fight environmental stresses that their mobile animal cousins can avoid. A range of these abiotic stresses initiate the production in plant cells of reactive oxygen and nitrogen species that ultimately lead to oxidative damage affecting the yield and quality of plant products. A complex network of enzyme systems, producing and quenching these reactive species operate in different organelles. It is the integration of these compartmented defense systems that coordinates an effective response to the various stresses. Future attempts to improve plant growth or yield must consider the complexity of inter-organelle signaling and protein targeting if they are to be successful in producing plants with resistance to a broad range of stresses. Here we highlight the role of pre-oxidant, antioxidant, and post-oxidant defense systems in plant mitochondria and the potential role of proteins targeted to both mitochondria and chloroplasts, in an integrated defense against oxidative damage in plants.

The expression of alternative oxidase and uncoupling protein during fruit ripening in mango.

The expression of alternative oxidase (Aox) and uncoupling proteins (Ucp) was investigated during ripening in mango (Mangifera indica) and compared with the expression of peroxisomal thiolase, a previously described ripening marker in mango. The multigene family for the Aox in mango was expressed differentially during ripening. Abundance of Aox message and protein both peaked at the ripe stage. Expression of the single gene for the Ucp peaked at the turning stage and the protein abundance peaked at the ripe stage. Proteins of the cytochrome chain peaked at the mature stage of ripening. The pattern of protein accumulation suggested that increases in cytochrome chain components played an important role in facilitating the climacteric burst of respiration and that the Aox and Ucp may play a role in post-climacteric senescent processes. Because both message and protein for the Aox and Ucp increased in a similar pattern, it suggests that their expression is not controlled in a reciprocal manner but may be active simultaneously.