Antisense-Mediated Depletion of Tomato Chloroplast Omega-3 Fatty Acid Desaturase Enhances Thermal Tolerance

A chloroplast-localized tomato （Lycopersicon esculentum Mill.） ω-3 fatty acid desaturase gene （LeFADT） was isolated and characterized with regard to its sequence, response to various temperatures, and function in antisense transgenic tomato plants. The deduced amino acid sequence had four histidine-rich regions, of which three regions were highly conserved throughout the whole ω-3 fatty acid desaturasegene family. Southern blotting analysis showed that LeFAD7was encoded by a single copy gene and had two homologous genes in the tomato genome. Northern blot showed that LeFAD7 was expressed in all organs and was especially abundant in leaf tissue. Meanwhile, expression of LeFAD7 was induced by chilling stress （4 ℃）, but was inhibited by high temperature （45 ℃）, in leaves. Transgenic tomato plants were produced by integration of the antisense LeFAD7DNA under the control of a CaMV35S promoter into the genome. Antisense transgenic plants with lower 18 ： 3 content could maintain a higher maximal photochemical efficiency （Fv/Fm） and O2 evolution rate than wild-type plants. These results suggested that silence of the LeFAD7 gene alleviated high-temperature stress. There was also a correlation between the low content of 18 ： 3 resulting from silence of the LeFAD7 gene and tolerance to high-temperature stress.

An efficient and coordinated mapping algorithm in virtualized SDN networks

Software-defined networking(SDN) enables the network virtualization through SDN hypervisors to share the underlying physical SDN network among multiple logically isolated virtual SDN networks(v SDNs),each with its own controller.The v SDN embedding,which refers to mapping a number of v SDNs to the same substrate SDN network,is a key problem in the SDN virtualization environment.However,due to the distinctions of the SDN,such as the logically centralized controller and different virtualization technologies,most of the existing embedding algorithms cannot be applied directly to SDN virtualization.In this paper,we consider controller placement and virtual network embedding as a joint vS DN embedding problem,and formulate it into an integer linear programming with objectives of minimizing the embedding cost and the controller-to-switch delay for each v SDN.Moreover,we propose a novel online vS DN embedding algorithm called CO-v SDNE,which consists of a node mapping stage and a link mapping stage.In the node mapping stage,CO-vS DNE maps the controller and the virtual nodes to the substrate nodes on the basis of the controller-to-switch delay and takes into account the subsequent link mapping at the same time.In the link mapping stage,CO-v SDNE adopts the k-shortest path algorithm to map the virtual links.The evaluation results with simulation and Mininet emulation show that the proposed CO-v SDNE not only significantly increases the long-term revenue to the cost ratio and acceptance ratio while guaranteeing low average and maximum controller-to-switch delay,but also achieves good v SDN performance in terms of end-to-end delay and throughput.

Antisense-mediated Depletion of Tomato Endoplasmic Reticulum Omega-3 Fatty Acid Desaturase Enhances Thermal Tolerance

An endoplasmic reticulum-localized tomato ω-3 fatty acid desaturase gene （LeFAD3） was isolated. The antisense tomato plants were obtained under the control of the cauliflower mosaic virus 35S promoter （35S-CaMV）. Northern blot analysis confirmed that the expression of LeFAD3 was inhibited in the tomato genome. Levels of 18：3 decreased and correspondingly levels of 18：2 increased in total lipids of leaves and roots. After heat stress, the fresh weight of the aerial parts of antisense transgenic plants was higher than that of the wild type （WT） plants. The membrane system ultrastructure of chloroplasts in leaf cells and all of the subcellular organelles in the root tips of transgenic plants remained more intact than those of WT. Relative electric conductivity increased less in transgenic plants than in WT. Under heat stress, the maximal photochemical efficiency of photosystem Ⅱ （Fv/Fm） and the O2 evolution rate decreased more in WT than in transgenic plants. These results suggested that the depletion of LeFAD3 increased the saturation of fatty acids and alleviated high temperature stress.

Reduction of methylviologen-mediated oxidative stress tolerance in antisense transgenic tobacco seedlings through restricted expression of StAPX

Ascorbate peroxidases are directly involved in reactive oxygen species (ROS) scavenging by reducing hydrogen peroxide to water. The tomato thylakoid-bound ascorbate peroxidase gene (StAPX) was introduced into tobacco. RNA gel blot analysis confirmed that StAPX in tomato leaves was induced by methylviologen-mediated oxidative stress. The sense transgenic seedlings exhibited higher tAPX activity than that of the wild type (WT) plants under oxidative stress conditions, while the antisense seedlings exhibited lower tAPX activity. Lower APX activities of antisense transgenic seedlings caused higher malondialdehyde contents and relative electrical conductivity. The sense transgenic seedlings with higher tAPX activity maintained higher chlorophyll content and showed the importance of tAPX in maintaining the optimal chloroplast development under methylviologen stress conditions, whereas the antisense lines maintained lower chlorophyll content than WT seedlings. Results indicated that the over-expression of StAPX enhanced tolerance to methylviologen-mediated oxidative stress in sense transgenic to- bacco early seedlings, whereas the suppression of StAPX in antisense transgenic seedlings showed high sensitivity to oxidative stress.