An investigation of phenolic glycoside and condensed tannin homeostasis in Populus by salicyl alcohol feeding to cell cultures and by transgenic manipulation of the sucrose transporter, PTSUT4, in planta

Secondary metabolites play an important role in plant protection against biotic and abiotic stress. In Populus, phenolic glycosides (PGs) and condensed tannins (CTs) are two such groups of compounds derived from the common phenylpropanoid pathway. The basal levels and the inducibility of PGs and CTs depend on genetic as well as environmental factors, such as soil nitrogen (N) level. Carbohydrate allocation, transport and sink strength also affect PG and CT levels. A negative correlation between the levels of PGs and CTs was observed in several studies. However, the molecular mechanism underlying such relation is not known. We used a cell culture system to understand negative correlation of PGs and CTs. Under normal culture conditions, neither salicin nor higher-order PGs accumulated in cell cultures. Several factors, such as hormones, light, organelles and precursors were discussed in the context of aspen suspension cells’ inability to synthesize PGs. Salicin and its isomer, isosalicin, were detected in cell cultures fed with salicyl alcohol, salicylaldehyde and helicin. At higher levels (5 mM) of salicyl alcohol feeding, accumulation of salicins led to reduced CT production in the cells. Based on metabolic and gene expression data, the CT reduction in salicin-accumulating cells is partly a result of regulatory changes at the transcriptional level affecting carbon partitioning between growth processes, and phenylpropanoid CT biosynthesis. Based on molecular studies, the glycosyltransferases, GT1-2 and GT1-246, may function in glycosylation of simple phenolics, such as salicyl alcohol in cell cultures. The uptake of such glycosides into vacuole may be mediated to some extent by tonoplast localized multidrug-resistance associated protein transporters, PtMRP1 and PtMRP6. In Populus, sucrose is the common transported carbohydrate and its transport is possibly regulated by sucrose transporters (SUTs). SUTs are also capable of transporting simple PGs, such as salicin. Therefore, we characterized the SUT gene family in Populus and investigated, by transgenic analysis, the possible role of the most abundantly expressed member, PtSUT4, in PG-CT homeostasis using plants grown under varying nitrogen regimes. PtSUT4 transgenic plants were phenotypically similar to the wildtype plants except that the leaf area-to-stem volume ratio was higher for transgenic plants. In SUT4 transgenics, levels of non-structural carbohydrates, such as sucrose and starch, were altered in mature leaves. The levels of PGs and CTs were lower in green tissues of transgenic plants under N-replete, but were higher under N-depleted conditions, compared to the levels in wildtype plants. Based on our results, SUT4 partly regulates N-level dependent PG-CT homeostasis by differential carbohydrate allocation.

Security Evaluation of Substation Network Architectures

In recent years, security of industrial control systems has been the main research focus due to the potential cyber-attacks that can impact the physical operations. As a result of these risks, there has been an urgent need to establish a stronger security protection against these threats. Conventional firewalls with stateful rules can be implemented in the critical cyberinfrastructure environment which might require constant updates. Despite the ongoing effort to maintain the rules, the protection mechanism does not restrict malicious data flows and it poses the greater risk of potential intrusion occurrence. The contributions of this thesis are motivated by the aforementioned issues which include a systematic investigation of attack-related scenarios within a substation network in a reliable sense. The proposed work is two-fold: (i) system architecture evaluation and (ii) construction of attack tree for a substation network. Cyber-system reliability remains one of the important factors in determining the system bottleneck for investment planning and maintenance. It determines the longevity of the system operational period with or without any disruption. First, a complete enumeration of existing implementation is exhaustively identified with existing communication architectures (bidirectional) and new ones with strictly unidirectional. A detailed modeling of the extended 10 system architectures has been evaluated. Next, attack tree modeling for potential substation threats is formulated. This quantifies the potential risks for possible attack scenarios within a network or from the external networks. The analytical models proposed in this thesis can serve as a fundamental development that can be further researched.