Cloning and characterization of the genes involved in cambial growth in response to elevated [CO2]

Aspen (Populus tremuloides) trees growing under elevated [CO2] at a free-air CO2 enrichment (FACE) site have produced significantly more biomass compared to control trees. The molecular mechanisms underlying the observed increase in biomass productivity was investigated by producing transcriptomic profiles of the vascular cambium zone (VCZ) and leaves, followed by a comparative study to identify genes and pathways that showed significant changes following long-term exposure to elevated [CO2]. This study is mainly to verify if genetic modification of a few selected candidate genes including CAP1, CKX6, and ASML2 that are expressed in vascular cambium in response to elevated [CO2] can cause the changes in plant growth and development. To this end, these three genes were cloned into both sense and antisense constructs. Then antisense and sense transgenic lines of above-mentioned genes were developed. 15 events were generated for 5 constructs, which were confirmed with regular PCR and RT-PCR. Confirmed plants were planted in greenhouse for growth and phenotypic characterization. The expression of CAP1, CKX6 and ASML2 in antisense plants was measured by real-time RT-PCR, and the changes caused by gene interference in cambial growth were studies by analyzing the microscopic sections made from the antisense transgenic plants. It has been found that 1) CAP1 is mainly expressed in xylem and root. 2) RNAi suppression of CAP1 significantly affected height and diameter. 3) CAP1, ASML2 and CKX6 affected xylem and phloem cell proliferation and elongation. Due to the delay in regenerating sense transgenic plants, the characterization of sense transgenic plants is limited to growth only.

Optical Access Engine Setup and Validation

The optical access engine integrated with the diagnostic and optical measurement techniques is a great platform for engine research because it provides clear visual access to the combustion chamber inside the engines. An optical access engine customized based on a 4-cylinder spark ignited direct injection (SIDI) production engine is located in the Advanced Power Systems Laboratories (APS LABS) at Michigan Technological University. This optical access engine inside the test cell has been set up for different engine research. In this report, two SAE papers in engine research utilizing the optical access engine are reviewed to gain basic understanding of the methodology. Though the optical engine in APS LABS is a little bit different from the engines used in the literature, the methodology in the papers provides guidelines for engine research through optical access engines. In addition, the optical access engine instrumentation including the test cell setup and the optical engine setup is described in detail in the report providing a solid record for later troubleshooting and reference. Finally, the motoring tests, firing tests and optical imaging experiment on the optical engine have been performed to validate the instrumentation. This report only describes so far the instrumentation of the optical engine in the APS LABS by April 2015.