Natural Resources, School of


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A DISSERTATION Presented to the Faculty of The Graduate College at the University of Nebraska In Partial Fulfillment of Requirements For the Degree of Doctor of Philosophy, Major: Natural Resource Sciences, Under the Supervision of Professor Tala N. Awada. Lincoln, Nebraska: December, 2011

Copyright (c) 2011 Joseph Msanne


Cellular and molecular aspects of abiotic stress responses in Arabidopsis thaliana subjected to cold, drought, and high salinity and in two photosynthetic green alga, Chlamydomonas reinhardtii and Coccomyxa sp. C-169, subjected to nitrogen deprivation were investigated. Cold, drought, and high salinity can negatively affect plant growth and crop production. The first research aimed at determining the physiological functions of the stress-responsive Arabidopsis thaliana RD29A and RD29B genes. Cold, drought, and salt induced both genes; the promoter of RD29Awas found to be more responsive to drought and cold stresses, whereas the promoter of RD29B was highly responsive to salt stress. Therefore, RD29A and RD29B gene sequences have the potential to confer abiotic stress resistance in crop species grown in arid and semi-arid regions. RD29A and RD29B proteins were also found to unlikely serve directly as protective molecules. The second study aimed at investigating the impacts of nitrogen deprivation in Chlamydomonas reinhardtii and Coccomyxa sp. C-169; results showed that these microalgae altered their lipid metabolism by synthesizing and accumulating the neutral lipid triacylglycerol (TAG). Since microalgae have emerged as suitable feedstocks for renewable biofuel production, the purpose of this analysis was to understand the genetic and biochemical mechanisms associated with the induction of TAG synthesis in Chlamydomonas and Coccomyxa subjected to nitrogen deprivation under photoautotrophic conditions. In addition to TAG accumulation, nitrogen depletion triggered an early synthesis of starch and up-regulation of several genes in Chlamydomonas, including some diacylglycerol:acyl-CoA acyltransferases, catalyzing the acylation of diacylglycerol to TAG. Protein degradation in nitrogen-deprived cells might provide carbon skeletons for TAG biosynthesis. In a related study, the effects of the autophagy-inducer rapamycin and the autophagy-inhibitor 3-methyladenine (3-MA) on the accumulation of TAG in Chlamydomonas cells subjected to nitrogen deprivation were investigated. 3-MA induced TAG accumulation in cells growing in both nitrogen-deprived and control media. The increase in TAG content in cells subjected to nitrogen deprivation might not be a direct response to an autophagic activity induced by nutrient depletion.

Advisor: Tala N. Awada