SEASONAL NITROGEN CYCLING AND HOMEOSTASIS IN POPULUS: SOURCE-SINK COMMUNICATION
Coleman, Gary D
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Nitrogen (N) is an essential nutrient for plant growth, development and reproduction. Seasonal N cycling is an adaption to nutrient limitation and a feature of the perennial lifestyle of trees. Poplar (Populus) is a model system used to study forest tree genetics and molecular biology, including seasonal N cycling. The accumulation of Bark storage proteins (BSP) is a central feature of seasonal N cycling in poplar, yet our understanding of the contribution of the BSP storage pool to N remobilization during growth and mechanisms that regulate BSP accumulation, catabolism and N remobilization is limited. The research presented in this dissertation is directed towards advancing knowledge of the regulation of seasonal N cycling in poplar using a combination of experimental approaches. The role of the N storage pool to N remobilization was examined through N sink manipulations and the specific role of BSP storage was investigated by N source reduction via RNAi mediated knockdown of BSP accumulation. From this it was found that N remobilization from BSP significantly contributes to shoot growth following dormancy and initial shoot growth is source limited. Poplar bark transcriptome analyses during regrowth following dormancy revealed an enrichment for up-regulated genes associated with auxin transport and signaling. Based on the transcriptome analysis experiments that manipulate auxin production or polar auxin transport were performed and the results indicate that BSP catabolism and N remobilization likely involves polar auxin transport from expanding buds and shoots and auxin-mediated regulation of protease gene expression. Analysis of DNA microarrays of bark RNA during short-day (SD) induction of BSP gene expression was used to identify putative regulatory factors that may play a role of BSP accumulation. The transcriptome analyses indicated that SD represses the expression of genes involved in ethylene production as well as a reduction in bark ethylene biosynthesis. Additionally, treatment of excised stems with ACC or ethephon repressed BSP gene expression while AVG induced BSP gene expression. This repression was reduced in ethylene-insensitive poplars expressing Arabidopsis dominant gain-of-function allele etr1-1. Furthermore, transient expression of ERF12 and ERF41, two transcription factors with the greatest induction in SD treatment, in transgenic tobacco stably transformed with BSPA promoter fused with GUS resulted in enhanced GUS activity suggesting ERF12 and ERF41 may act as positive regulators of BSP gene expression. Since glutamine plays a pivotal role in N partitioning during N storage and remobilization, the possible role of the PII glutamine sensor was studied and it was found that the transcript levels of PII increased in bark during SD-induced leaf senescence and BSP accumulation. PII knockdown poplars using RNAi showed reduced glutamine-induced BSP gene expression. Moreover, glutamine-induced BSP gene expression was also inhibited by 2-OG, a PII-NAGK interaction antagonist, suggesting a possible role of PII-glutamine sensing in BSP accumulation. Taken together, this study provides important insights into the mechanism of seasonal N accumulation and remobilization in poplar.