Functional analysis of NLP7 for optimising nitrate responsive growth

Lead Research Organisation: John Innes Centre
Department Name: Contracts Office


We have recently discovered that the NIN-like transcription factor NPL7 integrates both nitrate- and carbohydrate- mediated signals to control the expression of a set of nitrate uptake and assimilatory genes. This discovery may provide an approach to understanding mechanisms integrating the supplies of nitrate taken up from the soil and supplies of photosynthate from leaves. Such understanding will contribute to the analyses of nitrate uptake and use efficiency in crops aimed at optimising these processes.

The specific objectives of research in this proposal are to:
1. Use proteomics to identify post-translational modifications to NLP7 and interacting proteins in response to nitrate and glucose, and to test the effects of altered modifications and domain deletions and mutations on NLP7 localization and transcriptional activation of NRT2.1;
2. Identify genes influencing NLP7 activity by genetic screens and by screens for interacting proteins and testing their functions using genetics, biochemical assays and gene expression;
3. Use a high throughput screen to establish quantitative understanding of how NRT2.1 expression is influenced by of NLP7 domains and mutations, and by interacting proteins;
4. Perform re-iterative screens based on mutagenesis of NLP7 and interacting proteins and assessing combinations of mutations and components on quantitative transcription of NRT2.1;
5. Functional analysis of selected mutations of NLP7 and interacting proteins in stable transgenic Arabidopsis to determine nitrate uptake and utilization;
6. Maximize the impact of the work by working with industry and crop researchers, by promoting the role of basic research, generating understanding and resources, and by providing training opportunities.


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