FP7Individual fellowship2009–2011

TEMPIC · Temperature Perception in Crop Plants

FP7 — People (Marie Curie Actions)

Duration
2009-08-01 → 2011-07-31
EU contribution
€173,569
Participants
1
Scheme
MC-IIF

Lines connect the coordinator with its partners.

Results in brief

Temperature Perception in Crop Plants

The purpose of this project was to assess the impact that changes in ambient temperature, through phenomena such as climate change, are likely to have on monocot crop plants. Specifically, this was done using the model organism Brachypodium distachyon, which is a temperate grass closely related to wheat and barley. Three stages of plant development were assessed, including; young vegetative seedlings, vegetative to reproductive transition and the period of endosperm development and starch accumulation during grain filling, with the later being addressed to increase our understanding of how climate change will effect yield production in major crops, which has strong socio-economic impacts. Firstly, to address the effect that temperature changes have on young seedlings, transcript levels were compared between plants grown at 12ºC to those shifted to either 22ºC or 27ºC, to identify genes that displayed an altered expression pattern proportional to the rise in temperature and to determine the overall response of the transcriptome. Through this analysis, two genes called Heat Shock Protein 70 (HSP70) and Heat Shock Factor 23 (HSF23) were identified as having an increase in gene transcription that was proportional to the rise in temperature. Subsequent analysis showed this result was consistent with plants grown at constant temperatures of 17ºC, 22ºC or 27ºC. This analysis was performed on a genome wide scale using the Brachypodium microarray, with the expression of candidate genes confirmed by quantitative PCR. The microarray revealed 323 genes were unregulated at 27ºC compared to 12ºC, and 344 genes were down-regulated. While this response is robust, the number of genes with altered expression is greatly reduced compared to corresponding experiments in Arabidopsis, suggesting that vegetative tissue of temperate grasses is less responsive than that in dicot plants. This response was further characterised by analysing the behaviour of nucleosomes surrounding the transcription start site (TSS), which had been shown in Arabidopsis to coordinate the transcriptional events that occur as temperature changes. The sensitivity of these particular nucleosomes to changes in temperature is affected by the presence of the histone 2A variant, H2A.Z. Using transgenic lines containing a tagged version of H2A.Z, which were developed during this project, H2A.Z levels and overall nucleosome occupancy at sites surrounding the TSS of these two genes were assessed at 17ºC and 27ºC. These results supported the transcript data, as they suggested that the nucleosomes were not very responsive within this temperature range, which is contrary to results obtained in Arabidopsis. Experiments including tissue from plants grown at 32ºC indicated that a higher ambient temperature did significantly reduce the levels of nucleosomes and H2A.Z. Included in the analysis of the ambient temperature response in vegetative tissue was an assessment of how temperature effects the transition of Brachypodium from vegetative to reproductive life-cycles. Unlike the well-characterised response of Arabidopsis where temperature has been show to compensate for photoperiod in the initiation of flowering, growth in short-days at 27ºC was not able to promote flowering.

Data: CORDIS, © European Union

Project objective

The objective of this proposal is to discover components of the ambient temperature perception pathway in Brachypodium distachyon. This will help us understand the way in which crop plants perceive and respond to changes in ambient temperature, including climate change. A gene expression atlas for the major ambient temperature responses in Brachypodium will be created using microarrays, to identify genes that show a continually variable expression across the ambient (i.e. non-stress inducing) temperature range. These experiments will be compared to data from Arabidopsis to identify the level of conservation in temperature perception pathways between monocots and dicots. The promoters of temperature responsive genes identified through this analysis will be fused to the firefly Luciferase gene to develop ambient temperature reporter lines. Selected lines will be used in forward genetic screens (using fast neutron irradiation mutagenesis) to identify key components of the Brachypodium temperature sensing pathway. This analysis will be complemented by functional characterisation of temperature sensing genes identified in parallel screens in Arabidopsis that have been carried in the Wigge lab as well as using the expression atlas data. These reverse genetics approaches will use RNA interference technology to knock-down the activity of the genes of interest. The phenotypes of these plants will be assessed to determine the direct role of each candidate within the temperature perception pathway. Taken together, this project aims to provide novel information about the way in which our major crop plants will respond to changes in ambient temperature by continuing the development of Brachypodium as a model organism for temperate cereals, such as wheat and barley, and identifying genetic components within the monocot temperature perception pathway.

Original text from CORDIS.

Participants

Links

Data: CORDIS, © European Union