Optimising water use in wheat

Lead Research Organisation: Rothamsted Research
Department Name: Unlisted


Water is a major determinant of world-wide crop yield. In the UK yield losses due to drought, in wheat, the most widely grown UK crop, average 1-2 t ha-1 and cost approximately £60M per year. The predicted changes in climate patterns are projected to increase the losses (e.g. in the south and east of the UK, a 1-2 oC increase in mean summer temperatures and decrease summer rainfall as a result of global climate change). Drought tolerance is highly complex trait made up from a number of usually multigenic component traits. By integrating genetics, genomics, crop physiology, biochemistry and biomathematics and agronomy we aim to both understand and deconvolute the drought response, identify the component traits important in attaining high stable yield under drought, identify QTLs and the candidate genes. We will investigate natural variation in germplasm (mapping populations and accessions) for water use efficiency, integrative morpho-physiological traits and yield. We will establish the genotype X environment interactions under drought exploiting a large qtl datasets representing more than 30 environments.
Metabolic signalling networks and regulation of metabolism control responses and adaptation to drought (metabolism,growth, and physiology). Whilst the identities of some signalling elements are known the identities of the sensors and overall cross talk between pathways is less clear. Variation in the key genes involved in primary metabolism and metabolic signalling will be determined by wide germplasm screening and expression studies. The function of candidate genes will be established by transformation or TILLING. We will develop new conceptual models to describe physiological, molecular and biochemical response of genotypes to progressive drought.


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Araus JL (2014) Phenotyping and other breeding approaches for a New Green Revolution. in Journal of integrative plant biology

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Chen L (2014) Progress in TILLING as a tool for functional genomics and improvement of crops. in Journal of integrative plant biology

Description The C4 photosynthetic pathway involves the assimilation of CO2 by phosphoenolpyruvate carboxylase (PEPC) and the subsequent decarboxylation of C4 acids. The enzymes of the CO2 concentrating mechanism could be affected under water deficit and limit C4 photosynthesis. Three different C4 grasses were submitted to gradually induced drought stress conditions: Paspalum dilatatum (NADP-malic enzyme, NADP-ME), Cynodon dactylon (NAD-malic enzyme, NAD-ME) and Zoysia japonica (PEP carboxykinase, PEPCK). Moderate leaf dehydration affected the activity and regulation of PEPC in a similar manner in the three grasses but had species-specific effects on the C4 acid decarboxylases, NADP-ME, NAD-ME and PEPCK, although changes in the C4 enzyme activities were small. In all three species, the PEPC phosphorylation state, judged by the inhibitory effect of l-malate on PEPC activity, increased with water deficit and could promote increased assimilation of CO2 by the enzyme under stress conditions. Appreciable activity of PEPCK was observed in all three species suggesting that this enzyme may act as a supplementary decarboxylase to NADP-ME and NAD-ME in addition to its role in other metabolic pathways. Rubisco isolated from each of the three C4 grasses was characterized by smaller specificity factors (SC/O), larger Michaelis-Menten constants for CO2 (Kc) and O2 (Ko), and larger maximum carboxylation velocities (Vc) than Rubisco from wheat, which can be rationalized in terms of the CO2-rich environment of C4 Rubisco in the bundle sheath. During leaf dehydration the quantity and maximum activity of Rubisco remained unchanged but the initial and total activities declined slightly, possibly due to increased inhibition. Tight-binding inhibitors were present in the light but were more abundant in the dark, especially in Z. japonica, and increased in quantity with drought stress. The inhibitor from darkened leaves of Z. japonica was identified as 2-carboxyarabinitol-1-phosphate (CA1P). Consistent with the presence of CA1P, the total activity of Rubisco was decreased after 12 h darkness in Z. japonica. Ribulose-1,5-bisphosphate (RuBP) in the leaves decreased with drought stress, to quantities approximating those of Rubisco catalytic sites. The magnitude of the decrease in RuBP suggested that, at least in C. dactylon and Z. japonica, it could contribute to the drought-induced decrease in photosynthesis.
The genes for CA1Pase (2-carboxy-D-arabinitol-1-bisphosphate phosphatase) from French bean, wheat, Arabidopsis and tobacco were identified and cloned. The deduced protein sequence included an N-terminal motif identical with the PGM (phosphoglycerate mutase) active site sequence [LIVM]-x-R-H-G-[EQ]-x-x-[WN]. The corresponding gene from wheat coded for an enzyme with the properties published for CA1Pase. The expressed protein lacked PGM activity but rapidly dephosphorylated 2,3-DPG (2,3-diphosphoglycerate) to 2-phosphoglycerate. DTT (dithiothreitol) activation and GSSG inactivation of this enzyme was pH-sensitive, the greatest difference being apparent at pH 8. The presence of the expressed protein during in vitro measurement of Rubisco (ribulose-1,5-bisphosphate carboxylase/oxygenase) activity prevented a progressive decline in Rubisco turnover. This was due to the removal of an inhibitory bisphosphate that was present in the RuBP (ribulose-1,5-bisphosphate) preparation, and was found to be PDBP (D-glycero-2,3-pentodiulose-1,5-bisphosphate). The substrate specificity of the expressed protein indicates a role for CA1Pase in the removal of 'misfire' products of Rubisco.
Plants produce various compounds in response to water deficit. We identified, the presence and identification of a drought-inducible non-protein amino acid in the leaves of two C4 grasses. The soluble amino acids extracted from the leaves of three different species were measured by high-performance liquid chromatography of derivatives formed with o-phthaldialdehyde and ß-mercaptoethanol. One amino acid that increased in amount with drought stress had a retention time not corresponding to any common amino acid. Its identity was determined by metabolite profiling, using 1H NMR and GC-MS. This unusual amino acid was present in the dehydrated leaves of Cynodon dactylon (L.) Pers. and Zoysia japonica Steudel, but was absent from Paspalum dilatatum Poir. Its identity as 2-amino-5-hydroxypentanoic acid (5-hydroxynorvaline, 5-HNV) was confirmed by synthesis and co-chromatography of synthetic and naturally occurring compounds. The amount of 5-HNV in leaves of the more drought tolerant C4 grasses, C. dactylon and Z. japonica, increased with increasing water deficit; therefore, any benefits from this unusual non-protein amino acid for drought resistance should be further explored.
The yeast regulatory protein kinase, general control non-derepressible-2 (GCN2) plays a key role in general amino acid control. GCN2 phosphorylates the alpha subunit of the trimeric eukaryotic translation initiation factor-2 (eIF2), bringing about a decrease in the general rate of protein synthesis but an increase in the synthesis of GCN4, a transcription factor that promotes the expression of genes encoding enzymes for amino acid biosynthesis. The present study concerned the phosphorylation of Arabidopsis eIF2alpha (AteIF2alpha) by the Arabidopsis homologue of GCN2, AtGCN2, and the role of AtGCN2 in regulating genes encoding enzymes of amino acid biosynthesis and responding to virus infection. A null mutant for AtGCN2 called GT8359 was obtained and western analysis confirmed that it lacked AtGCN2 protein. GT8359 was more sensitive than wild-type Arabidopsis to herbicides that affect amino acid biosynthesis. Phosphorylation of AteIF2alpha occurred in response to herbicide treatment but only in wild-type Arabidopsis, not GT8359, showing it to be AtGCN2-dependent. Expression analysis of genes encoding key enzymes for amino acid biosynthesis and nitrate assimilation revealed little effect of loss of AtGCN2 function in GT8359 except that expression of a nitrate reductase gene, NIA1, was decreased. Analysis of wild-type and GT8359 plants infected with Turnip yellow mosaic virus or Turnip crinkle virus showed that AteIF2alpha was not phosphorylated
Sucrose nonfermenting-1 (SNF1)-related protein kinases (SnRKs) form subfamilies: SnRK1, SnRK2, and SnRK3. SnRK1 plays a major role in the regulation of carbon metabolism and energy status, while SnRKs 2 and 3 have been implicated in stress and abscisic acid (ABA)-mediated signalling pathways. We have shown that the levels of SnRK1 protein were shown to decline dramatically in wheat roots in response to ABA treatment, although the amount of phosphorylated (active) SnRK1 remained constant. Multiple SnRK2-type protein kinases were detectable in the root extracts and showed differential responses to ABA treatment. They included a 42 kDa protein that appeared to reduce in response to 3 h of ABA treatment but to recover after longer treatment. There was a clear increase in phosphorylation of this SnRK2 in response to the ABA treatment. Fractions containing this 42 kDa SnRK2 were shown to phosphorylate synthetic peptides with amino acid sequences based on those of conserved phosphorylation sites in AREBPs. The activity increased 8-fold with the addition of calcium chloride, indicating that it is calcium-dependent. The activity assigned to the 42 kDa SnRK2 also phosphorylated a heterologously expressed wheat AREBP.
Exploitation Route The results have increased our understanding of the underlying biochemistry and physiology. This new knowledge can be exploited by identifying natural variation that can be used in breeding approaches for crop improvement or through biotechnological approaches
Sectors Agriculture, Food and Drink

Description Food, Health and Environmental Security 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact New breeding approaches to increase the yield & quality of crops. Palma de Mallorca, February 20 - 22nd, 2018
Year(s) Of Engagement Activity 2018
Description 2nd Agriculture and Climate Change Conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact 2nd Agriculture and Climate Change Conference
Year(s) Of Engagement Activity 2017
Description DEFRA University Workshop 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact This workshop brought together a group of universities with Defra policy teams / evidence specialists to explore the opportunities available through academic partnerships (including fellowships, studentships and other grant processes) and the benefits of engaging with external academic experts through developing contacts and networks.
Year(s) Of Engagement Activity 2018