Wounding and methyl jasmonate increase cyanogenic glucoside concentrations in Sorghum bicolor via upregulation of biosynthesis.

The cyanogenic glucoside, dhurrin, present in Sorghum bicolor is thought to have multiple functions, including in defence against herbivory. The hormone methyl jasmonate (MeJA) is also induced by herbivory and is key to instigating defence processes in plants. To investigate whether dhurrin is induced in response to herbivore attack and also to the associated presence of MeJA, sorghum plants were either wounded or exogenous MeJA was applied. We show that specific wounding (pin board and perforation) and the application of MeJA increases dhurrin concentration in leaves and sheath tissue 12 h after treatment. Quantitative PCR shows that the expression of two genes, SbCYP79A1 and SbUGT85B1, involved in the synthesis of dhurrin are significantly induced by exogenous MeJA and by wounding. Analysis of 2 kb of sequence upstream of the start codon of SbCYP79A1 identifies several cis-acting elements that have been linked to MeJA induction. A promoter deletion series, coupled to GFP, and transiently expressed in Nicotiana benthamiana suggests that there are potentially three sequence motifs (~-925 to -976) involved in the binding of transcription factors that result in increased expression of SbCYP79A1 and the synthesis of dhurrin in response to MeJA.

Regulation of cyanogenic glucosides in wild and domesticated Eusorghum taxa.

Domesticated sorghum (Sorghum bicolor [L.] Moench subsp. bicolor) diverts significant amounts of nitrogen away from primary metabolism to the synthesis of cyanogenic glucosides (CNglc) - specialized metabolites that release toxic hydrogen cyanide (HCN). Our aim was to identify the point in the genus Sorghum Moench at which plants gained the ability to maintain hazardous concentrations of cyanogenic glucosides in their leaves into maturity (HCN potential >0.4 mg g-1 ). This ability occurs in domesticated sorghum (in the subgenus Eusorghum), but not in wild taxa in other Sorghum subgenera. Eight accessions from the subgenus Eusorghum were grown in a common garden: an improved sorghum line, five sorghum landraces, the crop's wild progenitor (S. bicolor subsp. verticilliflorum [Steud.] de Wet ex Wiersema & J. Dahlb.) and wild Sorghum propinquum (Kunth) Hitchc. HCN potential was measured in plants (n = 80) at the three-leaf stage and at 6 weeks old. All study accessions, including the wild taxa, had hazardous CNglc concentrations in the leaves at both the three-leaf stage (mean HCN potentials > = 2.5 mg g-1 ) and at 6 weeks old (mean HCN potentials > = 0.68 mg g-1 ), greatly contrasting the much lower mature leaf HCN potentials previously found in wild Sorghum taxa outside subgenus Eusorghum (generally <= 0.01 mg g-1 ). Our results suggest that the ability to maintain hazardous leaf HCN potentials into maturity might have arisen during the divergence of Eusorghum from other Sorghum subgenera, rather than during the speciation or domestication of S. bicolor, and highlights the value of utilizing Sorghum taxa outside Eusorghum in efforts to improve the crop safety of sorghum.

Dhurrin increases but does not mitigate oxidative stress in droughted Sorghum bicolor.

MAIN CONCLUSION: Droughted sorghum had higher concentrations of ROS in both wildtype and dhurrin-lacking mutants. Dhurrin increased in wildtype genotypes with drought. Dhurrin does not appear to mitigate oxidative stress in sorghum. Sorghum bicolor is tolerant of high temperatures and prolonged droughts. During droughts, concentrations of dhurrin, a cyanogenic glucoside, increase posing a risk to livestock of hydrogen cyanide poisoning. Dhurrin can also be recycled without the release of hydrogen cyanide presenting the possibility that it may have functions other than defence. It has been hypothesised that dhurrin may be able to mitigate oxidative stress by scavenging reactive oxygen species (ROS) during biosynthesis and recycling. To test this, we compared the growth and chemical composition of S. bicolor in total cyanide deficient sorghum mutants (tcd1) with wild-type plants that were either well-watered or left unwatered for 2 weeks. Plants from the adult cyanide deficient class of mutant (acdc1) were also included. Foliar dhurrin increased in response to drought in all lines except tcd1 and acdc1, but not in the roots or leaf sheaths. Foliar ROS concentration increased in drought-stressed plants in all genotypes. Phenolic concentrations were also measured but no differences were detected. The total amounts of dhurrin, ROS and phenolics on a whole plant basis were lower in droughted plants due to their smaller biomass, but there were no significant genotypic differences. Up until treatments began at the 3-leaf stage, tcd1 mutants grew more slowly than the other genotypes but after that they had higher relative growth rates, even when droughted. The findings presented here do not support the hypothesis that the increase in dhurrin commonly seen in drought-stressed sorghum plays a role in reducing oxidative stress by scavenging ROS.

Utilization of a high-throughput shoot imaging system to examine the dynamic phenotypic responses of a C4 cereal crop plant to nitrogen and water deficiency over time.

The use of high-throughput phenotyping systems and non-destructive imaging is widely regarded as a key technology allowing scientists and breeders to develop crops with the ability to perform well under diverse environmental conditions. However, many of these phenotyping studies have been optimized using the model plant Arabidopsis thaliana. In this study, The Plant Accelerator(®) at The University of Adelaide, Australia, was used to investigate the growth and phenotypic response of the important cereal crop, Sorghum bicolor L. Moench and related hybrids to water-limited conditions and different levels of fertilizer. Imaging in different spectral ranges was used to monitor plant composition, chlorophyll, and moisture content. Phenotypic image analysis accurately measured plant biomass. The data set obtained enabled the responses of the different sorghum varieties to the experimental treatments to be differentiated and modelled. Plant architectural instead of architecture elements were determined using imaging and found to correlate with an improved tolerance to stress, for example diurnal leaf curling and leaf area index. Analysis of colour images revealed that leaf 'greenness' correlated with foliar nitrogen and chlorophyll, while near infrared reflectance (NIR) analysis was a good predictor of water content and leaf thickness, and correlated with plant moisture content. It is shown that imaging sorghum using a high-throughput system can accurately identify and differentiate between growth and specific phenotypic traits. R scripts for robust, parsimonious models are provided to allow other users of phenomic imaging systems to extract useful data readily, and thus relieve a bottleneck in phenotypic screening of multiple genotypes of key crop plants.

Plant nutrient acquisition and utilisation in a high carbon dioxide world.

Producing enough food to meet the needs of an increasing global population is one of the greatest challenges we currently face. The issue of food security is further complicated by impacts of elevated CO2 and climate change. In this viewpoint article, we begin to explore the impacts of elevated CO2 on two specific aspects of plant nutrition and resource allocation that have traditionally been considered separately. First, we focus on arbuscular mycorrhizas, which play a major role in plant nutrient acquisition. We then turn our attention to the allocation of resources (specifically N and C) in planta, with an emphasis on the secondary metabolites involved in plant defence against herbivores. In doing so, we seek to encourage a more integrated approach to investigation of all aspects of plant responses to eCO2.