I. Responses of plant to trace metallic elements excess
- A. Cadmium accumulation and tolerance in plants
Several strategies are being developed in our laboratory to identify genes involved in Cd tolerance and hyperaccumulation.
Screening of cadmium resistant yeasts
Growth of S. cerevisiae (strain BY474)
Comparison of Noccaea caerulescens or Arabidopsis halleri populations
Populations originating from heavy metal polluted or non polluted (metalliferous) soils, have been studied in collaboration with Prof. A. Smith (University of Oxford). (Roosens et al. 2003)
This study has revealed that the growth of some metallicolous populations (originating from South of France, St Félix-de-Pallière) was enhanced by the addition of Cd concentrations that are toxic for other populations. The molecular mechanism behind this effect is not deciphered yet.
The second major QTL for Cd tolerance co-localizes with CAX1 encoding a Ca++/H+vacuolar antiporter (Baliardini et al. 2015). CAX1 seems to play an important role in Cd tolerance of plants by limiting oxidative stress. CAX1 activity is thought to disrupt the positive feedback between increasy in cytosolic Ca++ and ROS (Baliardini et al. 2015, Baliardini et al. 2015).
Using the same strategy with a Arabidopsis halleri non metallicolous population the QTL analysis revealed only HMA4 as sole major determinant for the Cd and Zn tolerance traits. This result provides evidence that preadaptation process in Arabidopsis halleri prior colonization of Zn and Cd contaminated sites included HMA4 duplication (Meyer et al. 2016)
Overexpression of selected genes in Arabidopsis thaliana
Selected genes are overexpressed in Arabidopsis thaliana, which is the easiest transformable plant since transformation can be done by flower dipping. As A. thaliana is very sensitive to cadmium, effect of gene overexpression on Cd tolerance can be easily measured. If enhanced cadmium accumulation and/or tolerance is observed, application will be foreseen in cultivated species.
- B. COPPER ACCUMULATION AND TOLERANCE IN PLANTS
We have studied Haumaniastrum katangense, called the copper flower, which shows extreme tolerance to copper and high accumulation capacity (PhD thesis of François Chipeng, Chipeng et al. 2009). We have demonstrated a higher requirement for copper for normal growth. Surplus copper was also required for cultivating H. katangense in sterile conditions, suggesting that Cu excess may be necessary for needs other than pathogen defence. Further studies have included the presence of cobalt excess in the soils of the copper arc. In particular, the endemic species and obligate metallophyte Haumaniastrum robertii was studied together with its facultative metallophyte congener H. katangense. H. robertii growth was conditional on addition of both copper and cobalt. Furthermore the beneficial effect of cobalt on the plant growth was conditional on simultaneous copper supplement. Co and Cu accumulate in different tissues of Haumaniastrum and they show different mobility in plants (PhD thesis of Francine Ilunga Kabeya, Kabeya et al. submitted)
The endophytic community of H. katangense and Crepidorhopalon tenuis, an other cuprophyte of the copper arc, was studied in collaboration with the Institut de Recherches Microbiologiques JMW/Laboratoire de Microbiologie and Max Mergeay (CEN/SCK), as a first step to evaluate their potential contribution to plant adaptation to copper excess (PhD thesis of Alfred Cubaka ; Cubaka et al. 2010). Bacteria with outstanding resistance to copper have been identified in the endophytes of both cuprophytes and in their rhizosphere (manuscript in preparation).
II. Contribution to the study of magnesium homeostasis in plants
- Plant nutrition
- Physiological characterization of Mg deficiency in plants
- Magnesium homeostasis in plants and interaction with the circadian clock
(i) The first approach uses the variation of Mg concentration in Arabidopsis mutants and natural accessions as a source of diversity to find new genes and alleles controlling root and shoot Mg homeostasis.
(ii) Because the knowledge about the impact of Mg deficiency on physiological processes was scarce, we proceeded to transcriptome analyses to provide non-biased hints about targets of Mg starvation. We published a thorough description of the transcriptomic responses (within hours or days before the outbreak of visual symptoms) of Mg deprivation (Hermans et al. 2010a & b). Unlike other mineral deficiencies, putative Mg transporters genes were not induced (post-transcriptional induction cannot be excluded). Interestingly, the rhythmic expression of circadian clock component genes was altered in roots, while abscisic acid signalling was triggered in leaves. The transcriptomic analysis confirmed the visual observation that Mg starvation affected the shoot more than the root in Arabidopsis. Further study of the circadian rhythm confirmed interaction between the circadian clock and Mg status. This aspect is currently further investigated.
Effects of Mg deficiency on photosystems
Visual symptoms of Mg deficiency in sugar beet. Mg deficiency appears first on the most recently developped leaves as chlorosis between the veins, which remind green. Brown spottings and necroses appear when deficiency is severe or under high light intensity.
The behaviour of PSII and PSI was assessed using direct and modulated fluorescence measurement, near-infrared absorbance changes and 77K fluorescence emission spectroscopy. An early effect on the maximum PSI oxidation rate could be identified before any decrease in the rate of PSII maximum quantum yield of primary photochemistry, PSII electron transport or any chlorophyll degradation. Also, a decrease in the amplitude fluorescence emission peak at 735 nm has been observed, suggesting an early effect of Mg deficiency on PSI. Concomitantly, sucrose accumulates in source leaves upon Mg deficiency as an early response. High sucrose levels are known to down-regulate the chloroplast electron chain transport. The delay of photosynthesis regulation in Mg-deficient plants may provoke an unbalance between light and dark reactions, so inducing oxidative stress and further leading to necrogenesis on leaves.
Effects of Mg deficiency on sucrose partitioning
Sucrose pathway from production to consumption and storage organs. In sugar beet, sucrose loading from the apoplasm to companion cell is operated trough sucrose/H+ symporter.
III. Nitrogen use efficiency
A sustained improvement in crop yield is required to meet the food demands of the rapidly growing world population. By 2050, a societal challenge will be to almost double food production from existing land areas in order to feed more than nine billion people, while facing yield-depressing consequences of climate change. The first research theme addresses the field of agriculture and the improvement of mineral element acquisition by crops. Nitrogen-inorganic fertilization has been used for decades to increase crop yield but excessive concentrations of nitrate are detrimental to the environment. One way to reduce nitrogen fertilizer input is to breed for crops with improved Nitrogen Use Efficiency (NUE). Our aim is to provide a genomic entry into enhanced plant productivity through optimized nitrogen acquisition by roots. The rationale is that NUE can be ameliorated by redesigning a more branched root system that explores a larger soil volume in order to prevent nitrate leaching (Fig. 1).
Nitrate-dependent lateral root development in the model species Arabidopsis and related crop
Arabidopsis thaliana is a genetically and molecularly plant model species. We are studying the genetic variation induced by mutagenesis (EMS screen) and the biodiversity (public collection and accessions collected in Belgium and bordering countries) of root morphology in response to nitrate. In parallel, we tackle the cross-talk between N nutrition and plant growth regulators (e.g. the gaseous hormone, ethylene). Natural variation of ethylene production is explored to improve NUE, in particular by manipulating features like the biomass production and the timing of senescence during N limitation.
Brassica napus (oilseed rape) is an increasingly important cash crop that diversifies cereal dominated crop rotations. However, that crop has a low ratio of seeds produced per N unit applied, around half that for cereals and the small recovery involves risks for N leaching. Brassica napus (2n=4x=38) has an allotetraploid genome and is closely related to Arabidopsis thaliana (2n=2x=10). Large diversity panels of double haploid lines are screened for root morphology in laboratory settings. Root RNA-seq analyses are carried to build regulatory networks of genes expressed during lateral root development and in response to nitrate supply. We are also challenging crop performance in field conditions (collaboration with CARAH).
Fig. 1. Premises on ideal root architectural attributes to optimize nitrogen acquisition in time and space. Nitrate (NO3-) can leach through the soil and quickly be depleted in surface strata. A root system with rapid exploitation of deep soil would optimize the capture of that mobile resource. A dual effect of external nitrate on lateral root development has been described: (i) a localized stimulation of N-starved roots elongation at the contact with rich nitrate source and (ii) a systemic inhibition of uniformly high nitrate concentrations on lateral root elongation. We define a root system ideotype as a profuse and deep lateral root branching and with unresponsiveness to local N. Figure adapted from Louvieaux et al. (2018)