Catalina Cabot

Lessons from crop plants struggling with salinity

Salinity is a persistent problem, causing important losses in irrigated agriculture. According to global climate change prediction models, salinity is expected to expand in the near future. Although intensive studies have been conducted on the mechanisms by which plants cope with saline conditions, the multi-component nature of salt stress tolerance has rendered most plant breeding efforts to improve the plants response to salinity unsuccessful. This occurs despite the extensive genetic diversity shown by higher plants for salt tolerance and the similar mechanisms found in salt-sensitive and salt-tolerant genotypes in response to the presence of excess of salts in the growth media. On the other hand, there is an urge to increase crop yield to the maximum to cope with the growing world population demands for food and fuel. Here, we examine some major elements and signaling mechanisms involved in the plants response to salinity following the pathway of salt-footprints from the soil environment to leaf. Some of the possible contrasting determinants for a better-balanced resource allocation between salt tolerance and plant growth and yield are considered.

Abscisic Acid Decreases Leaf Na+ Exclusion in Salt-Treated Phaseolus vulgaris L.

Previous results showed that in short-term NaCl-treated beans increased leaf abscisic acid (ABA) concentration was triggered by Na+ but not by Cl-. In this work, the specificity of ABA signaling for Na+ homeostasis was studied by comparing the plant’s responses to solutions that modified accumulation of ABA and/or Na+ uptake and distribution, such as supplemental Ca2+, increased nutrient strength, different isosmotic composition, application of exogenous ABA, fluridone (an ABA inhibitor) and aminooxiacetic acid (AOA, an ethylene inhibitor). After fluridone pretreatment, salt-treated beans had lower Na+ uptake and higher leaf Na+ exclusion capacity than non-pretreated plants. Moreover, Na+ uptake was increased and leaf Na+ exclusion was decreased by AOA and ABA. NaCl and KCl similarly increased leaf ABA and decreased transpiration rates, whereas supplemental Ca2+ and increased strength nutrient solution decreased leaf ABA and leaf Na+. These results show (1) a non-ion-specific increase in ABA that probably signaled the osmotic component of salt, and (2) increased ABA levels that resulted in higher leaf Na+ concentrations due to lower Na+ exclusion or increased root-shoot Na+ translocation.

Ion allocation in two different salt-tolerant MediterraneanMedicagospecies

The relationship between Na+, major cation concentrations and salt tolerance under long-term saline conditions of Medicago arborea and Medicago citrina was studied. Plants were grown in solution culture in 1, 50, 100, or 200 mmol/L NaCl for 30 days in a climate-controlled greenhouse. Stem and petiole growth was the most affected by salt in both species. Leaf growth was inhibited in M. arborea, with increased salt, while only the 200 mmol/L NaCl-treated M. citrina plants were significantly affected. Both species had the highest Na+ concentrations in the shoots, however, the allocation pattern was different; M. arborea showed the highest concentrations in the leaf blades, whereas M. citrina distributed the salt into the petioles. K+/Na+ ratio decreased with salt in both species; however, leaf K+ use efficiency (g leaf DW mg-1 leaf K+) was higher in M. citrina. The difference in Na+ allocation and cation concentrations found in these medic species and their importance is discussed in relation to their response to NaCl salinity.

Relationship between carbon partitioning and Na+, Cl- and ABA allocation in fruits of salt-stressed bean.

Summary Ion toxicity is considered one of the most important limiting factors affecting plants growing under long-term saline conditions. Abscisic acid participates in the plants adaptation to different environmental stresses. However, most of the information available on this subject is for plants exposed to water deficit during the vegetative growth phase. In this study, we examine the role of ABA and ion allocation in the control of fruit growth in salt-stressed bean. Plants of Phaseolus vulgaris were grown in solution culture with 1, 25, 50, or 75 mmol/L NaCl. After 40 days, growth, photosynthetic parameters and ion, protein and ABA concentrations were recorded in the different plant organs. Leaf area and instantaneous C02 assimilation rates were inhibited by salt. Salinity induced a decrease in harvest index (fruit dry weight/total plant dry weight) simultaneously with an increase in root index (root dry weight/total plant dry weight). The changes observed in carbon allocation were greatly related to sodium index (fruit Na+/total plant Na+). No differences among treatments were found either in chloride index (fruit Cl-/total plant Cl-) or ABA index (fruit ABA/total plant ABA). Since bean is a Na+ excluder species, an adaptation mechanism that promotes root over fruit growth would enhace Na+ exclusion and favor plant survival. The signalling mechanism may be triggered by a possible effect of apoplastic Na+ on the plasma membrane, but seems not to be mediated by an increase in ABA.

Characterization of Zinc and Cadmium Hyperaccumulation in Three Noccaea (Brassicaceae) Populations from Non-metalliferous Sites in the Eastern Pyrenees

The Southern slope of the Pyrenees is the meridional limit for the distribution of several Noccaea populations. However, the systematic description of these populations and their hyperaccumulation mechanisms are not well established. Morphological and genetic analysis (ITS and 3 chloroplast regions) were used to identify Noccaea populations localized on non-metallicolous soils during a survey in the Catalonian Pyrenees. Cd and Zn concentrations were analyzed in soils and plants both sampled in the field and grown hydroponically. The expression of selected metal transporter genes was assessed by quantitative PCR. The populations were identified as Noccaea brachypetala (Jord.) F.K. Mey by conspicuous morphological traits. Principal component analysis provided a clear separation among N. brachypetala, Noccaea caerulescens J. Presl & C. Presl and Noccaea occitanica (Jord.) F.K. Mey., three Noccaea species reported in the Pyrenees. Contrastingly, ITS and cpDNA analyses were unable to clearly differentiate these taxa. Differences in the expression of the metal transporter genes HMA3, HMA4, and MTP1 between N. caerulescens and N. brachypetala, and those amongst the N. brachypetala populations suggest differences in the strategies for handling enhanced Cd and Zn availability. This is the first report demonstrating Cd and Zn hyperaccumulation by N. brachypetala both in the field and in hydroponics. This comprehensive study based on taxonomic, molecular, and physiological data allows both the correct identification of this species and the characterization of population differences in hyperaccumulation and tolerance of Zn and Cd.

Cytokinin activity of disubstituted aminopurines in Amaranthus

Cytokinin (CK) receptors have different affinities for certain ligands, and consequently, studies of the plants response to CK analogues constitute a good approach to identify active compounds that trigger specific plant responses. In this study, N(6) and N(6),N(6)-substituted CK analogues were synthesized and their CK-like activity was examined in the Amaranthus betacyanin and the bacterial receptor assay. The compounds showed CK-like activities that were not always associated with their binding affinity to the Arabidopsis receptors AHK3 and CRE1/AHK4. The highest level of activity in both bioassays was obtained for the N(6)-alkylaminopurines, which showed an especially high binding affinity to AHK3. In contrast to previously published data, we found remarkable activity of N(6),N(6)-alkylbenzylaminopurines in the Amaranthus betacyanin bioassay, which was not associated with their binding affinity to the tested receptors. The N(6),N(6)-substituted CK that showed the highest activity at the lowest concentration, N(6),N(6)-methylbenzylaminopurine (BAP-C1), was studied to determine its effect on different leaf parameters of whole Amaranthus plants, with benzylaminopurine (BAP) used as standard compound. The interaction with ethylene was examined in plants supplied with the ethylene-synthesis inhibitor aminooxiacetic acid (AOA). After 3d, the CKs supplied in the solution culture exerted effects on leaf dry weight and gas-exchange parameters. These effects of exogenous CKs are suggested to be ethylene-synthesis dependent.

Structural insights on the molecular recognition patterns between N(6)-substituted adenines and N-(aryl-methyl)iminodiacetate copper(II) chelates.

For a better understanding of the metal binding pattern of N(6)-substituted adenines, six novel ternary Cu(II) complexes have been structurally characterized by single crystal X-ray diffraction: [Cu(NBzIDA)(HCy5ade)(H2O)]·H2O (1), [Cu(NBzIDA)(HCy6ade)(H2O)]·H2O (2), [Cu(FurIDA)(HCy6ade)(H2O)]·H2O (3), [Cu(MEBIDA)(HBAP)(H2O)]·H2O (4), [Cu(FurIDA)(HBAP)]n (5) and {[Cu(NBzIDA)(HdimAP)]·H2O}n (6). In these compounds NBzIDA, FurIDA and MEBIDA are N-substituted iminodiacetates with a non-coordinating aryl-methyl pendant arm (benzyl in NBzIDA, p-tolyl in MEBIDA and furfuryl in FurIDA) whereas HBAP, HCy5ade, HCy6ade and HdimAP are N(6)-substituted adenine derivatives with a N-benzyl, N-cyclopentyl, N-cyclohexyl or two N-methyl groups, respectively. Regardless of the molecular (1-4) or polymeric (5-6) nature of the studied compounds, the Cu(II) centre exhibits a type 4+1 coordination where the tridentate IDA-like chelators adopt a mer-conformation. In 1-5 the N(6)-R-adenines use their most stable tautomer H(N9)adenine-like, and molecular recognition consists of the cooperation of the CuN3(purine) bond and the intra-molecular interligand N9H···O(coordinated carboxy) interaction. In contrast, N(6),N(6)-dimethyl-adenine shows the rare tautomer H(N3)dimAP in 6, so that the molecular recognition with the Cu(NBzIDA) chelate consist of the CuN9 bond and the N3H···O intra-molecular interligand interaction. Contrastingly to the cytokinin activity found in the free ligands HBAP (natural cytokinin), HCy5ade and HCy6ade, the corresponding Cu(II) ternary complexes did not show any activity.

Zinc triggers signaling mechanisms and defense responses promoting resistance to Alternaria brassicicola in Arabidopsis thaliana.

According to the elemental defense hypothesis the accumulation of trace elements by plants may substitute for organic defenses, while the joint effects hypothesis proposes that trace elements and organic defenses can have additive or synergistic effects against pathogens or herbivores. To evaluate these hypotheses the response of the pathosystem Alternaria brassicicola-Arabidopsis thaliana to control (2μM) and surplus (12μM) Zn was evaluated using the camalexin deficient mutant pad3-1 and mtp1-1, a mutant with impaired Zn vacuolar storage, along with the corresponding wildtypes. In vitro, a 50% inhibition of fungal growth was achieved by 440μM Zn. A. thaliana leaves could accumulate equivalent concentrations without harm. In fact, surplus Zn enhanced the resistance of A. thaliana to fungal attack in Columbia (Col-0), Wassilewskija (WS), and mtp1-1. However, surplus Zn was unable to protect pad3-1 demonstrating that Zn cannot substitute for camalexin, the main organic defense in A. thaliana. High, non phytotoxic leaf Zn concentrations enhanced the resistance to A. brassicicola of A. thaliana genotypes able to produce camalexin. This was mainly due to Zn-induced enhancement of the JA/ETH signaling pathway leading to enhanced PAD3 expression. These results support the joint effects hypothesis and highlight the importance of adequate Zn supply for reinforced pathogen resistance.

Zinc hyperaccumulation substitutes for defense failures beyond salicylate and jasmonate signaling pathways of Alternaria brassicicola attack in Noccaea caerulescens

The hypothesis of metal defense as a substitute for a defective biotic stress signaling system in metal hyperaccumulators was tested using the pathosystem Alternaria brassicicola-Noccaea caerulescens under low (2 µM), medium (12 µM) and high (102 µM) Zn supply. Regardless the Zn supply, N. caerulescens responded to fungal attack with the activation of both HMA4 coding for a Zn transporter, and biotic stress signaling pathways. Salicylate, jasmonate, abscisic acid and indoleacetic acid concentrations, as well as biotic stress marker genes (PDF1.2, CHIB, LOX2, PR1 and BGL2) were activated 24 h upon inoculation. Based on the activation of defense genes 24 h after the inoculation an incompatible fungal-plant interaction could be predicted. Nonetheless, in the longer term (7 days) no effective protection against A. brassicicola was achieved in plants exposed to low and medium Zn supply. After 1 week the biotic stress markers were even further increased in these plants, and this compatible interaction was apparently not caused by a failure in the signaling of the fungal attack, but due to the lack of specificity in the type of the activated defense mechanisms. Only plants receiving high Zn exhibited an incompatible fungal interaction. High Zn accumulation in these plants, possibly in cooperation with high glucosinolate concentrations, substituted for the ineffective defense system and the interaction turned into incompatible. In a threshold-type response, these joint effects efficiently hampered fungal spread and, consequently decreased the biotic stress signaling.