Metabolic responses to arbuscular mycorrhizal fungi are shifted in roots of contrasting soybean genotypes.

Modern breeding programs have reduced genetic variability and might have caused a reduction in plant colonization by arbuscular mycorrhizal fungi (AM). In our previous studies, mycorrhizal colonization was affected in improved soybean genotypes, mainly arbuscule formation. Despite substantial knowledge of the symbiosis-related changes of the transcriptome and proteome, only sparse clues regarding metabolite alterations are available. Here, we evaluated metabolite changes between improved (I-1) and unimproved (UI-4) soybean genotypes and also compare their metabolic responses after AM root colonization. Soybean genotypes inoculated or not with AM were grown in a chamber under controlled light and temperature conditions. At 20 days after inoculation, we evaluated soluble metabolites of each genotype and treatment measured by GC-MS. In this analysis, when comparing non-AM roots between genotypes, I-1 had a lower amount of 31 and higher amount of only 4 metabolites than the UI-4 genotype. When comparing AM roots, I-1 had a lower amount of 36 and higher amount of 4 metabolites than UI-4 (different to those found altered in non-AM treated plants). Lastly, comparing the AM vs non-AM treatments, I-1 had increased levels of three and reduced levels of 24 metabolites, while UI-4 only had levels of 12 metabolites reduced by the effect of mycorrhizas. We found the major changes in sugars, polyols, amino acids, and carboxylic acids. In a targeted analysis, we found lower levels of isoflavonoids and alpha-tocopherol and higher levels of malondialdehyde in the I-1 genotype that can affect soybean-AM symbiosis. Our studies have the potential to support improving soybean with a greater capacity to be colonized and responsive to AM interaction.

An In Vitro Method for Studying the Three-Way Interaction between Soybean, Rhizophagus irregularis and the Soil-Borne Pathogen Fusarium virguliforme.

In this work, we described an in vitro system adequate for investigating the pathosystem soybean/arbuscular mycorrhizal fungi (AMF)/Fusarium virguliforme. Pre-mycorrhized plantlets with Rhizophagus irregularis were infected by F. virguliforme either locally via a plug of gel supporting mycelium (Method 1) or via a macroconidia suspension applied to the medium surface (Method 2). Root colonization by the AMF and infection by the pathogen were similar to the usual observations in pot experiments. Within a period of 18 days, more than 20% of the roots were colonized by the AMF and infection by the pathogen was observed in all the plants. In presence of AMF, a decrease in symptoms and in the level of root tissue infection was noticed. With Method 1, smaller necrotic lesions were observed in the pre-mycorrhized plantlets. In Method 2, pathogen infection was slower but more homogenous. These results demonstrated the suitability of the in vitro cultivation system to study the pathosystem soybean/AMF/F. virguliforme. We propose this in vitro cultivation system for studying the mechanisms involved in the biocontrol conferred by AMF against F. virguliforme in soybean.

Mycorrhizal fungi symbiosis as a strategy against oxidative stress in soybean plants.

Oxidative stress responses generated by paraquat (PQ), an herbicide that triggers an oxidative stress reaction in leaves, were studied in non-arbuscular mycorrhizal (non-AM) and in arbuscular mycorrhizal (AM) soybean plants inoculated with Glomus mosseae (Gm) or Glomus intraradices (Gi). Some oxidative stress symptoms were evident in non-AM after 6 d of PQ application on leaves. Oxidative damage, measured as malondialdehyde content (MDA), was significantly higher, and although no changes were evident in total catalase (CAT, EC 1.11.1.6) and total superoxide dismutase (SOD, EC 1.15.1.1) activity, total ascorbate peroxidase (APX, EC 1.11.1.11) activity was significantly reduced. These effects were correlated with a significant decrease in growth parameters. By contrast, in both AM plants, foliar MDA content was reduced or unaltered and, interestingly, after PQ stress, its level was unchanged and significantly lower than in PQ non-AM plants. Unlike PQ stress in non-AM plants, total APX activity was unaltered or induced by AM plants, while total SOD activity was unchanged and no consistent effects were detected in total CAT activity. All these events coincided with no changes or a significant increase in growth parameters. Since oxidative stress is a common phenomenon triggered by several environmental stresses, these results highlight the importance of mycorrhizal fungi in oxidative stress regulation as a general strategy to protect plants from abiotic and biotic stress.

The maize Activator/Dissociation system is functional in hexaploid wheat through successive generations.

The aim of the present study was to provide useful background information and evidence of the functionality of the maize Activator/Dissociation (Ac/Ds) system in hexaploid wheat. Two transgenic parental wheat lines, one harbouring the immobilised Ac element (iAc) and the other the Ds element (pUbi[Ds-uidA]bar), were crossed. Transient GUS assays confirmed that the iAc transposase is active in hexaploid wheat. Selected F1 and F2 lines were analysed by PCR using primers specific to Ac, uidA and bar genes. The primer pair Ubi/bar-tag was used to detect excision of the Ds-uidA sequence, which occurred at a frequency of 39% in the F1 generation. Lines free of Ac and showing evidence of Ds excision were subject to Southern analysis, which indicated that at least one transposition event might have occurred in these lines. Although more evidence is required to unequivocally support the reintegration of the Ds element in the wheat genome, the evidence presented here nevertheless demonstrates the effectiveness and potential value of using this system to tag genes in wheat.

Drought controls on H2O2 accumulation, catalase (CAT) activity and CAT gene expression in wheat.

Plants co-ordinate information derived from many diverse external and internal signals to ensure appropriate control of gene expression under optimal and stress conditions. In this work, the relationships between catalase (CAT) and H2O2 during drought in wheat (Triticum aestivum L.) are studied. Drought-induced H2O2 accumulation correlated with decreases in soil water content and CO2 assimilation. Leaf H2O2 content increased even though total CAT activity doubled under severe drought conditions. Diurnal regulation of CAT1 and CAT2 mRNA abundance was apparent in all conditions and day/night CAT1 and CAT2 expression patterns were modified by mild and severe drought. The abundance of CAT1 transcripts was regulated by circadian controls that persisted in continuous darkness, while CAT2 was modulated by light. Drought decreased abundance, and modified the pattern, of CAT1 and CAT2 mRNAs. It was concluded that the complex regulation of CAT mRNA, particularly at the level of translation, allows precise control of leaf H2O2 accumulation.