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1.
Plant Signal Behav ; 10(2): e988076, 2015.
Article in English | MEDLINE | ID: mdl-25723847

ABSTRACT

Colonization of plant rhizosphere/roots by beneficial microorganisms (e.g. plant growth promoting rhizobacteria - PGPR, arbuscular mycorrhizal fungi - AMF) confers broad-spectrum resistance to virulent pathogens and is known as induced systemic resistance (ISR) and mycorrhizal-induced resistance (MIR). ISR or MIR, an indirect mechanism for biocontrol, involves complex signaling networks that are regulated by several plant hormones, the most important of which are salicylic acid (SA), jasmonic acid (JA) and ethylene (ET). In the present study, we investigated if inoculation of potato plantlets with an AMF (Rhizophagus irregularis MUCL 41833) and a PGPR (Pseudomonas sp R41805) either alone or in combination, could elicit host defense response genes in the presence or absence of Rhizoctonia Solani EC-1, a major potato pathogen. RT-qPCR revealed the significant expression of ethylene response factor 3 (EFR3) in mycorrhized potato plantlets inoculated with Pseudomonas sp R41805 and also in mycorrhized potato plantlets inoculated with Pseudomonas sp R41805 and challenged with R. solani. The significance of ethylene response factors (ERFs) in pathogen defense has been well documented in the literature. The results of the present study suggest that the dual inoculation of potato with PGPR and AMF may play a part in the activation of plant systemic defense systems via ERF3.


Subject(s)
Ethylenes/metabolism , Glomeromycota/physiology , Plant Proteins/metabolism , Pseudomonas/physiology , Solanum tuberosum/immunology , Solanum tuberosum/microbiology , Gene Expression Regulation, Plant , Mycorrhizae/physiology , Plant Proteins/genetics , Solanum tuberosum/genetics
2.
Microb Ecol ; 69(3): 652-67, 2015 Apr.
Article in English | MEDLINE | ID: mdl-25339308

ABSTRACT

Food security (a pressing issue for all nations) faces a threat due to population growth, land availability for growing crops, a changing climate (leading to increases in both abiotic and biotic stresses), heightened consumer awareness of the risks related to the use of agrichemicals, and also the reliance on depleting fossil fuel reserves for their production. Legislative changes in Europe mean that fewer agrichemicals will be available in the future for the control of crop pests and pathogens. The need for the implementation of a more sustainable agricultural system globally, incorporating an integrated approach to disease management, has never been more urgent. To that end, the Valorizing Andean Microbial Diversity (VALORAM) project (http://valoram.ucc.ie), funded under FP7, examined the role of microbial communities in crop production and protection to improve the sustainability, food security, environmental protection, and productivity for rural Andean farmers. During this work, microbial volatile organic compounds (mVOCs) of 27 rhizobacterial isolates were identified using gas chromatography/mass spectrometry (GC/MS), and their antifungal activity against Rhizoctonia solani was determined in vitro and compared to the activity of a selection of pure volatile compounds. Five of these isolates, Pseudomonas palleroniana R43631, Bacillus sp. R47065, R47131, Paenibacillus sp. B3a R49541, and Bacillus simplex M3-4 R49538 trialled in the field in their respective countries of origin, i.e., Bolivia, Peru, and Ecuador, showed significant increase in the yield of potato. The strategy followed in the VALORAM project may offer a template for the future isolation and determination of putative biocontrol and plant growth-promoting agents, useful as part of a low-input integrated pest management system.


Subject(s)
Bacteria/chemistry , Mycorrhizae/chemistry , Soil Microbiology , Solanum tuberosum/growth & development , Solanum tuberosum/microbiology , Volatile Organic Compounds/pharmacology , Bacteria/isolation & purification , Bolivia , Ecuador , Fungicides, Industrial/isolation & purification , Fungicides, Industrial/pharmacology , Gas Chromatography-Mass Spectrometry , Peru , Plant Roots/chemistry , Plant Roots/microbiology , Rhizoctonia/drug effects , Solanum tuberosum/chemistry , Solid Phase Microextraction , Volatile Organic Compounds/isolation & purification
3.
Environ Microbiol ; 16(8): 2389-407, 2014 Aug.
Article in English | MEDLINE | ID: mdl-25229098

ABSTRACT

Quantitative PCR (qPCR) and community fingerprinting methods, such as the Terminal Restriction Fragment Length Polymorphism (T-RFLP) analysis,are well-suited techniques for the examination of microbial community structures. The use of phylum and class-specific primers can provide enhanced sensitivity and phylogenetic resolution as compared with domain-specific primers. To date, several phylum- and class-specific primers targeting the 16S ribosomal RNA gene have been published. However, many of these primers exhibit low discriminatory power against non-target bacteria in PCR. In this study, we evaluated the precision of certain published primers in silico and via specific PCR. We designed new qPCR and T-RFLP primer pairs (for the classes Alphaproteobacteria and Betaproteobacteria, and the phyla Bacteroidetes, Firmicutes and Actinobacteria) by combining the sequence information from a public dataset (SILVA SSU Ref 102 NR) with manual primer design. We evaluated the primer pairs via PCR using isolates of the above-mentioned groups and via screening of clone libraries from environmental soil samples and human faecal samples. As observed through theoretical and practical evaluation, the primers developed in this study showed a higher level of precision than previously published primers, thus allowing a deeper insight into microbial community dynamics.


Subject(s)
Bacteria/genetics , DNA Primers/chemistry , Genes, rRNA , Phylogeny , RNA, Ribosomal, 16S , Soil Microbiology , Bacteria/classification , Base Sequence , Computer Simulation , DNA Primers/genetics , Databases, Genetic , Feces/microbiology , Humans , Molecular Sequence Data , Polymerase Chain Reaction/methods , Polymorphism, Restriction Fragment Length , RNA, Ribosomal, 16S/genetics
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