Addition of Trichoderma consortia to Chilean endemic flora compost teas strongly enhances in vitro and in vivo biocontrol of phytopathogenic fungi.

AIM: To evaluate the in vitro and in vivo antifungal capability of diverse compost teas of endemic Chilean flora inoculated with a consortium of fungal strains of Trichoderma spp. (biocontrol agent) against three important phytopathogens: Botrytis cinerea, Fusarium oxysporum, andLasiodiplodia theobromae. METHODS AND RESULTS: Compost teas were obtained from the endemic flora of Chile (Azara celastrina, Citronella mucronate, Cryptocarya alba, Peumus boldus, and Quillaja saponaria). Eleven Trichoderma strains were isolated, and antagonism tests were performed to develop fungal consortiums with biocontrol properties. The biocontrol effect of compost teas inoculated with Trichoderma consortia was also analyzed. The results showed that the teas possess antifungal activity against B. cinerea and F. oxysporum and, to a lower degree, against L. theobromae. In vitro tests showed that Trichoderma consortiums improved the suppressive effect against B. cinerea (94-97%), F. oxysporum (89-92%), and L. theobromae (51-73%). Peumus boldus tea showed the highest suppressive effect against the plant pathogen L. theobromae. In addition, the in vivo assay showed that tomato plants treated only with Trichoderma or compost tea did not show differences in height with regard to control plants. However, when these two treatments were combined, the best performance in plant height and protection against pathogens was observed. CONCLUSIONS: This study indicates that the addition of a consortium of Trichoderma strains with intra- and interspecific incompatibilities significantly improves the inhibitory effect of compost teas in in vitro tests against the plant pathogenic fungi, while in vivo it enhances tomato plant growth and reduces plant disease symptoms.

Responses of Vitis vinifera cv. Cabernet Sauvignon roots to the arbuscular mycorrhizal fungus Funneliformis mosseae and the plant growth-promoting rhizobacterium Ensifer meliloti include changes in volatile organic compounds.

Arbuscular mycorrhizal (AM) fungi and plant growth-promoting rhizobacteria (PGPR) are beneficial microorganisms that may associate with grapevine roots, improving stress tolerance, growth, and nutrition. AM fungi and PGPR enhance the production of plant secondary metabolites, including volatile organic compounds (VOCs) that play a key role in the interaction of plants with the environment and are involved in defence mechanisms. The aim of this study was to analyse the effects of an AM fungus and a rhizobacterium on plant growth and VOCs in Vitis vinifera cv. Cabernet Sauvignon roots to gain insight into the potential role of plant-rhizosphere microorganisms in vine growth and defence. Grapevines were inoculated or not with the AM fungus Funneliformis mosseae IN101 and/or the plant growth-promoting rhizobacterium Ensifer meliloti TSA41. Both microbial strains enhanced plant growth. Fifty-eight VOCs extracted from ground roots were identified using headspace solid-phase microextraction coupled to gas chromatography/mass spectrometry. VOCs were induced by F. mosseae IN101, increasing up to 87% compared with control plants. Monoterpenes were strongly enhanced by F. mosseae IN101, increasing up to 113% compared with control plants. Interestingly, monoterpene alcohols related to plant defence, such as myrtenol, p-cymen-7-ol, and p-mentha-1.8-dien-7-ol were increased. By contrast, E. meliloti TSA41 did not significantly affect VOCs. The knowledge of the effects of AM fungi and PGPR on grapevine VOCs may contribute to an integrated and sustainable management of vineyards.

Microbial Diversity of Psychrotolerant Bacteria Isolated from Wild Flora of Andes Mountains and Patagonia of Chile towards the Selection of Plant Growth-Promoting Bacterial Consortia to Alleviate Cold Stress in Plants.

Cold stress decreases the growth and productivity of agricultural crops. Psychrotolerant plant growth-promoting bacteria (PGPB) may protect and promote plant growth at low temperatures. The aims of this study were to isolate and characterize psychrotolerant PGPB from wild flora of Andes Mountains and Patagonia of Chile and to formulate PGPB consortia. Psychrotolerant strains were isolated from 11 wild plants (rhizosphere and phyllosphere) during winter of 2015. For the first time, bacteria associated with Calycera, Orites, and Chusquea plant genera were reported. More than 50% of the 130 isolates showed ≥33% bacterial cell survival at temperatures below zero. Seventy strains of Pseudomonas, Curtobacterium, Janthinobacterium, Stenotrophomonas, Serratia, Brevundimonas, Xanthomonas, Frondihabitans, Arthrobacter, Pseudarthrobacter, Paenarthrobacter, Brachybacterium, Clavibacter, Sporosarcina, Bacillus, Solibacillus, Flavobacterium, and Pedobacter genera were identified by 16S rRNA gene sequence analyses. Ten strains were selected based on psychrotolerance, auxin production, phosphate solubilization, presence of nifH (nitrogenase reductase) and acdS (1-aminocyclopropane-1-carboxylate (ACC) deaminase) genes, and anti-phytopathogenic activities. Two of the three bacterial consortia formulated promoted tomato plant growth under normal and cold stress conditions. The bacterial consortium composed of Pseudomonas sp. TmR5a & Curtobacterium sp. BmP22c that possesses ACC deaminase and ice recrystallization inhibition activities is a promising candidate for future cold stress studies.

Analyses of Virulence Genes of Clavibacter michiganensis subsp. michiganensis Strains Reveal Heterogeneity and Deletions That Correlate with Pathogenicity.

Clavibacter michiganensis subsp. michiganensis (Cmm) is the causal agent of bacterial canker of tomato. Differences in virulence between Cmm strains have been reported. The aim of this study was the characterization of nine Cmm strains isolated in Chile to reveal the causes of their differences in virulence. The virulence assays in tomato seedlings revealed different levels of severity associated with the strains, with two highly virulent strains and one causing only mild symptoms. The two most virulent showed increased cellulase activity, and no cellulase activity was observed in the strain causing mild symptoms. In three strains, including the two most virulent strains, PCR amplification of the 10 virulence genes analyzed was observed. In the strain causing mild symptoms, no amplification was observed for five genes, including celA. Sequence and cluster analyses of six virulence genes grouped the strains, as has been previously reported, except for gene pelA1. Gene sequence analysis from the genomes of five Chilean strains revealed the presence of deletions in the virulence genes, celB, xysA, pat-1, and phpA. The results of this study allow us to establish correlations between the differences observed in disease severity and the presence/absence of genes and deletions not previously reported.

The arbuscular mycorrhizal fungus Funneliformis mosseae induces changes and increases the concentration of volatile organic compounds in Vitis vinifera cv. Sangiovese leaf tissue.

Arbuscular mycorrhizal fungi (AMF) are beneficial obligate symbionts of plant roots. Volatile organic compounds (VOCs) participate in plant communication and defence. The aim of this study was to analyse the effects of the arbuscular mycorrhizal fungus Funneliformis mosseae IMA1 on VOCs in Vitis vinifera cv. Sangiovese leaf tissue. Grapevine plants inoculated with F. mosseae IMA1 were incubated for 23 weeks. VOCs were extracted from leaves and identified using headspace solid-phase microextraction (HS-SPME) coupled to GC-MS. VOCs in leaf tissue were strongly enhanced (85%) by F. mosseae IMA1. The mycorrhizal fungus IMA1 modified the levels of specific VOCs synthesised in different anabolic pathways. An increase in volatiles that have been related to plant defences under pathogen/herbivore attack or linked to water stress, such as (E)-2-hexenal, 3-hexenal, geraniol, benzaldehyde and methyl salicylate, was observed in mycorrhizal plants. In contrast, some C13-norisoprenoids decreased strongly in mycorrhizal plants. The study of the effects of AMF on VOCs in grapevine plants may provide useful information to establish sustainable viticultural practices.