Xiphinema index is the most aggressive root parasite affecting vines in Chile, controlled mainly chemically. The aim of this study was to identify rhizobacteria in grapevine roots growing in infested soils and to determine if some of these affect the parasitism caused by the nematode. Rhizobacteria of ungrafted grapevine cultivars from 11 vineyards were isolated using TSBA and identified with FAMEs analysis. The antagonistic effect to X. index of those from soils 1 to 4 was assessed in 500 ml pots treating 2 months old in vitro plants with a bacterial suspension containing 1×10 6 c.f.u./ml and 400 nematodes, determining after 4 months growth nematode populations and root damages. Also culture filtrates were assessed. 400 isolates in 25 genera were obtained from the 11 vineyards. The most frequent species were Pseudomonas putida (35.1%), Escherichia coli (7.6%) and Pseudomonas fluorescens (6.1%). Isolates of soils 1 to 4, show that Bacillus megaterium, B. brevis, Pseudomonas corrugata, P. savastanoi, Stenotrophomonas maltophilia and Serratia plymuthica reduced root damage and suppressed populations, meanwhile strains of B. brevis and Comamonas acidovorans increased plant growth but did not control nematodes. It is concluded that these isolates may be useful in biological control programmes in vineyards.
The use of rhizobacteria to control plant parasitic nematodes has been widely studied. Currently, the research focuses on bacteria-nematode interactions that can mitigate this complex microbiome in agriculture. Various enzymes, toxins and metabolic by-products from rhizobacteria antagonize plant parasitic nematodes, and many different modes of action have been proposed. Hydrolytic enzymes, primarily proteases, collagenases and chitinases, have been related to the nematicide effect in rhizobacteria, proving to be an important factor involved in the degradation of different chemical constituents of nematodes at distinct developmental stages. Exuded metabolites may also alter the nematode-plant recognition process or create a hostile environment for nematodes in the rhizosphere. Specific bacteria strains responsible for the production of toxins, such as Cry proteins, are one of the strategies used by rhizobacteria. Characterization of the rhizobacteria mode of action could strengthen the development of commercial products to control populations of plant parasitic nematodes. This review aims to provide an overview of different enzymes and compounds produced by rhizobacteria related to the process of antagonism to plant-parasitic nematodes.
Enzyme phenotypes, specifically esterases (EST) and malate dehydrogenase (MDH), were used to characterise different species of Meloidogyne from Chile. Esterase activity was highly polymorphic and was the most useful in the identification of the different species. Using this enzyme it is possible to characterise and identify M. ethiopica in about 80% of samples on grapevine, kiwi and tomatoes. Another three species, M. javanica, M. hapla and M. arenaria, were identified on tomatoes, kiwi and pomegranate with only one or a few populations. It was possible to detect minor atypical (unidentified) phenotypes, generally in mixed populations with M. ethiopica. Only the profiles N1 and H1 of MDH were detected. N1 was not specific and H1 allowed identification of M. hapla. Contaminated nursery stock has probably resulted in serious infestation by M. ethiopica in vineyards in various localities in Chile.
The action of metabolites and exoenzymes from rhizobacteria on different plant-parasitic nematodes has an influence on the nematicidal efficacy of the microbe. Seven rhizobacteria, divided into two bacterial groups, were evaluated in vitro for nematicidal activity on Meloidogyne ethiopica and Xiphinema index. The direct effect of their filtrates on egg hatching and juveniles of M. ethiopica as well as mobile stages of X. index was evaluated during a 72-h period. The production of four exoenzymes and two metabolites associated with nematode mortality was investigated. Molecular characterization of three isolates was performed, and the physiological profiles and lipase activity of all isolates were obtained using the BIOLOG EcoPlate system. While chitinase and collagenase were measured using the BIOLOG MT2 plate system, protease, hydrogen cyanide and hydrogen sulphide were directly determined in Petri dishes. Nematode mobile stages exposure to the bacterial filtrate revealed a nematicidal effect up to 93.7% on X. Index and up to 83.3% on M. ethiopica. The control of egg hatching varied between 35 and 85%. A positive correlation was found between the mortality of both nematode mobile stages and the concerted activities of the bacterial enzymes as well as the level of the volatile metabolites. The nematicidal effect of rhizobacteria strains varies by nematode genera and among the developmental stages evaluated.
Xenorhabdus is a symbiotic group of bacteria associated with entomopathogenic nematodes of the family Steinernematidae. Although the described Steirnernema species list is extensive, not all their symbiotic bacteria have been identified. One single motile, Gram-negative and non-spore-forming rod-shaped symbiotic bacterium, strain VLST, was isolated from the entomopathogenic nematode Steinernema unicornum. Analyses of the 16S rRNA gene determined that the VLST isolate belongs to the genus Xenorhabdus , and its closest related species is Xenorhabdus szentirmaii DSM 16338T (98.2 %). Deeper analyses using the whole genome for phylogenetic reconstruction indicate that VLST exhibits a unique clade in the genus. Genomic comparisons considering digital DNA–DNA hybridization (dDDH) values confirms this result, showing that the VLST values are distant enough from the 70 % threshold suggested for new species, sharing 30.7, 30.5 and 30.3 % dDDH with Xenorhabdus khoisanae MCB, Xenorhabdus koppenhoeferi DSM 18168T and Xenorhabdus miraniensis DSM 18168T, respectively, as the closest species. Detailed physiological, biochemical and chemotaxonomic tests of the VLST isolate reveal consistent differences from previously described Xenorhabdus species. Phylogenetic, physiological, biochemical and chemotaxonomic approaches show that VLST represents a new species of the genus Xenorhabdus , for which the name Xenorhabdus lircayensis sp. nov. (type strain VLST=CCCT 20.04T=DSM 111583T) is proposed.
Artículo de publicación ISIMeloidogyne ethiopica is one of the most important plant-parasitic nematodes affecting vines in Chile and is very aggressive and difficult to control. This study evaluated 16 strains of rhizobacteria, originally isolated from roots of grapevines, for their effects on parasitism and nematode damage to potted vine plants. The antagonistic effect of rhizobacteria was assessed by treating 2-month-old plants of cv. Chardonnay in 3-l pots with a suspension containing 1×106cfuml−1 of the bacteria and 1,000 nematode eggs. After 6 months of growth, the plants were cut and root and canopy weights, nematode populations and root damage determined. The effect of rhizobacterial culture filtrate on hatching of nematode eggs was also assessed in vitro. Seven strains of rhizobacteria proved effective in inhibiting damage or reproduction of the nematode. These were strains of Serratia marcescens, Comamonas acidovorans, Pantoea agglomerans, Sphingobacterium spiritivorum, Bacillus mycoides, Alcaligenes piechaudii and Serratia plymuthica. A further three strains, of Bacillus megaterium, P. agglomerans and Pseudomonas savastanoi, significantly increased root weight, but did not decrease nematode damage or population density. The supernatant of all strains significantly decreased hatching of juvenile nematodes after 24 h of immersion, with isolates of P. putida and B. megaterium being the most effective
Vineyards in the Atacama region in Chile were surveyed from 2007 to 2009 for the presence of viruses. This region is an important area of table grape production, supplying international markets with its fruits in the off season of the Northern Hemisphere. Reverse transcription‐polymerase chain reaction (RT‐PCR) assay was used to detect the most economically important grapevine viruses in 1000 samples, including symptomatic and asymptomatic plants. The rate of positive samples was 8.8% for Grapevine leafroll‐associated virus 1 (GLRaV‐1), 46.8% for Grapevine leafroll‐associated virus 2 (GLRaV‐2), 9.1% for Grapevine leafroll‐associated virus 3 (GLRaV‐3), 12.3% for Grapevine virus A (GVA), 30.7% for Grapevine fleck virus (GFkV) and 9.6% for Grapevine fanleaf virus (GFLV). Overall virus infection was 68.7%. DNA sequencing confirmed the identification of viruses in selected samples, and comparative analysis indicated that Chilean isolates have moderate‐to‐high molecular identities with corresponding virus reference strains selected from GenBank. The high level of viral infection observed indicates that viruses are involved in decreasing table grape production in the region. This is the first extensive virus survey performed in the Atacama region, is also the first study of genetic comparison of grapevine viruses developed in South America with a wide spectrum of viruses and isolates and provides an assessment of grapevine viruses on table grape.
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