Dinophysis acuta and D. acuminata are associated with lipophilic toxins in Southern Chile. Blooms of the two species coincided during summer 2019 in a highly stratified fjord system (Puyuhuapi, Chilean Patagonia). High vertical resolution measurements of physical parameters were carried out during 48 h sampling to i) explore physiological status (e.g., division rates, toxin content) and ii) illustrate the fine scale distribution of D. acuta and D. acuminata populations with a focus on water column structure and cooccurring plastid-bearing ciliates. The species-specific resources and regulators defining the realized niches (sensu Hutchinson) of the two species were identified. Differences in vertical distribution, daily vertical migration and in situ division rates (with record values, 0.76 d−1, in D. acuta), in response to the environmental conditions and potential prey availability, revealed their niche differences. The Outlying Mean Index (OMI) analysis showed that the realized niche of D. acuta (cell maximum 7 × 103 cells L−1 within the pycnocline) was characterized by sub-surface estuarine waters (salinity 23 -25), lower values of turbulence and PAR, and a narrow niche breath. In contrast, the realized niche of D. acuminata (cell maximum 6.8 × 103 cells L−1 just above the pycnocline) was characterized by fresher (salinity 17 -20) outflowing surface waters, with higher turbulence and light intensity and a wider niche breadth. Results from OMI and PERMANOVA analyses of co-occurring microplanktonic ciliates were compatible with the hypothesis of species such as those from genera Pseudotontonia and Strombidium constituting an alternative ciliate prey to Mesodinium. The D. acuta cell maximum was associated with DSP (OA and Please note that this is an author-produced PDF of an article accepted for publication following peer review. The definitive publisher-authenticated version is available on the publisher Web site.DTX-1) toxins and pectenotoxins; that of D. acuminata only with pectenotoxins. Results presented here contribute to a better understanding of the environmental drivers of species-specific blooms of Dinophysis and management of their distinct effects in Southern Chile. Previous article Highlights► 48 h of high frequency physical data for co-occurring blooms of 2 Dinophysis species. ► D. acuta (exceptional µ) thin layer briefly disrupted by an increase in turbulence. ► Co-occurring D. acuminata and D. acuta blooms showed a clear niche differentiation. ► Niche analysis results compatible with putative ciliate prey other than Mesodinium. ► D. acuta maximum associated with DSP toxins (OA, DTX1), D. acuminata with PTX2 only.
In late February 2016, a harmful algal bloom (HAB) of Alexandrium catenella was detected in southern Chiloé, leading to the banning of shellfish harvesting in an extended geographical area (~500 km). On April 24, 2016, this bloom produced a massive beaching (an accumulation on the beach surface of dead or impaired organisms which were drifted ashore) of surf clams Mesodesma donacium in Cucao Bay, Chiloé. To determine the effect of paralytic shellfish poisoning (PSP) toxins in M. donacium, samples were taken from Cucao during the third massive beaching detected on May 3, 2016. Whole tissue toxicity evidence a high interindividual variability with values which ranged from 1008 to 8763 μg STX eq 100 g−1 and with a toxin profile dominated by GTX3, GTX1, GTX2, GTX4, and neoSTX. Individuals were dissected into digestive gland (DG), foot (FT), adductor muscle (MU), and other body fractions (OBF), and histopathological and toxin analyses were carried out on the obtained fractions. Some pathological conditions were observed in gill and digestive gland of 40–50% of the individuals that correspond to hemocyte aggregation and haemocytic infiltration, respectively. The most toxic tissue was DG (2221 μg STX eq 100 g−1), followed by OBF (710 μg STX eq 100 g−1), FT (297 μg STX eq 100 g−1), and MU (314 μg STX eq 100 g−1). The observed surf clam mortality seems to have been mainly due to the desiccation caused by the incapability of the clams to burrow. Considering the available information of the monitoring program and taking into account that this episode was the first detected along the open coast of the Pacific Ocean in southern Chiloé, it is very likely that the M. donacium population from Cucao Bay has not had a recurrent exposition to A. catenella and, consequently, that it has not been subjected to high selective pressure for PSP resistance. However, more research is needed to determine the effects of PSP toxins on behavioral and physiological responses, nerve sensitivity, and genetic/molecular basis for the resistance or sensitivity of M. donacium.
An understanding of biocontrol activities is important when developing microorganism‐based alternatives to conventional fungicides. From our bacterial collection, we selected two strains (BBC023 and BBC047) for their outstanding antagonistic capacity against fungal phytopathogens and growth‐promoting abilities towards Arabidopsis thaliana. According to physiological and molecular characterizations, both strains were classified as Bacillus amyloliquefaciens and were tested against Botrytis cinerea in vitro and in a tomato. Both strains secrete lipopeptide‐like compounds that contribute to their in vitro antagonism. SEM‐images showed altered B. cinerea mycelial structures that were consistent with previous reports of the direct action of lipopeptides against fungal hyphae. The strains were applied to the roots (R), leaves (foliar ‐ F) or root/leaves (R/F) on tomato plants. All treatments significantly reduced the severity of B. cinerea infection (measured as a control index). However, only root applications (R and R/F) led to growth promotion in the tomato plants. We detected the production of indole acetic acid (IAA) and 2,3‐butanediol as growth promotion traits in the two strains. For both strains, the R/F treatment showed the highest control index, suggesting a synergic effect of direct antagonism against B. cinerea and resistance induction in the plant. In addition, in vitro antagonism of BBC023 and BBC047 against B. cinerea was similar; whereas in the F application, strain BBC047 significantly improved plant resistance and maintained a higher population density over time on tomato leaves, compared to BBC023. BBC047 was also able to produce a complex and robust biofilm in Msgg medium compared with that of BBC023. We linked the reduced biocontrol of BBC023 on leaves with its limited ability to generate robust biofilms and colonize the phylloplane. At last, we highlight the potential of the native Bacillus strains as promising alternatives for the development of bioproducts for sustainable agriculture.
Six aristolochic acids were identified in the Chilean species Aristolochia chilensis using high-performance liquid chromatography coupled to a diode array detector (HPLC-DAD) and subsequent confirmation with mass spectrometry (MS). The fractions of each signal were collected and injected directly into an Orbitrap mass detector model Q Exactive Focus (Thermo Scientific). The acids extraction was done with 0.10–0.50 g of lyophilized and pulverized sample and concentrated in Soxhlet extraction equipment. The liquid-liquid separations and a subsequent solid phase extraction (SPE) C18 were performed using 100 µL of the extract that contains the aristolochic acids present in the Aristolochia chilensis plant. The HPLC conditions used a single mobile phase acetonitrile : water (1 : 1) acidified with 0.1% acetic acid and an isocratic elution to 1 mL·min−1. The column InertSustain C18 250 × 4.6 mm and 3 µm was used, the injection volume was 20 µL, and the time of run was reduced to 15 min. Calibration curves were constructed with r 2 being 0.9997. The quantification limit for AAI was 0.138 ± 0.010 µg/mL, and for AAII, it was 0.558 ± 0.042 µg/mL.
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