A simple and efficient method for isolating small RNAs from different plant species

Background  

Small RNAs emerged over the last decade as key regulators in diverse biological processes in eukaryotic organisms. To identify and study small RNAs, good and efficient protocols are necessary to isolate them, which sometimes may be challenging due to the composition of specific tissues of certain plant species. Here we describe a simple and efficient method to isolate small RNAs from different plant species.     

New morphological data on the first-stage larvae of two Procamallanus species (Nematoda: Camallanidae) based on SEM studies

First-stage larvae of camallanid nematodes Procamallanus (Procamallanus) laeviconchus (Wedl, 1862) and Procamallanus (Procamallanus) sp. from naturally infected Distichodus niloticus (Hasselquist) and Clarias gariepinus (Burchell), respectively, from Lake Turkana, Kenya (new geographical records) are described, being for the first time studied by scanning electron microscopy. Larvae of both species are characterised by the presence of a dorsal cephalic tooth, four submedian cephalic papillae and a pair of amphids, and by the elongate tail with several terminal digit-like processes. The latter formations probably serve for the attachment of larvae to the substrate in water when the larvae attract copepod intermediate hosts by their movements; these structures, especially their numbers, may be of taxonomic importance in camallanid nematodes.     

A new species of Acanthochondria (Copepoda: Chondracanthidae) parasitizing the flounder Xystreurys rasile (Pleuronectiformes: Paralichthyidae) from Argentina

A new copepod species, Acanthochondria sagitta sp. n., is described based on specimens collected from the flounder Xystreurys rasile (Jordan) (Pleuronectiformes, Paralichthyidae), caught in the coastal waters off Necochea, Buenos Aires Province, Argentina. The new species differs from its congeners by the following combination of characters: presence of three pairs of cephalic outgrowths; the triangular shape of the trunk with diverging postero-lateral processes; leg 2 of an intermediate shape between Types C and D, which projects laterally from the trunk, and Type B-V antennule bearing two proximal processes (one ventral and one dorsal) on the swollen basal portion.     

Convergence of developmental mutants into a single tomato model system: 'Micro-Tom' as an effective toolkit for plant development research

Background

The tomato (Solanum lycopersicum L.) plant is both an economically important food crop and an ideal dicot model to investigate various physiological phenomena not possible in Arabidopsis thaliana. Due to the great diversity of tomato cultivars used by the research community, it is often difficult to reliably compare phenotypes. The lack of tomato developmental mutants in a single genetic background prevents the stacking of mutations to facilitate analysis of double and multiple mutants, often required for elucidating developmental pathways.

Results

We took advantage of the small size and rapid life cycle of the tomato cultivar Micro-Tom (MT) to create near-isogenic lines (NILs) by introgressing a suite of hormonal and photomorphogenetic mutations (altered sensitivity or endogenous levels of auxin, ethylene, abscisic acid, gibberellin, brassinosteroid, and light response) into this genetic background. To demonstrate the usefulness of this collection, we compared developmental traits between the produced NILs. All expected mutant phenotypes were expressed in the NILs. We also created NILs harboring the wild type alleles for dwarf, self-pruning and uniform fruit, which are mutations characteristic of MT. This amplified both the applications of the mutant collection presented here and of MT as a genetic model system.

Conclusions

The community resource presented here is a useful toolkit for plant research, particularly for future studies in plant development, which will require the simultaneous observation of the effect of various hormones, signaling pathways and crosstalk.     

<Emphasis Type="Italic">Erwinia aphidicola</Emphasis> isolated from commercial bean seeds (<Emphasis Type="Italic">Phaseolus vulgaris)</Emphasis>

In late 2003, a new disease appeared in protected bean crops in southeastern Spain, causing a decrease of over 50% in production. Several samples of affected plants were collected and analyzed and the agent of this disease was identified as the bacterium Erwinia aphidicola, which had never been described as a pathogen previously. We attempted to determine the possible bacterium transmission through seeds, using 120 commercial bean seeds from the same batch as that used in an affected farm, and 120 seeds from the fruiting plants of the same farm. Seed coats, cotyledons and leaves of plants originating from them, were taken and analyzed. Several of the developed symptoms on plants from commercial and fruiting plant seeds were internervial chlorosis, necrotic pits and rough roots and they coincided with those observed on affected crops. Bacteria present in commercial seed cotyledons were isolated and analyzed by biochemical and molecular tests. Results confirmed the presence of Erwinia aphidicola in four analyzed seeds; moreover, Bacillus simplex/Bacillus muralis, Pseudomonas mendocina, Pseudomonas putida and Paenibacillus polymyxa were also identified.     

Effect of cropping systems and crop successions on fumonisin levels in corn from Northern Paraná State,Brazil

In this study the effect of different cropping systems and crop successions was evaluated on natural Fusarium sp. contamination and fumonisin levels in corn. The cropping systems consisted of a conventional and no-tillage area cultivated with corn in the summer following either oats or fallow in the winter (2006 and 2007 growing seasons). In addition, the effect of applying nitrogen fertilizer (0, 22.5, 45.0, 90.0 and 90.0 kg ha⁻¹ nitrogen supplemented with potassium oxide) on fumonisin contamination was evaluated in the 2006 growing season. Fusarium sp. was detected in 90% samples in 2006 and in 100% samples in 2007. In both growing seasons, no-till corn following oats showed the highest mean fumonisin levels and differed significantly (P < 0.05) from all the others (2006) and from conventional till corn following either oats or fallow in the winter (2007). Fumonisin levels ranged from 0.13 to 19.52 μg g⁻¹ (mean 6.97 μg g⁻¹) and from 3.70 to 7.75 μg g⁻¹ (mean 6.29 μg g⁻¹) in no-till corn following oats from the 2006 and 2007 growing seasons, respectively. Plots treated with 0 kg ha⁻¹ and 22.5 kg ha⁻¹ nitrogen showed the highest mean fumonisin levels and differed significantly from those with 45.0 and 90 kg ha⁻¹ nitrogen. Fumonisin levels correlated negatively (P < 0.05) with the nitrogen fertilization rates. Although no-till is advantageous from a soil conservation standpoint, it may enhance the potential for fumonisin contamination in corn.     

Reduced metribuzin pollution with phosphatidylcholine-clay formulations

BACKGROUND: Metribuzin is a widely used herbicide that has been identified as a groundwater contaminant. In this study, slow‐release formulations of metribuzin were designed by encapsulating the active ingredient in phosphatidylcholine (PC) vesicles and adsorbing the vesicles onto montmorillonite. RESULTS: The maximum active ingredient content in the slow‐release formulations was 246 g kg^?1. Infrared spectroscopy results revealed that the hydrophobic interactions between metribuzin and the alkyl chains on PC were necessary for encapsulation. In addition, water bridges connecting the herbicide and the PC headgroup enhanced the solubility of metribuzin in PC. Adsorption experiments in soils were performed to evaluate the relationship between sorption and leaching. Funnel experiments in a sandy soil revealed that the herbicide was not irreversibly retained in the formulation matrix. In soil column experiments, PC–clay formulations enhanced herbicide accumulation and biological activity in the top soil layer relative to a commercial formulation. PC–clay formulations also reduced the dissipation of metribuzin by a factor of 1.6–2.5. CONCLUSIONS: A reduction in the recommended dose of metribuzin can be achieved by employing PC–clay formulations, which reduces the environmental risk associated with herbicide applications. Moreover, PC and montmorillonite are non‐toxic and do not negatively affect the environment. Copyright © 2010 Society of Chemical Industry     

Insecticide resistance status of Aedes aegypti (L.) from Colombia

BACKGROUND: To evaluate the insecticide susceptibility status of Aedes aegypti (L.) in Colombia, and as part of the National Network of Insecticide Resistance Surveillance, 12 mosquito populations were assessed for resistance to pyrethroids, organophosphates and DDT. Bioassays were performed using WHO and CDC methodologies. The underlying resistance mechanisms were investigated through biochemical assays and RT‐PCR. RESULTS: All mosquito populations were susceptible to malathion, deltamethrin and cyfluthrin, and highly resistant to DDT and etofenprox. Resistance to lambda‐cyhalothrin, permethrin and fenitrothion ranged from moderate to high in some populations from Chocó and Putumayo states. In Antioquia state, the Santa Fe population was resistant to fenitrothion. Biochemical assays showed high levels of both cytochrome P450 monooxygenases (CYP) and non‐specific esterases (NSE) in some of the fenitrothion‐ and pyrethroid‐resistant populations. All populations showed high levels of glutathione‐S‐transferase (GST) activity. GSTe2 gene was found overexpressed in DDT‐resistant populations compared with Rockefeller susceptible strain. CONCLUSIONS: Differences in insecticide resistance status were observed between insecticides and localities. Although the biochemical assay results suggest that CYP and NSE could play an important role in the pyrethroid and fenitrothion resistance detected, other mechanisms remain to be investigated, including knockdown resistance. Resistance to DDT was high in all populations, and GST activity is probably the main enzymatic mechanism associated with this resistance. The results of this study provide baseline data on insecticide resistance in Colombian A. aegypti populations, and will allow comparison of changes in susceptibility status in this vector over time. Copyright © 2011 Society of Chemical Industry     

Possibility of Bean yellow mosaic virus detection in Gladiolus plants by different methods

Journal of Plant Diseases and Protection - Serological and molecular methods were compared for the detection of Bean yellow mosaic virus (BYMV) in gladiolus plants. Plants showing mosaic symptoms...     

Partial replacement of Fe(o,o‐EDDHA) by humic substances for Fe nutrition and fruit quality of citrus

The most widely used Iron (Fe) fertilizer in calcareous soils is the synthetic chelate Fe(o,o‐EDDHA). However, humic substances are occasionally combined with Fe chelates in drip irrigation systems in order to lower costs. We investigated the effect of various mixtures of Fe(o,o‐EDDHA) and a commercially available humic substance on Fe availability in a calcareous soil from Murcia, Spain (in vitro experiment) and on leaf Fe content and fruit‐quality attributes of Citrus macrophylla (field experiment). In the in vitro experiment, a calcareous soil was incubated for 15 d with solutions of sole Fe(o,o‐EDDHA) and humic substance and of a mixture of humic substance and Fe(o,o‐EDDHA) to determine the dynamics of available Fe. While the mixture did not significantly increase the available soil Fe, it did decrease the rate of Fe retention in the surface soil compared to sole Fe(o,o‐EDDHA). In the field experiment, the substitution in the application solution of 67% of Fe(o,o‐EDDHA) by commercial humic substance increased leaf P in lemon trees from 0.19% with sole Fe(o,o‐EDDHA) to 0.30% and leaf Fe from 94 mg kg^–1 to 115 mg kg^–1. Some quality parameters like vitamin C content and peel thickness were also improved with a partial substitution of Fe(o,o‐EDDHA) by humic substances. We conclude that a partial substitution of commercial Fe chelates by humic substance can improve crop Fe uptake and may thus be economically attractive. The underlying physiological mechanisms and ecological implications require further studies.