Differential expression of defence-related proteins in Vigna unguiculata (L. Walp.) seedlings after infection with Fusarium oxysporum

This work aimed to study the induction of defence proteins in cowpea seedlings during the first days after infection with the fungi Fusarium oxysporum f. sp. cubenses and F. oxysporum f. sp. phaseoli. Cowpea seeds, after disinfection, were transferred to Petri dishes containing 0.5% agar and, after germination, were infected with a drop of a suspension containing 0.5 × 10⁴ spores ml⁻¹. Seedlings were collected at 24, 48, 72 and 96 h after infection and were dissected into leaves, hypocotyls, roots, cotyledons and teguments, which were measured and weighed for morphometric analysis. The agar medium was also analyzed. Enzymatic assays of proteic extracts yielded antimicrobial peptides detected by Western blotting. The cowpea seedlings showed a complex pattern of induction and repression of defence proteins in response to infection by both pathogens. Furthermore, morphometric analysis showed differences between infected and control seedlings. Infected samples did not at any time exhibit chitinase activity, but did exhibit different β-1,3-glucanase and peroxidase activities. Western blotting for lipid transfer protein (LTP) demonstrated its presence in all parts of the infected seedlings. Exuded proteins, also obtained from cowpea seeds in the germination medium, were separated by SDS-PAGE and tricine gel electrophoresis. The analysis showed that some proteins were exuded from moistened cowpea seeds, particularly after F. oxysporum infection.     

Expression of conserved mucin domains by epithelial tissues in various mammalian species

Mucins are related to infectious and non-infectious diseases in Veterinary and Human Medicine. MUC1 mucin is a transmembrane glycoprotein expressed on the apical surface of human epithelia while MUC5AC is the predominant secreted mucin expressed in human gastric epithelium and goblet cells of lung and eyes. MUC5AC C-terminus cysteine rich regions and the cytoplasmic tail of MUC1 domains are conserved among several mammalian species. Objective: to compare the expression of MUC1 and MUC5AC mucins in mammalian epithelia. CT33 anti-MUC1 cytoplasmic tail (MUC1CT) polyclonal antibody and 45M1 anti-MUC5AC monoclonal antibody were employed. By immunohistochemistry, MUC1CT was expressed in most tissues while MUC5AC was restricted to gastric surface epithelium and goblet cells from trachea and lung. By western blot, MUC1CT showed a band at approximately 35 kDa in most tissues; MUC5AC revealed bands at >180 kDa in stomach and lung secretions from rat, cat, pig and cow. When rat MUC5AC was immunoprecipitated, a band at about 180 kDa was obtained.     

Yield map generation of perennial crops for fresh consumption

Yield mapping technologies can help to increase the quantity and quality of agricultural production. Current systems only focus on the quantification of the harvest, but the quality has equal or greater importance in some perennial crops and impacts directly on the financial profitability. Therefore, a system was developed to quantify and relate the quality obtained in the classification line with the plants of the orchard and for decision-making. The system is comprised of hardware, which obtains the location of the harvester bag during harvesting and unloading at the unloading site, and software that processes the collected data. The cloud of real-time data contributed from the different collectors (bins) allows the construction of yield maps, considering the multi-stage harvesting system. Further, the system enables the creation of a detailed map of the plants and fruits harvested. As the harvest focuses on quality, it takes place in stages, depending on the ripening of the fruits. In addition to the yield maps, the system allows identification of the efficiency of each worker undertaking the harvest by the number of performed discharges and by the time spent. The system was developed in partnership with the Federal Technological University of Paraná and Embrapa Uva & Vinho and was tested in apple orchards in southern Brazil. Although the system was evaluated with only data from apple cultivation, monitoring the quality and quantifying other orchard fruits can positively impact the fruit sector.

     

Feeding of yeast (Candida spp.) improves in vitro ruminal fermentation of fibrous substrates

In vitro gas production technique (IVGPT) was used with the objective of determining the inclusion effect of live cells of two strains of Candida yeast on in vitro ruminal fermentation of two fibrous substrates. In order to achieve this, two experiments were performed: A) using oat straw (Avena sativa) as substrate; B) using alfalfa hay (Medicabo sativa) as substrate, comparing the effect of two different strains of Candida genre, both isolated from the rumen, on the mentioned substrates. Levica 25 (Candida tropicalis) yeast belongs to the culture collection of the Institute of Animal Science, Cuba, and Levazoot 15 (Candida norvegensis) yeast is part of the collection of the Faculty of Zootechnology and Ecology of the Autonomous University of Chihuahua, Mexico. Both strains demonstrated their potential in activating the ruminal fermentation. They stimulated (P<0.0001) the ruminal fermentation of the substrates under study. However, the Levazoot strain stimulated the dry matter (DM) fermentation of alfalfa in 21.43%, more than Levica 25. It is concluded that there is an influence of yeast strain and diet on the rumen environment and, therefore, it is important to select the appropriate strain in every production condition.     

Increase in Iron and Zinc Concentrations in Rice Grains Via the Introduction of Barley Genes Involved in Phytosiderophore Synthesis

Increasing the iron (Fe) and zinc (Zn) concentrations of staple foods, such as rice, could solve Fe and Zn deficiencies, which are two of the most serious nutritional problems affecting humans. Mugineic acid family phytosiderophores (MAs) play a very important role in the uptake of Fe from the soil and Fe transport within the plant in graminaceous plants. To explore the possibility of MAs increasing the Fe concentration in grains, we cultivated three transgenic rice lines possessing barley genome fragments containing genes for MAs synthesis (i.e., HvNAS1, HvNAS1, and HvNAAT-A and HvNAAT-B or IDS3) in a paddy field with Andosol soils. Polished rice seeds with IDS3 inserts had up to 1.40 and 1.35 times higher Fe and Zn concentrations, respectively, compared to non-transgenic rice seeds. Enhanced MAs production due to the introduced barley genes is suggested to be effective for increasing Fe and Zn concentrations in rice grains.     

Giardia duodenalis in colony stray cats from Italy

Giardia duodenalis is the most common intestinal protozoan in humans and animals worldwide, including eight morphologically identical assemblages, infecting pets, livestock, wildlife and human beings. Assemblages A and B are those with the higher zoonotic potential, and they have been detected in several mammals other than humans; the others (C to H) show a higher host specificity. Cats can harbour both the specific Assemblage F and the zoonotic ones A and B. Several studies have been carried out on G. duodenalis genotypes in cats; however, the role of this species in the epidemiology of giardiasis is still poorly understood. In this scenario, the present study carried out the detection and genetic characterization at sub-assemblage level of G. duodenalis from colony stray cats in central Italy. In the period 2018–2019, 133 cat faecal samples were analysed for the presence of G. duodenalis cysts by a direct immunofluorescence assay. Positive samples were subsequently subjected to molecular analyses for assemblage/sub-assemblage identification. Forty-seven samples (35.3%) were positive for G. duodenalis cysts by immunofluorescence. G. duodenalis DNA was amplified at SSU-rDNA locus from 39 isolates: 37 were positive for zoonotic Assemblage A and 2 showed a mixed infection (A + B). Positive results for the β-giardin gene were achieved for 25 isolates. Sequence analysis revealed 16 isolates belonging to Sub-assemblage AII and 8 to Sub-assemblage AIII. One isolate resulted as ambiguous AI/AIII. Large sequence variability at the sub-assemblage level was detected, with several double peaks and mutations, making complex a proper isolate allocation. When compared with previous studies, the 35.3% prevalence of G. duodenalis in cats reported in the present article was surprisingly high. Moreover, all positive cats resulted to be infected with zoonotic assemblages/sub-assemblages, thus indicating stray cats as a possible source of human giardiasis and highlighting the sanitary relevance of cat colonies in the study area.     

A new record of the freshwater pearl mussel Margaritifera margaritifera L. (Bivalvia,Unionoida) from the River Narcea (Asturias,north‐western Spain)

1. A population of the freshwater pearl mussel Margaritifera margaritifera L. was found in the River Narcea in the province of Asturias (north‐western Spain). Primary analysis of the population shows that it is well distributed along the river and is particularly abundant in shady river banks with a gravel substrate. 2. The length of living mussels ranges between a minimum of 22 mm and a maximum of 116 mm, with most in the size class between 65 mm and 70 mm. There is a good proportion of young mussels, although no fertile females were found in the course of the investigation. 3. Although there is no specific legislation in Spain for the protection of M. margaritifera, it has recently been proposed to include M. margaritifera in the Spanish List of Threatened Species as ‘vulnerable’. 4. Although the River Narcea seems to provide suitable conditions for the pearl mussel (water quality, available salmonid hosts, etc.), the main conservation measures must be directed at avoiding river fragmentation from dams, channelization and organic enrichment from individual farms or towns situated close to the river bank. Copyright © 2000 John Wiley & Sons, Ltd.     

Not the one,but the only one: about <Emphasis Type="Italic">Cannabis cryptic virus</Emphasis> in plants showing ‘hemp streak’ disease symptoms

Interveinal chlorosis and leaf margin wrinkling are widespread symptoms of Cannabis sativa. They are traditionally attributed to the so-called hemp streak virus (HSV), but its existence has not been demonstrated yet. To our knowledge, no molecular investigation has so far been performed in order to identify the causal agent of this symptomatology, we therefore decided to use traditional and molecular virology techniques to better characterize symptoms and pursue the etiological agent. No pathogenic virus was found by using targeted PCR reactions and by RNA sequencing, whereas we were able to detect the Cannabis cryptic virus (CanCV) with both techniques. We, therefore, developed an RT-qPCR assay based on a CanCV-specific TaqMan probe and applied it to a wide range of symptomatic and symptomless plants, using a two-step (for quantification), or a one-step (for fast detection) protocol. Both symptoms and the virus were only shown to be transmitted vertically and did not pass via mechanical inoculation or grafting, though we could not find any cause-effect correlation between them. In fact, the virus was found in all the tested hemp samples, and its abundance varied greatly between different accessions and individuals, independently from the presence and severity of symptoms. The suggestion that hemp streak is caused by a virus is therefore questioned. Some abiotic stresses seem to play a role in triggering the symptoms but this aspect needs further investigation. For breeding purposes, a selection of parental plants based on the absence of symptoms proved to be efficient in containment of the disease.     

Glyphosate-resistant weeds of South American cropping systems: an overview

Herbicide resistance is an evolutionary event resulting from intense herbicide selection over genetically diverse weed populations. In South America, orchard, cereal and legume cropping systems show a strong dependence on glyphosate to control weeds. The goal of this report is to review the current knowledge on cases of evolved glyphosate-resistant weeds in South American agriculture. The first reports of glyphosate resistance include populations of highly diverse taxa (Lolium multiflorum Lam., Conyza bonariensis L., C. canadensis L.). In all instances, resistance evolution followed intense glyphosate use in fruit fields of Chile and Brazil. In fruit orchards from Colombia, Parthenium hysterophorus L. has shown the ability to withstand high glyphosate rates. The recent appearance of glyphosate-resistant Sorghum halepense L. and Euphorbia heterophylla L. in glyphosate-resistant soybean fields of Argentina and Brazil, respectively, is of major concern. The evolution of glyphosate resistance has clearly taken place in those agroecosystems where glyphosate exerts a strong and continuous selection pressure on weeds. The massive adoption of no-till practices together with the utilization of glyphosate-resistant soybean crops are factors encouraging increase in glyphosate use. This phenomenon has been more evident in Argentina and Brazil. The exclusive reliance on glyphosate as the main tool for weed management results in agroecosystems biologically more prone to glyphosate resistance evolution.