Purification and some properties of Papaya meleira virus, a novel virus infecting papayas in Brazil

'Meleira', or 'sticky disease', is currently the most damaging papaya disease in the mid-eastern Brazilian growing regions. Consistent disease transmission via latex injection, presence of similar isometric particles in the laticiferous vessels of diseased plants, and detection of double-stranded DNA in naturally and experimentally infected papaya trees suggest that a virus is the causal agent. Conclusive evidence for viral aetiology was previously lacking, mostly because every attempt to purify the putative virus from infected papayas had failed. Following the successful purification and partial characterization of the meleira virus, healthy papaya seedlings injected with purified virus particles later developed typical symptoms of the disease. Negatively stained, isometric, full and 'empty' purified virus particles measured 42 and 38 nm, respectively. The viral genome was a single dsRNA molecule of about 12 kbp. Several capsid proteins, ranging in size from 14·4 to 45 kDa, were consistently revealed by PAGE. Papaya meleira virus (PMeV) appears to represent a novel group of viruses, with no known similar counterpart among known plant-, vertebrate-, invertebrate- or prokaryote-infecting viruses.     

Anastomosis Groups, Pathogenicity and Sensitivity to Fungicides of Rhizoctonia solani Isolates Collected on Potato Crops in France

A collection of 241 isolates of Rhizoctonia solani obtained from potato plants grown in different areas in France was characterized for anastomosis grouping, symptomatology on tubers of different cultivars and sensitivity to three fungicides. Most isolates collected belonged to (anastomosis groups (AGs)) AG 3, but 2% and 4% of the isolates were AG 5 and AG 2-1. AG 3 and AG 2-1 isolates were mostly obtained from sclerotia on tubers, but all AG 5, some AG 3 and some AG 2-1 isolates were recovered from superficial tuber alterations, like deformations, corky or scabby lesions. Sclerotia were formed on tubers produced by healthy stem cuttings grown in soil artificially infested with AG 3, but not on tubers grown in soil infested with either AG 5 or AG 2-1. No variation in susceptibility to sclerotial formation was observed among five potato cultivars. In all cases, a large proportion of tubers showed superficial corky lesions, often associated with deformations. The proportion of tubers with lesions and deformations was highest in soil infested with AG 2-1 and significantly lower on cv. Samba in all treatments. All isolates were highly sensitive to flutolanil, iprodione and pencycuron, except the AG 5 isolates, moderately sensitive to pencycuron. These results show that, although AG 3 is the most common R. solani group on potato in France, AG 5 and AG 2-1 may be present. Isolates differed for pathogenicity. In vitro sensitivity to fungicides varied among AGs.     

Frequency of transmission of artichoke Italian latent nepovirus by Longidorus fasciatus (Nematoda: Longidoridae) from artichoke fields in the Iria and Kandia areas of Argolis in northeast Peloponnesus, Greece

Artichoke Italian latent nepovirus (AILV) transmitted by Longidorus fasciatus is a causative agent of artichoke patchy chlorotic stunting (APCS) in northeast Peloponnesus in southern Greece. Populations of L. fasciatus collected from the Iria and Kandia areas of Argolis in northeast Peloponnesus were used in laboratory experiments to determine the frequency of transmission of the virus by its natural vector. One tenth to almost one half of the L. fasciatus specimens recovered from soil collected in two artichoke fields showing APCS transmitted AILV. Allowing nematodes access for 4 wk to Nicotiana clevelandii mechanically infected with AILV did not increase the number of individual specimens able to transmit virus. The total number of specimens transmitting virus in an experiment did not exceed fifty percent of the individuals tested. Virus-like particles were only observed adsorbed to the inner surface of the odontostyle and it is suggested that the high frequency of transmission of AILV by L. fasciatus is a result of efficient dissociation of virus particles from the specific sites of retention in the vector.     

Preliminary investigation of the causal agent(s) of a disease causing leaf curl of tobacco in South Africa

A high incidence (86%) of potyvirus infection was noted in tobacco plants exhibiting a form of leaf curl in South Africa. Despite leaf curl being reported in the literature to be of geminiviral aetiology, no geminiviruses were detected. Furthermore, no other virus particles were detected by virus purification, TEM and serology. Twelve species of dsRNA were consistently isolated from these tobacco plants, but were absent from other forms of leaf curl-affected and healthy tobacco. Aphid and mechanical inoculation demonstrated that the purified potyvirus(es) did not cause leaf curl symptoms, but rather mild mottle and mosaic symptoms in tobacco. Partial characterization of the potyvirus preparation showed a possible relationship to a South African strain of potato virus Y. Because potyvirus-inoculated plants did not manifest leaf curl symptoms, and because leaf curl symptoms were noted in some plants not infected with a potyvirus, it was concluded that the potyvirus is not involved in the leaf curl aetiology, but causes a latent infection, the symptoms of which are masked. The pattern of the dsRNA banding, induction of enations and lack of mechanical and seed transmission are common to plant reoviruses. The possibility of a phytoreovirus involvement in this form of leaf curl is currently being investigated. The results from this study suggest that tobacco leaf curl disease worldwide, with regard to geminiviruses, be re-evaluated.     

Role of acquisition and inoculation time in the expression of clonal variation for BYDV-PAV transmission in the aphid species Rhopalosiphum padi

Vector efficiency of 20 Rhopalosiphum padi clones, originating from Europe, North America and North Africa and exhibiting different types of life cycle, was evaluated by transmitting a French BYDV-PAV isolate to barley plants under five different acquisition/inoculation sequences (AAP/IAP). Differences between clones in transmission efficiency were found only when a short AAP was followed by a long IAP (6 h/120 h) and, to some extent, when a long AAP (48 h) was followed by a short IAP (6 h), but no differences were found when the conditions for virus transmission were optimal, i.e. long AAP followed by long IAP (48 h/120 h). There were no differences in transmission rates by clones of different geographical origins and with different life cycles. As a consequence, clonal variation is probably of little importance in the vector aspects of the epidemiology of PAV serotypes transmitted by R. padi , but the availability of a range of clones exhibiting transmission differences under limiting AAP or IAP conditions could be of interest for studies of virus–vector relationships.     

Weed communities of winter wheat as influenced by input level and rotation

The influence of cropping systems constituted by the combination of three input levels (low, intermediate and high) and four 2-year rotations between a spring summer crop and winter wheat ( Triticum aestivum L.) on the density and composition of the winter wheat weed flora was evaluated from 1989 to 1992. Spring summer crops were maize ( Zea mays L.). soyabean ( Glycine max (L.) Merr.). sugar beet ( Beta vulgaris L.) and sunflower ( Helianthus and L.) Total weed density at the end of the wheat tillering stage ranged from 2.5 to 128.7 plants m^-2 and generally decreased with cropping intensity, although to a differing extent, depending upon rotation. Differences among weed communities of the 12 cropping systems were mostly dependent upon input level. The effect of the preceding crop on the weed flora composition of winter wheat was evident only with reduced herbicide use (low-input systems). Weed species could be ascribed to four categories: (a) species sporadically associated with cropping systems (most of the weeds); (b) species associated with a specific input level (e.g. Papaver rhoeas L. to low and intermediate input levels and Veronica persica Poiret to the high input ievel): (c) species associated with a specific rotation (e.g. Anm majus L. to sugar beet winter wheat): and (d) species associated with specitie plots that become important every other year (e.g. Convolvulus arvensis L. and Stellaria media (L.) Vill.).     

Footsteps from Insect Larvae Damage Leaf Surfaces and Initiate Rapid Responses

Plant resistance to insect herbivory involves gene expression in response to wounding and the detection of insect elicitors in oral secretions (Kessler and Baldwin, 2002, Ann. Rev. Plant/ Biol. 53: 299–328). However, crawling insect larvae stimulate the synthesis of 4-aminobutyrate within minutes and imprints of larval footsteps can be visualized within seconds through superoxide production or transient increases in chlorophyll fluorescence (Bown et al., 2002, Plant Physiol. 129: 1430–1434). Here cryo-scanning electron microscopy was used to demonstrate that larval feet, which are equipped with a perimeter row of hook-like crochets, damage leaf tissue and result in larval footprints. Staining for cell death shows that areas of wounding correspond to footsteps detected through increased chlorophyll fluorescence. Superoxide production in response to footsteps was inhibited by diphenyleneiodonium, an inhibitor of the plasma membrane NADPH oxidase enzyme. Inhibition of superoxide production, however, did not eliminate the detection of cell death. The results demonstrate that larval footsteps damage leaf tissue, and initiate rapid local responses which are not dependent on herbivory or oral secretions. It is proposed that superoxide production at the wound site prevents opportunistic pathogen infection.     

The Impact of Natural Deep Eutectic Solvents and Extraction Method on the Co-Extraction of Trace Metals from Fucus vesiculosus

In recent years, natural deep eutectic solvents (NADES) have been widely investigated for the extraction of food and medicinal plants as well as seaweeds. However, the ability of NADES for trace elements co-extraction from natural sources is not well investigated. The aim of this study was to investigate the ability of common NADES for trace elements co-extraction from Fucus vesiculosus. All of the tested NADES did not recover As and Co (concentration <LOQ). Moreover, all of the tested NADES provided a low recovery (<9%) of Ba, Ca, Fe, Mg, Mn, Sr, and Zn. The method of extraction had not shown a statistically significant effect on the co-extraction of all elements (excluding Ba and Ca). In contrast, the water content in NADES was significantly affected on the recovery of Ba, Ca, Mg, Mn, Sr, and Zn. The recovery of Al and Cr was relatively high and considerably varied (from 1.5 to 59.9%). NADES comprising lactic acid:glucose:H₂O (5:3:1) provided the lowest contents of all elements, and the highest extracted amounts were obtained employing water contents of 60–80%. The calculated daily intake of all the elements contained in NADES extracts were less than the daily dose risk estimators. The hazard quotients, hazard indexes, and carcinogenic risk calculated for all trace elements and their combination were considerably less than 1. This evidences no health risk, and carcinogenic risk after topical application of all studied NADES. For the first time, the results of the current study demonstrated that NADES extracts of F. vesiculosus contain a lower amount of trace metals and are safer than the extracts obtained with water and 70% acetone. This indicates a significant advantage for NADES compared with the other solvents.