Triploid Induction in the Yellowtail Tetra,Astyanax altiparanae,Using Temperature Shock: Tools for Conservation and Aquaculture

Triploidization is an interesting tool to produce sterile fish. In the yellowtail tetra, Astyanax altiparanae, this can be applied for aquaculture and surrogate technologies. In this study, we compared the efficacy of cold (2 C) or heat shock (38 C, 40 C, and 42 C) on triploid induction in the yellowtail tetra. The eggs were treated with cold or heat shock, 2 min postfertilization (30 min in cold shock or 2 min in heat shock). Intact embryos served as the control group. Ploidy status was confirmed by karyotyping, flow cytometry, and nuclear diameter of erythrocytes. The hatching rate decreased after cold shock (12.69 ± 15.76%) and heat shock at 42 C (0.35 ± 0.69%) in comparison with the control group (63.19 ± 16.82%). At 38 C and 40 C, hatching rates (61.29 ± 17.73% and 61.75 ± 22.1%, respectively) were not decreased. Only one triploid arose at 38 C (1/80). At 40 C, a high number of triploids arose (72/78). At 42 C, very few embryos developed into the hatching stage. A large number of haploid individuals arose after cold shock (61/75), with only one triploid. Our results indicate that heat shocking of embryos at 40 C is optimum for triploid production in the yellowtail tetra.     

Replacement of Fishmeal by a Mixture of Soybean Meal and Chlorella Meal in Practical Diets for Juvenile Crucian Carp,Carassius auratus

A growth trial was conducted to evaluate the effect of a mixture of soybean meal and Chlorella meal (SCM) as a dietary fishmeal (FM) substitute on growth performance, apparent digestibility coefficients (ADCs), digestive enzymatic activities, and histology of juvenile crucian carp, Carassius auratus. Five isonitrogenous and isoenergetic diets were formulated to replace 0 (SCM0), 25 (SCM25), 50 (SCM50), 75 (SCM75), and 100% (SCM100) of protein from FM with SCM, respectively. The diets were fed to triplicate groups of juvenile crucian carp for 6 wk. Weight gain, specific growth rate, feed intake, protein efficiency ratio, and intestinal digestive enzymatic activities (amylase, trypsin, and lipase) tended to decline with increasing FM replacement levels (P > 0.05). Dietary SCM substitution significantly influenced dry matter content in muscle, and crude protein and lipid contents in liver (P < 0.05). ADCs for dry matter, protein, lipid, energy, and most amino acids showed no significant differences between the control and SCM25 group, but tended to decline with replacement levels over 25%. Higher SCM substitution (50–100%) caused karyopyknosis and necrosis in liver, but intestinal histology did not show noticeable pathological changes. The present study indicated that FM could be replaced by 25% of SCM, without significant adverse growth performance, feed utilization, and histology of crucian carp.     

Management strategies for the control of Tuta absoluta (Lepidoptera: Gelechiidae) damage in open‐field cultivations of processing tomato in Tuscany (Italy)

Since its recent introduction into the Mediterranean area, Tuta absoluta (Meyrick) has become widespread and an important pest throughout the region, including in Tuscany, where it was first recorded in 2009. Although several studies have been conducted within controlled environments on the ecology and management of T. absoluta, limited information is currently available on the importance of this pest in open‐field cultivations, which may lead to ineffective management strategies. This study was carried out to monitor fluctuations in adult populations of T. absoluta in organic and conventional cultivations of processing tomatoes using pheromone lures, and to collect inter‐annual data on direct yield loss and fruit damage from larval feeding in two separate trials in Grosseto (Tuscany). The first trial included eight conventionally managed fields; the second trial was carried out in four organically managed fields. Results show relatively higher crop damage for tomatoes transplanted later in the season and harvested towards the end of August to early September, and limited fruit damage from this pest during the second year of the study for both conventional and organically managed fields. Finally, biological and chemical pest management strategies for the control of T. absoluta adopted by farmers within the region are reviewed and discussed.     

Comparative spatial analysis of the sooty blotch/flyspeck disease complex,bull’s eye and bitter rots of apples

The incidence of sooty blotch/flyspeck (SBFS) and bitter and bull’s eye rots were assessed in a Fuji apple orchard during two seasons. Using a regular sampling design, 252 trees were selected and 20 fruits per tree were sampled at harvest and scored for disease incidence. For bitter and bull’s eye rots, additional assessments were made on all symptomless fruit after a 30‐day period of storage. Randomness in the spatial pattern was assessed using beta‐binomial analysis of incidence data for three sampling scales (one, three or six adjacent trees as sampling units) and using Spatial Analysis by Distance Indices (sadie ) for disease counts for the 3‐tree sampling scale. sadie was also used for testing spatial associations between a pair of diseases, between years for the same disease or between rotted and latently infected fruit. Using a toroidal‐shifts procedure, 360 maps of disease counts were created based on the observed data, which were further analysed using sadie . Most datasets showed an aggregated spatial pattern, which was more consistent for the two fruit rots than SBFS, which showed distinct patterns depending on the year or method of analysis. The two fruit rots were spatially associated in most situations but SBFS and bull’s eye rot were dissociated in one season. Results from virtual orchards showed that the patterns observed in the original maps may accurately represent those in similar apple‐growing areas. Hypotheses regarding aspects of ecology and epidemiology of pathogens studied and potential efficacy of control measures in the region are discussed.     

The significance of guttation in the secondary spread of Clavibacter michiganensis subsp. michiganensis in tomato greenhouses

Bacterial canker, caused by Clavibacter michiganensis subsp. michiganensis (Cmm), can spread in commercial tomato greenhouses causing epidemics. Results of greenhouse experiments with Cmm‐contaminated tools demonstrated disease spread for only a limited distance (<4 plants) from infected plants. However, touching symptomless infected plants bearing guttation droplets prior to touching nearby plants spread the pathogen over longer distances within rows (>22 plants). The pathogen was exuded in large numbers in the guttation fluid of infected plants; its presence in the guttation fluid was not influenced by the inoculation procedures, leaf age or the volume of the guttation droplets. Population size of Cmm and the incidence of leaflets with epiphytic bacteria were significantly higher in plants placed in a guttation‐induction chamber than in those kept in a growth chamber with high humidity, suggesting exudation through guttation contributed to the formation of epiphytic populations on leaflets. This new knowledge may provide a simple and environmentally friendly means for decreasing the spread of the disease by avoiding contact with plants during periods when they bear guttation droplets.     

Biological control of botrytis bunch rot in organic wine grapes with the yeast antagonist Candida sake CPA‐1

The aim of this research was to confirm the efficacy of the yeast antagonist Candida sake CPA‐1 in suppressing botrytis bunch rot development, in an organic vineyard under Mediterranean conditions for two seasons, and compare its performance with that of two biologically based products currently registered for botrytis bunch rot control in New Zealand. In 2009, treatments applied were: commercial formulations of Ulocladium oudemansii (BOTRY‐Zen^®) and chitosan (ARMOUR‐Zen^®), C. sake CPA‐1 combined with the fatty acid‐based additive Fungicover^® and combinations of these products. All treatments were applied six times between early flowering and harvest and compared with an unsprayed control. In 2010, the treatments focused on C. sake and Fungicover and the number of applications was reduced from six to four. The population dynamics of U. oudemansii and C. sake were measured and wine quality tests were carried out in both seasons. Disease control achieved by C. sake treatments in 2009 were comparable to those achieved by BOTRY‐Zen and ARMOUR‐Zen. Applications of C. sake plus Fungicover between flowering and harvest significantly (P < 0·05) reduced botrytis bunch rot incidence and severity by 64% and 90%, respectively, compared with the untreated control in 2009, and by 67% and 89%, respectively, in 2010. Treatments did not adversely affect wine quality parameters after treated grapes were processed. Candida sake consistently provided effective control of botrytis bunch rot in grapes under different meteorological and disease pressure conditions, thereby improving its potential for future commercial applications.     

Co‐infection by Botryosphaeriaceae and Ilyonectria spp. fungi during propagation causes decline of young grafted grapevines

Decline of newly planted, grafted grapevines is a serious viticultural problem worldwide. In the Riverina (New South Wales, Australia), characteristic symptoms include low fruit yields, very short shoots and severely stunted roots with black, sunken, necrotic lesions. To determine the cause, roots and wood tissue from affected plants in 20 vineyards (Vitis vinifera cv. Chardonnay grafted to V. champini cv. Ramsey rootstock) were assayed for microbial pathogens. Ilyonectria spp. (I. macrodidyma or I. liriodendra, producers of phytotoxin brefeldin A, BFA, and cause of black foot disease of grapevines) and Botryosphaeriaceae spp. (predominantly Diplodia seriata) were isolated from rootstocks of 100 and 95% of the plants, respectively. Togninia minima and Phaeomoniella chlamydospora (cause of grapevine Petri disease) were isolated from 13 and 7% of affected plants, respectively. All Ramsey rootstock stems of grafted plants sampled from a supplier nursery were infected with Ilyonectria spp. and D. seriata. Diplodia seriata, but not Ilyonectria spp., was also isolated from 25% of canes sampled from the rootstock source block. Root inoculation of potted, disease‐free Chardonnay plants with Ilyonectria isolates from diseased vineyards caused typical disease symptoms, while co‐inoculation with Botryosphaeriaceae spp. increased disease severity. This is the first study to show that a major cause of young grapevine decline can be sequential infection by Botryosphaeriaceae from rootstock cuttings and Ilyonectria spp. from nursery soil. Although the Petri disease fungi were less common in young declining grafted grapevines in the Riverina, they are likely to contribute to the decline of surviving plants as they mature.     

A SNaPshot assay for the rapid and simple detection of known point mutations conferring resistance to ACCase‐inhibiting herbicides in Lolium spp.

Evolution of resistance to herbicides in weeds is becoming an increasing problem worldwide. To develop effective strategies for weed control, a thorough knowledge of the basis of resistance is required. Although non‐target‐site‐based resistance is widespread, target site resistance, often caused by a single nucleotide change in the gene encoding the target enzyme, is also a common factor affecting the efficacies of key herbicides. Therefore, fast and relatively simple high‐throughput screening methods to detect target site resistance mutations will represent important tools for monitoring the distribution and evolution of resistant alleles within weed populations. Here, we present a simple and quick method that can be used to simultaneously screen for up to 10 mutations from several target site resistance‐associated codons in a single reaction. As a proof of concept, this SNaPshot multiplex method was successfully applied to the genotyping of nine variable nucleotide positions in the CT domain of the chloroplastic ACCase gene from Lolium multiflorum plants from 54 populations. A total of 10 nucleotide substitutions at seven of these nine positions (namely codons 1781, 1999, 2027, 2041 2078, 2088 and 2096) are known to confer resistance to ACCase‐inhibiting herbicides. This assay has several advantages when compared with other methods currently in use in weed science. It can discriminate between different nucleotide changes at a single locus, as well as screening for SNPs from different target sites by pooling multiple PCR products within a single reaction. The method is scalable, allowing reactions to be carried out in either 96‐ or 384‐well plate formats, thus reducing work time and cost.     

Nouveau signalement de Hypocryphalus scabricollis (Eichhoff, 1878) en Tunisie,un ravageur émergent sur figuier (Ficus carica L.)

Hypocryphalus scabricollis (Eichhoff) (Coleoptera, Curculionidae, Scolytinae) is a new pest on fig trees (Ficus carica L.). The severe damage observed on this host in Tunisia is described and illustrated. This paper describes the morphological identification of the species and gives information on the biology of the pest and initial recommendations for its control. This is a first report for Tunisia.