The inheritance of chemical phenotype in <Emphasis Type="Italic">Cannabis sativa</Emphasis> L. (III): variation in cannabichromene proportion

The mechanism that controls the proportion of cannabichromene (CBC), a potential pharmaceutical, in the cannabinoid fraction of Cannabis sativa L. is explored. As with tetrahydrocannabinol (THC) and cannabidiol (CBD), CBC is an enzymatic conversion product of the precursor cannabigerol (CBG). CBC is reported to dominate the cannabinoid fraction of juveniles and to decline with maturation. This ontogeny was confirmed in inbred lines with different mature chemotypes. A consistent CBC presence was found in early leaves from a diverse clone collection, suggesting that CBC synthase is encoded by a fixed locus. Morphological variants possessing a ‘prolonged juvenile chemotype’ (PJC), a substantial proportion of CBC persisting up to maturity, are presented. PJC is associated with a reduced presence of floral bracts, bracteoles, and capitate-stalked trichomes. Genetic factors causing these features were independent of the allelic chemotype locus B that was previously postulated and regulates THC and CBD synthesis and CBG accumulation. In contrast to previously described Cannabis chemotypes, the cannabinoid composition of PJCs showed plasticity in that reduced light levels increased the CBC proportion. The ability of PJC plants to enable the production of pharmaceutical raw material with high CBC purity is demonstrated.     

Resistance of 57 greenhouse-grown accessions of Lycopersicon esculentum and three cultivars to Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae)

The aim of this study was to evaluate resistance to Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) by antixenosis on 57 Lycopersicon esculentum Mill. accessions from the Horticultural Germplasm Bank (HGB) of Universidade Federal de Viçosa and by the three commercial cultivars (Santa Clara, Moneymaker and TOM-601) under greenhouse conditions. A randomized complete block design was used with three replications. Infestations with T. absoluta adults were performed weekly and the following characteristics were evaluated: number of small, large and total mines/leaf and % of leaves mined at days 60, 75 and 90 after planting. Low infestation occurred at days 60 and 75 but at day 90, infestation was sufficient to evaluate insect damage. Based on these data it was concluded that only accessions HGB-674 and HGB-1497 appeared to be the most promising. In addition, to determine possible chemical causes of resistance, hexane extracts were analyzed at day 90 by gas chromatography/mass spectrometry and the major peaks identified by a mass spectral database using similarity index. Nine hydrocarbons, viz., hexadecane, heptadecane, eicosane, tricosane, 2-methyltricosane, tetracosane, hexacosane, octacosane and triacontane were identified in the hexane extracts in many samples. Tricosane, tetracosane and hexacosane presented significant correlations with the leaves mined. Only tricosane presented a negative correlation with the number of small mines (r = −0.28), total number of mines (r = −0.27) and % of leaves mined (r = −0.22). However, tetracosane and hexacosane presented significant positive correlations (r = 0.25 and 0.24, respectively) with the % of leaves mined.     

Evaluation of tangerine hybrid resistance to Alternaria alternata

The Alternaria Brown Spot, caused by Alternaria alternata, is a major fungal disease in some kinds of tangerines, tangor, mandarins and pomelos. In Brazil as well as worldwide, A. alternata can cause necrosis in fruits, branches and leaves, causing substantial profit loss. In the present research, in laboratory conditions and in the field, we evaluated the resistance to the fungus, in leaves and fruits, for 22 varieties and hybrids of tangerines. To this end, we evaluated genotypes belonging to the Germplasm Bank of the Estação Experimental de Citricultura de Bebedouro. The resistant genetic materials (found in leaves and fruits) represented four varieties of clementines (Citrus clementina); six varieties of mandarins (two belonging to C. reticulata, two to C. tangerina, one to C. deliciosa and one to C. nobilis); one tangelo (C. tangerina × C. paradisi); two mandarin hybrids (one resulting from crossing C. nobilis × C. deliciosa and the other from crossing C. clementina × C. reticulata); one tangor hybrid (C. clementina) and two satsuma hybrids (C. unshiu × C. deliciosa). We also determined a relation between the inoculation of leaves and fruits. The resistance and susceptibility following inoculation in leaves and fruits supports a relationship between these organs and the physiological responses observed for the evaluated genotypes.     

Implementation of two high through-put techniques in a novel application: detecting point mutations in large EMS mutated plant populations

Background  

The establishment of mutant populations together with the strategies for targeted mutation detection has been applied successfully to a large number of organisms including many species in the plant kingdom. Considerable efforts have been invested into research on tomato as a model for berry-fruit plants. With the progress of the tomato sequencing project, reverse genetics becomes an obvious and achievable goal.     

Effects of density on growth and survival of juvenile Red Swamp Crayfish, Procambarus clarkii (Girard), reared under laboratory conditions

The aim of this study was to quantify the importance of population density on the growth and survival of juvenile red swamp crayfish, Procambarus clarkii (Girard), and to understand the effects of density on both the period between moults and the increment per moult at high densities without food limitation. A laboratory experiment was performed to determine growth and survival of juvenile crayfish at five densities (20, 40, 60, 80 and 100 crayfish m^?2). Density clearly affected crayfish growth. The impact of density on final weight was higher than on total length increase, growth rate and final length (FL) (by decreasing order of effect). When comparing minimum and maximum densities, we observed that FL suffered a 34% reduction. Number of moults, mean intermoult period (IP) and survival were not significantly affected by density. Our results indicated that the IP and the percentage of increment per moult are affected by the size of crayfish before moult and we provide equations for these relations. Our findings are relevant both for crayfish aquaculture management and for the management of wild or harvested rice‐field crayfish populations.     

Feeding responses of juvenile shrimp Litopenaeus vannamei (Boone) fed at different frequencies under laboratory conditions

In shrimp farms, food partitioning during the course of the day is contradictory, ethology studies may help in determining the proper frequency. Litopenaeus vannamei juveniles were placed in 30 L aquariums (41 m⁻²), exposed to a 12:12 h light–dark cycle. Feeding was provided at 10% of biomass three, four and seven times per day, with observations made in 15 min h⁻¹ windows, initiated after the ration was offered. Latency to access the feeding tray and to start eating was recorded using instantaneous focal sampling and digestive tract filling (DTF) by the continuous focal method. Weight gain was recorded at the end of the experiment. We performed seven repetitions, with 28 individuals observed for 33 days for each treatment (490 h of observation). The three-times offering induced less latency for accessing the tray and for ingesting the feed as well as a higher DTF when compared with the other frequencies. Animals fed three and four times had similar weight gains, and were greater than those fed seven times. Our results indicate that a more spaced food offering stimulates the search for and ingestion of feed. As compared with other frequencies, the three-times-per-day option assumes lower labour costs and a more efficient use of the feed.     

Growth,absorption and assimilation efficiency by mature cuttlefish (<Emphasis Type="Italic">Sepia officinalis</Emphasis>) fed with alternative and artificial diets

The effects of feeding three natural frozen diets, grass shrimp (Palaemonetes sp.), crayfish (Procambarus clarkii) and fish (Sardina pilchardus) and two semi-humid artificial diets (based on fish powder) to mature cuttlefish, Sepia officinalis, were analysed. Growth and feeding rates (GR and FR, % BW day⁻¹), food conversions (FC, %), and total protein and lipid composition of the diets were determined. Digestive gland to body weight ratio and absorption efficiency were calculated for each diet. Cuttlefish fed shrimp and crayfish grew larger (1.5 and 1.1% BW day⁻¹, respectively) compared to the other diets. Shrimp promoted the highest FC, followed by crayfish, and sardine. The highest FR was obtained for cuttlefish fed crayfish (8.4% BW day⁻¹). Although both artificial diets were accepted, none produced growth. A positive correlation (r = 0.96) between cuttlefish ingestion rate and digestive gland weight was obtained. Some cannibalism occurred among cuttlefish fed the artificial diets during the last week of the experiment. According to the results obtained, P. clarkii could be used as an alternative prey to shrimp for rearing adult mature (>50 g) S. officinalis.     

Salmonid fish viruses and cell interactions at early steps of the infective cycle

A flow cytometric virus-binding assay that directly visualizes the binding and entry of infectious pancreatic necrosis virus (IPNV), infectious haematopoietic necrosis virus (IHNV) and virus haemorrhagic septicaemia virus (VHSV) to several cell lines was established. The highest efficiency of binding was shown by the BF-2 cell line and this was used to study, at the attachment level, the interactions of these cells with salmonid fish viruses in coinfections, and to further determine if the earliest stage of the viral growth cycle could explain the previously described loss of infectivity of IHNV when IPNV is present. Our results demonstrated that IPNV binds to around 88% of cells either in single or dual infections, whereas IHNV attachment always decreased in the presence of any of the other viruses. VHSV binding was not affected by IPNV, but coinfection with IHNV reduced the percentage of virus-binding cells, which suggests competition for viral receptors or co-receptors. Internalization of the adsorbed IHNV was not decreased by coinfection with IPNV, so the hypothetical competence could be restricted to the binding step. Treatment of the cells with antiviral agents, such as amantadine or chloroquine, did not affect the binding of IPNV and VHSV, but reduced IHNV binding by more than 30%. Tributylamine affected viral binding of the three viruses to different degrees and inhibited IPNV or IHNV entry in a large percentage of cells treated for 30 min. Tributylamine also inhibited IHNV cytopathic effects in a dose-dependent manner, decreasing the virus yield by 4 log of the 50% endpoint titre, at 10 mm concentration. IPNV was also inhibited, but at a lower level. The results of this study support the hypothesis that IHNV, in contrast to VHSV or IPNV, is less efficient at completing its growth cycle in cells with a simultaneous infection with IPNV. It can be affected at several stages of viral infection and is more sensitive to the action of antiviral compounds.