Complete nucleotide sequence, genomic organization and phylogenetic analysis of Citrus leprosis virus cytoplasmic type

The complete nucleotide sequence of the genomic RNA 1 (8745 nt) and RNA 2 (4986 nt) of Citrus leprosis virus cytoplasmic type (CiLV-C) was determined using cloned cDNA. RNA 1 contains two open reading frames (ORFs), which correspond to 286 and 29 kDa proteins. The 286 kDa protein is a polyprotein putatively involved in virus replication, which contains four conserved domains: methyltransferase, protease, helicase and polymerase. RNA 2 contains four ORFs corresponding to 15, 61, 32 and 24 kDa proteins, respectively. The 32 kDa protein is apparently involved in cell-to-cell movement of the virus, but none of the other putative proteins exhibit any conserved domain. The 5' regions of the two genomic RNAs contain a 'cap' structure and poly(A) tails were identified in the 3'-terminals. Sequence analyses and searches for structural and non-structural protein similarities revealed conserved domains with members of the genera Furovirus, Bromovirus, Tobravirus and Tobamovirus, although phylogenetic analyses strongly suggest that CiLV-C is a member of a distinct, novel virus genus and family, and definitely demonstrate that it does not belong to the family Rhabdoviridae, as previously proposed. Based on these results it was proposed that Citrus leprosis virus be considered as the type member of a new genus of viruses, Cilevirus.

Pectin lyase from Aspergillus giganteus: Comparative study of productivity of submerged fermentation on citrus pectin and orange waste

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

A new phytophagous Bracon Fabricius (Hymenoptera, Braconidae) associated with Protium ovatum Engl. (Burseraceae) fruits from Brazilian savannah

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Nutrient digestibility of degermed, dehulled corn, citrus pulp, and soy protein concentrate by barrows

This study was carried out to determine apparent ileal digestibility (AID) and apparent total tract digestibility (ATTD) of DM, CP, GE, and their respective digestible content of degermed dehulled corn (Zea mays), citrus pulp, and soy (Glycine max) protein concentrate by pigs using the difference method. Thirty-two barrows (28.1 +/- 1.6 kg of BW) were fed a corn-soybean meal basal diet or 1 of 3 diets formulated by replacing 30% of the basal diet with 30% of 1 of the test feedstuffs for 11 d. Chromic oxide (0.3%) was included in the diets. Feces were collected from days 7 to 11 by grab sampling and ileal digesta were collected after pigs were slaughtered on day 12. The AID of DM and AID and ATTD of GE of degermed corn (77.4, 88.7, and 77.7%) were greater (P < 0.05) than those observed in citrus pulp (50.3, 86.5, and 55.8%) and in soy protein concentrate (63.5, 85.1, and 59.4%), which did not differ (P > 0.05). The ATTD of CP, total digestible CP, and total DE of soy protein concentrate (87.5%, 500 g/kg, and 3739 kcal/kg) were higher (P < 0.05) than the values in degermed corn (81.7%, 57.5 g/kg, and 3330 kcal/kg), which were greater (P < 0.05) than those in citrus pulp (60.5%, 39.5 g/kg, and 3223 kcal/kg). Total and ileal digestible DM, AID of CP, and ileal DE of degermed corn (782 g/kg, 673 g/kg, 70.7%, and 2913 kcal/kg) and soy protein concentrate (778 g/kg, 570 g/kg, 78.7%, and 2878 kcal/kg) were similar (P > 0.05) and greater (P < 0.05) than those in citrus pulp (737 g/kg, 436 g/kg, 50.6%, and 2081 kcal/kg). Ileal digestible CP of degermed corn (49.8 g/kg) and citrus pulp (33.0 g/kg) did not differ (P > 0.05) but were smaller (P < 0.05) than the value found in soy protein concentrate (434 g/kg). The DM and energy from degermed corn are more efficiently digested by the pig than those from soy protein concentrate and citrus pulp. Soy protein concentrate was the best protein source evaluated in this study.

Seed Dispersal Anachronisms: Rethinking the Fruits Extinct Megafauna Ate

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Ascomycetous Yeasts Associated with Naturally Occurring Fruits in a Tropical Rain Forest

Fruits from twenty different species of angiosperms were collected during the period from November, 1991 to January, 1992. Two hundred and two strains of yeasts and yeast-like fungi were isolated, of which 74 % showed ascomycetic affinity. Candida was the predominant genus, followed by (in descending order of occurrence): Cryptococcus, Klœckera, Sporobolomyces, Pichia, Hanseniaspora and Bullera. Black yeasts and other strains showing basidiomycetic affinity were also isolated. The genus Candida represented the highest number of identified species and the greatest variety of associated substrates. Among the ascomycetes and their anamorphs, 38 species were identified, with Klœckera apiculata being the most frequent among the isolates and the one which occurred in the largest variety of substrates. Some of the biotypes designated as Candida sp. A, B, C, D, E, F, G, H, I, and Pichia sp. did not correspond to the standard species description found in the literature, and may represent new species. The strains of yeasts isolated in this study were characterized and incorporated into the Tropical Culture Collection of the Fundação Tropical de Pesquisas e Tecnologia André Tosello, Campinas, São Paulo.

Effects of G a3 dosis associated with organosilicone on sweet oranges in Brazil

GA3 was tested in sweet oranges 'Pera' and 'Hamlin' for delay the picking time without loosening of fruit quality for processing. Hamlin is the firths cv processed in Brazil and Pera is a mid season cv and extending their period of processing is important. Two experiments were made at the Citrus Experimental Station during 1996 season. The treatments are 5 ppm of GA 3 + 0,05% Silwett L-77® (organosilicone), 10 ppm of GA 3 + 0,05% of Silwet L-77®, 20 ppm of GA3 + 0,1% Herbitensil® (Noniphenoloxietilate 40%m/v + isopropilic alcohol 15% m/v) and control, repeated 7 times for Hamlin and 8 times for Pera, with one tree each parcel. The treatments were applied in May 1996, at the stage of greenish yellow colour of the fruits. Evaluations were made each 20 days interval till the final picking, It was analysed fruit quality and retention force for picking and puncture resistance. The results showed no differences for fruit quality of Hamlin from July to mid September and for Pera till September. After some differences occurred. The GA3 treatments were effectives in maintain the fruit retention force for both cvs for 120 days after application. In relation to fruit puncture resistance the treatments with GA3 differed of the control for both cvs, accordingly with the doses and mixtures. The colour index was better maintained with 5ppm of GA3 plus 0,05% of Silwet L-77®. The total fruit production did not differ for both cultivars.

Incidence of patulin in fruits and fruit juices marketed in Campinas, Brazil

One hundred and eleven samples of processed fruit juices (apple, grape, pineapple, papaya, guava, banana and mango) and 38 samples of sound fruits (apple, papaya, mango, pear and peach) produced and marketed in Brazil, were analysed for patulin by HPLC. Only one out of 30 samples of apple juice was found positive at 17 μg/l. Patulin was not detected in the other foodstuffs. It was found in 14 samples of spoiled fruit samples of apple (150-267 μg/kg), pear (134-245 μg/kg) and peach (92-174 μg/kg). Confirmation of the identity of patulin was based on the UV spectrum obtained by the HPLC diode array detector, compared with that of standard patulin, TLC developed by several solvent systems and sprayed with 3-methyl-2-benzothiazolinone hydrazone, and by acetylation with acetic anhydride.

Distribution of Xylella fastidiosa in citrus rootstocks and transmission of citrus variegated chlorosis between sweet orange plants through natural root grafts

To study translocation of Xylella fastidiosa to citrus rootstocks, budsticks from citrus variegated chlorosis (CVC)-affected cv. Pera sweet orange (Citrus sinenesis (L.) Osb.) were top grafted on 15 citrus rootstocks. Disease symptoms were conspicuous 3 months later on all 15 rootstocks tested. The presence of X. fastidiosa was confirmed by light microscopy, double-antibody sandwich enzyme-linked immunosorbent assays, and polymerase chain reaction in rootlets and main roots of CVC-symptomatic Pera sweet orange in 11 of the 15 rootstocks tested. These results suggest that bacterial translocation from the aerial plant parts to the root system occurs but is not essential for X. fastidiosa to induce symptoms in the aerial parts. Bacterial translocation to the roots was not correlated with CVC leaf-symptom severity in the Pera scion. To determine if CVC disease could be transmitted by natural root grafts, two matched seedlings of each of four sweet orange cultivars (Pera, Natal, Valencia, and Caipira) were transplanted into single pots. One seedling rootstock of each pair was inoculated by top grafting with a CVC-contaminated budstick while the other seedling rootstock was cut but not graft inoculated. Transmission of X. fastidiosa from an inoculated plant to a noninoculated plant sharing the same pot was observed in all four sweet orange cultivars tested. Transmission was confirmed by observation of natural roots grafts between the two plants, presence of X. fastidiosa in the root grafts, and disease development in the uninoculated plants. This is the first report of transmission of CVC disease through natural root grafts.

Aspects of IFP for citrus production in Brazil

Brazilian citriculture represents about 25% of the total world citrus production with an area of 851,518 ha and a total production of more than 17 million tons in 1996. Besides its importance to the brazilian economy, represented by more than 1 billion US $ by year from FCOJ exportation, the citriculture has problems related to low productivity, due to several cultural practices and management. The productivity would be improved by an IFP system. The main problems are related to soil, as poor conservation, use of poor drained soils and bad preparation for planting; diseases-canker, CVC, leprosis, Phytophtora gummosis and other fungus diseases; pests - mites, scales, nematodes and others; the use of disease free and improved scion and rootstocks propagative material is usual; cultural practices - as nutrition, irrigation, wind breaks, weed control, pruning, replant and others, as density planting could be improved. Some possible solutions will be discussed for improving the brazilian citrus productivity and quality by IFP, based on research made.

Susceptibility of tangerines to citrus variegated chlorosis

Citrus variegated chlorosis (CVC), a citrus disease first discovered in Brazil in 1987, is caused by the bacterium Xylella fastidiosa and transmitted by sharpshooters and budwood. Since the disease affects almost all sweet orange cultivars, it has become one of the most serious problems for Brazilian citriculture. To evaluate their resistance to CVC disease, fifteen tangerines or mandarins (C. reticulata Blanco) and their hybrids were grafted on Rangpur lime (C. limonia Osb.) and inoculated with CVC-contaminated Pera sweet orange (C. sinensis (L.) Osb.) by twig grafting in a greenhouse. Tangerines and their hybrids Wilking, Fortune, Sunki, Ellendale, Orlando tangelo, Nunes clementine, Nova, Sun Shu Sha Kat, Suenkat, and Batangas showed CVC leaf symptoms and gave positive results on enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) (with specific primers for X. fastidiosa), indicating that they are susceptible to CVC. Although X. fastidiosa bacteria were detected by ELISA and PCR in inoculated plants of tangerines Cravo and Oneco, no CVC leaf symptoms were observed on these two cultivars, suggesting that they are tolerant to the disease. CVC leaf symptoms were not observed and X. fastidiosa was not detected in tangerine Dancy and mandarins Okitsu satsuma and Ponkan after inoculation, showing that they are resistant to the disease.

Evaluation of microsatellite markers in cultivars of sweet orange (Citrus sinensis [L.] Osbeck)

Sweet orange is considered a very important species in the citrus world market and presents wide morphological variability. However, its characterization at the molecular level by random amplified polymorphic DNA (RAPD) and isozyme markers is not appropriate. Microsatellite or simple sequence repeats (SSRs) have been suggested as ideal for studies in cultures of vegetative propagation and as value markers for mapping in several species. However, information on microsatellite polymorphism in citrus species is scarce. In this work, microsatellite markers (AG-repeats) were developed from an enrichment library of genomic DNA of sweet orange cv. Pera (Citrus sinensis [L.] Osbeck), and 31 cultivars of sweet orange were evaluated. Evaluation of 18 microsatellite primers did not permit differentiation of the varieties studied. New microsatellite primers are being evaluated with the aim of detecting polymorphisms among the cultivars and closely related species to be used in genetic mapping programs.

Adubação verde em Citrus: Influência na porosidade e retenção de água de um latossolo vermelho-escuro do noroeste do estado de São Paulo

The objective of this research was to study the porosity, bulk density and retention of water of an Oxisol, located in the Northwestern region of Sn̄o Paulo state, Brazil. The soil was cultivated with Citrus sp., to which green manure was applied between rows for three years. Each of six species of green manure crops (Crotalaria juncea L., Mucuna deeringiana Steph. & Bart., Canavalia ensiformis L. DC., Cajanus cajan L., Lablab purpureum L. and Ricinus communis L.) were seeded for three years (1995, 1996 and 1997) between Citrus rows, plus a treatment with a mix of all six species and a control (natural regrowth af vegetation). The experimental design was a randomized complete block design, with four replications for each of the eight treatments. Water retention, microporosity, macroporosity, total porosity and bulk density were analyzed in the beginning (1995) and end (1997) of the experiment, at three depth ranges (0-0.10; 0.10-0.20 and 0.20-0.40m). We concluded that there were statistically significant differences for bulk density, macroporosity, total porosity and retention of water among the different soil depth ranges; there were no significant differences among treatments though.