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.

Diurnal and seasonal variations of CWSI and non-water-stressed baseline with nectarine trees

The work described was part of the programme, Innovative biological indicators to improve the efficiency of water and nitrogen use and the fruit quality in tree crops Project, a partnership between ISA and INRA. Field studies were conducted in Portugal on different irrigated plots of nectarine trees; a fully irrigated (unstressed plot) and a plot that was not irrigated for some days (stressed plot). The aim of this work was to investigate the effects of plant water stress on canopy temperature, to determine the nonwater-stressed baseline and to observe diurnal and seasonal variations of Crop Water Stress Index (CWSI). Canopy temperature, psychrometric and wind speed data were taken each half-hour, between 9:30 and 15:30 h. Results showed that canopy temperature was higher during the daytime, for both unstressed and stressed plots. A linear regression of canopy-air temperature differential and the vapor pressure deficit (non-water-stress baseline) showed a r2= 0.65. During the stress period, the average canopy temperature of the stressed plot was up to 5.4°C higher than the unstressed plot. Diurnal and seasonal average of CWSI values showed differences between unstressed and stressed plots, during the stress period.

Phenology of Atlantic rain forest trees: A comparative study

This paper describes the phenology of leaf, flower, and fruit phenology in the Atlantic rain forests of southeastern Brazil. For 17 months, we observed the phenological patterns of trees from two Atlantic forest types at four sites: premontane forest (Sites I and IV; the typical Atlantic rain forest) and coastal plain forest (Sites II and III). All sites experience a nonseasonal, tropical wet climate, characterized by an annual rainfall usually > 2000 mm and lacking a dry season. We tested for the occurrence (or absence) of seasonal phenological patterns within each site and compared the patterns detected among the four different forest sites using circular statistics. The expected weakly seasonal phenological patterns were not observed for these forests. Flowering and leaf flush patterns of Atlantic rain forest trees were significantly seasonal, concentrated at the beginning of the wettest season, and were significantly correlated with day length and temperature. These results stress the influence that seasonal variation in day length has on ever-wet forest tree phenology. Fruiting phenologies were aseasonal in all four forests. Flowering patterns did not differ significantly among three of the four forest sites analyzed, suggesting the occurrence of a general flowering pattern for Atlantic rain forest trees.

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.

Activity of benomyl for control of postbloom fruit drop of citrus caused by Colletotrichum acutatum

Postbloom fruit drop (PFD) of citrus caused by Colletotrichum acutatum produces orange-brown lesions on petals and induces the abscission of young fruitlets and the retention of the calyces. Despite the fact that C. acutatum is not highly sensitive to benomyl in culture, this fungicide provides good control of the disease under field conditions. This study was undertaken to determine the effect of benomyl on various stages of disease development to understand the basis for its effectiveness in the field. We found that benomyl at 1.0 μg/ml reduced colony area of C. acutatum by about 75% and completely inhibited growth of C. gloeosporioides. Benomyl did not prevent conidial germination even at 100 μg/ml, but reduced germ tube elongation at 10 and 100 μg/ml. When benomyl was applied to flower clusters on screen-house-grown plants before inoculation, disease severity was greatly reduced. Applications at 24 and 48 h, but not at 72 h, after inoculation reduced PFD severity. Application of benomyl to symptomatic petals not bearing conidia did not prevent or reduce production of inoculum. Application to petals bearing conidia reduced viability of these fungal propagules by only about 50%. The viability of appressoria on mature leaves was not affected by benomyl application. Even when appressoria on mature leaves were stimulated to germinate by treatment with flower extracts, subsequent application of benomyl did not reduce propagule numbers below original levels. Benomyl appears to act by preventing infection and early development of the fungus in petals. However, once symptoms have developed, this fungicide has only minimal effects on further disease development and spread.

Phosphorus foliar fertilization in guava trees

Currently there is very little information on the response of fruiting perennial plants to applied P. This is especially true for tropical production areas where soils have a high capacity of P fixation, and are poor in native phosphorus. An alternative to soil P fertilization, which is inefficient in fixing soils, is to apply phosphorus as a foliar spray. P is quickly absorbed by leaves, and is redistributed quite well through the plants because its phloem mobility, and foliar application may be a viable practice. The purpose of this present work, is to determine the effectiveness of foliar P application on the nutritional status and yield of guava. The experiment was done in a Typic Hapludox, for three consecutive agricultural years, in an adult orchard of 'Paluma' guava. Five treatments were tested: four rates leaf applications of P (0-0.5-1.0 and 2.0% of P2O5) and a control where P was applied to soil (200 g of P2O5/plant). Through the results it was verified that the foliar application of P altered the concentration of the nutrient in the soil (13 to 48 mg dm-3 P-resin), and in the guava leaves (1.2 to 1.8 g of P kg-1), but did not affect the production of fruits. In conclusion, in field conditions, it is viable to combine the phosphorus foliar fertilization with disease control, without increasing the operations and, consequently, the production cost.

Benomyl sensitivity of isolates of Colletotrichum acutatum and C. gloeosporioides from citrus

Postbloom fruit drop (PFD) of citrus, caused by Colletotrichum acutatum, produces orange-brown lesions on petals and results in premature fruit drop and the retention of calyces. C. gloeosporioides is common in groves and causes postharvest anthracnose on fruit. Both diseases are controlled effectively by the fungicide benomyl in research fields and commercial orchards. Highly sensitive and resistant isolates of C. gloeosporioides were found, whereas all isolates of C. acutatum tested were moderately resistant. In preliminary studies conducted in vitro with three isolates of each, mycelial growth of sensitive isolates of C. gloeosporioides was inhibited completely by benomyl (Benlate 50 WP) at 1.0 μg/ml, whereas resistant isolates grew well at 10 μg/ml. Growth of all isolates of C. acutatum was inhibited by about 55% at 0.1 μg/ml and by 80% at 1.0 μg/ml. Spore germination of C. acutatum was inhibited more at 0.1 μg/ml than at 1.0 μg/ml or higher concentrations. In all, 20 isolates of C. acutatum from 17 groves and 20 isolates of C. gloeosporioides from 7 groves were collected from locations with different histories of benomyl usage in São Paulo, Brazil, and Florida, United States. Benomyl at 1.0 μ.g/ml completely inhibited growth of 133 isolates of C. gloeosporioides, with the exception of 7 isolates that were highly resistant to the fungicide, whereas all isolates of C. acutatum were only partially inhibited at 0.1 and 1.0 μg/ml. Analysis of variance indicated that the sensitivity of the isolates of C. acutatum was not affected by benomyl usage or grove of origin, and country of origin had only minor effects. No highly resistant or sensitive isolate of C. acutatum was recovered. Partial sequencing of the β-tubulin gene did not reveal nucleotide substitutions in codons 198 or 200 in C. acutatum that usually are associated with benomyl resistance in other fungi.

Genetic variation in growth traits and yield of rubber trees (Hevea brasiliensis) growing in the Brazilian state of São Paulo

Analysis of variance and covariance was preformed on growth traits (stem girth, bark thickness, total height gain and rubber yield) of 22 open-pollinated progenies of the rubber tree Hevea brasiliensis from an Asian Hevea collection introduced to Agronomic Institute (Instituto Agronômico, Campinas, São Paulo, Brazil; IAC) in 1952. This progeny trial was replicated at three sites in São Paulo state and it was found that at three years from sowing there was statistically significant variation for girth, bark thickness, height and rubber yield. An individual test sites, values of individual plant heritability for girth ranged from ĥ i 2 = 0.36 to ĥ i 2 = 0.89 whereas values for heritability for progeny means ranged from ĥ i 2 = 0.77 to ĥ i 2 = 0.87. These moderate and high heritabilities suggest that a combination of progeny and within-progeny selection would be effective at increasing girth in this population at individual sites. Across sites, values of individual-plant heritability for girth ranged from ĥ i 2 = 0.36 to ĥ i 2 = 0.47, whereas values for heritability of progeny means girth ranged from ĥ x̄ 2 = 0.77 to ĥ x̄ 2 = 0.87. There were high positive genetic correlations between increased girth and bark thickness suggesting that breeding aimed at increasing girth would also increase bark thickness and possibly height. Copyright by the Brazilian Society of Genetics.

Effect of air humidity on the egg viability of predatory mites (Acari: Phytoseiidae, Stigmaeidae) common on rubber trees in Brazil

Rubber pest mites, Calacarus heveae and Tenuipalpus heveae, reach economic damage levels at the end of the rainy season and the beginning of the dry season in Brazil. Therefore, low humidity adaptation might be an important characteristic for predatory mites to successfully control pest organisms. This study determined the effect of the relative humidity (RH) levels of 30-100% on the hatching of larvae of Amblyseius acalyphus, Euseius citrifolius, Iphiseiodes zuluagai, Metaseiulus camelliae, Agistemus floridanus and Zetzellia malvinae at 25 ± 0.5°C. These predatory mites are common on rubber trees in the state of São Paulo and might be used for introduction in the major rubber tree production regions in the state of Mato Grosso. At 70% RH or higher, viability was 70% or higher for all species, indicating that their performance might be higher during the rainy season than during the dry season. Eggs of E. citrifolius and M. camelliae presented higher viability at the lower relative humidity levels than those of other species, indicating that these species might have higher chance to persist in the dry season. It is suggested that M. camelliae should be further evaluated for introduction in the state of Mato Grosso, considering that this mite is not yet present in that area. © Springer 2006.

Attractiveness of citrus pulp and orange albedo extracts to Atta sexdens rubropilosa (Hymenoptera: Formicidae)

The present study aimed to compare the attractiveness of industrial citrus pulp with the handmade orange albedo to the workers of Atta sexdens rubropilosa. For this, filter paper fragments were impregnated with organic extracts obtained through chemical extraction and sequential fractioning with hexane and dichloromethane and offered to different field nests. It was verified that the industrial citrus pulp extract is as good as the handmade orange albedo extract. This preference is discussed keeping in mind the chemical, behavioral and nutritional factors.

A preliminary investigation of pre-dispersal seed predation by Acanthoscelides schrankiae horn (Coleoptera: Bruchidae) in Mimosa bimucronata (DC.) Kuntze trees

This is the first record of Acanthoscelides schrankiae Horn, feeding in seeds of Mimosa bimucronata (DC.) Kuntze. We investigated the pattern of oviposition and seed exploitation by A. schrankiae, and the distribution of mature fruits and seed predation in the inflorescences. We also compared the percentage of predated seeds, the total dry weight of fruits and non-predated seeds, the percentage of aborted seeds, and the percentage of non-emergent insects, among different quadrants of the M. bimucronata canopy. To determine the occurring species, the emergence of bruchids and parasitoids was observed in the laboratory, resulting altogether, only in individuals of A. schrankiae and Horismenus sp. (Hymenoptera: Eulophidae) species, respectively. Mean number of fruits produced in the median region of inflorescence was significantly higher than in the inferior and superior regions, and the frequencies (observed and expected) of predated and non-predated seeds differed among the different regions of inflorescence. Females of A. schrankiae laid their eggs on fruits, and larvae, after emergence, perforated the exocarp to reach the seeds. Most fruits presented one to three eggs and only one bruchid larva was observed in each seed. The highest value of the rate number of eggs/fruit and the highest percentage of predated seeds were recorded in April. Dry weight of fruits (total) and seeds (non-predated), proportions of predated seeds, seed abortions, and non-emergent seed predators, were evenly distributed in the canopy.

Oxidative stability and α-tocopherol retention in soybean oil with lemon seed extract (Citrus limon) under thermoxidation

The synergistic effect of lemon seed extract with tert-butylhydroquinone (TBHQ) in soybean oil subjected to thermoxidation by Rancimat was investigated, and the influence of these antioxidants on α-tocopherol degradation in thermoxidized soybean oil. Control, LSE (2,400 mg/kg Lemon Seed Extract), TBHQ (50 mg/kg), Mixture 1 (LSE + 50 mg/kg TBHQ) and Mixture 2 (LSE + 25 mg/kg TBHQ) were subjected to 180°C for 20 h. Samples were taken at time 0, 5, 10, 15 and 20 h intervals and analysed for oxidative stability and α-tocopherol content. LSE and Mixtures 1 and 2 showed the capacity of retarding lipid oxidation when added to soya oil and also contributed to α-tocopherol retention in oil heated at high temperatures. However, Mixtures 1 and 2 added to the oil presented a greater antioxidant power, consequently proving the antioxidants synergistic effect.