Efecto de dos técnicas físicas de captación de agua sobre el establecimiento de las especies Mangifera indica L, Citrus sinensis y Coco nucifera L, en la microcuenca Las Marías, municipio de Télica. León (2007 al 2008)

En la investigación realizada en la micro cuenca Las Marías se propuso conocer el efecto de dos técnicas físicas de captación de agua sobre el establecimiento de las especies Mangifera indica L, Citrus sinensis y Coco nucifera L. Para desarrollar el proceso de recolección de información se establecieron diseños experimentales de Bloques Completos Aleatorizados, con dos repeticiones (finca Montoya parte baja y finca Miguel parte media), dentro de cada ensayo experimental se establecieron técnicas de captación de agua (cáseo y bordes) y un testigo. En cada finca se establecieron de forma azarizada 36 plántulas de Mangifera indica L, Citrus sinensis y Coco nucifera L (12 de cada especie). A cada especie se monitorearon variables de sobrevivencias, crecimiento en diámetro y altura y humedad de suelo. De igual forma se realizaron muestreos de suelo para conocer el porcentaje de humedad, infiltración y calidad física y química. El periodo de evaluación consistió de Junio 2007 a Enero 2008. Al finalizar la investigación se encontró que la especie Coco nucifera fue la que obtuvo mayor sobrevivencia (100 %), con un incremento corriente mensual (ICM) en diámetro (0.33 cm) y altura (7.73 cm), en comparación con las especies Mangifera indica y Citrus sinencis. El ICM en diámetro y altura fue similar en los tratamientos (cáseo y bordes) y el testigo (no se encontró diferencias significativa). En el tratamiento cáseo fue donde se encontró el mayor porcentaje de humedad del suelo (22.25 %) en comparación con el tratamiento bordes y el testigo. Se encontró una correlación positiva en la finca Miguel entre los porcentajes de humedad del suelo y el ICM en diámetro y altura de las especies frutales, lo cual podría estar relacionado a la calidad física y química de los suelos.

Acaricidas utilizados na citricultura convencional e orgânica: manejo da leprose e populações de ácaros fitoseídeos

O controle da leprose dos citros no Estado de São Paulo é realizado quase que exclusivamente com aplicações de acaricidas para o controle do ácaro vetor Brevipalpus phoenicis, as quais contribuem para o aumento dos custos de produção e podem afetar negativamente as populações de organismos benéficos. Portanto, o objetivo do trabalho foi avaliar, durante quatro safras, os efeitos de acaricidas indicados para o controle do ácaro B. phoenicis em citros convencional e orgânico sobre a evolução da leprose dos citros e sobre ácaros fitoseídeos. O experimento foi instalado em outubro de 2003, em pomar de laranja localizado no município de Reginópolis-SP. O delineamento experimental foi em blocos casualizados, estabelecendo-se os tratamentos, expressos em mL de p.c./100 L de água: spirodiclofen a 20 mL e cyhexatin a 50 mL (aplicados em rotação), calda sulfocálcica a 4.000 mL e testemunha sem aplicação de acaricidas. Entretanto, a rotação entre spirodiclofen e cyhexatin iniciou-se em setembro de 2006 e, anteriormente a esse período, utilizou-se somente o spirodiclofen. A cada quinze dias, foram realizados levantamentos populacionais do ácaro B. phoenicis e dos ácaros predadores Iphiseiodes zuluagai e os do gênero Euseius. O nível de controle adotado para o B. phoenicis foi de 8,3%, sendo que as aplicações dos produtos foram realizadas com pulverizador de arrasto tratorizado munido com lanças manuais. Na safra de 2007-2008, coletaram-se 10 frutos caídos devido à leprose por parcela e quantificou-se o número de lesões de leprose presentes em cada fruto. Avaliaram-se, ao término da safra de 2007-2008, a produtividade, as perdas devido à leprose, bem como a incidência e a severidade da leprose. Constatou-se que o local das lesões no fruto é mais importante para determinar sua queda do que o número de lesões presentes. Quanto mais intensa a infestação do ácaro B. phoenicis, maior é o número de lesões de leprose, resultando em maior queda prematura de frutos. Os acaricidas spirodiclofen e cyhexatin e spirodiclofen em rotação proporcionaram controle mais eficiente de B. phoenicis, em relação à calda sulfocálcica, resultando em maior produtividade, menores perdas de frutos e nos menores níveis de severidade da leprose. As aplicações de calda sulfocálcica reduziram os níveis populacionais do ácaro B. phoenicis abaixo do nível de controle, porém não evitaram o surgimento de lesões de leprose. As aplicações dos acaricidas apresentaram efeito nocivo sobre os ácaros fitoseídeos, pois houve redução da densidade populacional.

Acaricidas utilizados na citricultura convencional e orgânica: manejo da leprose e populações de ácaros fitoseídeos

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

Characterization of electrical penetration graphs of the Asian citrus psyllid, Diaphorina citri, in sweet orange seedlings

Detailed information on probing behavior of the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is critical for understanding the transmission process of phloem-limited bacteria (Candidatus Liberibacter spp.) associated with citrus `huanglongbing` by this vector. In this study, we investigated stylet penetration activities of D. citri on seedlings of Citrus sinensis (L.) Osbeck cv. Pera (Rutaceae) by using the electrical penetration graph (EPG-DC system) technique. EPG waveforms were described based on amplitude, frequency, voltage level, and electrical origin of the observed traces during stylet penetration into plant tissues. The main waveforms were correlated with histological observations of salivary sheath termini in plant tissues, to determine the putative location of stylet tips. The behavioral activities were also inferred based on waveform similarities in relation to other Sternorrhyncha, particularly aphids and whiteflies. In addition, we correlated the occurrence of specific waveforms with the acquisition of the phloem-limited bacterium Ca. Liberibacter asiaticus by D. citri. The occurrence of a G-like xylem sap ingestion waveform in starved and unstarved psyllids was also compared. By analyzing 8-h EPGs of adult females, five waveforms were described: (C) salivary sheath secretion and other stylet pathway activities; (D) first contact with phloem (distinct from other waveforms reported for Sternorrhyncha); (E1) putative salivation in phloem sieve tubes; (E2) phloem sap ingestion; and (G) probably xylem sap ingestion. Diaphorina citri initiates a probe with stylet pathway through epidermis and parenchyma (C). Interestingly, no potential drops were observed during the stylet pathway phase, as are usually recorded in aphids and other Sternorrhyncha. Once in C, D. citri shows a higher propensity to return to non-probing than to start a phloem or xylem phase. Several probes are usually observed before the phloem phase; waveform D is observed upon phloem contact, always immediately followed by E1. After E1, D. citri either returns to pathway activity (C) or starts phloem sap ingestion, which was the longest activity observed.

Characterization of electrical penetration graphs of Bucephalogonia xanthophis, a vector of Xylella fastidiosa in citrus

The sharpshooter Bucephalogonia xanthophis (Berg) (Homoptera: Cicadellidae) is a vector of the xylem-limited bacterium, Xylella fastidiosa (Wells, Raju, Hung, Weisburg, Mandelco-Paul, and Brenner), which causes citrus variegated chlorosis. Despite the importance of citrus variegated chlorosis, the probing behavior of vectors on citrus and its implications for transmission of X. fastidiosa have not been studied. Here we studied electrical penetration graph (EPG-DC system) waveforms produced by B. xanthophis on Citrus sinensis (L.) Osbeck (Rutaceae), and their relationships with stylet activities and xylem ingestion. Electrical penetration graph waveforms were described based on amplitude, frequency, voltage level, and electrical origin of the observed traces during stylet penetration on plant tissues. The main waveforms were correlated with histological observations of salivary sheaths in plant tissues and excretion analysis, in order to determine stylet activities and their precise position. Six waveforms and associated activities are described: (S) secretion of salivary sheath and intracellular stylet pathway, (R) resting during stylet pathway, (Xc) contact of stylets with xylem vessels, (Xi) active xylem ingestion, (N) interruption within the xylem phase (during Xc or Xi), and (W) withdrawal of stylet from the plant. The sharpshooter spent 91.8% of its probing time with its stylet in the xylem, where the main activity was ingestion (Xi: 97.5%). During a probe, the most likely sequence of events is secretion of salivary sheath and pathway (S) through epidermal and parenchyma cells (all individuals), followed by contact with xylem (Xc) (67.6% of all individuals) and ingestion (Xi) (88.3% of those that exhibit waveform Xc). The mean time to contact the xylem (Xc) and initiate ingestion (Xi) after onset of the first probe was 27.8 and 34.2 min, respectively. However, sustained xylem ingestion (Xi > 5 min) was established after 39.8 min, on average. This information is basic for future studies on the transmission mechanisms of X. fastidiosa and in order to establish control strategies aimed at interfering with this process.

Transmission efficiency of Xylella fastidiosa by sharpshooters (Hemiptera : Cicadellidae) in coffee and citrus

Xylella fastidiosa (Wells, Raju, Hung, Weisburg, Mandelco-Paul, and Brenner) is a bacteria] pathogen transmitted by several Sharpshooters in two tribes of Cicadellinae (Proconiini and Cicadellini). Here, we compared the transmission efficiency of X. fastidiosa in coffee (Coffea arabica L) and citrus [Citrus sinensis (L) Osbeck] by Cicadellini [Bucephalogonia xanthophis (Berg) and Dilobopterus costalimai Young] and Proconiini [Homalodisca ignorata Melichar and Oncometopia facialis (Signoret) I sharpshooters that Occur in both crops. At different seasons, healthy adults of each species were submitted to a 48-h acquisition access period on citrus or coffee source plants infected with X. fastidiosa isolates that cause Citrus variegated chlorosis (CVC) and Coffee leaf scorch (CLS), respectively, and then confined on healthy seedlings of the corresponding host plant for a 48-h inoculation access period. No significant effect of inoculation season was observed when comparing infection rates of citrus or coffee plants inoculated by vectors at different times of the year. In Citrus, the transmission rate by single insects was significantly higher for H. ignorata (30%) in relation to B. xanthophis (5%) and O. facialis (1.1%) but there was no difference among vector species in coffee, whose transmission rates ranged from 1.2 to 7.2%. Comparing host plants, H. ignorata was more effective in transmitting X. fastidiosa to citrus (30%) in relation to coffee (2.2%), whereas the other vectors transmitted the bacterium to both hosts with similar efficiencies. Despite these variations. vector efficiency in coffee and Citrus is lower than that reported in other hosts.

Annual and polyetic progression of citrus canker on trees protected with copper sprays

The effects of copper sprays on annual and polyetic progress of citrus canker, caused by Xanthomonas citri subsp. citri, in the presence of the Asian citrus leafminer (Phyllocnistis citrella), were evaluated in a study conducted in a commercial orchard in northwest Parana state, Brazil, where citrus canker is endemic. Nonlinear monomolecular, logistic and Gompertz models were fitted to monthly disease incidence data (proportion of leaves with symptoms) for each treatment for three seasons. The logistic model provided the best estimate of disease progress for all years and treatments evaluated and logistic parameter estimates were used to describe polyetic disease dynamics. Although citrus canker incidence increased during each of the seasons studied, it decreased over the whole study period, more so in copper-treated trees than in water-sprayed controls. Copper treatment reduced disease incidence compared with controls in every year, especially 2004-2005, when incidence was ca. 10-fold higher in controls than in treated plots (estimated asymptote values 0 center dot 82 and 0 center dot 07, respectively). Copper treatment also reduced estimated initial disease incidence and epidemic growth rates every year.

Spatial pattern of black spot incidence within citrus trees related to disease severity and pathogen dispersal

Guignardia citricarpa, the causal agent of citrus black spot, forms airborne ascospores on decomposing citrus leaves and water-spread conidia on fruits, leaves and twigs. The spatial pattern of diseased fruit in citrus tree canopies was used to assess the importance of ascospores and conidia in citrus black spot epidemics in Sao Paulo State, Brazil. The aggregation of diseased fruit in the citrus tree canopy was quantified by the binomial dispersion index (D) and the binary form of Taylor`s Power Law for 303 trees in six groves. D was significantly greater than 1 in 251 trees. The intercept of the regression line of Taylor`s Power Law was significantly greater than 0 and the slope was not different from 1, implying that diseased fruit was aggregated in the canopy independent of disease incidence. Disease incidence (p) and severity (S) were assessed in 2875 citrus trees. The incidence-severity relationship was described (R-2 = 88.7%) by the model ln(S) = ln(a) + bCLL(p) where CLL = complementary log-log transformation. The high severity at low incidence observed in many cases is also indicative of low distance spread of G. citricarpa spores. For the same level of disease incidence, some trees had most of the diseased fruit with many lesions and high disease severity, whereas other trees had most of the fruit with few lesions and low disease severity. Aggregation of diseased fruit in the trees suggests that splash-dispersed conidia have an important role in increasing the disease in citrus trees in Brazil.

Transmission of Methylobacterium mesophilicum by Bucephalogonia xanthophis for Paratransgenic Control Strategy of Citrus Variegated Chlorosis

Methylobacterium mesophilicum, originally isolated as an endophytic bacterium from citrus plants, was genetically transformed to express green fluorescent protein (GFP). The GFP-labeled strain of M. mesophilicum was inoculated into Catharanthus roseus (model plant) seedlings and further observed colonizing its xylem vessels. The transmission of this endophyte by Bucephalogonia xanthophis, one of the insect vectors that transmit Xylella fastidiosa subsp. pauca, was verified by insects feeding from fluids containing the GFP bacterium followed by transmission to plants and isolating the endophyte from C. roseus plants. Forty-five days after inoculation, the plants exhibited endophytic colonization by M. mesophilicum, confirming this bacterium as a nonpathogenic, xylem-associated endophyte. Our data demonstrate that M. mesophilicum not only occupy the same niche of X. fastidiosa subsp. pauca inside plants but also may be transmitted by B. xanthophis. The transmission, colonization, and genetic manipulation of M. mesophilicum is a prerequisite to examining the potential use of symbiotic control to interrupt the transmission of X. fastidiosa subsp. pauca, the bacterial pathogen causing Citrus variegated chlorosis by insect vectors.

`Swingle` citrumelo propagation by cuttings for citrus nursery tree production or inarching

`Swingle` citrumelo [Citrus paradisi MacFaden x Poncirus trifoliata (L.) Raf.] has been extensively used as a rootstock in several citrus growing regions of the World, including Southern Brazil where `Rangpur` lime (Citrus limonia Osbeck) is still the predominant variety despite being affected by several important pathogens. in this case, `Swingle` citrumelo is used to produce nursery trees to establish new orchards or to be inarched in adult and healthy groves in order to change the rootstock. We report herein a system to produce trees on `Swingle` citrumelo more rapidly by budding onto non-rooted cuttings, as well as assessing potential to rapidly multiply `Swingle` through rooting of non-budded cuttings. Therefore, two potential products are described: budded trees and rooted rootstock cuttings. `Valencia` sweet orange [Citrus sinensis (L.) Osbeck] was budded at different heights on cuttings derived from eight-month old rootstocks. Grafted and additional non-budded cuttings were then treated with indole-3-butyric acid (500 mg L(-1)) or left untreated before rooting. Three types of cuttings were evaluated: softwood, semi-hardwood and hardwood. The use of nursery trees derived from pre-budded hardwood cuttings of `Swingle` citrumelo is an alternative grafting method on this cultivar. Softwood cuttings with one leaf pair were considered the most adequate material for rapid multiplication of `Swingle` citrumelo by cutting. This could be particularly useful for inarching production or conventional budding after transplant of cutting-derived rootstocks. (C) 2008 Elsevier B.V. All rights reserved.

Evaluation of citrus somatic hybrids for tolerance to Phytophthora nicotianae and citrus tristeza virus

Somatic hybridization is a biotechnology tool that can be used in citrus breeding programs to produce somatic hybrids with the complete genetic combination of both parents. The goal of this work was to test the reaction of citrus somatic hybrids that may be useful as rootstocks to trunk and root infections caused by Phytophthora nicotianae van Breda de Haan (P parasitica Dastur) and to citrus tristeza virus (CTV). The somatic hybrids evaluated were `Caipira` sweet orange (Citrus sinensis L. Osbeck) + `Rangpur` lime (C. limonia Osbeck), `Caipira` sweet orange + `Cleopatra` mandarin (C. reshni hort. ex Tanaka), `Caipira` sweet orange + `Volkamer` lemon (C. volkameriana V Ten. & Pasq.), `Caipira` sweet orange + rough lemon (C. jambhiri Lush.), `Cleopatra` mandarin + `Volkamer` lemon, `Cleopatra` mandarin + sour orange (C. aurantium L.), `Rangpur` lime + `Sunki` mandarin (C. sunki (Hayata) hort. ex Tanaka), `Ruby Blood` sweet orange (C. sinensis L. Osbeck) + `Volkamer` lemon, `Rohde Red` sweet orange (C. sinensis L. Osbeck) + `Volkamer` lemon, and `Valencia` sweet orange + Fortunella obovata hort. ex Tanaka. For P. nicotianae trunk and root infection assays, plants of the somatic hybrids, obtained from 9-month semi-hardwood cuttings, were evaluated and compared with diploid citrus rootstock cultivars after mycelia inoculation in the trunk or spore infestation in the substrate, respectively. `Cleopatra` mandarin + sour orange, `Rangpur` lime + `Sunki` mandarin, `Cleopatra` mandarin + `Volkamer` lemon, `Ruby Blood` sweet orange + `Volkamer` lemon, `Rohde Red` sweet orange + `Volkamer` lemon, and `Caipira` sweet orange + `Volkamer` lemon had less trunk rot occurrence, whereas the somatic hybrids `Cleopatra` mandarin + `Volkamer` lemon, `Cleopatra` mandarin + sour orange, `Caipira` sweet orange + `Volkamer` lemon, and `Caipira` sweet orange + `Rangpur` lime were tolerant to root rot. For CTV assays, plants of the somatic hybrids along with tolerant and intolerant rootstocks were budded with a mild strain CTV-infected or healthy `Valencia` sweet orange budwood. Differences in average scion shoot length indicated that the hybrids `Cleopatra` mandarin + sour orange and `Valencia` sweet orange + Fortunella obovata were intolerant to CTV (c) 2007 Elsevier B.V. All rights reserved.

Selection processes in a citrus hybrid population using RAPD markers

The objective of this work was to evaluate the processes of selection in a citrus hybrid population using segregation analysis of RAPD markers. The segregation of 123 RAPD markers between 'Cravo' mandarin (Citrus reticulata Blanco) and 'Pêra' sweet orange (C. sinensis (L.) Osbeck) was analysed in a F1 progeny of 94 hybrids. Genetic composition, diversity, heterozygosity, differences in chromosomal structure and the presence of deleterious recessive genes are discussed based on the segregation ratios obtained. A high percentage of markers had a skeweness of the 1:1 expected segregation ratio in the F1 population. Many markers showed a 3:1 segregation ratio in both varieties and 1:3 in 'Pêra' sweet orange, probably due to directional selection processes. The distribution analysis of the frequencies of the segregant markers in a hybrid population is a simple method which allows a better understanding of the genetics of citrus group.

Stability of Citrus tristeza virus protective isolates in field conditions

The objective of this work was to monitor the maintenance of Citrus tristeza virus (CTV) protective isolates stability in selected clones of 'Pêra' sweet orange (Citrus sinensis), preimmunized or naturally infected by the virus, after successive clonal propagations. The work was carried out in field conditions in the north of Paraná State, Brazil. Coat protein gene (CPG) analysis of 33 isolates collected from 16 clones of 'Pêra' sweet orange was performed using single strand conformational polymorphism (SSCP). Initially, the isolates were characterized by symptoms of stem pitting observed in clones. Then viral genome was extracted and used as template for the amplification of CPG by reverse transcription polimerase chain reaction (RTPCR). RTPCR products electrophoretic profiles were analyzed using the Jaccard coefficient and the UPGMA method. The majority of the clones had weak to moderate stem pitting symptoms and its CTV isolates showed alterations in the SSCP profiles. However, the stability of the protective complex has been maintained, except for isolates from two analised clones. Low genetic variability was observed within the isolates during the studied years.

Genetic similarity of citrus fresh fruit market cultivars

The objective of this work was to assess the genetic similarity of the following citrus fresh fruit market seedless cultivars: Lane Late, Navelate, Navelina and Salustiana sweet oranges (Citrus sinensis (L.) Osbeck), Clemenules and Marisol mandarins (C. reticulata Blanco) and Okitsu satsuma mandarin (C. unshiu Marcovitch), and the hybrids Nova [C. clementina x (C. paradisi x C. tangerina)] and Ortanique (tangor probably derived from C. sinensis (L.) Osbeck x C. reticulata Blanco), utilizing isoenzymatic markers. Electrophoresis analysis of proteins extracted from leaf tissues was utilized to detect polymorphisms at ten isoenzymatic systems. Out of 30 alleles, 16 were polymorphic. The Jaccard coefficient was utilized to estimate the genetic similarity between the cultivars and the unweigthed pair-group method using an arithmetic average (UPGMA) was used to obtain the phenogram (NTSYS 1.7). The cultivars showed high genetic similarity (>72.5%), and were classified in five main groups: sweet oranges, 'Clemenules' and 'Marisol' mandarins, 'Nova', 'Ortanique', and 'Okitsu' satsuma mandarin.

Efeito da poda, raleio de frutos e uso de fitorreguladores na produção de tangerinas (Citrus deliciosa Tenore) cv. montenegrina

O experimento aqui relatado teve o propósito de estudar a poda de ramos, raleio de frutos e uso de fitorreguladores para diminuir a alternância de produção e melhorar a qualidade físico-química dos frutos de tangerineiras 'Montenegrina' (Citrus deliciosa Tenore). São plantas enxertadas sobre laranjeira 'Caipira'(Citrus sinensis L. Osbeck) e estão em um pomar comercial de 6 anos de idade, da empresa Panoramas Citrus, situada no município de Butiá-RS, na latitude 29º57'S e longitude 51º40'W. Neste experimento, foram realizados os seguintes tratamentos: A) Testemunha nº1: plantas com carga excessiva; B) Testemunha nº2: plantas sem carga; C) Raleio manual de 66% dos frutos em plantas com carga excessiva, em fevereiro; D) Pulverizações de plantas excessivamente carregadas, utilizando-se de 200 mg.L-1 de Ethrel (24% ethephon), em novembro; E) Poda de frutificação, em plantas excessivamente carregadas, em dezembro; F) Idem "E" acrescido de raleio manual de 33 % dos frutos, em fevereiro; G) Idem "E" acrescido de pulverização com 50 mg.L-1 de 2,4-DP (95% de 2,4 diclorofenoxipropiônico), no final da queda natural de frutos, em dezembro; H) Poda de plantas com alternância de produção, em dezembro; I) Idem "H" acrescido de pulverização com 15 mg.L-1 de ácido giberélico (10% de AG3), em maio. O delineamento experimental foi o de blocos casualizados, com parcelas subdivididas no tempo, tendo 4 repetições e 3 plantas úteis por parcela. A poda diminuiu a produção por planta no ano de excessiva carga de frutos e reduziu a alternância de produção, quando associada à prática do raleio manual de 33% dos frutos. Em plantas alternantes a poda foi mais eficiente para quebrar a alternância de produção. A poda e o raleio manual de 66% aumentaram a massa média e melhoraram a qualidade dos frutos, mas o raleio manual de 66% dos frutos, somente, foi insuficiente para quebrar a alternância de produção. A 200 mg L-1 , o ethephon não exerceu ação de raleio de frutos. O 2,4-DP com pulverização de 50 mg L-1 não modificou o tamanho dos frutos. O AG3, na concentração de 10 mg L-1, aplicado em maio, não inibiu a diferenciação floral.