Azteca barbifex Forel (Hymenoptera: Formicidae): potential pest of citrus crops in eastern Amazon

Damage caused by the ant Azteca barbifex (Forel) was identified in orange trees (Citrus sinensis), in Capitão Poço County, Guamá microregion, Pará State. The damage caused by the scraping of stems and branches lead to reduction in yield with subsequent death of the plant. These characteristics indicate A. barbifex as a potential pest of citrus crops in the eastern region of Amazon.

CARACTERIZAÇÃO PÓS-COLHEITA DE SEIS CULTIVARES DE CARAMBOLA (Averrhoa carambola L.)

Avaliaram-se as qualidades física e química dos frutos de seis cultivares de carambola (Averrhoa carambola L.), propagadas pela Estação Experimental de Citricultura de Bebedouro, São Paulo, objetivando a obtenção de informações sobre a qualidade dos frutos produzidos nesta região. Foram utilizados frutos das cultivares Tean-ma, Fwang Tung, Arkin, Mal��sia, Golden Star e Nota-10. Os frutos foram analisados quanto à massa fresca, comprimento, largura, volume, porcentagem de suco, polpa e semente, número de sementes, coloração (L, Cromaticidade e ângulo Hue). Na polpa, determinou-se o pH, assim como o teor de umidade, sólidos sol��veis totais (SST), acidez total titul��vel (ATT), vitamina C, açúcares sol��veis totais e redutores, cinzas, proteína, pectina total e sol��vel, e minerais (Ca, P, K, Mg, S, Cu, Fe, Mn e Zn). Os resultados foram avaliados estatisticamente através de um delineamento inteiramente casualizado e revelaram que os melhores frutos são os originários das cultivares Arkin, Tean-ma e Nota-10, pois são os maiores, os mais pesados e apresentam os maiores teores de sólidos sol��veis totais (SST) e açúcares sol��veis (AST), assim como boa relação SST/ATT. Os frutos da 'Arkin' mostraram coloração mais amarela, enquanto os da 'Golden Star' foram considerados de coloração mais clara, ou com maior translucidez. A textura foi menor nos frutos da 'Arkin', e se relacionou com a maior solubilização de suas pectinas

Fruit quality of 'Tahiti' lime (Citrus latifolia Tanak) grafted on twelve different rootstocks

No presente trabalho, avaliou-se a influência de doze porta-enxertos sobre a qualidade dos frutos da lima-ácida 'Tahiti' (Citrus latifolia Tanaka), clone 'IAC-5', amostrados em duas posições nas plantas, em experimento conduzido na Estação de Citricultura de Bebedouro, em um pomar de três anos. O espaçamento utilizado foi de 8.0 x 5.0m. Os porta-enxertos utilizados foram: citrangeiro 'Carrizo' (C. sinensis (L.) Osbeck x Poncirus trifoliata (L.) Raf.); os híbridos 'Cravo' x 'Swingle' (C. limonia Osbeck x P. trifoliata (L.) Raf. x C. paradisi Macf.) e 'Changsha' x 'English Small' (C. sunki Hort. ex Tan. x P. trifoliata Raf.); as tangerineiras 'Sun Chu Sha Kat' (C. reticulata Blanco) e 'Sunki' (C. sunki Hort. ex Tanaka); os limoeiros 'Cravo Limeira' e 'Cravo FCAV' (C. limonia Osbeck); o citrumeleiro 'Swingle' (P. trifoliata Raf. x C. paradisi Macf.), o tangeleiro 'Orlando' (C. reticulata Blanco x C. paradisi Macf.) e os trifoliateiros 'Rubidoux', 'FCAV' e 'Flying Dragon' (Poncirus trifoliata Raf.). Foi utilizado um delineamento em blocos casualizados, com doze tratamentos e seis repetições. Os distintos porta-enxertos induziram diferenças na qualidade dos frutos, entretanto todas as características de qualidade foram consideradas aceitáveis para a variedade, sendo bons substitutos para o limão 'Cravo'.

EFEITOS DE GIBERELINAS, CITOCININAS E do NITRATO DE POTÁSSIO, NO PROCESSO GERMINATIVO DE SEMENTES DE LIMOEIRO `CRAVO' (Citrus limonia OSBECK)

Foram estudados os efeitos de reguladores vegetais do grupo das giberelinas e citocininas, bem como do nitrato de potássio na germinação de sementes do limoeiro `Cravo' (Citrus limonia Osbeck). O experimento foi realizado, contendo papel de filtro como substrato para a germinação das sementes, regulado à temperatura de 25oC. As sementes foram retiradas de frutos maduros no final da safra do limoeiro `Cravo', lavadas, secas à sombra e armazenadas durante 11 dias em câmara fria. em seguida, receberam tratamento com os fitorreguladores e KNO3 por 24 horas, de acordo com os tratamentos: KNO3 0,1% e 0,2%; GA3 50, 100 e 250 mg.L-1; GA4 + GA7 + fenilmetilaminopurina 100 mg.L-1; fenilmetilaminopurina 20 mg.L-1 e água destilada (testemunha). As avaliações foram iniciadas 15 dias após a semeadura, em intervalos de 5 dias. Conclui-se que os reguladores vegetais utilizados não afetaram o processo germinativo das sementes e que os tratamentos com nitrato de potássio 0,1% e 0,2% exerceram efeito inibitório sobre a germinação.

ÁCIDO GIBERÉLICO E O DESENVOLVIMENTO DE PL��NTULAS DE TANGERINA 'CLEÓPATRA' (Citrus reshni hort. ex. Tanaka)

Pl��ntulas de tangerina 'Cleópatra'(Citrus reshni hort. ex. Tanaka), foram utilizadas em delineamento inteiramente casualizado, com três repetições no esquema de parcelas sub-divididas, onde os tratamentos principais foram 5 concentrações de ácido giberélico: T1 = 0, T2 = 25, T3 = 50, T4 = 100 e T5 = 150 mg.L-1, parceladas em quatro pulverizações. As coletas de material foram realizadas dos 70 aos 210 dias, em intervalos de 14 dias. Pelos resultados obtidos, o fitorregulador promoveu efeito favorável com relação ao incremento do comprimento do caule, sendo que aplicações de 100 e 150 mg.L-1 foram superiores as demais. O diâmetro do caule não foi influenciado pela aplicação de ácido giberélico.

Biologia de Polyphagotarsonemus latus (Banks) (Acari: Tarsonemidae) em limão siciliano (Citrus limon Burm)

No estudo da biologia de Polyphagotarsonemus latus em limão Siciliano, foram utilizados potes pl��sticos circulares com capacidade de 250 ml, contendo areia esterilizada como suporte para dois frutos novos com aproximadamente 2,0 cm de diâmetro. O ensaio foi conduzido a 27,1 ± 0,5°C, umidade relativa de 67,6 ± 1,3% e fotofase contínua. O período de ovo a adulto durou 3,7 ± 0,1 dias para fêmeas e 3,6 ± 0,1 dias para machos, com sobrevivência de 100%. Após um período de pré-oviposição de 1,0 ± 0,2 dias, as fêmeas depositaram 5,6 ± 0,5 ovos por dia durante 10,5 ± 0,9 dias, totalizando 58,9 ± 6,7 ovos por fêmea. A longevidade foi de 13,4 ± 1,0 dias para fêmeas e 12,0 ± 2,4 dias para machos. A razão intrínseca de aumento (rm) foi de 0,359, a razão finita de aumento (l) de 1,43 indivíduos por fêmea por dia, o tempo médio de uma geração (T) de 10,34 dias e a taxa l��quida de reprodução (Ro) de 41,0.

A triply cloned strain of Xylella fastidiosa multiplies and induces symptoms of citrus variegated chlorosis in sweet orange

Xylella fastidiosa isolate 8.1,b obtained from a sweet orange tree affected by citrus variegated chlorosis in the state of Sb Paulo, Brazil, and shown in 1993 to be the causal agent of the disease, was cloned by repeated culture in liquid and on solid PW medium, yielding triply cloned strain 9a5c. The eighth and the 16th passages of strain 9a5c were mechanically inoculated into sweet orange plants. Presence of X. fastidiosa in sweet orange leaves of shoots having grown after inoculation (first-flush shoots) was detected by DAS-ELISA and PCR. Thirty-eight days after inoculation, 70% of the 20 inoculated plants rested positive, and all plants gave strong positive reactions 90 days after inoculation. Symptoms first appeared after 3 months and were conspicuous after 5 months. X. fastidiosa was reisolated from sweet orange leaves, 44 days after inoculation. These results indicate that X. fastidiosa strain 9a5c, derived from pathogenic isolate 8.1.b by triply cloning, is also pathogenic, Strain 9a5c is now used for the X. fastidiosa genome sequencing project undertaken on a large scale in Brazil.

Pectinolytic activity secreted by yeasts isolated from fermented citrus molasses

Aims: the aim of this study was to obtain improved strains of pectinolytic yeasts adapted to the conditions of an industrial fermentation process, which was continuously operated to convert citrus molasses into ethanol.Methods and Results: the starter yeast of the industrial fermentation process was a commercial baker's yeast, which was capable of growing without forming any secretion halo of pectinase activity on solid medium. Nevertheless, isolates showing secretion of pectinolytic activity on plates were obtained from the fermentation process. The secretion of pectin-degrading activity by isolates on plates was repressed by galactose and improved as the result of colony aging on polygalacturonic acid plates at 30 degrees C. Liquefaction of polygalacturonate gels as well as the splitting of the pectin-degrading activity into a wall-linked and a supernatant fraction were also observed when the starter yeast was propagated under agitation in liquid medium containing pectin.Conclusions: Isolates capable of secreting pectinolytic activity on plates were predominant at the end of the citrus molasses fermentation. Nevertheless, the sizes of the secretion haloes on plates were not necessarily an indication of the levels of pectinolytic activity secreted in the liquid medium.Significance and Impact of the Study: Improved pectinolytic strains of Saccharomyces can be used as a source of pectinases for a variety of applications. This organism also participates in plant deterioration processes.

Morphological and molecular characterization of Colletotrichum spp. from citrus orchards affected by postbloom fruit drop in Brazil

Brazilian isolates of Colletotrichum spp. from citrus orchards affected by postbloom fruit drop were examined for colony colour, mycelial growth, benomyl-resistance, pathogenicity, and genetic variability by random amplified polymorphic DNA (RAPD) analysis. All isolates were obtained from flowers and persistent calyxes from different citrus hosts from São Paulo, Brazil. DNA polymorphisms detected after amplification with random 10-mer primers were used to classify the isolates into two groups. Group I isolates grew rapidly on potato-dextrose agar (PDA) and were sensitive to benomyl, and group II isolates grew slowly on PDA and were benomyl-resistant. Colletotrichum acutatum was analyzed by RAPD and had high genetic similarity with group II isolates of Colletotrichum from citrus. Probably, the group I is C, gloeosporioides and group II is C. acutatum.

Efficacy of pre- and postinoculation application of Fungicides to expanding young citrus leaves for control of melanose, scab, and alternarial brown spot

In greenhouse trials, copper hydroxide, pyraclostrobin, and famoxadone were applied to actively crowing young citrus seedlings to determine the duration of protection of young leaves provided by these fungicides against melanose, caused by Diaporthe citri, citrus scab, caused by Elsinoe fawcettii, and Alternaria brown spot, caused by Alternaria alternata. Fungicides were applied to different sets of potted plants of grapefruit for control of melanose, of rough lemon for control of scab, and of Dancy tangerine for control of Afternaria brown spot 1 to 6 days prior to inoculation. as well as on the day of inoculation. Leaf area of treated shoots was estimated on the day of fungicide application and the day of inoculation and disease severity evaluated subsequently. In most cases. copper hydroxide and famoxadone provided at least 50% control of all three diseases for only about 2 days after application. Generally, there was little or no disease control when the products were applied 4 or more days before inoculation. In contrast, pyraclostrobin usually provided a high level of control of all three diseases when applied up to 5 days prior to inoculation. The level of disease control decreased as the interval between a fungicide application and inoculation increased and the relationship between disease control and leaf expansion best fit a quadratic equation. Effective disease control was observed with copper hydroxide and famoxadone until leaf area had increased by 100 to 200%, whereas control with pyraclostrobin was observed up to 400 to 500% increase in leaf area. In postinoculation tests with scab and melanose, pyraclostrobin provided high levels of disease control (>75%) when applied up to 2 days after inoculation. whereas copper hydroxide and famoxadone had minimal postinoculation activity. Applications of pyraclostrobin to the spring flush growth of citrus trees are much more likely to provide control of melanose, scab, and Alternaria brown spot than those of famoxadone or copper hydroxide.

Candidatus Liberibacter americanus, associated with citrus huanglongbing (greening disease) in São Paulo State, Brazil

Symptoms of huanglongbing (HLB) were reported in São Paulo State (SPS), Brazil, in March 2004. In Asia, HLB is caused by 'Candidatus Liberibacterasiaticus'and in Africa by 'Candidatus Liberibacter africanus'. Detection of the liberibacters is based on PCR amplification of their 16S rRNA gene with specific primers. Leaves with blotchy mottle symptoms characteristic of HLB were sampled in several farms of SIPS and tested for the presence of liberibacters. 'Ca. L. asiaticus' was detected in a small number of samples but most samples gave negative PCR results. Therefore, a new HLB pathogen was suspected. Evidence for an SPS-HLB bacterium in symptomatic leaves was obtained by PCR amplification with universal primers for prokaryotic 16S rRNA gene sequences. The amplified 16S rRNA gene was cloned and sequenced. Sequence analysis and phylogeny studies showed that the 16S rRNA gene possessed the oligonucleotide signatures and the secondary loop structure characteristic of the alpha-Proteobacteria, including the liberibacters. The 16S rRNA gene sequence phylogenetic tree showed that the SPS-HLB bacterium clustered within the a-Proteobacteria, the liberibacters being its closest relatives. For these reasons, the SPS-HLB bacterium is considered a member of the genus 'Ca. Liberibacter'. However, while the 16S rRNA gene sequences of 'Ca. L. asiaticus' and 'Ca. L. africanus' had 98-4% similarity, the 16S rRNA gene sequence of the SPS-HLB liberibacter had only 96(.)0% similarity with the 16S rRNA gene sequences of 'Ca. L. asiaticus'or'Ca. L. africanus'. This lower similarity was reflected in the phylogenetic tree, where the SPS-HLB liberibacter did not cluster within the 'Ca. L asiaticus'/'Ca. L. africanus group', but as a separate branch. Within the genus 'Candidatus Liberibacter' and for a given species, the 16S/23S intergenic region does not vary greatly. The intergenic regions of three strains of 'Ca. L. asiaticus', from India, the People's Republic of China and Japan, were found to have identical or almost identical sequences. In contrast, the intergenic regions of the SPS-HLB liberibacter, 'Ca. L. asiaticus' and 'Ca. L. africanus' had quite different sequences, with similarity between 66(.)0 and 79(.)5%. These results confirm that the SPS-HLB liberibacter is a novel species for which the name 'Candidatus Liberibacter americanus' is proposed. Like the African and the Asian liberibacters, the 'American' liberibacter is restricted to the sieve tubes of the citrus host. The liberibacter could also be detected by PCR amplification of the 16S rRNA gene in Diaphorina citri, the psyllid vector of 'Ca. L. asiaticus', suggesting that this psyllid is also a vector of 'Ca. L. americanus' in SPS. 'Ca. L. americanus' was detected in 216 of 218 symptomatic leaf samples from 47 farms in 35 municipalities, while 'Ca. L. asiaticus' was detected in only 4 of the 218 samples, indicating that 'Ca. L. americanus' is the major cause of HLB in SIPS.

Comparative analyses of the complete genome sequences of Pierce's disease and citrus variegated chlorosis strains of Xylella fastidiosa

Xylella fastidiosa is a xylem-dwelling, insect-transmitted, gamma-proteobacterium that causes diseases in many plants, including grapevine, citrus, periwinkle, almond, oleander, and coffee. X. fastidiosa has an unusually broad host range, has an extensive geographical distribution throughout the American continent, and induces diverse disease phenotypes. Previous molecular analyses indicated three distinct groups of X.fastidiosa isolates that were expected to be genetically divergent. Here we report the genome sequence of X. fastidiosa (Temecula strain), isolated from a naturally infected grapevine with Pierce's disease (PD) in a wine-grape-growing region of California. Comparative analyses with a previously sequenced X.fastidiosa strain responsible for citrus variegated chlorosis (CVC) revealed that 98% of the PD X.fastidiosa Temecula genes are shared with the CVC X. fastidiosa strain 9a5c genes. Furthermore, the average amino acid identity of the open reading frames in the strains is 95.7%. Genomic differences are limited to phage-associated chromosomal rearrangements and deletions that also account for the strain-specific genes present in each genome. Genomic islands, one in each genome, were identified, and their presence in other X.fastidiosa strains was analyzed. We conclude that these two organisms have identical metabolic functions and are likely to use a common set of genes in plant colonization and pathogenesis, permitting convergence of functional genomic strategies.

Identification and analysis of single nucleotide polymorphisms (SNPs) in citrus

Most of the cultivated species of citrus have narrow genetic basis. Relationships among species and cultivars are obscured by sexual compatibility, polyembryony, apomixis and a high incidence of somatic mutations. DNA analysis is crucial in genetic studies not only for citrus breeding programs but also for characterization of hybrids and species. In this paper, single nucleotide polymorphisms ( SNPs) were investigated in 58 accessions of Citrus, hybrids and related genera. Genomic sequences of 'Pera IAC' sweet orange ( Citrus sinensis L. Osbeck) were used for primer design and selection of sequence tagged sites (STSs) for identification of SNPs. Analysis of 36 STSs showed identical sequences among 40 of the 41 sweet orange accessions studied. However, these accessions were heterozygous for many SNPs. Ten selected STSs were analyzed in 17 additional accessions from 13 species and hybrids. Comparing to the 'Pera IAC' sweet orange accession, a total of 150 polymorphic nucleotides were identified and most of the alterations were transitions ( 52.7%). The greatest number of SNPs was observed in Poncirus trifoliata ( L.) Raf. and the smallest in 'Ponkan' mandarin ( Citrus reticulata Blanco). At the intra-specific level, 'Bafa Gigante' ( Citrus sinensis L. Osbeck) was the only sweet orange accession with a divergent SNPs genotype, which corroborates the hypothesis of a hybrid origin for this accession. Although the STSs analyzed represent randomly sampled genomic sequences, they provided consistent information about the level of polymorphism and showed the potential of SNPs markers for characterization and phylogenetic studies.

Pratylenchus jaehni sp n. from citrus in Brazil and its relationship with P-coffeae and P-loosi (Nematoda : Pratylenchidae)

A lesion nematode population infecting citrus in the state of São Paulo, Brazil is described and named Pratylenchus jaehni sp. n. Biological, molecular and morphological characteristics of this new species are compared with those of the morphologically similar P. coffeae and P. loosi. Results of mating experiments showed that R jaehni is reproductively isolated from P coffeae. Molecular (D2/D3 DNA sequences) dissimilarities among P. jaehni sp, n., P. coffeae and P. loosi were documented in a previous study. The morphology of seven R coffeae populations from tropical America and eastern Java and a P loosi population from Sri Lanka is used for comparison with the morphology of P. jaehni sp. n. Pratylenchus jaehni differs from R coffeae and P. loosi by only a few morphological ;characters of the females. The mean values of stylet length, stylet knob height, and vulva position are smaller (less than or equal to15 vs greater than or equal to15 mum, less than or equal to2.7 vs greater than or equal to2.7 mum, less than or equal to79 vs greater than or equal to79%) than those in P coffeae and P loosi. The tail terminus is usually subhemispherical and smooth in P jaehni sp. n.. whereas it is commonly truncate and indented in most P. coffeae populations and bluntly or finely pointed in P. loosi. Because of the morphological similarities among P. jaehni sp. n., P. coffeae and P. loosi, examination of at least ten specimens is required to obtain a reliable diagnosis based on morphology. Nineteen morphometric parameters for P. jaehni sp. n. and P. coffeae ranged from 0-13% smaller in fixed than in live specimens.

Determination of buprofezin, pyridaben, and tebufenpyrad residues by gas chromatography mass-selective detection in clementine citrus

A gas chromatography-mass-selective (GC-MS) detection method to determine buprofezin, pyridaben, and tebufenpyrad on the pulp, peel, and whole fruit of clementines is described. The extraction/partition procedure was performed in one step and no cleanup was necessary with the GC-MS in the SIM-mode pesticide determination. Recovery ranged from 75 to 124% with coefficients of variance ranging between 1 and 13%. The limit of determination was 0.01 mg/kg for all pesticides. The field trials showed a similar degradative behavior for all active ingredients (AI), with a great residue decrease during the first week and stability in the second. Just after treatment buprofezin and tebufenpyrad showed lower residues than the maximum residue limit (MRL) fixed in Italy, while pyridaben was below the MRL after a week.