Aspectos biológicos de Chrysoperla externa(Hagen, 1861)(Neuroptera: Chrysopidae) alimentada com Planococcus citri (Risso, 1813)(Hemiptera: Pseudococcidae) e Toxoptera citricida (Kirkaldy, 1907) (Hemiptera: Aphididae)

The citrus-growing ecosystem houses a great deal of species of natural enemies, among them Chrysoperla externa (Hagen, 1861), which presents a great potential for use in the biological control of pests. The aim of this work was to evaluate the effects of the preys Planococcus citri (Risso, 1813) and Toxoptera citricida (Kirkaldy, 1907) on the development of larva and pupa of C. externa. The trial was carried out in an acclimatized room at 25 +/- 1 degrees C, 70 +/- 10% RH and 12-hour photophase, the treatments consisting of the continuous and alternate supply of those two preys in each instar of the predator, in addition to eggs of Anagasta kuehniella (Zeller, 1879) utilized for comparison purposes. Both the white mealybug and the aphids were given in sufficient amounts to the C. externa larvae to be able to feed ad libitum. The lifetime and the survival rate of the phases of larva, pupa and the larval-adult period of C. externa were evaluated. It was found that T. citricida given either singly along the larval development or in two consecutive instars of the predator was not an adequate prey, 100% of mortality over the larval-adult period being found. When the larvae fed on P. citri, given in two instars of the predator, the survival rate in the larval-adult period was similar to the diet constituted of only A. kuehniella eggs, except when fed with T. citricida in the first instar.

Desenvolvimento de Planococcus citri (Risso, 1813) (Hemiptera: Pseudococcidae) em cafeeiros

A cochonilha Planococcus citri (Risso, 1813) suga a seiva dos botões florais e frutos do cafeeiro, atacando as rosetas desde a floração até a colheita. Embora seja relatada há alguns anos na cafeicultura, são escassas as informações sobre o desenvolvimento dessa cochonilha em cafeeiros. Assim, o presente trabalho teve por objetivos avaliar alguns aspectos biológicos da fase ninfal de P. citri em plantas de café. Ovos dessa cochonilha foram retirados de uma criação em laboratório, isolados em placas de Petri contendo discos foliares de Coffea arabica L., das cultivares Acaiá Cerrado, Mundo Novo e Catuaí Vermelho e de C. canephora Pierre & Froenher, cultivar Apoatã. As placas foram mantidas em câmara climatizada a 25 ± 1ºC, 70 ± 10% de umidade relativa e 12h de fotofase. Constatou-se que a cultivar Catuaí Vermelho foi a que proporcionou maior duração do período ninfal das fêmeas, porém, não foram constatadas diferenças na mortalidade. Essa cochonilha se desenvolveu satisfatoriamente em todas as cultivares de café estudadas e os resultados não mostraram diferenças claras de susceptibilidade.

Caracterização molecular através da técnica fAFLP de isolados de Diaporthe citri

A citricultura é um mercado em expansão, principalmente no Estado de São Paulo, cuja importância na balança comercial já é reconhecida. Como em qualquer espécie cultivada, o crescimento das áreas de cultivo favorecem também o crescimento de problemas fitossanitários. Desta forma, as espécies de citros são afetadas por diversas doenças destacando-se entre elas a melanose, causada por Diaporthe citri (Wolf.), à qual a grande maioria das variedades comerciais são suscetíveis. O conhecimento da diversidade intra-específica é de grande importância, já que esta poderá auxiliar na seleção de variedades com resistência. O objetivo deste trabalho foi avaliar a variabilidade genética em isolados de Diaporthe citri, originários de diferentes locais, variedades e partes da planta, utilizando marcadores moleculares. Marcadores do tipo AFLP (Amplified Fragment Length Polymorphism) foram utilizados para caracterização de dez isolados do patógeno. Os DNAs genômicos extraídos da massa micelial foram utilizados nas reações de amplificação. A técnica fluorescent AFLP permitiu a distinção dos isolados estudados, tendo sido classificados em quatro grupos distintos. Contudo, estes grupos não foram formados em razão da região geográfica, parte da planta ou variedade.

Desenvolvimento da cochonilha-branca planococcus citri (risso, 1813) (hemiptera: pseudococcidae) em frutíferas

The citrus mealybug, Planococcus citri is an important pest attacking several crops. Biotic and abiotic factors, as well as the substrate they feed on influence its population. The present study was aimed to study the effect of some feeding substrates on mealybug development in laboratory conditions. Leaf disks of guava, fig and cherimoya were placed individually on an agar film inside a Petri dish. One egg of the insect was confined inside each container, replicated 60 times for each treatment. Containers were kept in a climatized chamber at 25[degree] C, 70 [plus or minus] 10% RH and 12 hours photophase. Insects reared on guava leaves showed a longer nymphal development, while the female longevity was longer on fig leaves. However, no statistical differences were found in the survival index, which showed values higher than 83%, suggesting that all substrates studied here were adequate for the development of P. citri.

Selection and validation of reference genes for gene expression studies by reverse transcription quantitative PCR in Xanthomonas citri subsp citri during infection of Citrus sinensis

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

Genetic Diversity of a Brazilian Strain Collection of Xanthomonas citri subsp citri Based on the Type III Effector Protein Genes

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

New genes of Xanthomonas citri subsp citri involved in pathogenesis and adaptation revealed by a transposon-based mutant library

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

Proteome of the phytopathogen Xanthomonas citri subsp citri: a global expression profile

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

Produção e sensibilidade de isolados brasileiros de Xanthomonas axonopodis pv. citri à bacteriocinas

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

Subcellular localization of proteins labeled with GFP in Xanthomonas citri ssp citri: targeting the division septum

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

The copper resistance operon copAB from Xanthomonas axonopodis pathovar citri: gene inactivation results in copper sensitivity

Xanthomonas axonopodis pv. citri (Xac) causes citrus canker and the completion of the Xac genome sequence has opened up the possibility of investigating basic cellular mechanisms at the genomic level. Copper compounds have been extensively used in agriculture to control plant diseases. The copA and copB genes, identified by annotation of the Xac genome, encode homologues of proteins involved in copper resistance. A gene expression assay by Northern blotting revealed that copA and copB are expressed as a unique transcript specifically induced by copper. Synthesis of the gene products was also induced by copper, reaching a maximum level at 4 h after addition of copper to the culture medium. CopA was a cytosolic protein and CopB was detected in the cytoplasmic membrane. The gene encoding CopA was disrupted by the insertion of a transposon, leading to mutant strains that were unable to grow in culture medium containing copper, even at the lowest CUSO4 concentration tested (0.25 mM), whereas the wild-type strain was able to grow in the presence of 1 mM copper. Cell suspensions of the wild-type and mutant strains in different copper concentrations were inoculated in lemon leaves to analyse their ability to induce citrus canker symptoms. Cells of mutant strains showed higher sensitivity than the wild-type strain in the presence of copper, i.e. they were not able to induce citrus canker symptoms at high copper concentrations and exhibited a more retarded growth in planta.

Crystallization and preliminary X-ray diffraction analysis of XAC1151, a small heat-shock protein from Xanthomonas axonopodis pv. citri belonging to the alpha-crystallin family

The hspA gene (XAC1151) from Xanthomonas axonopodis pv. citri encodes a protein of 158 amino acids that belongs to the small heat-shock protein ( sHSP) family of proteins. These proteins function as molecular chaperones by preventing protein aggregation. The protein was crystallized using the sitting-drop vapour-diffusion method in the presence of ammonium phosphate. X-ray diffraction data were collected to 1.65 angstrom resolution using a synchrotron-radiation source. The crystal belongs to the rhombohedral space group R3, with unit-cell parameters a = b = 128.7, c = 55.3 angstrom. The crystal structure was solved by molecular-replacement methods. Structure refinement is in progress.

Saprophytic colonization of citrus twigs by Diaporthe citri and factors affecting pycnidial production and conidial survival

Melanose, caused by Diaporthe citri, produces reddish brown lesions on the fruit, leaves, and twigs of citrus trees, and greatly reduces the marketability of fresh fruit. Most of the inoculum is produced in pycnidia on dead twigs in the tree canopy, which exude large numbers of conidia in slimy masses. In this study, detached twigs inoculated with conidia were readily colonized and produced large numbers of pycnidia within 30 to 40 days when they were soaked 3 to 4 h on alternate days. Conidial production was measured by wetting twigs in a rain tower periodically and collecting the conidia in the runoff water. Production began after 80 days and continued for nearly 300 days. In other experiments, production of mature pycnidia on detached twigs was greatest at 94 to 100% relative humidity (RH) and at 28 degrees C. Low RH and temperature, however, favored survival of conidia in exuded masses on twigs. In the field, colonization of detached twigs by D. citri was high in rainy season, moderate in spring and early fall, and minimal in late fall and winter. Twig colonization was positively related to the number of rain days and average temperature, but not to total rainfall. In another experiment, inoculated twigs placed in the tree canopy developed pycnidia and then produced conidial masses for about 200 days. D. citri is a serious pathogen, but a weak parasite, that survives primarily by colonization and reproduction on dead twigs.

Development and validation of a Xanthomonas axonopodis pv. citri DNA microarray platform (XACarray) generated from the shotgun libraries previously used in the sequencing of this bacterial genome

Background. From shotgun libraries used for the genomic sequencing of the phytopathogenic bacterium Xanthomonas axonopodis pv. citri (XAC), clones that were representative of the largest possible number of coding sequences (CDSs) were selected to create a DNA microarray platform on glass slides (XACarray). The creation of the XACarray allowed for the establishment of a tool that is capable of providing data for the analysis of global genome expression in this organism. Findings. The inserts from the selected clones were amplified by PCR with the universal oligonucleotide primers M13R and M13F. The obtained products were purified and fixed in duplicate on glass slides specific for use in DNA microarrays. The number of spots on the microarray totaled 6,144 and included 768 positive controls and 624 negative controls per slide. Validation of the platform was performed through hybridization of total DNA probes from XAC labeled with different fluorophores, Cy3 and Cy5. In this validation assay, 86% of all PCR products fixed on the glass slides were confirmed to present a hybridization signal greater than twice the standard deviation of the deviation of the global median signal-to-noise ration. Conclusions. Our validation of the XACArray platform using DNA-DNA hybridization revealed that it can be used to evaluate the expression of 2,365 individual CDSs from all major functional categories, which corresponds to 52.7% of the annotated CDSs of the XAC genome. As a proof of concept, we used this platform in a previously work to verify the absence of genomic regions that could not be detected by sequencing in related strains of Xanthomonas. © 2010 Moreira et al; licensee BioMed Central Ltd.

Antibacterial activity of alkyl gallates against Xanthomonas citri subsp. citri

The plant-pathogenic bacterium Xanthomonas citri subsp. citri is the causal agent of Asiatic citrus canker, a seriousdisease that affects all the cultivars of citrus in subtropical citrus-producing areas worldwide. There is no curative treatment for citrus canker; thus, the eradication of infected plants constitutes the only effective control of the spread ofX. citri subsp. citri. Since the eradication program in the state of São Paulo, Brazil, is under threat, there is a clear risk of X. citri subsp. citri becoming endemic in the main orange-producing area in the world. Here we evaluated the potential use of alkyl gallates to prevent X. citri subsp. citri growth. These esters displayed a potent anti-X. citri subsp. citri activity similar to that of kanamycin (positive control), as evaluated by the resazurin microtiter assay (REMA). Thetreatment of X. citri subsp. citri cells with these compounds induced altered cell morphology, and investigations of the possible intracellular targets using X. citri subsp. citri strains labeled for the septum and centromere pointed to a commontarget involved in chromosome segregation and cell division. Finally, the artificial inoculation of citrus with X. citri subsp. citri cells pretreated with alkyl gallates showed that the bacterium loses the ability to colonize its host, which indicates the potential of these esters to protect citrus plants against X. citri subsp. citri infection. © 2013, American Society for Microbiology.