Breakdown of resistance in sweet pepper against Pepper yellow mosaic virus in Brazil

Plantas de Capsicum annuum cv. Magali R, resistentes ao Pepper yellow mosaic virus (PepYMV), exibindo sintomas severos de mosaico amarelo, malformação foliar e subdesenvolvimento foram encontradas em plantios na região de Lins, SP, Brasil, em 2003/04. Partículas semelhantes àquelas do gênero Potyvirus foram observadas em extrato foliar de planta infectada examinado em microscópio eletrônico de transmissão. O extrato foliar também reagiu com anti-soro contra o PepYMV em PTA-ELISA. Além de C. annuum cv. Magali R, esse potyvirus também infectou sistemicamente C. annuum cv. Rubia R, que é resistente ao PepYMV. A seqüência de nucleotídeos de parte do gene da proteína capsidial (CP) desse potyvirus apresentou 96-98% de identidade com a de outros isolados do PepYMV. A seqüência parcial de nucleotídeos da região 3' não traduzida (3' NTR) apresentou 94-96% de identidade com a do PepYMV. Esses resultados são indicativos de que o potyvirus que quebrou a resistência em pimentão é um isolado do PepYMV.

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 plásticos 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 líquida de reprodução (Ro) de 41,0.

Cultivo de embriões imaturos de citros em diferentes concentrações de carvão ativado e ácido giberélico

Adição de carvão ativado e giberelina no meio de cultura podem proporcionar melhores condições no desenvolvimento de embriões imaturos de citros. Objetivou-se avaliar o efeito de carvão ativado e GA3 (ácido giberélico) no cultivo de embriões imaturos provenientes do cruzamento entre laranjeira 'Pêra Rio' x tangerineira 'Poncã'. Após 118 dias da polinização, frutos imaturos, com 3 a 4 cm de diâmetro, foram coletados, suas sementes removidas e tratadas com álcool (70%) por cinco minutos, hipoclorito de sódio (2%) por 20 minutos e, posteriormente, lavadas três vezes em água destilada e autoclavada. em condições assépticas, os tegumentos das sementes foram separados, os embriões globulares excisados e inoculados em tubos de ensaio contendo 15 mL do meio MT, acrescido de carvão ativado (0; 0,5; 1; 1,5 e 2 g L-1) e GA3 (0; 0,01; 0,1; 1 e 10 mg L-1). Após a inoculação, os embriões permaneceram por 90 dias em sala de crescimento a 27+1ºC, fotoperíodo de 16 horas e irradiância de 32 mmol m-2 s-1. Maior comprimento da parte aérea foi obtido em meio MT, acrescido de 0,1 e 1 mg L-1 de GA3, combinado com 2 g L-1 de carvão ativado. Maior comprimento do sistema radicular, massa da matéria fresca e número de folhas de plântulas foram obtidos em meio MT, acrescido de 0,01 mg L-1 de GA3, na ausência de carvão ativado. A adição de carvão ativado influenciou na concentração de ácido giberélico acrescido no meio de cultura.

Nitrogen assimilation in Citrus based on CitEST data mining

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

PR gene families of citrus: their organ specific-biotic and abiotic inducible expression profiles based on ESTs approach

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Phytophthora parasitica transcriptome, a new concept in the understanding of the citrus gummosis

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

Distribution and biological role of the oligopeptide-binding protein (OppA) in Xanthomonas species

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

Dry matter production of shoots and root density of two cultivars of Lablab purpureus (L.) Sweet

This experiment was conducted in green house conditions to evaluate the DM accumulation in the shoots and in the roots of two cultivars of Lablab purpureus (L.) Sweet. A 2x3 factorial (two cultivars and three evaluation dates) was conducted according to a randomized complete block design with four replications, being the cultivars Highworth and Rongai evaluated at 42, 56, and 70 days after seedling emergence (DASE). The results indicated that the cvs. Highworth and Rongai have the same pattern of DM accumulation in the shoots. In the upper layer of the soil (0-0.20 in) it was found 38.83% and 43.64% of the DM accumulated in the roots down to 2.00 in depth, in the cvs. Highworth and Rongai, respectively. In the deepest layer (1.80-2.00 in) it was found 3.02% and 1.5% of the DM accumulated in the roots of the cvs. Highworth and Rongai, respectively. The root density showed a striking decrease upper layer from the soil (0-0.2 m) down to the depth of 0.60 0.80 in (from 10.83 to 1.75 cm.cm(-3) in the cv. Highworth and from 10.76 to 1.28 cm.cm(-3) in the cv. Rongai). At the bottom layer (1.80-2.00 in) the root density values were 0.98 cm.cm(-3) and 0.59 cm.cm(-3), respectively for the cvs. Highworth and Rongai. The root/shoot ratios were similar in both cvs. and decreased from 42 to 70 DASE showing that the cvs. evaluated had the same dynamics of DM accumulation.

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.

Identification of QTLs associated with citrus resistance to Phytophthora gummosis

Citrus gummosis, caused by Phylophthora spp., is an important citrus disease in Brazil. Almost all citrus rootstock varieties are susceptible to it to some degree, whereas resistance is present in Poncirus trifoliata, a closely related species. The objective of this study was to detect QTLs linked to citrus Phylophthora gummosis resistance. Eighty individuals of the F, progeny, obtained by controlled crosses between Sunki mandarin Citrus sunki (susceptible) and Poncirus trifoliata cv. Rubidoux (resistant), were evaluated. Resistance to Phytophthora parasitica was evaluated by inoculating stems of young plants with a disc of fungal mycelia and measuring lesion lengths a month later. Two QTLs linked to gummosis resistance were detected in linkage groups I and 5 of the P. trifoliala map, and one QTL in linkage group 2 of the C sunki map. The phenotypic variation explained by individual QTLs was 14% for C sunki and ranged from 16 to 24% for P. trifoliala. The low character heritability (h(2) = 18.7%) and the detection of more than one QTL associated with citrus Phytophthora gummosis resistance showed that inheritance of the resistance is quantitative.

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.

Evaluation of the genetic diversity of Xylella fastidiosa strains from citrus and coffee hosts by single-nucleotide polymorphism markers

The aim of this study was to obtain information about genetic diversity and make some inferences about the relationship of 27 strains of Xylella fastidiosa from different hosts and distinct geographical areas. Single-nucleotide polymorphism (SNP) molecular markers were identified in DNA sequences from 16 distinct regions of the genome of 24 strains of X. fastidiosa from coffee and citrus plants. Among the Brazilian strains, coffee-dependent strains have a greater number of SNPs (10 to 24 SNPs) than the citrus-based strains (2 to 12 SNPs); all the strains were compared with the sequenced strain 9a5c. The identified SNP markers were able to distinguish, for the first time, strains from citrus plants and coffee and showed that strains from coffee present higher genetic diversity than the others. These markers also have proven to be efficient for discriminating strains from the same host obtained from different geographic regions. X. fastidiosa, the causal agent of citrus variegated chlorosis, possesses genetic diversity, and the SNP markers were highly efficient for discriminating genetically close organisms.