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.

Produtividade e resistência à ferrugem do cafeeiro (Hemileia vastatrix BERK. ET BR.) de progênies F5 de catuí amarelo com o híbrido de timor

Foram estudadas 18 progênies na geração F5 do híbrido H 419, provenientes do cruzamento entre o Híbrido de Timor (UFV 445-46) e o Catuaí Amarelo IAC 30 (UFV 2143), 5 progênies na geração F5 do híbrido H 516, proveniente do cruzamento entre o Híbrido de Timor (UFV 446-08) e o Catuaí Amarelo IAC 86 (UFV 2154) e como controle o Catuaí Vermelho IAC 44 ( UFV 2144 ) com e sem controle da ferrugem. em relação a produção de café cereja por planta e ao ataque da ferrugem as progênies H419-3-1-1-14 e a H516-2-1-1-18, foram as que apresentaram maiores produtividades e resistência a ferrugem ( nota 1 ), seguidas da H419-6-3-6-12, porém com resistência parcial à ferrugem. Essas progênies também foram as que apresentaram maior comprimento do ramo plagiotrópico, que influenciou em 37,73% (R²) na produção de café cereja por planta. O comprimento do ramo plagiotrópico, diâmetro do tronco e altura das plantas foram os atributos que mais correlacionaram r= 0.5977, r= 0.3316 e r= 0.2848, respectivamente p< 0.01, com as produtividades dessas progênies, concordando com resultados obtidos por Dhaliwal (1968). em relação a resistência ao fungo Hemileia vastatrix Berk. et Br.,as progênies apresentaram herdabilidade no sentido amplo elevada (h a² = 0,80 entre e h a² = 0,96 entre e dentro das progênies), mostrando variabilidade genética alta para seleção de material resistentes à ferrugem do cafeeiro.

Adubação nitrogenada e substratos na produção de mudas de maracujazeiro doce

Com o objetivo de avaliar diferentes substratos e doses de nitrogênio na produção de mudas de maracujazeiro doce, conduziu-se um experimento no Setor de Fruticultura do Departamento de Agricultura da Universidade Federal de Lavras (UFLA). Foram estudadas cinco doses de nitrogênio 0; 400; 800; 1600 e 3200 mg de N/dm³ e duas composições de substratos: A (composto orgânico + areia + solo na proporção de 1:1:3 em volume) e B (Plantmax® + areia + solo na proporção de 1:1:3 em volume ). Foi utilizado o delineamento experimental em blocos casualizados em esquema fatorial 5 x 2, com quatro repetições e cinco plantas por parcela. Foi observado que doses elevadas de adubação de cobertura com N podem promover efeitos depressivos nas mudas. E que o melhor substrato foi o B, contendo: Plantmax® + areia + solo na proporção de 1:1:3 em volume.

Polinização em cultura de laranja (Citrus sinensis L. Osbeck, var. Pera-Rio)

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

Isoenzymatic polymorphism in Citrus spp. and Poncirus trifoliata (L.) Raf. (Rutaceae)

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)

Avaliação de fontes de amônia para o tratamento de fenos de gramíneas tropicais. 1. Constituintes da parede celular, poder tampão e atividade ureática

Avaliaram-se as alterações da fração fibrosa e as características químicas dos fenos de braquiária decumbens (Brachiaria decumbnes Stapf) e jaraguá (Hyparrhenia rufa Ness Stapf) não-tratados, tratados com uréia (U-5,4% da MS), uréia (UL-5,4% da MS) mais labe-labe (Lablab purpureus L. Sweet, cv. Highworth-3,0% da MS) ou amônia anidra (NH3 -3,0% da MS). O tratamento químico com uréia ou NH3 aumentou o pH e a digestibilidade in vitro verdadeira dos fenos. A amonização não alterou os teores de fibra em detergente ácido e celulose, mas diminuiu os de fibra em detergente neutro, hemicelulose e lignina. O uso do labe-labe como fonte adicional de urease não aumentou a eficiência da uréia no tratamento dos volumosos. As avaliações do conteúdo de umidade, do poder tampão e da atividade ureática são técnicas que podem auxiliar na previsão das respostas dos volumosos à amonização com o uso de uréia.

Avaliação de fontes de amônia para o tratamento de fenos de gramíneas tropicais. 2. Compostos nitrogenados

Este estudo foi desenvolvido para se avaliarem as alterações nos conteúdos de compostos nitrogenados dos fenos de braquiária decumbens (Brachiaria decumbnes Stapf) e jaraguá (Hyparrhenia rufa Ness Stapf) não-tratados, tratados com uréia (U - 5,4% da MS), uréia (UL - 5,4% da MS) mais labe-labe (Lablab purpureus L. Sweet, cv. Highworth-3,0% da MS) ou amônia anidra (NH3 -3,0% da MS). A aplicação de amônia anidra ou de uréia aumentou os teores de N total, N insolúvel em detergente neutro, N insolúvel em detergente ácido, N não-protéico e N amoniacal. A amonização diminuiu as relações N insolúvel em detergente neutro/N total e N insolúvel em detergente ácido/N total e aumentou as relações N não-protéico/N total, N amoniacal/N total e os teores de PB. O N aplicado foi retido, principalmente, nas formas de NNP e N amoniacal.

Mites (Arachnida, Acari) on Citrus sinensis L. Osbeck orange trees in the state of Amazonas, Northern Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Influence of two methods of CaCl2 application and concentrations used to maintain passion fruit firmness

This is a report on the influence of the methods of postharvest application of CaCl2 in several concentrations, on the maintenance of the firmness of the sweet passion fruit (Passiflora alata Dryander). The fruits, picked in the pre-climateric stage, were selected desinfection with thiabendazol and submitted to CaCl2 in concentrations of 0% (control), 1%, 2%, 3% or 4% with two methods: immersion (for 2 h) and infiltration under vacuum and pressure (- 25 vacuum kPa, for 1 minute and, + 25 pressure kPa, for 1 min). All fruits were stored under refrigeration (9 degreesC, 85-90% RH), for 30 d. The fruits were evaluated for texture every 3 d. From the results, it is concluded that application of CaCl2 by infiltration favored the maintenance of fruit consistency during the first 18 days of storage. Fruits treated with 1% and 2% of CaCl2 were firmer than the others.

Ineffectiveness of pruning to control citrus huanglongbing caused by Candidatus Liberibacter americanus

The huanglongbing (HLB) disease of citrus trees, caused by Candidatus Liberibacter asiaticus and Ca. Liberibacter americanus, was first reported in Brazil in March, 2004. The presence of the disease has caused serious concerns among growers. Pruning experiments were conducted to determine if removal of symptomatic branches or the entire canopy (decapitation) would eliminate infected tissues and save HLB-affected trees. Pruning was done in five blocks on a total of 592 3- to 16 year-old 'Valencia', 'Hamlin' or 'Pera' sweet orange trees showing no symptoms or with two levels of symptom severity. Ten decapitated trees per block were caged and all trees were treated with insecticides to control the psyllid vector, Diaphorina citri. Mottled leaves reappeared on most symptomatic (69.2%) as well on some asymptomatic (7.6%) pruned trees, regardless of age, variety, and pruning procedure. Presence of the pathogen (Ca. Liberibacter americanus) in all symptomatic trees was confirmed by PCR. In general, the greater the symptom severity before pruning the lower the percentage of trees that remained asymptomatic after pruning.

An experimental inoculation system to study citrus-Xylella fastidiosa interactions

Difficulties in reproducing the citrus variegated chlorosis (CVC) disease symptoms in expertmental plants have delayed implementation of studies to better understand the essential aspects of this important disease. In an extensive Study, cultivars of sweet orange (Citrus sinensis) were inoculated with Xylella fastidiosa using procedures that included root immersion, and stein absorption, pricking, or infiltration of the inoculum into plants of different ages. Inoculum consisted of 5-day-old cultures or cell suspensions of CVC strain 9a5c diluted in phosphate-buffered saline. Inoculated plants and controls were grown, or transferred just after inoculation, to 5-liter pots or 72-cell foam trays. Approximately 4, 5, 9, and 12 months after inoculation, leaves were collected and processed for polymerase chain reaction analysis or X. fastidiosa isolation on BCYE agar medium. Root immersion and stem inoculation of 4- and 6-month-old plants resulted in low percentages of symptomatic (0 to 7%) and plants positive by isolation (0 to 9%). Pinpricked or injected stems of I-month-old seedlings resulted in high percentages of plants symptomatic (29 and 90% in Pera Rio, 75, 59, and 83% in Valencia, and 77% in Natal) or positive by isolation (26 and 93% in Pera Rio, 98, 96, and 83% in Valencia, and 77% in Natal), In foam trays, the seedlings grew less, the incubation period was shorter. and disease severity was higher than in pots. This system allows testing of higher numbers of plants in a reduced space with a more precise reproduction of the experimental conditions.

An evaluation of the genetic diversity of Xylella fastidiosa isolated from diseased citrus and coffee in São Paulo, Brazil

Strains of Xylella fastidiosa, isolated from sweet orange trees (Citrus sinensis) and coffee trees (Coffea arabica) with symptoms of citrus variegated chlorosis and Requeima do Cafe, respectively, were indistinguish able based on repetitive extragenic palindromic polymerase chain reaction (PCR) and enterobacterial repetitive intergenic consensus PCR assays. These strains were also indistinguishable with a previously described PCR assay that distinguished the citrus strains from all other strains of Xylella fastidiosa. Because we were not able to document any genomic diversity in our collection of Xylella fastidiosa strains isolated from diseased citrus, the observed gradient of increasing disease severity from southern to northern regions of São Paulo State is unlikely due to the presence of significantly different strains of the pathogen in the different regions. When comparisons were made to reference strains of Xylella fastidiosa isolated from other hosts using these methods, four groups were consistently identified consistent with the hosts and regions from which the strains originated: citrus and coffee, grapevine and almond, mulberry, and elm, plum, and oak. Independent results from random amplified polymorphic DNA (RAPD) PCR assays were also consistent with these results; however, two of the primers tested in RAPD-PCR were able to distinguish the coffee and citrus strains. Sequence comparisons of a PCR product amplified from all strains of Xylella fastidiosa confirmed the presence of a CfoI polymorphism that can be used to distinguish the citrus strains from all others. The ability to distinguish Xylella fastidiosa strains from citrus and coffee with a PCR-based assay will be useful in epidemiological and etiological studies of this pathogen.

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.

Recombinant expression and characterization of a Xylella fastidiosa cysteine protease differentially expressed in a nonpathogenic strain

Xylella fastidiosa is a xylem-limited, Gram-negative bacterium responsible for citrus variegated chlorosis (CVC) in sweet oranges. In the present study, we present the recombinant expression, purification and characterization of an X. fastidiosa cysteine protease (dubbed Xylellain). The recombinant Xylellain ((HIS)Xylellain) was able to hydrolyze carbobenzoxy-Phe-Arg-7-amido-4-methylcoumarin (Z-FR-MCA) and carbobenzoxy-Arg-Arg-7-amido-4-methylcoumarin (Z-RR-MCA) with similar catalytic efficiencies, suggesting that this enzyme presents substrate specificity requirements similar to cathepsin B. The immunization of mice with (HIS)Xylellain provided us with antibodies, which recognized a protein of c. 31 kDa in the X. fastidiosa pathogenic strains 9a5c, and X. fastidiosa isolated from coffee plants. However, these antibodies recognized no protein in the nonpathogenic X. fastidiosa J1a12, suggesting the absence or low expression of this protein in the strain. These findings enabled us to identify Xylellain as a putative target for combating CVC and other diseases caused by X. fastidiosa strains.