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Abstract The objective of this work was to evaluate the photosynthetic response of citrus varieties to the inoculation of isolates T30 and T3 of Citrus tristeza virus (CTV) characterized as having potential to provide cross-protection against severe CTV isolates in citrus. Citrus plants of 'Campbell' orange, 'Persian' lime, and 'Key' lime were subjected to inoculations by both isolates by patch. Noninoculated plants were used as the control. Symptom expression, photosynthetic and transpiration rates, and stomatal conductance were evaluated by IRGA. Stomatal density and size were evaluated by epidermal impressions. The CTV isolates were also evaluated and molecularly characterized by RT-PCR and Sanger sequencing. 'Campbell' orange and 'Persian' lime plants remained asymptomatic after the inoculations. The inoculation treatments with both CTV isolates did not reduce photosynthetic capacity, transpiration rate, and stomatal conductance, in comparison with the control. Stomatal density and size varied according to the citrus species. T30 and T3 do not affect the photosynthetic responses of 'Campbell' orange and 'Persian' lime; therefore, these isolates have the potential to provide cross-protection to citrus varieties against severe isolates of CTV.

Various stresses are commonly encountered by probiotics in their ecological niches. Effects of adaptation treatments on the heat tolerance of Lactobacillus plantarum KLDS 1.0628 were evaluated. Cells pre-adapted to heat, cold, oxidative, acid, bile salt, and osmotic stresses were evaluated for tolerance and cross-protection against subsequent challenge. Except for cold, pre-adaptation to sublethal levels of stress significantly improved the cell viability after heat challenge compared to the control. Heat-adapted cells showed the highest heat stress tolerance, with their tolerance factor against heat challenge reaching 31.38-fold after pre-adaptation at 45°C for 1 h. Moreover, adaptation treatments improved tolerance to the same stress and evoked cross-protection against heterologous challenges, inducing a significantly lower ratio of saturated to unsaturated fatty acid content in the cell membrane of L. plantarum KLDS 1.0628. The results indicated that adaptation treatments are potential strategies for enhancing the survival rate of probiotics and exerting their functional properties.

The use of tolerant rootstocks and preimmunization has satisfactorily controlled losses associated with the Citrus tristeza virus (CTV). Several researchers have shown that CTV mild isolates that are selected in the same region where they are used are superior to isolates obtained from other areas. Thus, budwoods of 20 outstanding citrus trees were collected in north and northwestern Paraná state (Brazil) citrus-producing areas and established to be used in a preimmunization program. These budwoods were tested to evaluate the potential protection of the inherently present viral complex. Based on biological indexing and molecular characterization of the capsid protein gene by RFLP (restriction fragment length polymorphism, which indicated that the plants were infected with mild isolates of CTV), some of the selected plants could be used in a preimmunization program. These potentially mild and protective isolates were challenged with severe 'Rolândia' isolate inoculations by grafting and by the brown citrus aphid (Toxoptera citricida Kirkaldy) vector, which was faster in transmitting the virus. Some isolates had a better protective value than others, particularly when challenged with the severe CTV isolate. The SSCP (single strand conformational polymorphism) molecular analysis was an excellent complementary tool for monitoring the performance of the experiments and the stability of the viral complex present in the plants. Isolate number 1, collected in the municipality of Cruzeiro do Sul (CS-1), was the most promising for protecting commercial Pêra sweet orange (C. sinensis L.) orchards in northern and northwestern Paraná. The Rolândia severe CTV isolate was stable and had a high genetic divergence among the severe isolates used as a control (Capão Bonito and Barão B) and all of the isolates tested.

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.O objetivo deste trabalho foi monitorar a manutenção da estabilidade de isolados protetores contra Citrus tristeza virus (CTV) em clones selecionados de laranja 'Pêra' (Citrus sinensis) pré-imunizados ou infectados naturalmente pelo vírus, após sucessivas propagações clonais. O trabalho foi realizado em condições de campo, no norte do Estado do Paraná. A análise do gene da capa protéica (GPC) de 33 isolados, coletados de 16 clones de laranjeira 'Pêra', foi realizada com o uso da técnica polimorfismo conformacional da fita simples (SSCP). Inicialmente, os isolados foram caracterizados por meio de sintomas de caneluras observados nos clones. Em seguida, o genoma viral foi extraído e utilizado como molde para a amplificação do GCP com uso da transcrição reversa da reação em cadeia da polimerase (RTPCR). Os perfis eletroforéticos dos produtos da RTPCR foram analisados com emprego do coeficiente de Jaccard e do método UPGMA. A maioria dos clones apresentou sintomas fracos a moderados de caneluras, bem como alterações nos perfis dos isolados de CTV. Contudo, a estabilidade dos complexos protetores foi mantida, com exceção dos isolados presentes em dois dos clones analisados. Foi observada baixa variabilidade genética nos isolados durante os anos avaliados.