Seasonal effects on the relationship between photosynthesis and leaf carbohydrates in orange trees

To understand the effect of summer and winter on the relationships between leaf carbohydrate and photosynthesis in citrus trees growing in subtropical conditions, 'Valencia' orange trees were subjected to external manipulation of their carbohydrate concentration by exposing them to darkness and evaluating the maximal photosynthetic capacity. In addition, the relationships between carbohydrate and photosynthesis in the citrus leaves were studied under natural conditions. Exposing the leaves to dark conditions decreased the carbohydrate concentration and increased photosynthesis in both seasons, which is in accordance with the current model of carbohydrate regulation. Significant negative correlations were found between total non-structural carbohydrates and photosynthesis in both seasons. However, non-reducing sugars were the most important carbohydrate that apparently regulated photosynthesis on a typical summer day, whereas starch was important on a typical winter day. As a novelty, photosynthesis stimulation by carbohydrate consumption was approximately three times higher during the summer, i.e. the growing season. Under subtropical conditions, citrus leaves exhibited relatively high photosynthesis and high carbohydrate levels on the summer day, as well as a high nocturnal consumption of starch and soluble sugars. A positive association was determined between photosynthesis and photoassimilate consumption/exportation, even in leaves showing a high carbohydrate concentration. This paper provides evidence that photosynthesis in citrus leaves is regulated by an increase in sink demand rather than by the absolute carbohydrate concentration in leaves.

Genetic transformation of three sweet orange cultivars from explants of adult plants

The difficulty in adult tissue genetic transformation in woody species is still an obstacle to be overcome, including in most sweet orange cultivars of the Brazilian citrus industry. This work reports that, after in vitro culture adjustments, transgenic adventitious buds of 'Hamlin', 'Pra', and 'Valencia' sweet oranges (Citrus sinensis L. Osbeck) were recovered using adult material as explant source, in genetic transformation experiments via Agrobacterium tumefaciens. The transgenic buds were identified by the GUS histochemical analysis and confirmed by PCR analysis, which indicated the presence of an amplified fragment of 817 bp corresponding to the uidA gene sequence. The efficiencies of genetic transformation for 'Hamlin', 'Pra', and 'Valencia' sweet orange cultivars were 2.5, 1.4, and 3.7%, respectively. Media supplemented with auxins and cytokinins during co-culture, and media with high concentrations of cytokinins (3 mg L-1) during transgenic selection led to the transformation and, consequently, the regeneration of adequate number of adventitious buds for the three cultivars. The use of sonication during the explant disinfection was not effective to reduce endophytic contamination and reduced transformation efficiency.

Selection of early-maturing sweet orange cultivars by performance indexes

The objective of this work was to select adequate early-maturing sweet orange cultivars for the fresh fruit market and for industrial processing using performance indexes. Performance indexes for citrus were established from data collected in an experiment carried out in the southwest region of the state of Sao Paulo, involving 12 early-maturing sweet orange cultivars. New results were obtained by identifying cultivars with superior characteristics. In a comparison with 'Hamlin' sweet orange, a standard early-maturing cultivar, 'Valencia 2' and 'Salustiana' were considered better materials for the fresh fruit market, whereas 'Westin' sweet orange was identified as a superior cultivar for orange juice processing.

Somatic embryogenesis from ovaries of sweet orange cv. Tobias

Callogenesis, somatic embryogenesis, and regeneration were obtained from tissues of unfertilized ovaries of sweet orange (Citrus sinensis Osbeck.) cv. Tobias. The influence of two modified basal media, woody plant medium (WPM) and N6 medium, to induce callus formation from pistils was determined. Overall, high frequencies of callogenesis were observed when either medium was used. However, initial culture of explants in WPM medium followed by transfer of callus to N6 medium resulted in higher frequency of callus induction (of 2.30 callus per explant that were larger than 0.5 cm in size), and of subsequent development of embryogenic callus (10%). A total of 125 somatic embryos were obtained. After 6 months of culture, 72% of somatic embryos germinated into plantlets. These plantlets were subsequently micrografted in vitro, and then acclimatized. Ploidy of these plants were determined using flow cytometry and TRAPS molecular markers were used to confirm their maternal origin.

Período de incubação de Guignardia citricarpa em diferentes estádios fenológicos de frutos de laranjeira 'Valência'

A mancha preta dos citros (MPC), causada pelo fungo Guignardia citricarpa, produz lesões em frutos, os quais ficam depreciados para o mercado interno e os restringem para a exportação. O grande período de suscetibilidade dos frutos cítricos, em adição ao fato de G. citricarpa causar infecções latentes, dificulta o entendimento sobre o período de incubação da doença. O objetivo do trabalho foi determinar o período de incubação da MPC inoculando frutos de laranjeira 'Valência' em diferentes estádios fenológicos. Para a inoculação foram empregadas suspensões de conídios de G. citricarpa (10³, 10(4), 10(5) e 10(6) conídios mL-1) em diferentes diâmetros dos frutos (1,5; 2,0; 2,5; 3,0; 5,0 e 7,0 cm). O período de incubação da MPC para os diferentes diâmetros dos frutos inoculados apresentou uma relação polinomial negativa. Em frutos com até 3 cm de diâmetro o período de incubação médio foi superior a 200 dias, enquanto que em frutos com diâmetros superiores a 5 cm o período de incubação médio foi inferior a 84 dias. A MPC apresenta período de incubação variável dependente do estádio fenológico em que os frutos são infectados. A concentração de conídios de G. citricarpa, na infecção, não interfere no período de incubação da doença.

Elsinoe australis; Myriangiaceae

Elsinoe australis Bitanc. & Jenkins

Citricultura desenvolvida na agricultura de base familiar do município de Russas, Ceará.

Brasil é o maior produtor mundial de laranjas doces [Citrus sinensis (L.) Osbeck]. A citricultura distribui-se nacionalmente, enquanto o Nordeste constitui-se segundo maior produtor (IBGE, 2015). Russas sobresaiu-se historicamente pelo extrativismo da carnaúba (Copernícia prunífera) ecultivo da laranja doce, recebendo o título "Terra da Laranja Doce", devido ao dominio histórico de uma variedade nativa,chamada ?Laranja de Russas?. Porém, entre as cheias que ocorreram nos anos de 1974 e 1985, a ?gomose? (Phytophthora spp.) proliferou-se e atingiu 90% dos pomares citricos do município, constituídos da ?laranja de Russas? propagada seminalmente (pé franco), levando ao declínio da cadeia produtiva, que desenvolvia-se basicamente através da agricultura familiar. Recentemente, uma parceria entre Embrapa Mandioca e Fruticultura Tropical e Secretaria de Agricultura (SEAGRI-RUSSAS), inicou pesquisas voltadas a revitalização da citricultura no município, através da seleção de exemplares da laranja de ?Russas?, identificando-se oito clones de ?Russas? de elevada produtividade e fitossanidade (PASSOS et. al, 2013); introdução de porta-enxertos híbridos com tendências ao nanismo e resistentes as principais doenças, principalmente ?Gomose?; além da implantação de pomares na agricultura familiar e assistência técnica. Portanto, o presente trabalho objetivou-se alevantar e caracterizar os principais aspectos e técnicas empregadas pelos agricultores familiares de Russas na implantação e desenvolvimento inicial dos pomares cítricos.

Caracterização da rebentação da tangerineira ‘Setubalense’ em anos de safra e de contrassafra

A produção de citrinos encontra-se amplamente disseminada por todos os continentes, representando uma atividade económica geradora de milhões de euros. As crescentes exigências dos consumidores conduzem ao aparecimento de novas variedades, em detrimento das cultivares tradicionais. A produção de tangerineira ‘Setubalense’ tem vindo a sofrer um acentuado declínio, mas os consumidores mais exigentes em aromas cítricos e fragrâncias intensas continuam a apreciar esta cultivar. Por isso, esta cultivar tem algum espaço comercial para mercados diferenciados em que se valorizam as cultivares tradicionais. Estas características e a sua época de maturação, que implica ser pouco afetada pela Ceratitis capitata, fazem da ‘Setubalense’ uma cultivar recomendável para o modo de produção biológico. Com o objetivo de caracterizar o padrão produtivo alternante e tendo por objeto de estudo um pomar de 5 anos de tangerineira ‘Setubalense’ enxertado sobre citranjeira ‘Troyer’ [Citrus sinensis (L.) Osbeck x Poncirus trifoliata L.], foi avaliada a composição da rebentação e da floração nos anos de safra e de contrassafra. A intensidade e as características da rebentação dependem do quadrante da árvore e do ano (safra ou contrassafra). Nos anos de safra a percentagem de nós rebentados oscila entre 67,6 a 79,5 %, que contrasta com os 25,4 a 28,2 % dos anos de contrassafra, função do quadrante analisado. Nos anos produtivos os rebentos formados são essencialmente florais e nos improdutivos quase exclusivamente vegetativos. A formação de elevado número de flores nos anos de safra conduz a uma intensa abcisão de órgãos generativos. O número de folhas formada por cada 100 nós é similar nos anos de safra e de contrassafra. Os resultados obtidos refletem o padrão alternante desta cultivar, evidenciando o cuidado particular que esta cultura tem de ter na sua condução.

Endophytic population of Pantoea agglomerans in citrus plants and development of a cloning vector for endophytes

Harmless bacteria inhabiting inner plant tissues are termed endophytes. Population fluctuations in the endophytic bacterium Pantoea agglomerans associated with two species of field cultured citrus plants were monitored over a two-year period. The results demonstrated that populations of P. agglomerans fluctuated in Citrus reticulata but not C. sinensis. A cryptic plasmid pPA3.0 (2.9 kb) was identified in 35 out of 44 endophytic isolates of P. agglomerans and was subsequently sequenced. The origins of replication were identified and nine out of 18 open reading frames (ORFs) revealed homology with described proteins. Notably, two ORFs were related to cellular transport systems and plasmid maintenance. Plasmid pPA3.0 was cloned and the gfp gene inserted to generate the pPAGFP vector. The vector was introduced into P. agglomerans isolates and revealed stability was dependent on the isolate genotype, ninety-percent stability values were reached after 60 hours of bacterial cultivation in most evaluated isolates. In order to definitively establish P. agglomerans as an endophyte, the non-transformed bacterium was reintroduced into in vitro cultivated seedlings and the density of inner tissue colonization in inoculated plants was estimated by bacterium re-isolation, while the tissue niches preferred by the bacterium were investigated by scanning electronic microscopy (SEM). Cells from P. agglomerans (strain ARB18) at similar densities were re-isolated from roots, stems and leaves and colonization of parenchyma and xylem tissues were observed. Data suggested that P. agglomerans is a ubiquitous citrus endophyte harboring cryptic plasmids. These characteristics suggest the potential to use the bacterium as a vehicle to introduce new genes in host plants via endophytic bacterial transformation.

Citrus asymmetric somatic hybrids produced via fusion of gamma-irradiated and iodoacetamide-treated protoplasts

The objective of this study was to produce citrus somatic asymmetric hybrids by fusing gamma-irradiated protoplasts with iodoacetamide-treated protoplasts. Protoplasts were isolated from embryogenic suspension cells of grapefruit (Citrus paradisi Macfad.) cultivars Ruby Red and Flame, sweet oranges (C. sinensis Osbeck) 'Itaboraí', 'Natal', Valencia', and 'Succari', from 'Satsuma' (C. unshiu Marcow.) and 'Changsha' mandarin (C. reticulata Blanco) and 'Murcott' tangor (C. reticulata x C. sinensis). Donor protoplasts were exposed to gamma rays and receptor protoplasts were treated with 3 mmol L-1 iodoacetamide (IOA), and then they were fused for asymmetric hybridization. Asymmetric embryos were germinated, and the resulting shoots were either grafted onto sour orange, rough lemon or 'Swingle' (C. paradisi x Poncirus trifoliata) x 'Sunki' mandarin rootstock seedlings, or rooted after dipping their bases in indol-butyric acid (IBA) solution. The products were later acclimatized to greenhouse conditions. Ploidy was analyzed by flow cytometry, and hybridity was confirmed by amplified fragment length polymorphism (AFLP) analysis of plantlet DNAsamples. The best treatment was the donor-recipient fusion combination of 80 Gy-irradiated 'Ruby Red' protoplasts with 20 min IOA-treated 'Succari' protoplasts. Tetraploid and aneuploid plants were produced. Rooting recalcitrance was solved by dipping shoots' stems in 3,000 mg L-1 IBA solution for 10 min.

Growth regulators, culture media and antibiotics in the in vitro shoot regeneration from mature tissue of citrus cultivars

The objective of this study was to evaluate the effects of 6-benzylaminopurine (BAP) and α-naphthaleneacetic acid (NAA) combinations, basal media and beta-lactam antibiotics on in vitro organogenesis from mature stem segments of 'Pêra', 'Valência' and 'Bahia' sweet oranges and 'Cravo' rangpur lime. For induction of shoot regeneration, the segments of the four cultivars were placed on Murashige and Skoog (MS) medium containing the following BAP/NAA concentrations: 0.0/0.0; 0.25/0.0; 0.25/0.25; 0.5/0.0; 0.5/0.5; 1.0/0.0; 2.0/0.0; 2.0/0.25; 2.0/0.5; and 2.0/1.0 mg L-1. In order to test the influence of the culture media on shoot-bud induction, (MS), Murashige and Tucker (MT), and woody plant medium (WPM) formulations were evaluated, associated with the best combination of plant growth regulators obtained in the previous experiment. The influence of four beta-lactam antibiotics (timentin, cefotaxime sodium salt, meropenem trihydrate and augmentin) on shoot regeneration was determined. Better regeneration responses were achieved when internodal segments were cultured onto MS-based medium with 500 mg L-1 cefotaxime with the following BAP/NAA concentrations: 0.5 + 0.25 mg L-1 for 'Cravo', 1.0 + 0.25 mg L-1 for 'Valência' and 'Bahia', and 1.0 + 0.5 mg L-1 for 'Pêra'. Genotype, growth regulators, basal media and beta-lactam antibiotics affect the morphogenetic response in mature tissues of citrus.

Organogênese in vitro a partir de diferentes regiões do epicótilo de Citrus sp

O estabelecimento de protocolos para regeneração de plantas in vitro é essencial para o uso de técnicas de transformação genética no melhoramento de citros. Visando à obtenção de um protocolo eficiente de regeneração in vitro para laranja-azeda (Citrus aurantium), laranjas 'Natal' e 'Pêra' (C. sinensis), limão 'Volkameriano' (C. volkameriana) e citrange 'Carrizo' (Poncirus trifoliata x C. sinensis), avaliou-se a resposta morfogênica de diferentes regiões do epicótilo (basal, mediana e apical) em relação a distância do nó cotiledonar, na presença (1,0 mg/L-1) ou ausência de 6-BAP, em meio de cultura MT. Após 60 dias, avaliaram-se a porcentagem de explantes responsivos e o número de gemas adventícias por explante. A resposta morfogênica em função da região do epicótilo e da presença ou ausência da citocinina (6-BAP) foi influenciada pelo genótipo. A presença de 6-BAP no meio de cultura promoveu aumento na porcentagem de explantes responsivos para citrange 'Carrizo'. A suplementação do meio de cultura com a citocinina 6-BAP proporcionou aumento no número de brotos por explante para citrange 'Carrizo', laranja 'Natal' e limão 'Volkameriano'.

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

In the present work, the effect of twelve rootstocks on fruit quality of the nucellar clone IAC-5 of 'Tahiti' lime, (Citrus latifolia Tanaka) and the influence of fruit position on tree in fruit quality was evaluated in the Citrus Experimental Station of Bebedouro (EECB), located in the Bebedouro county, state of São Paulo, Brazil. A 8.0 x 5.0m planting frame was utilized. The evaluated rootstocks were: 'Carrizo' citrange (C. sinensis (L.) Osbeck x Poncirus trifoliata (L.) Raf.); the hybrids 'Rangpur' lime x 'Swingle' citrumello (C. limonia Osbeck x P. trifoliata Raf) and 'Changsha' x 'English Small'(C. sunki Hort. ex Tan. x P. trifoliata Raf.); the mandarins 'Sun Chu Sha Kat' (C. reticulata Blanco) and 'Sunki' (C. sunki Hort. ex Tanaka); the 'Rangpur' limes 'Cravo Limeira' and ' Cravo FCAV' (C. limonia Osbeck); the 'Swingle' citrumello (P. trifoliata Raf. x C. paradisi Macf.); the 'Orlando' tangelo (C. reticulata Blanco x C. paradisi Macf.) and the trifoliates cvs. 'Rubidoux', 'FCAV' and 'Flying Dragon' (P. trifoliata Raf.). The experiment was arranged in a randomized block design, with twelve treatments, six replicates and one plant per plot. The rootstocks induced differences in fruit quality; however, all the evaluated quality characteristics were within the values considered as normal and acceptable for the variety, constituting good alternative rootstocks for the 'Rangpur' lime. Additionally, the fruit position in the plant (northeastern or southwestern) had a significant influence on the external fruit color regardless of the rootstock.

Identification of reciprocal hybrids in citrus by the broadness of the leaf petiole wing

Broadness of leaf petiole wing (WB) was investigated as a morphological marker for screening hybrids of the very narrow-winged species Citrus limonia and C. sunki with broad-winged species C. aurantium and C. sinensis. Controlled polinizations produced over 500 reciprocal hybrids with potential in the ongoing rootstock breeding program identified by the Pgi-1 and Prxa-1 isozyme loci. Measurement ratios WB/leaf length, WB/leaf broadness and WB/petiole length identified 86 to 91% of the reciprocal hybrids produced. However, visual classification of WB was an equally efficient but much easier and faster method. It can be very useful in breeding programs when large number of plants have to be screened or when isozyme, RFLP or RAPD laboratories are not available.

Patogenicidade cruzada de Ceratobasidium spp. do caquizeiro (Diospyros kaki) e do chá(Camellia sinensis) e reação de cultivares de caqui ao patógeno

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