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.

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.

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.

Marine diatoms in deep sea sediments

Long-term evolution is thought to take opportunities that arise as a consequence of mass extinction (as argued, for example, by Gould, 2002) and the following biotic recovery, but there is absolutely no evidence for this being the case. However, our study shows that eutrophication by oceanic mixing also played a part in the enhancement of several evolutionary events amongst marine organisms, and these results could indicate that the rates of oceanic biodiversification may be slowed if upwelling becomes weakened by future global warming. This paper defines three distinct evolutionary events of resting spores of the marine diatom genus Chaetoceros, to reconstruct past upwelling through the analysis of several DSDP, ODP and land-based successions from the North, South and equatorial Pacific as well as the Atlantic Ocean during the past 40 million years. The Atlantic Chaetoceros Explosion (ACE) event occurred across the E/O boundary in the North Atlantic, and is characterized by resting spore diversification that occurred as a consequence of the onset of upwelling following changes in thermohaline circulation through global cooling in the early Oligocene. Pacific Chaetoceros Explosion events-1 and -2 (PACE-1 and PACE-2) are characterized by relatively higher occurrences of iron input following the Himalayan uplift and aridification at 8.5 Ma and ca. 2.5 Ma in the North Pacific region. These events not only enhanced the diversification and increased abundance of primary producers, including that of Chaetoceros, other diatoms and seaweeds, but also stimulated the evolution of zooplankton and larger predators, such as copepods and marine mammals, which ate these phytoplankton and plants. Current thinking suggests new evolutionary niches open up after a mass extinction, but our study finds that eutrophication can also stimulate evolutionary diversification. Moreover, in the opposite fashion, our results show that as thermohaline circulation abates, global warming progresses and the ocean surface becomes warmer, many marine organisms will be affected by the environmental degradation.

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.

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

No presente trabalho, avaliou-se a influência de doze porta-enxertos sobre a qualidade dos frutos da lima-ácida 'Tahiti' (Citrus latifolia Tanaka), clone 'IAC-5', amostrados em duas posições nas plantas, em experimento conduzido na Estação de Citricultura de Bebedouro, em um pomar de três anos. O espaçamento utilizado foi de 8.0 x 5.0m. Os porta-enxertos utilizados foram: citrangeiro 'Carrizo' (C. sinensis (L.) Osbeck x Poncirus trifoliata (L.) Raf.); os híbridos 'Cravo' x 'Swingle' (C. limonia Osbeck x P. trifoliata (L.) Raf. x C. paradisi Macf.) e 'Changsha' x 'English Small' (C. sunki Hort. ex Tan. x P. trifoliata Raf.); as tangerineiras 'Sun Chu Sha Kat' (C. reticulata Blanco) e 'Sunki' (C. sunki Hort. ex Tanaka); os limoeiros 'Cravo Limeira' e 'Cravo FCAV' (C. limonia Osbeck); o citrumeleiro 'Swingle' (P. trifoliata Raf. x C. paradisi Macf.), o tangeleiro 'Orlando' (C. reticulata Blanco x C. paradisi Macf.) e os trifoliateiros 'Rubidoux', 'FCAV' e 'Flying Dragon' (Poncirus trifoliata Raf.). Foi utilizado um delineamento em blocos casualizados, com doze tratamentos e seis repetições. Os distintos porta-enxertos induziram diferenças na qualidade dos frutos, entretanto todas as características de qualidade foram consideradas aceitáveis para a variedade, sendo bons substitutos para o limão 'Cravo'.

Susceptibility of tangerines to citrus variegated chlorosis

Citrus variegated chlorosis (CVC), a citrus disease first discovered in Brazil in 1987, is caused by the bacterium Xylella fastidiosa and transmitted by sharpshooters and budwood. Since the disease affects almost all sweet orange cultivars, it has become one of the most serious problems for Brazilian citriculture. To evaluate their resistance to CVC disease, fifteen tangerines or mandarins (C. reticulata Blanco) and their hybrids were grafted on Rangpur lime (C. limonia Osb.) and inoculated with CVC-contaminated Pera sweet orange (C. sinensis (L.) Osb.) by twig grafting in a greenhouse. Tangerines and their hybrids Wilking, Fortune, Sunki, Ellendale, Orlando tangelo, Nunes clementine, Nova, Sun Shu Sha Kat, Suenkat, and Batangas showed CVC leaf symptoms and gave positive results on enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) (with specific primers for X. fastidiosa), indicating that they are susceptible to CVC. Although X. fastidiosa bacteria were detected by ELISA and PCR in inoculated plants of tangerines Cravo and Oneco, no CVC leaf symptoms were observed on these two cultivars, suggesting that they are tolerant to the disease. CVC leaf symptoms were not observed and X. fastidiosa was not detected in tangerine Dancy and mandarins Okitsu satsuma and Ponkan after inoculation, showing that they are resistant to the disease.

Metabolite profiles of essential oils in citrus peels and their taxonomic implications

China is an important center of origin for the genus Citrus L. of the family Rutaceae and is rich in wild Citrus species. The taxonomy of Citrus has been a subject of controversy for more than a half century. We propose that the metabolite profiles of Chinese native Citrus species can be used for classification and understanding of the taxonomic relationships within the Citrus germplasm. In this study, triplicate gas chromatography-mass spectrometry (GC-MS) metabolite profiles of 20 Citrus species/varieties were acquired, including 10 native varieties originating in China. R-(+)-limonene, alpha-pinene, sabinene and alpha-terpinene were found to be major characteristic components of the essential oils analyzed in this study, and these compounds contributed greatly to the metabolic classification. The three basic species of the subgenus Eucitrus (Swingle's system), i.e., C. reticulata Blanco, C. medica L. and C. grandis Osb., were clearly differentiated based upon their metabolite profiles using hierarchical cluster analysis (HCA) and partial least square-discriminant analysis (PLS-DA). All the presumed hybrid genotypes, including sweet orange (C. sinensis Osb.), sour orange (C. aurantium L.), lemon (C. limon Burm.f.), rough lemon (C. jambhiri Lush.), rangpur lime (C. limonia Osb.) and grapefruit (C. paradisi Macf.), were grouped closely together with one of their suggested parent species in the HCA-dendrogram and the PLS-DA score plot. These results clearly demonstrated that the metabolite profiles of Citrus species could be utilized for the taxonomic classification of the genus and are complementary to the existing taxonomic evidence, especially for the identification and differentiation of hybrid species.

Genome Sequence Of Streptomyces Wadayamensis Strain A23, An Endophytic Actinobacterium From Citrus Reticulata.

The actinobacterium Streptomyces wadayamensis A23 is an endophyte of Citrus reticulata that produces the antimycin and mannopeptimycin antibiotics, among others. The strain has the capability to inhibit Xylella fastidiosa growth. The draft genome of S. wadayamensis A23 has ~7.0 Mb and 6,006 protein-coding sequences, with a 73.5% G+C content.

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.

Efficacy of Citrus reticulata and Mirazid in treatment of Schistosoma mansoni

This work has been carried out to investigate the effect of Schistosoma mansoni infection on mice livers after treatment with the ethanolic extract of Citrus reticulata root or the oleo-resin extract from Myrrh of Commiphora molmol tree (Mirazid), as a new antishistosomal drug. Marker enzymes for different cell organelles were measured; succinate dehydrogenase (SDH); lactate dehydrogenase (LDH) and its isoenzymes; glucose-6-phosphatase (G-6-Pase); acid phosphatase (AP) and 5'- nucleotidase. Liver function enzymes; aspartate aminotransferase (AST); alanine aminotransferase (ALT), and alkaline phosphatase (ALP) were also estimated. Parasitological studies through ova count and worm burden will also be taken into consideration. The results showed a marked reduction in SDH, LDH, AST, and ALT enzyme activities and a significant increase in G-6-Pase, AP, 5'- nucleotidase, and ALP after S. mansoni infection. A noticeable alteration in LDH subunits were also noticed. Treatment with C. reticulata or Mirazid improved all the previous enzyme activities with a noticeable reduction in ova count and worm burden.

CARACTERÍSTICAS DO FRUTO DA VARIEDADE SPAN AMERICANA (Citrus reticulata Blanco): UMA TANGERINA DO TIPO 'PONCÃ' DE MATURAÇÃO PRECOCE

A tangerina-'Poncã' é bastante apreciada pelo consumidor brasileiro. No Estado de São Paulo, a maturação de seus frutos ocorre nos meses de abril a agosto. Com o mercado ávido por frutas de mesa e agravado pelo problema da ocorrência da clorose variegada dos citros (CVC), que afeta principalmente as variedades de laranjas-doces, tem havido um aumento considerável no plantio dessa variedade. Dessa forma, um excesso de produção, num mesmo período, faz com que os preços caiam, desestimulando assim os produtores. O Centro de Citricultura Sylvio Moreira (CCSM) vem buscando outras alternativas no sentido de obter, durante o ano todo, esse tipo de tangerina. Assim sendo, foram realizados estudos de caracterização de frutos, em diversas épocas do ano, com o acompanhamento dos níveis de maturação, envolvendo acessos do Banco de Germoplasma do CCSM. Dentre as variedades estudadas, quanto à precocidade e qualidade dos frutos, destacou-se a Span Americana. A tangerina-'Poncã' tem seu período de maturação para as condições edafoclimáticas do CCSM, nos meses de maio e junho. Já a variedade Span Americana, que apresenta frutos similares à 'Poncã' tradicional, tem maturação bastante precoce, podendo ser iniciada a sua colheita no mês de março.

Indução do florescimento e crescimento de tangerineira 'Poncã' (Citrus reticulata Blanco) em função da irrigação e da aplicação de paclobutrazol

O experimento foi conduzido na Faculdade de Ciências Agronômicas - UNESP/Câmpus de Botucatu (SP), com o objetivo de avaliar o florescimento fora de época e o crescimento vegetativo da tangerineira 'Poncã'. Nesse contexto, adotou-se o delineamento estatístico em blocos casualizados, em parcelas subdivididas, com duas repetições, na instalação do ensaio. Os dois tratamentos de -0,03 e -0,05 MPa, como potenciais mínimos da água no solo, constituíram as parcelas e as quatro doses de paclobutrazol: 0; 4; 8 e 12 g por planta, nas subparcelas. No segundo ano de pesquisa, foram mantidas as mesmas parcelas e foram aplicadas nas plantas das subparcelas as doses de 0; 500; 1000 e 2000 mg l-1 de paclobutrazol, via foliar. Cada parcela foi constituída de 16 plantas, sendo oito destinadas à avaliação. Utilizou-se, em ambas as combinações, de um tratamento sem irrigação (-0,07 MPa) e sem aplicação de paclobutrazol, como testemunha. Foram avaliados parâmetros, como a altura, diâmetro médio, volume e área de projeção da copa e condutância estomática para caracterizar a resposta das plantas aos tratamentos empregados. Concluiu-se que a aplicação do paclobutrazol e a variação do potencial da água no solo não proporcionaram a indução do florescimento fora de época das tangerineiras, e que os níveis de paclobutrazol influenciaram no desenvolvimento das plantas.

Comportamento fisiológico de sementes de tangerina (Citrus reticulata Blanco) submetidas à desidratação

O objetivo desta pesquisa foi o de estudar os efeitos imediatos da desidratação sobre o comportamento fisiológico das sementes de tangerina (Citrus reticulata Blanco var. Cleópatra). Assim, sementes extraídas manualmente de frutos maduros foram lavadas em água corrente e drenadas à sombra. Em seguida, uma porção representativa de sementes foi removida a fim de constituir o tratamento com o maior grau de umidade (49%) a ser estudado. As sementes remanescentes foram submetidas à desidratação em equipamento com circulação forçada de ar (28±2°C), visando à obtenção dos tratamentos (39; 32; 24; 13; 10 e 7% H2O) referentes aos demais graus de umidade desejados. Foram realizadas avaliações do grau de umidade, da germinação, da taxa e da velocidade de emergência das plântulas, dos comprimentos da raiz, do hipocótilo e da plântula. Concluiu-se que o potencial fisiológico das sementes, considerando o intervalo de 49 a 7% para o grau de umidade, é favorecido pela desidratação até 39% H2O.