ANATOMICAL STUDIES OF IN VITRO ORGANOGENESIS INDUCED IN LEAF-DERIVED EXPLANTS OF PASSIONFRUIT

With the aim of studying the organogenesis in vitro in Passiflora edulis Sims f. flavicarpa Deg., the passionfruit, leaf-derived explants were cultured on media containing NAA or BAP and incubated either in continuous darkness or in light. The histological events leading to de novo organ formation were evaluated. Darkness induces rhizogenesis in the presence of NAA, whereas direct shoot regeneration is stimulated by light and BAP. This latter condition is recommended for passionfruit micropropagation as several adventitious shoot buds were formed from meristemoids of parenchymal origin.

Transgenic passionfruit expressing RNA derived from Cowpea aphid-borne mosaic virus is resistant to passionfruit woodiness disease

Sixteen transgenic yellow passionfruit (Passiflora spp.) plants (R0) were obtained which express a non-translatable transgenic RNA corresponding to the 3' region of the NIb gene and the 5' region of the CP gene, derived from the genome of a Brazilian isolate of Cowpea aphid-borne mosaic virus (CABMV). The transgenic plants were propagated by stem cuttings and challenged by sap inoculation with isolates CABMV-MG1 and CABMV-PE1. One transgenic plant (TE5-10) was resistant to the isolate CABMV-MG1, but susceptible to CABMV-PE1. The remaining transgenic plants developed systemic symptoms, equal to non-transformed plants, when inoculated with either isolate. The absence of virus in TE5-10 plants was confirmed by indirect ELISA. Transcription analysis of the transgene demonstrated that the TE5-10 plant did not accumulate transgenic mRNA, even before inoculation. After inoculation, viral RNA was only detected in plants inoculated with CABMV-PE1. These results confirm that the transgenic plant TE5-10 is resistant to isolate CABMV-MG1, and suggest that the resistance mechanism is post-transcriptional gene silencing, which is already activated in the transgenic plants before virus inoculation.

Histopathological features of infections caused by Fusarium oxysporum and F. solani in purple passionfruit plants (Passiflora edulis Sims)

The purple passionfruit plant, Passifloraedulis Sims, ranks second in fruit exportation in Colombia, and its main destination is the European market. However, its production is affected by several diseases, including fusariosis. This paper presents the histopathological features of different tissues affected by the pathogens Fusarium oxysporum and Fusarium solani. Both microorganisms produce similar responses on the plant: colonization of xylem vessels by hyphae and microconidia, hypertrophy and hyperplasia of the cambium, xylem and phloem; destruction of xylem fibers and amyloplasts in parenchymatous cells; and production of gels by the plant. However, there are differences in the colonization mechanism, F. solani penetrates and is concentrated especially at the collar zone, while F. oxysporum penetrates the roots and moves through the vascular system to colonize the plant.

Induction and Morpho-Ultrastructural Analysis of Organogenic Calli of a Wild Passionfruit

This work studied a new protocol for organogenic calli induction and characterization of the morphology and ultrastructure of callogenesis in leaf explants of Passiflora gibertii N. E. Brown, a native passion fruit species from Brazil. Calli induction was performed in different growth conditions (light and dark), different MS medium salt concentrations (MS and MS half strength) and the presence or absence of coconut water. The leaf explants maintained in the dark were more responsive to bud formation. In order to reduce spending on in vitro culture, the most suitable induction medium for P. gibertii organogenesis could, therefore be the MS half strength salt concentration medium maintained in the dark. The addition of coconut water to the culture medium was essential for both calli induction and bud formation. The morphological and ultrastructural features of the organogenic calli were isodiametric cells, characterized by an organized cellular system, nucleus with prominent nucleoli, presence of starch grains and dense cytoplasm rich in endoplasmic reticulum. The scanning electron microscopy demonstrated that buds were present on these calli.

In vitro organogenesis of Passiflora alata

Passiflora alata in vitro organogenesis was studied based on explant type, culture medium composition, and incubation conditions. The results indicated that the morphogenic process occurred more efficiently when hypocotyl segment-derived explants were cultured in media supplemented with cytokinin and AgNO(3) incubated under a 16-h photoperiod. The shoot bud elongation and plant development were obtained by transferring the material to MSM culture medium supplemented with GA(3) and incubated in flasks with vented lids. Histological analyses of the process revealed that the difficulties in obtaining plants could be related to the development of protuberances and leaf primordia structures, which did not contain shoot apical meristem. Roots developed easily by transferring elongated shoots to 1/2 MSM culture medium. Plant acclimatization occurred successfully, and somaclonal variation was not visually detected. The efficiency of this organogenesis protocol will be evaluated for genetic transformation of this species to obtain transgenic plants expressing genes that can influence the resistance to Cowpea aphid borne mosaic virus.

Genetic Transformation of Passionflower and Evaluation of R(1) and R(2) Generations for Resistance to Cowpea aphid borne mosaic virus

We report on the production and evaluation of passionflower transgenic lines for resistance to Cowpea aphid borne mosaic virus (CABMV). Genetic transformation was done using Agrobacterium tumefaciens and transgene integration was confirmed by Southern blot analyses, resulting in nine transgenic lines for `IAC 275` and three for `IAC 277`. Transgenic lines were clonally propagated and evaluated for resistance to CABMV After the third inoculation, under higher inoculum pressure, only propagated plants of the transgenic line T16 remained asymptomatic, indicating a high resistance to infection with CABMV. This transgenic line was self-pollinated and the RI generation was evaluated together with the RI generation of another resistant transgenic line (T2) identified previously. Plants were inoculated with CABMV by means of viruliferous Myzus nicotianae. All 524 T2R(1) plants became infected, whereas 13 of 279 T16R(1) remained asymptomatic after four successive inoculations. A TI6R(2) generation was obtained and plants were inoculated with CABMV mechanically or by aphids. After successive inoculations, 118 of 258 plants were symptomless, suggesting that the resistance to CABMV was maintained in the plant genome as the homozygous condition was achieved. Five selected resistant TI6R(2) plants which contained the capsid protein gene are being crossed for further analyses.

Análise filogenética de potyvírus causando endurecimento dos frutos do maracujazeiro no Nordeste do Brasil

Amostras foliares de plantas de maracujá-amarelo (Passiflora edulis f. flavicarpa) com sintomas típicos de endurecimento dos frutos foram coletadas nos estados de Pernambuco, Paraíba e Sergipe. A infecção viral foi comprovada por meio de teste sorológico e gama de hospedeiros. Os seis isolados virais obtidos foram capazes de infetar várias espécies testadas, porém apresentando diferenças na intensidade dos sintomas induzidos nessas hospedeiras. Teste de ELISA indireto demonstrou que os isolados obtidos a partir de plantas de maracujá são sorologicamente relacionados entre si e com o potyvírus Cowpea aphid-borne mosaic virus (CABMV). A seqüência de aminoácidos da proteína capsidial foi determinada para os seis isolados. A comparação dessas seqüências com as de outros potyvírus indicou uma identidade máxima com isolados de CABMV (86 a 94%). A identidade com isolados de Passionfruit woodiness virus (PWV) foi de 68 a 76%. Análise filogenética realizada a partir das seqüências de aminoácidos agrupou os isolados em estudo junto a isolados de CABMV, distante de isolados de PWV. Em conjunto, os resultados indicam que os isolados de maracujá analisados constituem na verdade uma estirpe do CABMV.

Ethylene and polyamine interactions in morphogenesis of Passiflora cincinnata: effects of ethylene biosynthesis and action modulators, as well as ethylene scavengers

Ethylene is a plant hormone that is of fundamental importance to in vitro morphogenesis, but in many species, it has not been thoroughly studied. Its relationship with polyamines has been studied mainly because the two classes of hormones share a common biosynthetic precursor, S-adenosylmethionine (SAM). In order to clarify whether competition between polyamines and ethylene influences in vitro morphogenetic responses of Passiflora cincinnata Mast., a climacteric species, different compounds were used that act on ethylene biosynthesis and action, or as ethylene scavengers. Treatment with the ethylene inhibitor, aminoethoxyvinylglycine (AVG) caused a greater regeneration frequency in P. cincinnata, whereas treatment with the ethylene precursor, 1-aminocyclopropane-1-carboxylic-acid (ACC) lessened regeneration frequencies. The data suggested that levels of polyamines and ethylene are not correlated with morphogenic responses in P. cincinnata. It was ascertained that neither the absolute ethylene and polyamine levels, nor competition between the compounds, correlated to the obtained morphogenic responses. However, sensitivity to, and signaling by, ethylene appears to play an important role in differentiation. This study reinforces previous reports regarding the requirement of critical concentrations and temporal regulation of ethylene levels for morphogenic responses. Temporal regulation also appeared to be a key factor in competition between the two biosynthetic pathways, without having any effects on morphogenesis. Further studies investigating the silencing or overexpression of genes related to ethylene perception, under the influence of polyamines in cell differentiation are extremely important for the complete understanding of this process.

Enraizamento de estacas de Passiflora nitida submetidas a diferentes concentrações de ácido indolbutírico (AIB)

O presente trabalho objetivou avaliar o enraizamento de estacas de P. nitida, utilizando dois tipos de estacas (com 1 e 2 gemas) e 4 doses de ácido indolbutírico (AIB) (0; 1.000; 3.000 e 5.000 mgL-1) com imersão lenta (5 segundos), com a finalidade de utilizá-las como porta-enxerto do maracujazeiro-azedo. O delineamento experimental utilizado foi inteiramente casualizado, em esquema fatorial 4x2 (concentrações de AIB x número de gemas na estaca), com quatro repetições de 10 estacas, totalizando 320 estacas. As estacas foram dispostas em bandejas plásticas, contendo vermiculita expandida de textura média, e mantidas sob sistema de nebulização intermitente, por 25 dias. As doses de AIB testadas influenciaram na sobrevivência, enraizamento das estacas e número e comprimento de raízes; e o número de gemas não influenciou no enraizamento de estacas.

Comportamento de maracujazeiros (Passiflora spp.) quanto à morte prematura

Na Universidade Estadual Paulista, Câmpus de Jaboticabal-SP, estudou-se o comportamento de Passifloráceas quanto à morte prematura de plantas, cultivadas em local com histórico da doença. O objetivo do trabalho foi avaliar o comportamento de diversos acessos de populações e espécies de maracujazeiros em relação a esta doença, sendo que as plantas resistentes deverão ser utilizadas como porta-enxertos de formas comerciais de maracujá-amarelo (Passiflora edulis Sims f. flavicarpa) e em programas de melhoramento genético. As espécies utilizadas foram P. edulis Sims, P. edulis Sims f. flavicarpa Degener, P. nitida H.B.K., P. cincinnata, P. giberti, P. laurifolia, P. morifolia, P. foetida e P. capsularis. em local com histórico da doença, plantaram-se mudas em número variável e em épocas distintas. A condução das plantas e os tratos culturais foram os recomendados para o maracujá-amarelo. A morte prematura das plantas ocorreu entre dois meses e dois anos da cultura no campo. P. giberti e P. nitida mostraram-se resistente à doença, independentemente do local de origem. Entre os demais acessos, não se encontraram fontes promissoras de resistência. Entretanto, novos acessos e novas espécies deverão ser estudadas na busca da resistência.

Incidência e severidade do vírus do endurecimento dos frutos em maracujazeiros enxertados em pé-franco

O objetivo deste trabalho foi avaliar a incidência e a severidade do vírus do endurecimento dos frutos em maracujazeiro-amarelo enxertado e pé-franco. O experimento foi conduzido no município de Adamantina-SP, no período de abril de 2006 a junho de 2007, adotando-se o delineamento em blocos ao acaso, com quatro tratamentos e oito repetições. Foram avaliados três porta-enxertos: Passiflora edulis, P. alata e P. gibertii, e plantas de pé-franco. Utilizou-se como copa o maracujazeiro-amarelo (Passiflora edulis Sims). Avaliaram-se a porcentagem de plantas com sintomas de virose e a severidade dos sintomas. As primeiras plantas com sintomas de virose ocorreram aos 90 dias do plantio das mudas no campo, atingindo, aos 180 dias, 100% de plantas com virose em P. alata e P. gibertii, e 97,5% em P. edulis e pé-franco.

Comportamento de diferentes maracujazeiros em relação ao nematóide formador de galhas

Estudou-se o comportamento de diferentes espécies de maracujazeiro Passiflora spp disponíveis em nosso meio, quando inoculadas com o nematóide formador de galhas Meloidogyne incognita. As avaliações foram feitas 80 dias após a inoculação do parasito, com base nos números de galhas e de ootecas observados nos sistemas radiculares das plantas. Verificou-se que Passiflora alata, P. giberti, P. maliformis e P. serrato digitata mostraram elevada suscetibilidade, enquanto que P. caerulea, P. edulis e P. edulis f. flavicarpa foram bastante resistentes.

produção de mudas de maracujazeiro amarelo submetidas à salinidade em solo com biofertilizante bovino

The experiment was conducted in a greenhouse from May 1 to July 31, 2008, in Areia county, Paraiba State, PB, Brazil, in order to evaluate the effects of irrigation water salinity on initial growth of the passionfruit seedlings in non-saline substrate with and without bovine biofertilizer. The treatments were distributed in a completely randomized design, with three replications and twelve plants per plots, in a factorial arrangement 5 × 2 × 2, corresponding the former to the levels of salinity in the irrigation water: 0.5; 1.0; 2.0; 3.0 and 4.0 dS m-1, in soils with and without bovine biofertilizer applied at two moments (25 and 65 days after seedling emergence). The growth of the seedlings and the soil electrical conductivity were evaluated at the end of the experiment. The biofertilizer was diluted in a low saline water at a 1:1 ratio and was applied once two days before sowing, corresponding to 10% of the substrates volume. The increase in water salinity inhibited the growth in height of plants, leaf area and root length, but always to a lesser extent in the treatments with bovine biofertilizer. The increase in electrical conductivity of the irrigation water elevated the soil salinity, independently of the addition of biofertilizer.

Efeitos de sistemas de condução sobre o crescimento, produção, qualidade dos frutos e custos de instalação de maracujazeiro amarelo (Passiflora edulis Sims, f. flavicarpa Deg)

Pós-graduação em Agronomia - FEIS

Epidemiologia do Cowpea aphid borne mosaic virus (CABMV) em maracujazeiros na região produtora da Alta Paulista, SP

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)