RESUMEN
Lasiodiplodia theobromae is a major pathogen of many different crop cultures, including cashew nut plants. This paper describes an efficient Agrobacterium tumefaciens-mediated transformation (ATMT) system for the successful delivery of T-DNA, transferring the genes of green fluorescent protein (gfp) and hygromycin B phosphotransferase (hph) to L. theobromae. When the fungal pycnidiospores were co-cultured with A. tumefaciens harboring the binary vector with hph-gfp gene, hygromycin-resistant fungus only developed with acetosyringone supplementation. The cashew plants inoculated with the fungus expressing GFP revealed characteristic pathogen colonization by epifluorescence microscopy. Intense and bright green hyphae were observed for transformants in all extensions of mycelium cultures. The penetration of parenchyma cells near to the inoculation site, beneath the epicuticle surface, was observed prior to 25 dpi. Penetration was followed by the development of hyphae within invaded host cells. These findings provide a rapid and reproducible ATMT method for L. theobromae transformation.
Asunto(s)
Agrobacterium tumefaciens/genética , Anacardium/genética , Ascomicetos/genética , Nueces/genética , Anacardium/crecimiento & desarrollo , Anacardium/microbiología , Ascomicetos/patogenicidad , Técnicas de Transferencia de Gen , Vectores Genéticos , Proteínas Fluorescentes Verdes , Hifa/genética , Hifa/patogenicidad , Nueces/crecimiento & desarrollo , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/microbiología , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/crecimiento & desarrollo , Transformación GenéticaRESUMEN
In this paper, we investigate the Casimir effect within a virus RNA, particularizing the study to the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Then, we discuss the possibility of occurring damage or mutation in its genome due to the presence of quantum vacuum fluctuations inside and around the RNA ribbon. For this, we consider the geometry and the nontrivial topology of the viral RNA as having a simple helical structure. We initially compute the non-thermal Casimir energy associated to that geometry, considering boundary conditions that constrain the zero point oscillations of a massless scalar field to the cylindrical cavity containing a helix pitch of RNA ribbon. Then we extend the obtained result to the electromagnetic field and, following, we calculate the probability of occurring damage or mutation in RNA by using the normalized inverse exponential distribution, which suppresses very low energies, and consider cutoff (threshold) energies corresponding to UV-A and UV-C rays, surely responsible by mutations. Then, by taking into account UV-A, we arrive at a mutation rate per base per infection cycle, which in the case of the SARS-CoV-2 is non-negligible. We find a maximum value of this mutation rate for an RNA ribbon radius, applying it for SARS-CoV-2, in particular. We also calculate a characteristic longitudinal oscillation frequency for the helix pitch value corresponding to the local minimum of the Casimir energy. Finally, we consider thermal fluctuations of classical and quantum nature and show that the corresponding probability of mutation is completely negligible for that virus. Therefore, we conclude that only the nontrivial topology and the geometric attributes of the RNA molecule contribute to the possible mutations caused by quantum vacuum fluctuations in the viral genome.
Asunto(s)
COVID-19 , SARS-CoV-2 , Humanos , SARS-CoV-2/genética , COVID-19/genética , Mutación , Tasa de Mutación , ARN Viral/genética , ARN Viral/químicaRESUMEN
Foliar uptake of dew is likely an important mechanism of water acquisition for plants from tropical dry environments. However, there is still limited experimental evidence describing the anatomical pathways involved in this process and the effects of this water subsidy on the maintenance of gas exchange and leaf lifespan of species from seasonally dry tropical vegetation such as the Brazilian caatinga. We performed scanning electron, bright-field and confocal microscopic analyses and used apoplastic tracers to examine the foliar water uptake (FWU) routes in four woody species with different foliar phenology and widely distributed in the caatinga. Leaves of plants subjected to water stress were exposed to dew simulation to evaluate the effects of the FWU on leaf water potentials, gas exchange and leaf lifespan. All species absorbed water through their leaf cuticles and/or peltate trichomes but FWU capacity differed among species. Leaf wetting by dew increased leaf lifespan duration up to 36 days compared to plants in the drought treatment. A positive effect on leaf gas exchange and new leaf production was only observed in the anisohydric and evergreen species. We showed that leaf wetting by dew is relevant for the physiology and leaf lifespan of plants from seasonally dry tropical vegetation, especially for evergreen species.
Asunto(s)
Hojas de la Planta/metabolismo , Fotosíntesis/fisiología , Hojas de la Planta/fisiología , Estomas de Plantas/metabolismo , Estomas de Plantas/fisiología , Transpiración de Plantas/fisiología , Agua/metabolismoRESUMEN
Naturalists Fritz and Hermann Müller hypothesised that heteranthery often leads to a division of labour into 'feeding' and 'pollinating' stamens; the latter often being as long as the pistil so as to promote successful pollination on the bees' back. In many buzz-pollinated species of Senna, however, the so-called pollinating stamens are short and not level with the stigma, raising the question of how pollen is shed on the bees' back. Here we explore a mechanism called 'ricochet pollination'. We test whether division of labour is achieved through the interaction between short lower stamens and strongly concave 'deflector petals'. We studied the arrangement and morphology of the floral organs involved in the ricochet pollination, functioning of the flowers through artificial sonication and observed the interactions between bees and flowers in the field. The middle stamens are adapted to eject pollen downwards, which can be readily collected on the bee mid legs. Most of the pollen is ejected towards the deflector petal(s). Pollen from this set of stamens is more likely to contribute to pollination. The pollen grains seem to ricochet multiple times against the deflector petals to eventually reach the bee's back. The pollen ricochet mechanism promotes a division of labour by involving additional floral organs, such as petals, reinforcing the Müllers' division-of-labour hypothesis. However, alternative, non-multiexclusive hypotheses could be explored in genus Senna and other angiosperm species.
Asunto(s)
Flores/fisiología , Polen/fisiología , Polinización/fisiología , Senna/fisiología , Animales , Abejas/fisiología , Flores/anatomía & histología , Polen/anatomía & histología , Senna/anatomía & histologíaRESUMEN
Lasiodiplodia theobromae is a phytopathogenic fungus causing gummosis, a threatening disease for cashew plants in Brazil. In an attempt to investigate the ultrastructural features of the pathogen colonization and its response to immunofluorescence labeling, light, confocal and electron microscope studies were conducted on different severity scale patterns of diseased plants. Lasiodiplodia-antisera was checked for cross reactivity against common cashew plants fungi. Optical microscopy analysis revealed a longitudinally sectioned hyphae located within the xylem vessels, showing an extensive hyphal development in the secondary xylem tissue. SEM images demonstrated that the fungus was found in some asymptomatic samples, particularly within the xylem vessels as confirmed by the optical images. Symptomatic sample images showed an extensive distribution of the fungus along the secondary xylem, within the vessels, infecting xylem parenchyma. A closer look in the secondary xylem parenchyma reveals a heavy and profuse invasion of the cells with a distinguishable cell wall disintegration and fully hyphae dispersal. There was no reactivity of Lasiodiplodia-antisera against mycelial extracts of Colletotrichum gloeosporioides, Phomopsis anardii and Pestalotiopsis guepinii. Following incubation of sections with the polyclonal antisera, the hyphae were intensely and regularly labeled. Rays, vessels and parenchyma cells were the preferred pathway for L. theobromae colonization. Artificial infection provides the information that the vascular cylinder is undoubtedly employed and used by the fungus for hyphae distribution. Immunofluorescence assay employed in situ was applied and the polyclonal antisera produced was able to recognize the fungus and proved to be a sensitive technique to detect it.
Asunto(s)
Anacardium/microbiología , Ascomicetos/crecimiento & desarrollo , Ascomicetos/patogenicidad , Enfermedades de las Plantas/microbiología , Brasil , MicroscopíaRESUMEN
The authors present a new technique for nipple reconstruction using muscle flaps obtained from muscles situated deep in the retromammary costal region. In the case illustrated, the muscle used was the rectus abdominal muscle, previously transposed for reconstruction in a patient mastectomized more than three years ago. The resulting nipple salience as well as the areolar area were covered with a skin graft obtained from the medial thigh donor site.