Genetically engineered bananas—From laboratory to deployment

Since the first two successful transformation events in banana were reported in 1995, considerable effort has been invested to develop new cultivars with improved tolerance to biotic and abiotic stresses and with enhanced nutrient levels, primarily using Agrobacterium‐mediated transformation and particle bombardment. In addition to many promising laboratory‐based studies, several genetically engineered banana cultivars have been trialled in the field. However, the deployment of genetically engineered varieties of bananas lags behind that of other major crops and there has been no commercial plantation. This article provides a review of advances in the genetic engineering of banana with an overview of noteworthy developments in several programmes that are being conducted worldwide. We identify the main challenges to translating the full potential of genetically engineered bananas for human consumption as the intellectual property issues surrounding the technology, public perceptions towards the adoption of the transformed bananas as well as various regulatory hurdles that hold the technology development from moving forward.     

A Method for Isolation of Total RNA from Fruit Tissues of Banana

We describe a rapid and efficient method for isolation of total RNA from banana fruit tissues. The RNA was extracted with a high ionic strength buffer at room temperature. The proteins, genomic DNA and secondary metabolites in the extract were then removed by precipitation with pre-cooled potassium acetate and repeated phenol/chloroform/isoamyl alcohol extractions. The RNA was recovered by ethanol precipitation. It was relatively free of ribonucleases and was suitable for RT-PCR and northern blot analysis. The procedure can be completed in less than 4 hours.     

Stormwater drainage pipes as a threat to a stream-dwelling amphipod of conservation significance, Austrogammarus australis, in southeastern Australia

The general hypothesis that catchment urbanization explained the distribution of the threatened, stream-dwelling amphipod Austrogammarus australis (listed under the Victorian Flora and Fauna Guarantee Act 1988) was tested using several surveys of 58 sites in streams draining the Dandenong Ranges on the eastern fringe of Melbourne, Victoria, Australia. More specifically, four catchment-scale elements of urban land, hypothesized as sources of stress to receiving streams, were separated: catchment imperviousness, drainage connection (proportion of impervious areas connected to streams by stormwater pipes), density of unsealed roads and density of septic tanks. The degree to which each attribute independently and jointly explained the occurrence of A. australis was assessed using hierarchical partitioning of logistic regression analyses. Drainage connection independently best explained the occurrence of A. australis, pointing to stormwater drainage design as the priority area of catchment management for the conservation of the species. The separation of urban land use into attributes that characterize likely stressor sources provides a useful framework for assessing and prioritizing the most appropriate management actions to minimize urban-related stresses to aquatic biota.     

Assessment of standing balance deficits in people who have undergone anterior cruciate ligament reconstruction using traditional and modern analysis methods

Modern methods of assessing standing balance such as wavelet and entropy analysis could provide insight into postural control mechanisms in clinical populations. The aim of this study was to examine what effect anterior cruciate ligament reconstruction (ACLR) has on traditional and modern measures of balance. Ninety subjects, 45 who had undergone ACLR and 45 matched controls, performed single leg static standing balance tests on their surgical or matched limb on a Nintendo Wii Balance Board. Data were analysed in the anterior–posterior axis of movement, which is known to be affected by ACLR. The traditional measures of path velocity, amplitude and standard deviation were calculated in this plane. Additionally, sample entropy and discrete wavelet transform derived assessment of path velocity in four distinct frequency bands related to (1) spinal reflexive loops and muscle activity, (2) cerebellar, (3) vestibular, and (4) visual mechanisms of postural control were derived. The ACLR group had significantly increased values in all traditional measures and all four frequency bands. No significant difference was observed for sample entropy. This indicated that whilst postural sway was amplified in the ACLR group, the overall mechanism used by the patient group to maintain balance was similar to that of the control group. In conclusion, modern methods of signal analysis may provide additional insight into standing balance mechanisms in clinical populations. Future research is required to determine if these results provide important and unique information which is of benefit to clinicians.     

Maintaining T lymphocyte homeostasis: another duty of autophagy

First identified as a pathway for nutrient recovery during periods of starvation, the role of autophagy has expanded to the clearance of "toxic" intracellular material including ubiquitin-positive protein aggregates, damaged organelles as well as microbial pathogens in various cell types. We have examined the role of autophagy in the development and function of the adaptive immune system. Genes encoding autophagy machinery are expressed in T lymphocytes, and autophagy occurs in primary CD4+ and CD8+ T cells. By generating fetal liver chimeric mice, we found that thymocyte development is largely normal but the mature T cell compartment is severely reduced in the absence of the essential autophagy gene Atg5. Consistent with a critical role for autophagy in promoting T cell survival, Atg5-/- CD8+ T cells display high levels of apoptosis. Surprisingly, Atg5-deficient T cells were also unable to efficiently proliferate after T-cell receptor (TCR) stimulation. These findings suggest that autophagy regulates T lymphocyte homeostasis by promoting both survival and proliferation. In addition, T cells offer a new, physiologically relevant system to study the regulation and function of autophagy pathways in vivo.     

Choriocapillaris atrophy after experimental destruction of the retinal pigment epithelium in the rat. A study in thin sections and vascular casts

Rats that receive intravenous injections of sodium iodate develop a retinopathy characterized by the partial loss of the retinal pigment epithelium (RPE). In thin sections examined by transmission electron microscopy the choriocapillaris atrophied adjacent to areas of RPE destruction. The endothelial cells thickened and lost their fenestrae and the lumen of the capillary was reduced. At sites where the RPE remained normal in appearance the choriocapillaris did not atrophy. Scanning electron microscopy of vascular casts of the choriocapillaris showed the coexistence of atrophic and normal choriocapillaris throughout the retina, presumably adjacent to sites where the RPE was destroyed or spared, respectively. Our observations support the concept that the RPE exerts some control over the structure and function of the choriocapillaris.     

High frequency plant regeneration from stem explants of Brassica alboglabra Bailey in vitro

Culture conditions for shoot regeneration and proliferation, and rooting of Brassica alboglabra Bailey were optimized by a judicious selection of explants and manipulation of hormonal combinations in the culture medium. Both half and whole stem explants were more regenerative than cotyledons and hypocotyls. The highest shoot regeneration frequency (100%) accompanied by high number of shoots was obtained using half stem explants grown on Murashige & Skoog [14] medium supplemented with 2 mgl^-1 BA in combination with 1 mgl^-1 NAA, or 4 mgl^-1 2iP with 0.5 mgl^-1 NAA. For shoot proliferation, 4 mgl^-1 kinetin was most effective. The presence of auxin reduced shoot proliferation significantly. Maximum rooting (100%) of shoot cuttings was obtained either in the absence or in the presence of 0.5 mgl^-1 NAA, or IBA or IAA ranging from 0.1 to 8 mgl^-1.