Agrobacterium-mediated transformation of the CrDAT gene and selection of transgenic periwinkle lines with a high vincristine accumulation

Catharanthus roseus contains vincristine and vinblastine, which are outstanding drugs for cancer. In the biosynthetic pathways of terpenoid indole alkaloids (TIAs) in C. roseus, deacetylvindoline 4-O-acetyltransferase (DAT) is a key enzyme that catalyses the last reaction of vindoline biosynthesis to form vinblastine and vincristine. In this study, the CrDAT transgene was transferred into the periwinkle by Agrobacterium-mediated transformation and generated transgenic periwinkle lines with an increase in vincristine accumulation. The C. roseus DAT gene was introduced into C. roseus plants and it was confirmed that CrDAT was successfully transferred into the genome of periwinkle plants and efficiently translated to synthesise recombinant DAT protein. Four transgenic periwinkle lines in T1 generation, T1-1, T1-3, T1-6, and T1-7, expressed recombinant DAT protein with the total protein content in the range of 2.86 μg.mg^?1 to 5.12 μg.mg^?1. Moreover, the vincristine contents of four transgenic lines increased by 1.63?2.48-fold compared to non-transgenic plants, ranging from 6.91 µg.g^?1 (fresh weight) to 10.53 µg.g^?1 (fresh weight). The T1-1 line had the highest vincristine content. Hence, the overexpression of the recombinant DAT protein can improve the vincristine accumulation of transgenic C. roseus plants.

Abbreviation: CrDAT - Catharanthus roseus Deacetylvindoline-4-O-Acetyl Transferase; D4H - Deacetoxyvindoline 4-hydroxylase; ELISA - Enzyme-Linked Immunosorbent Assay Monoterpene indole alkaloid; T0, T1 - Generations of transgenic plants; TIAs - Terpenoid indole alkaloids; WT- The wild-type tobacco plants (non transgenic plant); 35S - Cauliflower mosaic virus 35S promoter     

High vegetative compatibility diversity of Cryphonectria parasitica infecting sweet chestnut (Castanea sativa) in Britain indicates multiple pathogen introductions

Chestnut blight, caused by Cryphonectria parasitica, was identified in Devon, UK, in December 2016. Intensive surveys detected the disease at further sites in Devon (seven), Berkshire (one), Dorset (one), Derbyshire (four) and a cluster of eight sites in southeast London. Over 570 survey samples were tested, and 227 were positive for C. parasitica by isolation and real-time PCR. A total of 227 isolates were tested for mating type, and 197 screened for vegetative compatibility group (VCG) and compared with VCGs known from mainland Europe. The same isolates were also screened for the presence of Cryphonectria hypovirus 1 (CHV-1). Eleven VCGs were identified within the UK population. Five corresponded to already known European VCGs but six were unique. The European VCGs mainly came from the Devon, Dorset, Berkshire and Derbyshire disease outbreaks, whilst unique VCGs were almost exclusively from the southeast London cluster. Both mating types were detected, but only one mating type was present at each site, with the exception of a single Devon site. Perithecia of C. parasitica were never observed at any site. CHV-1 was found in seven isolates from three different locations and was always subtype-I, which has limited hypovirulence. Therefore, although CHV-1 is associated with C. parasitica at some outbreaks, it probably has limited impact on virulence. The diversity of VCGs and their distribution at outbreak sites, together with findings of CHV-1, suggests C. parasitica has been introduced to the UK multiple times over at least two decades through international plant trade.     

Genetic variation in root development responses to salt stresses of quinoa

Soil salinity has become a serious environmental abiotic stress limiting crop productivity and quality. The root system is the first organ sensing the changes in salinity. Root development under elevated salinity is therefore an important indicator for saline tolerance in plants. Previous studies focused on varietal differences in morphological traits of quinoa under saline stresses; however, variation in root development responses to salinity remains largely unknown. To understand the genetic variation in root development responses to salt stress of quinoa, we conducted a preliminary screening for salinity response at two salinity levels of a diverse set of 52 quinoa genotypes and microsatellite markers were used to link molecular variation to that in root development responses to salt stresses of represented genotypes. The frequency distribution of saline tolerance index showed continuous variation in the quinoa collection. Cluster analysis of salinity responses divided the 52 quinoa genotypes into six major groups. Based on these results, six genotypes representative of groups I to VI including Black quinoa, 2-Want, Atlas, Riobamba, NL-6 and Sayaña, respectively, were selected to evaluate root development under four saline stress levels: 0, 100, 200 and 300 mM NaCl. Contrasts in root development responses to saline stress levels were observed in the six genotypes. At 100 mM NaCl, significant differences were not observed in root length development (RLD) and root surface development (RSAD) of most genotypes except Black quinoa; a significant reduction was observed in this genotype as compared to controls. At 200 mM NaCl, significant reduction was detected in RLD and RSAD in all genotypes showing this as the best concentration to discriminate among genotypes. The strongest inhibition of root development was found for all genotypes at 300 mM NaCl as compared to lower saline levels. Among genotypes, Atlas of group III shows as a saline-tolerant genotype confirming previous reports. Variation in root responses to salinity stresses is also discussed in relation to climate conditions of origins of the genotypes and reveal interesting guidelines for further studies exploring the mechanisms behind this aspect of saline adaptation.     

A comparison of popular approaches to optimize landscape resistance surfaces

Landscape Ecology - Landscape resistance surfaces are often used to address questions related to movement, dispersal, or population connectivity. However, modeling landscape resistance is...     

Individual-based modeling highlights the importance of mortality and landscape structure in measures of functional connectivity

Landscape Ecology - Functional landscape connectivity is vital for the conservation of wildlife species. Landscape connectivity models often overlook factors such as mortality and asymmetry in...     

Movement behaviour of large female yellow European eel (Anguilla anguilla L.) in a freshwater polder area

Due to a recruitment decline of more than 90% in 30 years, the European eel (Anguilla anguilla L.) has been classified by IUCN as critically endangered. Although the species has been studied intensively to obtain knowledge to improve management, studies about the resident yellow stage are relatively scarce. In this study, 52 large female yellow eels were tagged with acoustic transmitters in a Belgian polder system and tracked by a network of 23 automatic listening stations. We studied both circadian and seasonal movement patterns and the effect of environmental variables on these patterns. Large female yellow eels were most active at night in late summer and early autumn. A generalised linear mixed model showed that their movement is only slightly influenced by environmental variables. Moreover, as yellow eels show high site fidelity (i.e., the majority was detected only in the habitat type of their catch‐release location), they do not encounter many human‐induced connectivity problems in polder systems, which makes these systems highly suitable as eel growth habitat. These results can contribute to an effective eel management regarding habitat protection and restoration.     

Effect of an intensive mechanical removal effort on a population of non‐native rainbow trout Oncorhynchus mykiss in a South African headwater stream

1. Invasions by non‐native species can compromise the conservation value of otherwise pristine headwater streams. While both developed and developing countries recognize this threat, few of the latter have suitable budgets to implement control programmes.
2. This study assessed the effectiveness of a mechanical project to remove non‐native rainbow trout Oncorhynchus mykiss from a 6 km section of the upper Krom River, a small headwater stream in the Cederberg Mountains in South Africa's Cape Floristic Region (CFR).
3. From October 2013 to February 2014, 354 O. mykiss were removed by angling (58%), fyke netting (28%) and gill netting (14%). This resulted in a marked reduction, but not eradication, of the O. mykiss population (fish relative abundance decreased from 0.53 ± 0.09 fish per net per night in October 2013 to 0.21 ± 0.09 fish per net per night in February 2014). Following the cessation of manual removals, the relative abundance of O. mykiss had increased to 0.56 ± 0.18 fish per net per night by March 2016, suggesting that without sustained removal effort, the population will rapidly return to its pre‐removal abundance level.
4. Further work is needed to refine the methodology and test the effectiveness of mechanical removal of non‐native freshwater fish in a variety of ecological settings in the CFR. This approach holds potential for meeting the dual goals of reducing the ecological impacts of non‐native fishes and generating employment opportunities in line with the policy objectives of developing nations.

     

Potential application of baited remote underwater video to survey abundance of west coast rock lobster Jasus lalandii

The first assessment of the baited remote underwater video system (BRUVs) for monitoring rock lobster Jasus lalandii (Milne‐Edwards) in South Africa was compared to annual ring net and trap survey data. Count data from 58 BRUVs, 36 ring net and 95 trap samples were fitted with generalised additive models using depth, habitat, time and spatial gradient as independent variables. A power analysis was used to determine the number of samples needed per annum to detect a 5 and 10% increase and decrease in population size. The BRUVs outperformed the nets and traps in terms of sampling effort and were better at detecting changes in abundance of lobster than abundance of three fish species. Considering factors such as cost, analysis time, additional data and environmental impact, traps are the most feasible method in deeper waters but BRUVs could replace nets and traps in depths less than 60 m, low‐density areas and marine reserves.