Synthesis of enzymatically active human alpha-L-iduronidase in Arabidopsis cgl (complex glycan-deficient) seeds

As an initial step to develop plants as systems to produce enzymes for the treatment of lysosomal storage disorders, Arabidopsis thaliana wild-type (Col-0) plants were transformed with a construct to express human alpha-l-iduronidase (IDUA; EC 3.2.1.76) in seeds using the promoter and other regulatory sequences of the Phaseolus vulgaris arcelin 5-I gene. IDUA protein was easily detected on Western blots of extracts from the T(2) seeds, and extracts contained IDUA activity as high as 2.9 nmol 4-methylumbelliferone (4 MU)/min/mg total soluble protein (TSP), corresponding to approximately 0.06 microg IDUA/mg TSP. The purified protein reacted with an antibody specific for xylose-containing plant complex glycans, indicating its transit through the Golgi complex. In an attempt to avoid maturation of the N-linked glycans of IDUA, the same IDUA transgene was introduced into the Arabidopsis cgl background, which is deficient in the activity of N-acetylglucosaminyl transferase I (EC 2.4.1.101), the first enzyme in the pathway of complex glycan biosynthesis. IDUA activity and protein levels were significantly higher in transgenic cgl vs. wild-type seeds (e.g. maximum levels were 820 nmol 4 MU/min/mg TSP, or 18 microg IDUA/mg TSP). Affinity-purified IDUA derived from cgl mutant seeds showed a markedly reduced reaction with the antibody specific for plant complex glycans, despite transit of the protein to the apoplast. Furthermore, gel mobility changes indicated that a greater proportion of its N-linked glycans were susceptible to digestion by Streptomyces endoglycosidase H, as compared to IDUA derived from seeds of wild-type Arabidopsis plants. The combined results indicate that IDUA produced in cgl mutant seeds contains glycans primarily in the high-mannose form. This work clearly supports the viability of using plants for the production of human therapeutics with high-mannose glycans.     

Research advances in plant-made flavivirus antigens

Outbreaks of flaviviruses such as dengue (DV), yellow fever (YFV), Japanese encephalitis (JEV), tick-borne encephalitis (TBEV) and West Nile (WNV) affect numerous countries around the world. The fast spread of these viruses is the result of increases in the human population, rapid urbanisation and globalisation. While vector control is an important preventive measure against vector-borne diseases, it has failed to prevent the spread of these diseases, particularly in developing countries where the implementation of control measures is intermittent. As antiviral drugs against flaviviruses are not yet available, vaccination remains the most important tool for prevention. Although human vaccines for YFV, TBEV and JEV are available, on-going vaccination efforts are insufficient to prevent infection. No vaccines against DENV and WNV are available. Research advances have provided important tools for flavivirus vaccine development, such as the use of plants as a recombinant antigen production platform. This review summarises the research efforts in this area and highlights why a plant system is considered a necessary alternative production platform for high-tech subunit vaccines.     

Ground beetle species (Carabidae,Coleoptera) activity and richness in relation to crop type,fertility management and crop protection in a farm management comparison trial

Ground beetle activity and species richness was monitored using pitfall traps in a plot trial system on a farm in northern England where the effects of organic and conventional fertility and crop protection management were separated within different crop types between 2005 and 2008. As well as analyses on species activity the beetles were split into small, medium‐sized and large groups, and into groups of herbivores and specific Collembola feeders. Crop type had significant effects on the activity of the 20 most abundant species and all groups, generally with most in beans and winter barley and least in vegetables and spring barley. Most significant reactions to crop protection and fertility management were in cereals and grass/clover. Activity of small species was highest in conventionally crop‐protected cereals but not in vegetables, with more medium‐sized and herbivorous species in organic plots, but there was little influence of crop protection management on large and Collembola feeding species. However, large species were significantly more active in organically fertilised cereals and grass/clover, but not in vegetables, and there were more Collembola feeders in conventional cereals but not in grass/clover. Small species were more abundant in conventionally fertilised grass/clover but there were more in organic cereals and vegetables. These inconsistent activity reactions to management were also observed with individual species but most preferred organically managed plots. There were few significant crop protection : fertility management interactions. Species richness was also significantly affected by crop type and where management had an influence, more species were found in organically managed plots. Constrained ordination emphasised that ground beetle activity was influenced more by crops than by management. Given the diverse nature of organic crop rotations, crop type should be considered a major influence in any environmental manipulation aimed at increasing ground beetle activity for provision of ecosystem services.     

Orgenic plants: Gene‐manipulated plants compatible with organic farming

Based on recent advances in plant gene technology, I propose to develop a new category of GM plants, orgenic plants, that are compatible with organic farming. These orgenic plants do not contain herbicide resistance genes to avoid herbicide application in agriculture. Furthermore, they either contain genes that are naturally exchanged between species, or are sterile to avoid outcrossing if they received a transgene from a different species. These GM plants are likely to be acceptable to most sceptics of GM plants and facilitate the use of innovative new crops.     

Plant bioreactors - the taste of sweet success

Thaumatins are intensely sweet proteins (3000 times sweeter than the same weight of sucrose) that are found in the arils of the tropical perennial plant Thaumatococcus daniellii Benth and are produced commercially by aqueous extraction from the fruits. The proteins are widely used as sweeteners and flavor enhancers in the food industry, and the European Food Safety Association (EFSA) has recently confirmed that their use as feed additive (1 to 5 mg/kg complete feed) is safe for all animal species. Given the large market for sweeteners and flavor enhancers, thaumatins could become increasingly important in the food and feed additives sector. In this issue of Biotechnology Journal, a study examines the production of thaumatin in tobacco hairy root cultures.     

Genetic diversity in commercial wineries: effects of the farming system and vinification management on wine yeasts

Aims: Analysis of the diversity and distribution of wine yeasts isolated from organically and conventionally grown grapes, and during the subsequent fermentation with or without starter cultures in six different commercial wineries. Methods and Results: PCR‐RFLP screening of isolates revealed the involvement of ten different species. Saccharomyces cerevisiae, scarcely isolated from grapes, was the dominant species during the latter phases of fermentation, identifying 108 different genotypes by means of SSR analysis. Species and strains’ diversity and presence were strongly influenced by the farming system used to grow the grapes and the system of vinification. Conclusions: Organic farming management was more beneficial in terms of diversity and abundance than the conventional one. Induced fermentation generated a great replacement of native yeasts. Although winery‐resident yeasts resulted to be predominant in the process, some noncommercial strains originally in the vineyard were found in final stages of the fermentation, confirming that autochthonous strains of S. cerevisiae are capable to conduct the fermentation process up to its end. Significance and Impact of the Study: The study of natural yeast communities from commercial vineyards and wineries is an important step towards the preservation of native genetic resources. Our results have special relevance because it is the first time that the real situation of the yeast ecology of alcoholic fermentation in commercial wineries belonging to the relevant wine‐producing Appellation of Origin ‘Vinos de Madrid’ is shown.     

Early Holocene cultivation before domestication in northern Syria

Charred plant remains from the sites of Tell Qaramel, Jerf el Ahmar, Dja’de and Tell ‘Abr situated in northern Syria and dated to the tenth and ninth millennia cal b.c. demonstrate that a wide variety of wild pulses, cereals, fruits and nuts was exploited. Five lines of evidence suggest that cultivation was practised at three of the sites. (1) Wild einkorn, wild rye and lentils occur outside their natural habitats. (2) The founder crops barley, emmer and single-grained einkorn appear at different times. (3) An assemblage of weeds of cultivation was identified. (4) There is a gradual decrease in gathered plants such as small seeded grasses and Polygonum/Rumex. (5) Barley grains increase in breadth and thickness. Morphological domestication did not become established, perhaps because seed stock was regularly collected from wild stands. Charred rodent droppings indicate large-scale grain storage.     

Managing private,commercial rangelands for agricultural production and wildlife diversity in Namibia and Zambia

The private game industry has grown across Africa since the mid-20th century. While considerable research has documented wildlife production on commercial land in many eastern and southern African countries, few studies have focused specifically on the integration of livestock and game production in Namibia and Zambia. This paper reports a survey of 43 commercial conservancy members in Namibia and 23 game farmers in Zambia conducted between September 2004 and June 2005. The survey was based on inductive sampling theory and queried farmers on how they have integrated wildlife production into their management practices. Farmers in each country reported considerable integration of wildlife conservation and agricultural production. Namibian farmers reported substantial problems with bush encroachment, whereas none of their Zambian counterparts raised similar complaints. This paper describes the state of rangeland management on commercial farms in Namibia and Zambia and identifies important areas where further research can contribute to the enhancement of this conservation-production system.     

Detecting Karenia brevis blooms and algal resuspension in the western Gulf of Mexico with satellite ocean color imagery

Blooms of the toxic dinoflagellate, Karenia brevis, have had detrimental impacts on the coastal Gulf of Mexico for decades. Detection of Karenia brevis blooms uses an ecological approach based on anomalies derived from ocean color imagery. The same anomaly product used in Florida produces frequent false positives on the Texas coast. These failures occurred during wind-driven resuspension events. During these events resuspension of benthic algae significantly increases chlorophyll concentrations in the water, resulting in confusion with normal water column phytoplankton, such as Karenia. A method was developed to separate the resuspended chlorophyll from the water column chlorophyll, decreasing the false positives used with the detection method.     

Development of tolerance against toxic cyanobacteria in Daphnia

We tested whether previous exposure to a toxic strain of cyanobacteria (Microcystis) affects survival, growth, and reproduction of a common herbivore, Daphnia magna. Samples from three natural populations of D. magna were each divided into two parts; one part was fed a mixture of toxic Microcystis and the non-toxic green alga Scenedesmus whereas the other part was fed only Scenedesmus. After four weeks, we compared the ability of these two populations to withstand the toxic Microcystis by assessing survivorship, growth, and reproduction. We found that the ability of D. magna to cope successfully with toxic Microcystis is improved if the animals have experienced previous exposure to toxic Microcystis. This suggests that the toxin may less affect the D. magna populations that are repeatedly exposed to toxic cyanobacteria in their natural habitat than populations lacking prior exposure. Since the ability to tolerate toxins is manifested in both improved survival and larger size of the animals, it may have considerable impact on zooplankton community composition in fresh-waters.