PROTEIN PRODUCTS FOR FUTURE GLOBAL GOOD
http://www.checkbiotech.org article on Aresa and GM Plant mine detection.Oct.05
Danish scientists have made a scientific discovery with significant humanitarian and environmental potential. They have shown that it is possible to produce plants which change colour in the presence of specific compounds within the soil, opening the way for the first bomb and land-mine detection plant.
IS FROM THE UNIVERSITY OF WISCONSIN-MADISON ©
Production Of Industrial And Animal Feed Enzymes In Transgenic Alfalfa
S. Austin-Phillips , E. T. Bingham , R. G. Koegel , J. Rausch, R.J. Straub , J. Will , T. Zeigelhoffer , P. Zeigellhoffer , R.R. Burgess1
Currently, most industrial enzymes are produced from microorganisms by large-scale fermentation. An alternative approach to the production of such enzymes would be to express and recover these enzymes from transgenic plants. We have been conducting a multidisciplinary feasibility Study for the production of industrial enzymes in transgenic alfalfa for the last 7-8 years, Alfalfa has certain advantages for this purpose. It is a perennial, widely-grown crop capable of three or more harvests a year. It is beneficial environmentally since it does not require annual tilling and planting, and needs fewer applications of fertilizer and pesticides than conventional row crops. Furthermore, technology for extracting protein from alfalfa while leaving a valuable residue for animal feed is well developed. The overall goal of the research is to develop genetically-engineered alfalfa that produces high levels of industrially-important enzymes, to develop rapid methods for extracting and purifying these enzymes from alfalfa, and thus provide a high value product which takes advantage of existing agricultural productivity.
As part of our feasibility study we have demonstrated the production of manganese-dependent lignin peroxidase (Mn-P) from the fungus Phanerochaete chrysosporium in transgenic plants (Austin et al., 1994, 1995). This enzyme has potential for large-scale industrial usage for lignin degradation and/or as a bleaching agent in blopulping processes. Expression of this enzyme in alfalfa may also have the potential for increasing the fiber digestibility of forages for ruminants. In addition we have expressed alpha-amylase from Bacillus licheniformis in alfalfa. It was chosen as a model enzyme for the overall feasibility study to test recovery protocols since it is a robust enzyme that is active within a wide pH and temperature range. We have published reports which outline the production arid characterization of transgenic alfalfa expressing alpha-amiylase or Mn-P and also presented data from field tests of transgenic plants (Austin et al., 1995). More recently we have produced and field-tested transgenic alfalfa expressing the animal feed enzyme phytase (from Aspergillus niger) and two different T. fusca cellulases (Ziegelhoffer et al., 1999). Phytase was expressed at levels up to 2% total soluble protein in some transgenic lines. This makes it an economically viable alternative to microbially produced phytase (Koegel et al, 1998; Austin et al.; 1999). Agronomic performance was not affected by expression of this heterologous protein and phytase expression levels remained constant over 3 growing seasons. The production of animal feed supplements in transgenic alfalfa is particularly appealing since the expressed juice or leaf meal preparations can be added directly to animal rations with no intervening extraction or purification necessary.
This from © NSF.GOV.
Making Plastics (and Other
Fine Things) from Plants
An important first step in designing better foods was made a few years ago when Chris Somerville and coworkers, then at Michigan State University in East Lansing and now at the Carnegie Institution of Washington at Stanford, California, found and cloned the Arabidopsis desaturase gene which codes for an enzyme that catalyzes the synthesis of polyunsaturated fatty acids. Dietary polyunsaturated acids have a role in lowering blood cholesterol and are needed for normal human growth.
Since the discovery of this gene in l992, scientists working with Somerville, including John Browse at Washington State University in Pullman and others at DuPont in Wilmington, Delaware, and at Monsanto in St. Louis, have isolated most of the eight different desaturase genes from Arabidopsis that control the polyunsaturation of plant oils. Plant breeders have then used these Arabidopsis genes to isolate the corresponding genes from crop species. What's more, they put copies of these genes into some plants, such as soybeans, canola, and flax, that typically make more saturated oils. This resulted in the production of nutritionally improved oils in these crops. These genetically transformed plants were field tested in the fall of l994, only 2 years after the key discoveries were made. "This gives an indication of how quickly industry can move to apply basic research," says Somerville. "We think we can tailor plant oils to specific nutritional needs." Somerville adds that "Most of the work that led up to this came from basic studies of Arabidopsis."
Related research focuses on developing custom-designed plants that will reduce our reliance on nonrenewable sources of petrochemicals, which are needed to make plastics and related materials. The goal is to modify plants genetically so that instead of producing edible oils, the plants will produce industrial oils and polymers, which are now made from petroleum stocks.
Somerville and his colleagues have shown that it is possible to genetically engineer Arabidopsis to produce granules of polyhydroxybutyrate (PHB), a polyester used for biodegradable plastic containers which is usually obtained from a bacterium, Alcaligenes eutrophus. The researchers did this by taking two genes the bacterium uses to make PHB and putting them into Arabidopsis. At first, the genetically transformed plants produced only minute granules of this valuable plastic. However, after a few years of tinkering, researchers inserted modified genetic constructs into the plants and could increase production enough to attract considerable commercial interest.
No one is suggesting that tiny Arabidopsis plants be developed as a commercial crop for plastics production, but whatever is learned from the Arabidopsis model can be used with other, more practical, crops, such as canola or soybean. Ganesh Kishore of Monsanto in St. Louis, which is one of several major companies studying the commercial possibilities of this system, says, "Our goal is to produce biodegradable plastic from a renewable source. We will create a whole new paradigm for the plastics industry."
Somerville adds that the production of plastics in plants will have the added benefit of giving farmers new markets for their harvests. "American agriculture produces too much of too few products. And novel plant varieties, created with the aid of Arabidopsis genes, will give farmers new cash crops," he says. What's more, he suggests that plastics are just the beginning, and that croplands of genetically transformed plants may be devoted to the production of, for example, hydraulic oils, lubricants, nylons, drugs, and valuable enzymes.
A DESCRIPTION OF A CURRENT TRANSGENIC NON-FOOD SUNFLOWER OIL EXPERIMENTAL PROJECT © BioMatNet
Introduction The overall aim of the project is the production of transgenic sunflower plants producing oil of modified fatty acid composition. In particular, it was intend to produce plants with high oleic acid content and reduced content of stearic acid as well as plants producing short- and medium- chain fatty acids. This was to be achieved by incorporating genetic constructs carrying the appropriate promoter and coding sequences.
Objectives The objective for the period covered by this report were to:
analyse transgenic plants produced so far
Results First results with transgenic sunflowers had indicated that the effect of the ds10-C1FatB3 construct on the fatty acid composition was only minor. In order to very the validity of this observation, transgenic rapeseed plants have been produced harbouring the same construct. None of the 52 analysed transgenic rape seed lines showed the occurrence of capric acid when analysed by gas chromatography. The presence of the appropriate protein was also undetectable by Western analysis in 6 randomly chosen lines. In contrast, rape seed lines harbouring the C1FatB3 gene under the control of its own promoter resulted not only in the expression of the expected protein but also in accumulation of capric acid (C10:0) in seed storage oil.
A new chimeric ds10::GUS::ds10 construct has therefore been constructed that contains the GUS coding region cloned in the previously described expression cassette ds 10EC1. Transgenic tobacco plants containing the new chimeric gene have been produced. Analysis of 19 independent transgenic plants showed only a moderate reduction of GUS expression in immature (16dpa) embryos, compared to the previously characterised ds 10F2-delta plants. These results indicate that other genes cloned in ds10EC1 could also be efficiently expressed in transgenic plants, except for problems not directly related with the structure of the ds10EC1 cassette. This result makes it likely that the low levels of changes in the fatty acid profile observed in the transgenic sunflowers and rapeseed plants produced with the ds10-C1FatB3 construct are not due to differences in the ds10 sequences present in ds10EC1, compared to ds10F2-delta.
The observed absence of expression may find an explanation in the residual expression of the ds10 promoter in pollen grains. Accumulation of capric acid that is not immediately captured by storage tri-acylglycerides may prove deleterious for the developing pollen grain. This working hypothesis is currently being checked by a segregation analysis of the offspring of these plants.
With respect to the engineering of fatty acid diversity in the storage oil of sunflower seeds, the contribution of exon II of the FatB gene family has been investigated. Chimeric constructs combining the FATB3 gene with exon II sequences of divers origin and expression in E. coli led to the conclusion that exon II sequences are necessary but not sufficient for the chain length specificity of Cuphea FatB genes.
The binary vector pMH000-0 carrying both the sulfonamide and Basta resistance gene described previously proved to perform particularly well for rape seed transformation. Optimisation of the transformation protocol has continued. The routinely obtained overall efficiency to confirmed transgenic flowering plants is 0.4%. However, limitations still exist. For example type 2 plants proved to be sterile due to unfavourable environment (temperature too elevated during summer). Sunflower plants expressing the ClFatB3 gene under its own promoter are expected in the near future.
Field trials of transgenic plant type 5 have been performed with 22 previously selected lines (3 internal controls, 14 pure lines and 8 hybrids) at two locations. Seeds were harvested on an individual basis, and leaf samples were conserved for each plant in order to be able to establish a correlation between genotype and phenotype if necessary. Seed samples are currently being analysed.
Two selected 'high oleic' interspecific hybrid lines with good transformation potential were further multiplied in the field so that these genotypes will soon become available for transformation experiments. Two further interspecific hybrids with exceptionally high oleic acid contents have been field grown from selected half-seeds for the development of elite lines. This selection continues at present in winter sites in South Africa.
Conclusions Transgenic plants expressing the proposed genetic constructs have been produced and introduced into field trials (fifth generation) on schedule. Full characterisation of these plants, as well as of additional plants which are available in lower generations, is in progress and, once completed, will yield extensive information as to their genetic composition, the alterations of their physiology, the interactions between these levels, and their agronomic potential.
During the course of the production of the genetic constructs and the transgenic plants, a number of questions have arisen which required a faster answer than can be given using sunflower. In these cases, transgenic tobacco and rapeseed plants have been produced (as appropriate) in order to obtain the required information that can then be utilised for the choice of appropriate constructs for sunflower. In spite of the considerable progress made with the improvement of the transformation protocol, sunflower transformation still remains a heavy and time-consuming task.
To date, all proposed milestones have been reached and the performed experiments have yielded a number of interesting results but most investigations are not yet finally concluded. So far, the project has yielded one patent application. First manuscripts for publication in scientific journals are being prepared.
Transformation of chicory into a high value non-food crop © BioMatNet
Chicory (Cichorium intybus) is a biennial crop native in Europe, which is traditionally grown for use as a coffee substitute or chicory drink or for use in the sugar industry. The current area under production for "sugar chicory" is 15,000 ha concentrated in Belgium, France and the Netherlands. The tap roots contain inulin (a fructose polymer) that is used for food products, including use as the starting material for fructose syrup. Inulin also has a high potential for use in non-food applications. The non-food industry is in need of new ingredients such as inulin that has a great market potential. Of the various inulin-producing crops known, chicory has the best potential for agricultural and industrial exploitation. However, the current properties of the inulin in the native chicory crop do not yet meet the requirements related to processing and chemical modification, necessary for successful application in non-food industry. The main obstacle is the low degree of polymerisation (DP) of the native chicory inulin. This project aims to upgrade the quality and volume of chicory inulin through genotype improvement focusing on the increase of the mean degree of polymerisation from the current value of 10 (low DP) to a value between 20 and 100 (High DP).
By introducing into native chicory a set of three genes originating from the same Asteraceae plant family, inulin metabolism will be induced to produce the targeted high DP inulin. Research activities concentrate on the selection of appropriate (wild) Asteraceae species; the isolation of genes or a combination of genes followed by the actual transformation of the native chicory into transgenic chicory. The transformation will be done along two lines of which the first one will result in agronomically tested transgenic chicory seed during the third year of this project. The second line will be released for agronomic testing by the end of the fourth year.
In addition to the transformation activities the project will assess the functional properties of high DP inulin from both sub-fractioned HDP inulin as well as the transgenic chicory inulin for use in several markets in the non-food industry.
From an agronomic point of view, chicory is a welcome addition to crop rotation schemes. It does not require much nitrogen, is more drought resistant than sugar beet and grows well on a large variety of soil types ranging from clay to sand. It is a familiar crop to sugar beet growers because cultivation as well as the related mechanisation is similar to that of sugar beet. Processing facilities for roots are similar to that of the sugar industry. A major economic benefit connected to the introduction of chicory as a high value non-food crop is the fact that the product of non-food chicory inulin is not limited by the sugar or fructose related quota agreements. This, therefore, opens new possibilities for (sugar beet) farmers to diversify as well as to expand their farming activities. Development of rural communities in traditional chicory growing areas, as well as in new European countries, is foreseen. Furthermore employment opportunities in the chicory seed and processing industries will increase.
The participants in the research consortium represent the European chicory seed industry (processing industry and growers as well as research institutes specialised in chicory, fructan and inulin research). By upgrading chicory from a currently marginal food crop to a multi-functional high value crop, this project will give a new impulse to both European agriculture and agro-industry. These experiments will result from the third year onwards in the decision by the chicory processing industry to make major investments in this new crop and the processing facilities.
Results To Date
This LINK describes an experiment from Purdue to express JoJoba wax in a transgenic Rape crop
Transgenic Trees Hold Promise for Pulp and Paper Industries © LifeSciencesWorld 2003
The expensive, energy-intensive process of turning wood into paper costs the pulp and paper industries more than $6 billion a year. Much of that expense involves separating woods cellulose from lignin, the glue that binds a trees fibers, by using an alkali solution and high temperatures and pressures. Although the lignin so removed is reused as fuel, wood with less lignin and more cellulose would save the industry millions of dollars a year in processing and chemical costs. Research at North Carolina State University shows promise of achieving that goal.
By genetically modifying aspen trees, Dr. Vincent L. Chiang, professor of forest biotechnology, and his colleagues have reduced the trees lignin content by 45 to 50 percent and accomplished the first successful dual-gene alteration in forestry science. Their results are described in the current issue of the Proceedings of the National Academy of Sciences (PNAS). According to Chiang, the NC State research shows not only a decrease in lignin but also an increase in cellulose in the transgenic aspens. And their work demonstrates another benefit: the trees grow faster.
That is very good news for the wood, paper and pulp industries, which do multibillion-dollar business worldwide. Fast-growing, low-lignin trees offer both economic and environmental advantages, because separating lignin from cellulose using harsh alkaline chemicals and high heat is costly and environmentally unfriendly. Harvesting such trees, using them as crops with desirable traits, would also reduce pressure on existing forests.
Chiang and his team chose aspens because, he says, theyre the lab rats of forestry research. The scientists scratch the leaves and expose the wound to bacteria carrying the beneficial genes. Treated leaf-disks, with their enhanced genomic structure, are then cloned, producing trees with predictable qualities.
As with any research involving genetic engineering, Chiangs modified aspens have faced questions of real-world properties, resistance to insects and diseases, and the possibility of unforeseen ecological impacts. There is a need for more data concerning the environmental effects and field performance of transgenic trees, said Chiang, but four-year field trials of such trees in France and the United Kingdom show that lignin-modified transgenic trees do not have detrimental or unusual ecological impacts in the areas tested.
In previous work, Chiang and his team had successfully reduced lignin in aspens by inhibiting the influence of a gene called 4CL. The current research modifies the expression of both 4CL and a second gene, CAld5H, in the trees. This dual-gene engineering alters the lignin structure, and produces the favorable characteristics of lower and more degradable lignin, higher cellulose and accelerated maturation of the aspens xylem cells.
The research is described in the paper Combinatorial modification of multiple lignin traits in trees through multigene co-transformation, published online by PNAS on March 31.
Chiang is co-director of the Department of Forestrys Forest Biotechnology Group in the College of Natural Resources at NC State. Headed by Chiang and Dr. Ron Sederoff, Edwin F. Conger and Distinguished University Professor of Forestry and a member of the National Academy of Sciences, the group is one of the worlds leading research organizations studying the molecular genetics of forest trees. The Forest Biotechnology Group is a key part of NC States research strength in genomics, an important new area of scientific research focused on identifying and mapping all the genes of living organisms. Its work is leading to a better understanding of the genetic basis of biological diversity, improved disease resistance in important tree species, and increased commercial forest productivity.
According to Dr. Bailian Li, associate professor of forestry at NC State, Dr. Chiangs results in this aspen model species are very significant and will have dramatic impacts on the future genetic improvement of forest trees for pulp and paper production. The improved tree growth and high cellulose content will increase pulp-yield production, while the reduced lignin content will reduce the pulping cost and energy consumption in the pulping process, he said. The ability to produce high-yield plantations with these desirable characteristics will enable us to produce wood more efficiently on less land, allowing natural forests to be managed less intensively for habitat conservation, aesthetics and recreational uses.
Citing the Forestry Departments Industry-Cooperative Tree Improvement Program working to improve plantation productivity, adaptation and disease-resistance in North Carolinas loblolly pines Li said, Results from Dr. Chiangs research are very encouraging to our research. Although his research is on aspen, the valuable information on genetic regulation of wood formation should be useful for our efforts in producing pine plantations with lower lignin, higher cellulose, and faster growth rates.
In the October issue of Nature Biotechnology, researchers from the giant agricultural company, Monsanto, report engineering transgenic plants to produce a biodegradable plastic. Kenneth Gruys and colleagues have successfully produced the plastic, PHBV, in thale cress plants and oil seed rape.
The researchers obtained cress and oil seed rape plants that produced poly(3-hydroxybuyrate-co-3-hydroxyvalerate) PHBV as approximately 3% of dry weight. The genetic engineering involved expression of four bacterial genes and manipulation of metabolic pathways in both amino acid and fatty acid synthesis. According to Yves Poirer, University of Luasanne, "one of the most complex feats of metabolic engineering yet performed in plants". But the resulting yield is still relatively low and would have to be improved before commercialization.
Although bacteria can also produce the environmentally friendly plastic, the process is currently five times more expensive than conventional plastics made from petroleum. Plants are much more efficient at converting carbon into plastic and could ultimately make the production of biodegradable plastic economically competitive.
Certain bacteria make plastics naturally, but the fermentation requires the addition of a carbon source, such as glucose, which must be first extracted from crops such as corn. This makes bacterial plastic production much more expensive than conventional plastic manufacture. As plants fix carbon directly from the atmosphere, crops engineered with the bacterial genes for producing the plastics should be much more efficient. To date, however, scientists have only succeeded in engineering plants to produce plastics with poor physical properties, such as brittleness, unsuitable for use as commodities.
Recombinant Proteins from Plants: Methods & Protocols - 2008. editors Loic Faye & Véronique Gomord - currently the best, most up to date book on Molecular Farming