Review of the biotechnology projects in the USA

Review of the biotechnology projects in the USA

Biotechnology development systematically supported by the state first of all started in the USA. In 2000 the Senate of this country passed a law encouraging industrial application of biomass. In October 2002 a vision was announced of the industrial biotechnology development in the USA for the period until year 2030, later on it was supplemented by more detailed plans. A special unit was established under the Federal Government of the USA for fostering this field (Office of the Biomass Program). It was envisaged until 2030 to create in the USA a competitive biotechnology industry which would produce 25 percent of the chemical products from the biomass and 20 percent of fuel. Long-lasting nature of the industrial biotechnology programs is highlighted, as well as the necessity for the state support for both the scientific researches and initial production organisation stages. Transition from the oil to renewing raw materials is a very radical change, which will have influence upon the whole global system of economy; therefore, it should be slow and gradual. A lot of projects dedicated to the development of biotechnology are implemented within this country.

The industrial biotechnology program, composed of the development of bio-energy and alternative energy application technologies, was implemented in the USA in 1999 – 2006. Currently it is finished and all the projects implemented under it can be found in the final report on the implementation of the program on the internet: http://www.ars.usda.gov/SP2UserFiles/Program/307/NP307AccomplishmentReportFinal-web.pdf. While developing industrial biotechnology receiving of bio-ethanol, bio-diesel and biomass was improved. An enzyme was identified which increases the production of sugar from the biomass, two new enzymes of micro organisms were patented, which are dedicated to the processing of cellulose, genetic libraries were created for the search on new enzyme biomass processing, dry fractioning wild pea transition into the plant enriched with proteins and starch was created as well as fermentation of pea starch into ethanol, new method for biochemical and chemical processing of wheat straw into butanol, new azeotropic ethanol purification technology was created, improved esterase of ferulic acid was identified, the enzyme hydrolising the bonds between lignin and hemicellulose, other works envisaged within the projects were performed. The next cycle of national program of industrial biotechnology was started by the meeting of consumers and intermediaries on September 2007. Due to high national and Congress interest in the renewing resources an increase in the financing is envisaged.

Further this database contains descriptions of the main green biotechnology projects of national importance of the USA, seeking to reveal as much as possible the spectrum of the issues solved with the help of biotechnology methods as well as that of the applied methods. If it is necessary, a more detailed description of the mentioned projects and the provided annual reports can be found on the indicated internet addresses.

In green plant biotechnology genetic modifications are widely spread, when seeking genetic improvement of the cultures, related to growth, sterility, blossoming or harvest, improvement of decorative characters of the flowers, increasing the nutrition and resistance against biotic and abiotic stress, inducting the neutrality to for the length of the day in a larger amount of strawberry cultivations as well as prolonging the production season of the strawberries which do not react to the length of the day. Moreover, new strategies and expression systems within the plants are also created, using plant viruses for the insertion of genetic material and transgene plants for the expression of borrowed proteins. Vectors based on the plant viruses are created for the production of bio-pharmaceutical substances within the plants. New technologies and products are developed for prevention, treatment and control of various human and animal diseases. Rhabdoviruses are used for the creation of vectors based on plant viruses. BVDV tests and diagnostic reagents are created. New technologies and products are developed within the plants, with the help of which it is sought to control such diseases as cancer of HIV.

Gene engineering is applied both for the practical purposes and for the implementation of scientific researches, when cloning the dominant genes. Especially big attention is paid to the markers related to gender, amplified in the strawberries, seeking to find out, if there are similar markers in peaches and to determine the presence of the loci, related to the sterility, blossoming or harvest. In another project dedicated for characterisation of the most important sequences of the genome, related to the increase of nutrition of the tomatoes, the objective was to determine the protein factors related to it, therefore, cis elements were analysed, in which the main nutrition genes are accumulated into a cluster, their functionality taking into consideration the transcription factors, which respond in a differentiated way according to the polyamines, metal-jasmonate and ethylene. Chimerical genes, which contain GUS/ GFP/ luciferase and complete sequences of promoters as well as sequences with deletions, are constructed and applying the method of bombing transferred into the tomatoes.

Molecular markers are very widely used for the genetic improvement of the plants; the most popular among them are SNP and SSR. The field of genomics and genetic variety are widely investigated. There are a little bit less works dedicated to plant proteomics and epigenetics as well as bioinformatics. A project is being implemented, the purpose of which is to create a computer program, which would be able to process the data of proteomics and to provide statistical solutions, processing the data on protein mass spectrum of soy, infected with scabs. The program will be adapted for processing and interpreting the data on protein mass spectrum of soy, infected with scabs, the results ill be provided in a form suitable for other analyses, for analysing the peptides and their grouping into proteins as well as for performing statistical analysis necessary for providing the results for evaluation of their reliability.

Plants of very different species are improved genetically, for example, tomatoes, sugar beets, petunias, lilies, strawberries, raspberries, blackberries, peaches, citrus plants. Investigations of separate cultures are characterised by different levels. For example, in soy investigations, using the methods of proteomics, the quality and safety of transgene plants is already analysed, while for lilies only the genetic transformation methods are developed so far. Biotechnology researches of soy are the most advanced among the researches of plants of high practical value. Genomic investigations are widely used, for example, analysis of EST sequences, composition of genetic maps, determination of qualitative character loci (QTL), related with various economically valuable characters. An increase of the variety of EST sequences is carried out, when sequencing five separate encoding DNA libraries affected by various abiotic stress. The processed sequencing results are put on GenBank databases. The data on SNP alleles in the cultivated and wild soy genotypes are allocated for the evaluation of genetic variety and defining population structure of the cultivated and wild soy genotypes as well as to determine sub-populations of the cultivated soy genotypes.

Comparing the biotechnology researches of cultures which have practical significance, soy is the most advanced. Especially a lot of work is performed with soy, investigating SNP genetic markers and composing the genetic maps of them. An integrated soy single nucleotide polymorphism (SNP) DNA marker genetic map is created, in which together with large amount of SNP markers together with previously investigated SSR markers would be listed. More complicated methods of investigation are also improved, seeking to investigate and apply the overlapping hybridisation system OVERGO of, allowing identifying the clones of soy artificial bacterium chromosome (BAC), which have specific DNA sequences. DNA sequences are taken from DNA markers known in the soy genetic maps: SSR and SNP (found in GenBank). The selected OVERGO nucleotides (approximately 1000) are located on the plates following specific order and used for hybridisation.

Among the solved issues, according to their nature, most of the works are dedicated to the control of diseases and pests as well as increase of the immunity. For this purpose the works are not confined only to the transfer of the immunity genes into the plants. Attempts are also made to create and to evaluate new methods for ensuring immunity, for that purpose using also phenomena of epigenetics, such as gene “silencing”, too high expression, opposition of the proteins and chemical inhibition of the processes of the keeper and the pathogen and the like. New genes of resistance are also sought or molecular markers related to them and the genetic maps of the latter are composed. Under the flowering culture improvement project, the pigmentation of anthocyans is investigated as well as immunity to the virus. A specific objective is set to establish genetic control of the production and regulation of anthocyans, using Solanaceous plants, to create genetically improved genetic lines of petunia and capsicum, characterised by new characters in looks and/ or increased resistance against insects, diseases and stress, to derive transgene flowering cultures which have immunity to viral diseases, to determine, how viruses can regulate the biosynthesis of anthocyans during the occurrence of mosaic phenomenon.

The newest found pathogens can be detected and analysed with the help of microscopy and molecular methods. Therefore molecular methods are widely used in the investigations of plant pathogens. Molecular markers are created for the detection and identification of evolutionary extraction of spiroplasma and phytoplasms. Attempts are made to identify as many as possible genes suitable for identification, and also to identify genes of the plant pathogen molecules, related to pathogenesis and occurrences of the characters, allocated for the identification of pathogen types, to define the correlation of the repeated sequences integrated into the chromosomes and non-chromosome DNA genome, plasticity and biological variety of the phytoplasms. A new practical strategy is developed for increasing the defence of the plant against the diseases caused by spiroplasma and phytoplasms. It is sought to find out signals crucial for the interaction of the plant/ pathogen and to use them increasing the immunity to diseases. The transmittance of the signal at the early stage is investigated, it is sought to identify, which structural characters of the viroid serve as a signal for replication, movement and pathogenicity. During the implementation of the project the transmittance of the signal based on redox and RNA is evaluated as well as mechanisms are established, which would increase the protection of the plant against the diseases caused by the bacteria and viroids. Modelling system of cell suspensions is used for the investigations as well as many other experiments such as agro-infiltration and in situ hybridization, yeast systems and field experiments with citrus plants. Phylogenetic relationships between important disease causing fungi basidiomycetes are described and analysed. Phylogenetic relationships among ascomycetous pathogens are characterised and analysed, based on the morphological characters and molecular characteristics. Especially big attention is given to cancer and fungi related to it, damaging the fruit trees and forest trees. During the implementation of the project PGR primers characteristic to that zone are composed, PGR is performed by amplifying the genes encoding ribosomes and proteins, including ITS, LSU, SSU, EF1-α and RNA polymerase gene regions. Efficient control of biologic cacao diseases is composed. For this purpose molecular mechanism crucial for the evolution of the disease, caused by Moniliophthora spp., is described by the influence and the interaction of cacao pathogens Moniliophthora spp. and Trichoderma fungi, which determines suppression of the diseases and increase of the harvest.

The project is also dedicated for the creation of database on the distribution of the spectrum of plant pathogen mass as well as the strategy and computer program, which using the above mentioned database would identify the pathogens according to the distribution of the mass spectrum. The data on mass spectrum, collected from the plant pathogens, is put on the search database. It is investigated, whether that spectrum is a unique bio-marker for the pathogen from which it has been taken. In the investigations of pathogens and diseases which have quarantine meaning, little covered pathogens and diseases are described, checking of the quarantine cultures is improved by improving their detection methods, increasing the sensitiveness, speed and reliability of such methods.