Solutions for modern agriculture and plant biotechnology.

Name: Commercial Laboratories of Plant Production, New Technologies

Technology Transfer:

The Know-How of construction of the Commercial Laboratories makes it possible to take advantage of all the advantages offered by the specific lighting conditions for each region, on the other hand the use of the Temporary Immersion Bioreactors (BIT) reduces the costs and increases the production volumes in limited physical spaces, the combination of these factors constitute a guarantee in obtaining high quality plants at low cost. The Center for Bioplants of the University of Ciego de Ávila has worked on the development, implementation and transfer of conventional micropropagation technologies since 1982. Prototypes of Temporary Immersion Bioreactors have been established as an important complement and superior technology in the Massive propagation of several botanical families of ornamental plants, fruit trees, ornamental plants and of agricultural interest in general. The transfer of technologies related to the massive propagation of plants under these premises present as fundamental characteristics that of reducing costs and increasing production volumes of propagules of high quality. For this purpose, natural light is used in growth chambers, simple and economical culture media, chemical sterilization, the use of temporary immersion techniques, as well as the qualification and professional improvement of specialists, together with international quality standards that make this a distinctive process. Characteristics and advantages of the project. The designed Commercial Laboratories present a construction know-how that makes it possible to take advantage of all the advantages offered by specific lighting conditions for each region. This allows costs to be reduced by the use of solar energy. They also present simple designs that are appropriate to the demands of the work to allow an inter-relationship or flow between the premises or departments that have a direct participation in the production process. This allows an adequate comfort to the workers as well as an easy handling of the specialists during the commercial exploitation of the same. The infrastructure of the Laboratory allows to achieve productive increases in the short term, as well as products with a quality comparable to those demanded by the international market. The plants can be certified in terms of genetic stability (true-to-type) and disease-free. It also includes a Department of establishment and scaling, meristem culture and in vitro conservation of germplasm, which will allow the work of mass propagation of plants in Temporary Immersion Bioreactors, somatic embryogenesis, conservation of cauline apices, plant material sanitation . Additionally, the cryoconservation of plant tissues and the obtaining of new varieties of ornamental plants can be implemented as a distinctive product in the productions of the Commercial Laboratory. Technical foundation. The design, construction and start-up of the Commercial Laboratory for the massive production of plants through the use of Temporary Immersion Bioreactors will be carried out in the following stages of work: Design of the Commercial Laboratory of Massive Plant Propagation: advising is included during detailed engineering, as well as advice for the purchase and assembly of equipment and furniture. Start-up of the Commercial Laboratory of Massive Propagation of Plants: it includes the consultancy for the validation of the start-up of the Laboratory, the transfers of technologies, specialized trainings of the managerial staff or specialists or of the workers in general of the Laboratory. Exploitation of the Commercial Laboratory of Massive Propagation of Plants: it includes the improvement and training of the personnel of the Commercial Laboratory of Massive Plant Propagation. Expected benefits. Increase of the production, of the productivity and quality of the product that makes it possible to reach a bigger national and international market for the commercialization of the seedlings in any of its phases of growth. Obtaining and introducing new varieties of plants that are more attractive to the market as those of a high value for human consumption due to their quality, yield or resistance to pests and / or diseases, as well as for crops that show ornamental interest, forestry or horticultural. Additionally, variations or variability in the crops of interest can be obtained and improved varieties can be established in the market, which will allow increasing the biodiversity of the plants or products that will be offered in the new Commercial Laboratory in order to reach a position of relevance in the market. National and international. Application experiences. The technology transfers of our products are used successfully in the following national and international institutions: National: Commercial laboratory of massive propagation of ornamental plants. University of Ciego de Avila, CUBA. Commercial laboratory of massive propagation of fruit plants. Bioplant Center. University of Ciego de Avila, CUBA. Biofactory of banana production. National Seed Company. Camagüey, CUBA. Biofactory of banana production. National Seed Company. Ciego de Avila, CUBA. Commercial laboratory of massive propagation of ornamental plants. Provincial Community Company. Ciego de Avila, CUBA. Tissue and forage tissue culture laboratory. Sancti Spíritus, CUBA. Cultures and tissues laboratory. Las Tunas, CUBA. TROPIFLORA culture and tissue laboratory, Havana City, CUBA. International: Federal University of Santa Catarina. BRAZIL. CENARGEN / EMBRAPA. Brasilia, BRAZIL CNPMF / EMBRAPA. Cruz das Almas, BRAZIL. CPERZUCAR. Pirazicaba, BRAZIL. Production Center of Peñaflor. San Gil? Bucaramanga, COLOMBIA. Laboratory of Plant Biotechnology. Eastern University. Black river ? Medellin Colombia. Laboratory of Plant Biotechnology. Agronomy faculty ? University of Montería, COLOMBIA. CBIOCA. ESPOL? Guayaquil, Ecuador. Technical University of Esmeralda (UTEC), ECUADOR. Laboratory of Plant Biotechnology. Technical Institute of Costa Rica (ITCR). Seat San Carlos, COSTA RICA. Laboratory of Plant Biotechnology. Fitotécnia Area? Autonomous University of Chapingo, MEXICO. Laboratory of Plant Biotechnology. La Molina University? Lima Peru. Scientific Research Council (SRC). Kingston, JAMAICA. Tissue culture laboratory. University of Belize, BELIZE. Agricultural University of Godollo. HUNGARY. SBW International, BV. THE NETHERLANDS. PLANTECK. Biotechnologies. (Québec), CANADA.

Name: Protocols of Massive Plant Propagation

Technology Transfer:

Each crop presents its specifications in terms of handling and care in each of its phases, from implantation to hardening, our Center offers the technical instructions for propagation either by conventional techniques or through the use of temporary immersion bioreactors ( BIT) of the following species: Forestry: The rescue of forest species of commercial interest is one of the priority lines of biotechnology as a basic tool in supporting the improvement of trees plus. In this way, it accelerates multiplication rates and participates in reforestation programs in areas with large deforestation problems as a way to maintain the balance of the natural ecosystem. (See forest products) Ornamentals: It is the ornamental plants that have been the subject of the greatest commercial micropropagation in the world. It is noted that the number of commercial laboratories in the plant tissue culture industry in Europe increases at a rate of 3.5 times per year. Some prestigious firms in Western Europe recognize that Cuba can obtain close to 300 million USD annually with the sale of ornamental plants, which is why it constitutes a powerful market for our country. It is for this reason that since its creation in 1991, the Center for Bioplants of Ciego de Avila has been working on the propagation and marketing of ornamental species of high commercial value on a national and international scale, among which are (See ornamental products). Fruit trees: Within the plantation crops, are the fruit trees of the plants with the greatest interest for micropropagation laboratories. Phytosanitary sanitation and the introduction of new varieties are the most important aspects when working fruit trees. In the Cuban fruit industry; bananas, citrus fruits and pineapple are among the most important items where micropropagation plays an essential role (See ornamental products).

Name: Temporary Immersion Bioreactors (BIT)

Technology Transfer:

The micropropagation of plants requires the periodic transfer of the crop to fresh medium due to the depletion and / or alteration of the nutrients, as well as the continued growth or proliferation of the tissue, which exceeds the capacity of the culture flask. Generally the cultures are kept in individual flasks and transferred to fresh medium in an interval of four to six weeks. The use of the liquid medium for in vitro propagation has some advantages and is considered an ideal technique for mass propagation of plants, because it reduces handling and is an indispensable requirement for the automation of the process. Some alternative micropropagation systems, with the use of liquid medium, have been developed with the purpose of automating the process and consequently reducing costs. More recently, Teisson and Alvard (1995) developed an apparatus for the micropropagation of plants from the modification of a 250 mL capacity Nalgene filtration unit whose trade name is RITA. In Israel, a group of researchers have designed a disposable bioreactor for organ culture that is currently marketed by OSMOTEK. In Cuba, from the development of the first semi-automated system of temporary immersion in 1997 developed by a team of researchers from the Bioplant Center, research has been conducted on the application of this technique in the proliferation of meristems of several species of agricultural interest, ornamental and forestry. Technological description of the Temporary Immersion Bioreactor (BIT) designed in the Bioplant Center. The system consists of two translucent autoclaveable plastic containers, sterilizable autoclavable silicone hoses, 0.2 * m hydrophobic filters, electrovalves and an air compressor. One bottle is the culture vessel for the explants and the other, the container for the culture medium. Both bottles are communicated through silicone hoses. The sterility conditions are achieved through the use of hydrophobic filters 0.2 * m in diameter. The culture batteries (formed by a vial for the culture of the explants and a bottle for the storage of medium) are placed on aluminum shelves with dimensions of 0.7 m wide x 2.3 m long x 2.5 m high. The distribution of air from the compressor to the bottles is made through a PVC connection. Each floor on the shelf has a light panel (fluorescent white) that guarantees a DFFF between 30 and 50 * mol. m-2 s-1. The frequency and the time of immersion are controlled through a programmable automaton. According to the programmed time, a valve is opened and the air coming from the compressor drives the culture medium towards the bottle containing the explants. During the immersion period, the air flow allows the bubbling of the medium, removes the explants and changes the atmosphere inside the culture vessel. After the immersion time, a second valve is activated and the medium is returned to the storage container. The frequency and the time of immersion are regulated by the automatic controller. Results for crops obtained with the BIT. Comparison with other methods of micropropagation The BIT has two possibilities of application: to obtain explants and their subsequent proliferation in conventional bottles of micropropagation or to obtain shoots suitable for rooting ex vitro and acclimatization. The following table summarizes the multiplication coefficients that have been reached in the different crops with the use of the BIT. Variety Cultivation Microp. Conv. Inm. Temp. Variety Cultivation Microp. Conv. Inm. Temp. Piña Cayena lisa 8.0 68.8 Malanga INIVIT 3.0 10.44 MD2 5.8 26.8 Mexico 1 2.8 7.71 Sugar cane C 91-301 3.7 34.1 Banana FHIA-18 3.8 7.4 C120-78 4.1 58.0 FHIA-01 3.4 10.4 C1051-73 3.9 30.2 Great dwarf 4.0 16.6 C323-68 4.3 39.5 Syngonium W. butterfly 7.3 28.0 C85-212 3.8 31.6 Pixie 2.2 18.4 Anthurium Tropical 4.0 79.8 Spathyphyllum Sensaion 3.7 61.5 Rose Miniature 3.5 15.9 Phylodendrom Xanadu 2.0 102.9 Graphic design of the Temporary Immersion Bioreactor (BIT). Components Description of the automatic temporary immersion system. a) A solenoid valve opens and the air pressure from the compressor drives the culture medium to the bottle containing the plants. b) Waiting bath time. c) After a selected period of time, a second valve opens and the air pressure returns the culture medium to the reservoir of medium. Air compressor. Solenoid valve. Hydrophobic filters (0.2 μm). Our specialists provide advice on the assembly, commissioning and operation of temporary immersion bioreactors. We have more than twenty technologies for different crops of agricultural, ornamental and forestry interest, while research is done on request for crops of specific interest. This technology is especially useful for the rapid proliferation of new varieties.

Name: Obtaining Bromelain

Technology Transfer:

The production of proteases, worldwide, represents 38% of the total volume of enzymes marketed. Bromelain (EC 3.4.22.32), of preferential use, for its broad specificity of action; It has been isolated and purified from plants of the Bromeliaceae family. We designed and optimized an original technology (Cuban Patent C12N 9/50) and economic, in which by protecting the groups? SH of the active center of the enzyme and the proper manipulation of the medium (nature, pH, ionic strength), high yields are achieved in terms of protein and specific activity. The functional characterization of the preparation allowed to evaluate the optimal kinetic parameters of activity. Its operational and conservation stability was demonstrated. The purification of the enzyme by combining chromatographic methods allowed its molecular characterization. The process is repeatable and easy to scale, which was evidenced by achieving yields of 15 to 25 g of product / kg of stems at an industrial level. Advantages of the new technology: Simple and easy to extract extraction process that only requires the addition of two reagents in small quantities. The extraction conditions allow the enzyme to be obtained with the -SH groups of the protected active center and at a pH that guarantees stability, decreasing proteolysis. Expenditures for fresh material are minimal, as pineapple stems are used as a source for extraction of fields to be demolished or as a byproduct of in vitro propagation methods of pineapple in Temporary Immersion Systems. You can obtain product with different degrees of purity. The EC (raw extract) that are used in the Biotechnology Industry to obtain culture media. The highest purity preparations are used in the Pharmaceutical Medical industry with potential for use as an antitumor.