FUTURE PROSPECTIVES FOR COTTON BIOTECHNOLOGY IN EGYPT OSAMA A. MOMTAZ Deputy Director for Research, Agricultural Genetic Engineering Research Institute Agriculture Research Center- EGYPT The Government of Egypt places great importance on the significant role the agricultural sector plays in the national economy. Agriculture accounts for 20 percent of total exports, and 34 percent of the total labor force. The agricultural sector contributes to the overall food needs of the country, and provides the domestic industry with agricultural raw materials. One of the major targets for biotechnology in Egypt is the production of transgenic plants conferring resistance to biotic stresses resulting from pathogenic viruses, bacteria, fungi, and insect pests, and abiotic stresses such as salinity, drought, and high temperature. These biotic and abiotic constraints are major agricultural problems leading to serious yield losses in many economically important crops in Egypt. AGERI (Agricultural Genetic Engineering Research Institute) was established in 1990 at the Agricultural Research Center (ARC) to promote the transfer and application of this technology. AGERI aims to adopt the most recent technologies available worldwide to address problems facing agricultural development (Table 1). Opportunities for Developing Egyptian Cotton Biotechnological Approaches 1. Producing transgenic plants resistant to indigenous biotic and abiotic stress 2.Gene mining and the use of omics technologies for Egyptian Cotton improvement 3. Improving the oil content and fiber qualities for Egyptian cotton. 4. Reducing the dependency on imported cotton byproducts (oil-fiber). Table 1 Examples of current plant genetic engineering research at AGERI/EGYPT Discipline Potato Tomato Cotton Maize Rice Faba bean Cucurbits Wheat Banana Date palm Virus resistance @ @ @ @ @ Insect resistance @ @ @ @ Stress tolerance @ @ @ @ Genome mapping @ @ @ @ @ @ @ and fingerprinting Fungal resistance @ @
Monsanto /AGERI; a Private/Public Partnership The private sector has access to biotechnology, and has invested heavily in research and development (R&D) of technology and the necessary ancillary expertise to bring a product to market. The competitive edge of a private company depends on the proprietary nature of its R&D and the protection offered by intellectual property laws. A private company might engage in development of a product in conjunction with a developing country because (a) it addresses a technical problem critical to its own product development, (b) it presents an opportunity to enhance its public relations, and/or (c) it provides a window to an important market, technology, or germplasm of interest. Developing country institutions may be interested in working with private companies to gain access to important technology, develop managerial and business expertise, build intellectual capacity, or form a partnership with an entity that has an existing capability of bringing a product to market. The relationship between AGERI, an Egyptian public sector institution, and Monsanto, a U.S. private company, was forged through a relationship that involved common business interests to develop Giza- Bollgard II varieties. The importance of codevelopment of technology as opposed to technology transfer is especially pertinent in the case of Monsanto s relationship with AGERI. In this partnership, a public sector institution was able to bring a significant contribution to the table. AGERI has a state of the art biocontainment facility, and a team of trained scientists. AGERI can provide access to the local Egyptian market and the broader Middle East market, both of which are sufficiently developed to be attractive. In turn, Monsanto came to the discussion table with technology as well as with marketing, regulatory, and legal expertise of value to AGERI. AGERI has therefore initiated a partnership with Monsanto R&D to achieve the following: 1. Research and training for AGERI scientists to be trained at Monsanto in methodologies relating to development recent technologies in detecting Bt bollgard II gene incorporated in G. barbedense Giza varieties. 2. Potential for product development; Monsanto was granted access to develop molecular markers for the G. barbedense Gize varieties used in the back crossing program for Bollgard II gene cassette. Three parties have signed this agreement: Monsanto, USA, AGERI-Egypt, Cotton Research Institute, Egypt, Agricultural Research Center president. Collaborators have provided inputs to this research and training effort. Joint discoveries resulting from the work will be shared and patent rights will be sought according to terms of agreement. Monsanto will retain sole ownership of its proprietary Bt gene(s) and ARC will retain sole ownership of its proprietary germplasm. Under a separate agreement, ARC granted material transfer agreements (MTAs) for Monsanto to transfer Egyptian germplasm for backcrossing experimentation at Monsanto USA. Options for possible commercial development of transgenic cotton have been also considered.
Omics and Gene Mining Future In Egyptian Cotton: OMICs technologies such as functional, structure and comparative genomics, proteomics, metabolomics and transcriptomics are the future addition in the cotton biotechnology program at AGERI. One of the impediments in the genetic improvement of cotton fiber is the paucity of information about genes associated with fiber development (Momtaz O. A. 1998). Identification and isolation of Gossypium barbadense fiber quality gene families provide a novel opportunity to study fiber-associated genes because previous studies revealed this line was associated with some superior fiber quality traits compared to other barbadense varieties. We used an integrated approach of suppression subtractive hybridization (SSH), microarray, and real-time reverse transcription-polymerase chain reaction (RT-PCR) technologies to identify the potential genes associated with fiber development (Momtaz et. al., 2007 b & c, 2008a). At the functional genomics level, isolation and characterizations of transcriptomes and how these genes are expressed and function in plant is a very challenging and promising future tasks in Egyptian cotton biotechnology program at AGERI.. Comparative studies for these genes expression performance in cotton and other plants took place, identification and characterization of some important fiber quality related gene families (Actin, expansin and tubulin) lead to emphasize on how much the identified sequences are important and related to other functions in other species and if it was related or linked to a specific genetic markers. Moreover, these studies showed how much was the genome of Egyptian varieties very divergent than that of the American. This could narrow or widen the relationship between the two species (barbadense or hirsutum) (Momtaz et al, 2007b). Another challenge is to characterize some agronomic traits in the cotton genome and diagnose it at the RNA and protein levels. Such studies reflect the copy number of the gene and the level of gene expression and if some changes had occurred at the transcription level or even after the transcription. If the change has affected the protein and its function or its release inside the cell (place or movement) then the proteomics will reveal all the difference. One of the leading studies in the field of functional gene expression was achieved in the Egyptian cotton was the success to develop fiber on a media plates by controlling the concentrations of the growth regulators in the media rather than the whole plant (Momtaz et al, 1998a). Moreover, some agronomic gene families such as heat shock and LEA were identified at the level of RNA and protein in addition to sequencing some of these genes (Momtaz et al, 1998b). MAS Application In Egyptian Cotton Breeding : Marker-aided selection (MAS) is the application of molecular landmarks-usually DNA markers near target genes-to assist the accumulation of desirable genes in plant varieties. There are many reasons why molecular markers are useful in plant breeding. The improvement of plant host resistance and substantial agronomic traits for egyptian cotton is a good example. Using conventional approaches the plant breeder must be continually adding and changing genes just to maintain their target. Breeding effort spent in maintenance is a potential loss of gains in other traits. A more sustainable system can be developed by deploying more than one gene at the same time. The challenge is to find the right
combination of genes and put them into varieties most suitable for local production. When two or more genes are incorporated into the variety it is called gene pyramiding. The impact of MAS will continue to be significant particularly in an increasingly intellectual property (IP)-conscious environment. Marker technology is based on knowledge of endogenous DNA sequences; this has important practical implications as the cotton genome will be completely sequenced by an international effort. As long as there is a public commitment to maintain a cotton genome sequence in the public domain, useful genes for MAS should be readily accessible to national and international cotton breeding programs. Thus, because of their relative simplicity, easy integration into conventional breeding, and minimal background intellectual property, DNA marker technology and MAS are expected to be strong driving forces in cotton improvement in the future. Molecular markers has developed in Egypt for cotton since 1997, yet still need more work to landmark the genome of Egyptian varieties which in a unique way were very different from that of American cotton (upland). The comparative sequence analysis study has shown it very clearly and triggered forward for more specific studies on the Egyptian commercial varieties. As a worldwide concern of facing poverty and hanger, the public health regulatory systems raised a concern in every country to monitor and identify any potential adverse human health effects of transgenic plants as well as environmental concern in a formal risk-assessment process. GE cotton is now facing the same umbrella of biosafety and risk assessment because of the cotton seed oil that the oil production is depending on its compartment in Egypt side by side with other compartments (corn and sunflower seed oils). Not only oil production for its direct effect on human health but also other GE cotton with Bt, drought and salinity tolerance ability. That was a good initiator for the arising of the Egyptian biosafety and risk assessment committee. Although much of the discussion about molecular biology applications in cotton biotechnology were focused on the opportunity of crop improvement by gene transfer through transformation Momtaz et al. 1998c, 2000, 2001a & b), the same science brings new tools to assist plant breeders transfer genes through more conventional approaches. The complex traits for adaptation to abiotic stress are often difficult to identify. These are often difficult to identify and utilize in a breeding program without the additional help of modern science. Plant genomics is the engine to drive trait discovery and help solve intractable problems in crop production. To fully exploit the wealth of structural information obtained from the genome we must understand the specific biological functions encoded by a DNA sequence through detailed genetic and phenotypic analyses. Thus functional genomics requires diversity of scientific expertise as well as biological resources. In many important food crops the public sector has a large investment in biological resources, in plant breeding programs, and a long and skilled history of understanding biological function through national variety evaluation networks, as well as the global crop networks of the international agricultural research centers. These biological resources, scientific knowledge and expertise will become increasingly important in gaining knowledge about the function of genes and in developing markers for assisting the breeding process.
Intellectual Property Management Many R&D programs face the challenge and opportunities of managing intellectual property. Partnerships are critical to effective management and investment in intellectual property protection. Learning to manage intellectual property is a critical issue for many countries and institutions. Intellectual property management includes clarifying the role of institutions, developing an inventory of IP, developing ownership of intellectual property where appropriate, undertaking technology transfer, and marketing of the intellectual property. Human resource development is a major need in this area. Benefit sharing with holders of indigenous knowledge and genetic resources is an important issue that must be addressed. It is most important to build up human resource capacity in intellectual property for scientists, managers, policymakers, and society as a whole. Societal changes are reflected in changing IPR requirements, and further changes are likely to result from further strengthening of IP protection and finding ways to reflect the contribution of indigenous knowledge. REFERENCES Momtaz, O. A. 1998a. Effect Of Plant Growth Regulators On InVitro Fiber Development From Unfertilized And Fertilized Egyptian Cotton Ovules. Plant Growth Regulation Journal, 25: 159-164. Momtaz, O. A. 1998b. Identification and Induction of D7, D11, and D19 LEA Protein Genes in Some Egyptian Cotton Varieties. International Congress on Molecular Biology, Ain Shams University, Cairo, Egypt. Vol 1, pp 19-28. Momtaz O.A., A.A. Diab, M.R.Abu Shady, and M.A. Madkour. 1998c. Transformation of Egyptian cotton tissue (Gossypium barbadense) using Agrobacterium tumefaciens. Proceedings of World Cotton Research Conference-2 New Frontiers in Cotton Research, Athens, Greece. pp 314-319 Momtaz, O.A. A.A. Ibrahim,, I. A. Kamel, and M. A. Madkour. 1999. Construction And Characterization Of Bacterial Artificial Chromosome Library Of Egyptian Cotton (Gossypium barbadense ). Egyptian Journal of Agricultural Research, 77 (4), 1769-1787. Momtaz, O..A. A.A. Diab,, and M. A. Madkour. 2000. Development of Transgenic Egyptian Cotton (Gossypium barbadense) Varieties from Meristematic Tissue. Proceedings of the Beltwide Cotton Conferences, Cotton Improvement Conference, San Antonio, TX, USA. Vol. 1: 513-516. Momtaz O.A., Ahmed Barakat, Ahmed Bahieldin, Mona Sadek and Magdy Madkour.. 2001a. Transformation of Egyptian Cotton Varieties Using a Bacterial mtld Gene. Proceedings of the Beltwide Cotton Conferences, Cotton Improvement Conference, Anaheim, 9-13 January, CA, USA, vol. 2, pp: 1441-1430. Momtaz, O.A., Hossam. Z. Hassan, Hoda M. Barakat, Nahla A. El Sherif, A. Bahieldin and M. A. Madkour. 2001b. Development of Transgenic Egyptian Cotton Varieties Expressing Bacterial Fructosyl Transferase Gene Coding For Fructan Accumulation. Proceedings of the Beltwide Cotton Conferences, Cotton Improvement Conference, 9-13 January, Anaheim, CA, USA. Vol. (2): 1423-1429. Momtaz, O.A, Mohammed, S.I.M., El-Khodary, S., Zaki, El-Din H. 2006. Identification and Isolation of smhsp Gene From Gossypium barbadense L. cv. Giza 90 Long Stable Egyptian Cotton Variety. Arab Journal of Biotechnology. Vol 9, No/(1).:203-216.
.Momtaz, O.A., Fahmy Ashraf.H, Ayman A. Diab, and A. Hanafy. 2007a. Comparative Analysis of Amino Acid between Transgenic and non Transgenic Egyptian Cotton (Gossypium barbadense) Lines under Different Salt Stress Conditions. American-Euroasian journal of Agricultural and Environmental Sciences, 2 (1)06-15. Momtaz O.A., A. ElFateh. A. Bahi El-Din, S. Hassanin.. 2007b. Identification of actin-related gene family from G. barbadense Giza88 cultivar using PCR-based positional cloning. Arab Journal of Biotechnology. Vol 10 No/(1).117-129. Momtaz O.A., A. El-Fatih, A. H. Fahmy, A.A. Diab. 2007c. Comparative Sequence Analysis of Actin Related Gene Family Isolated from Gossypium barbadense. World journal of Agricultural Sciences. Vol 3 (1). 130-139. Momtaz O.A.. 2008a. Identification, characterization, and sequencing of gene families responsible for Egyptian cotton Fiber length and strength traits (tubulin, and Expansin genes) (In Preparation) Momtaz O.A.. 2008b. Identification, characterization, and sequencing of gene families responsible for Egyptian cotton salt and drought tolerant traits (In Preparation)