Development OutlookIn the past 10 years, due to the increasing prominence of the strategic position of the ocean in the sustainable development of coastal countries, as well as mankind's understanding of the particularity of the marine environment and the characteristics of marine biodiversity continue to deepen, the development and utilization of marine biological resources on multiple levels has greatly contributed to the rapid development of marine biotechnology research and application. 1989, the first International Conference on Marine Biotechnology (hereinafter referred to as MPS Conference) was held in Japan with only a few dozen participants. In 1989, the first International Marine Biotechnology Conference (hereinafter referred to as MPS Conference) was held in Japan with only a few dozens of participants, while in 1997, the fourth IMBC Conference was held in Italy with more than 1,000 participants. Now IMBC has become an important symbol of the global development of marine biotechnology, and the situation has become very popular. IMBC 2000 has just been held in Australia, preparations for IMBC 2003 have already begun in Japan, and Israel has made an early publicity campaign for the organization of IMBC 2006 and secured the right to host it. The IMBC, which is held once every three years, not only attracts many high-level experts and scholars to display and exchange their research results and discuss new research directions, but also greatly promotes the development of regional marine biotechnology research. In each continent, regional academic exchange organizations have been established, such as the Asia-Pacific Marine Biotechnology Society, the European Marine Biotechnology Society and the Pan-American Marine Biotechnology Association. Countries have also set up a number of research centers, of which the more famous for the United States University of Maryland Marine Biotechnology Center, the University of California, San Diego, Marine Biotechnology and Environmental Center, the University of Connecticut Marine Biotechnology Center, the University of Bergen, Norway, the International Research Center for Molecular Biology of the sea and the Japan Marine Biotechnology Research Institute and so on. These academic organizations or research centers have been organizing various symposia or working group meetings to study and discuss marine biotechnology issues with regional characteristics, and in 1998, with the support of the European Society for Marine Biotechnology, the Japan Society for Marine Biotechnology and the Pan American Marine Biotechnology Association, the former Journal of Marine Biotechnology and Molecular Marine Biology and Biotechnology were merged to form the Journal of Marine Biotechnology (hereinafter referred to as MB T). (hereinafter referred to as MB T), which has now become an authoritative international journal. Marine Biotechnology as a new subject area has been clearly defined as "the molecular biology of marine life, such as cell biology and its technological applications".
In order to adapt to the rapid development of the situation, the United States, Japan, Australia and other developed countries have formulated national development plans, marine biotechnology research as a priority area of development in the 21st century. 1996, China has also wasted no time to marine biotechnology into the national high-tech research and development program (863 program), for the future development of the foundation. It is self-evident that marine biotechnology has not only become a brand-new research field developed by the intersection of marine science and biotechnology, but also an important part of the scientific and technological development of all countries in the world in the 21st century, and it will show a strong momentum of development and great potential for application.
1. Development characteristics
1.1Strengthening basic biological research is an important cornerstone to promote the research and development of marine biotechnology Marine biotechnology involves a wide range of marine organisms, including molecular biology, cell biology, developmental biology, reproductive biology, genetics, biochemistry, microbiology, and even biodiversity and marine ecology, etc., and in order to make the development of a solid foundation, researchers attach great importance to the development of marine biotechnology. In order to have a solid foundation for its development, researchers attach great importance to relevant basic research. During the IMBC 2000 conference, when the author of this paper asked a senior participant: what is the main progress of this conference? He answered without hesitation: the increase in research results at the molecular biology level. This is true. Statistics of recent research results show that basic research in marine biotechnology is more focused on the molecular level, such as gene expression, molecular cloning, genomics, molecular markers, marine biomolecules, substance activities and their compounds. These oriented basic researches will have an important shadow on the future development.
1.2 Promote the traditional industry is the main aspect of the application of marine biotechnology At present, the application of marine biotechnology to promote the development of the marine industry is mainly focused on two aspects of aquaculture and the development of marine natural products, which is also the development of marine biotechnology research momentum is strong. The reason for this is that the development of marine biotechnology research is strong. In aquaculture, encouraging progress has been made in improving the reproduction, development, growth and health of important cultured species, especially in cultivating the excellent traits of species and improving disease resistance, such as the cultivation of fish with transgrowth hormone genes, polyploid nurseries of shellfish, gender control of fish and crustaceans, disease detection and prevention, DNA vaccines and nutritional enhancement, etc.; and in the development of marine natural products, the The use of the latest principles and methods of biotechnology to develop the separation of active substances of marine organisms, determination of molecular composition and structure and biosynthesis methods, testing biological activity, etc., has significantly contributed to the development of the industrialization of a new generation of marine new drugs, enzymes, polymer materials, diagnostic reagents, and other new-generation bioproducts and chemicals.
1.3 Ensure the sustainable use of the marine environment is another important aspect of the research and application of marine biotechnology to use biotechnology to protect the marine environment, control pollution, so that the marine ecosystem biological production process is more effective is a relatively new application of the field of development, therefore, both from the perspective of technological development, or industrial development, it has a huge potential to be tapped out. At present, the research involved mainly includes bioremediation (such as biodegradation and enrichment, fixation of toxic substances technology, etc.), anti-bio-adhesion, ecotoxicology, environmental adaptation and **** birth. The countries concerned to "bioremediation" as a marine ecological environment protection and sustainable development of its industry as an important means of bioengineering, the United States and Canada jointly formulated a marine environment bioremediation program to promote the application and development of this technology.
1.4 Marine policies related to the development of marine biotechnology have always been a matter of public concern, including the development strategy of marine biotechnology, patent protection of marine biotechnology, the importance of marine biotechnology to the development of aquaculture, the safety of genetically modified species and the control of the issue of the relationship between marine biotechnology and biodiversity, as well as marine environmental protection, etc., the formulation and implementation of policies, regulations and other areas of concern. The development and implementation of policies and regulations on marine biotechnology and biodiversity, as well as marine environmental protection, have received much attention.
2. Key Development Areas
Currently, the key research and development areas of international marine biotechnology mainly include the following aspects:
2.1 Developmental and Reproductive Biological BasisThe clarification of the physiological process of the development, metamorphosis, maturation, and reproduction of marine organisms in all aspects of the development of embryos, as well as their molecular regulatory mechanisms, is of great scientific significance not only for the elucidation of molecular regulatory rules of marine organisms in terms of growth, development, and reproduction. Molecular regulation of the law of scientific significance, but also for the application of biotechnological means to promote the growth and development of certain organisms and regulate their reproductive activities, improve the quality of aquaculture and yield has an important application value. Therefore, the research in this area is one of the research focuses in the field of marine biotechnology in recent years. Mainly includes: growth hormone, growth factor, thyroid hormone receptor, gonadotropin, gonadotropin-releasing hormone, growth a prolactin, osmolality-regulating hormone, reproductive inhibitory factor, oocyte final maturation-inducing factor, sex-determining factor and sex-specific genes, such as hormones and regulating factors, gene identification, cloning and expression analysis, as well as the fish embryo in cell culture and directional differentiation, and so on.
2.2 Genomics and gene transferWith the implementation of the global genome program, especially the human genome program, the study of structural and functional genomes of various organisms has become the focus of life sciences, and the study of genomes of marine organisms, especially functional genomics, has naturally become a hot spot for marine biologists. The current research focuses on full sequence determination of genomes of representative marine organisms (including fish, shrimp, shellfish and pathogenic microorganisms and viruses), and at the same time, the cloning and functional analysis of specific functional genes, such as drug genes, enzyme genes, hormone polypeptide genes, anti-disease genes, and salt-tolerance genes, and so on. On this basis, gene transfer, as an effective technical means for genetic improvement of marine organisms and cultivation of fast-growing and stress-resistant superior varieties, has become the focus of applied technology research and development in this field. In recent years, the research focuses on target gene screening, such as disease resistance genes, insulin-like growth factor genes and green fluorescent protein genes as target genes; high-volume, efficient gene transfer methods are also the focus of gene transfer research, in addition to the traditional microinjection method, gene gun method and sperm carrying method, the development of the retrovirus-mediated method, electroporation method, transposon-mediated method, and embryonic cell-mediated method.
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2.3 Pathogenic biology and immunity With the gradual deterioration of the marine environment and the large-scale development of mariculture, the disease problem has become one of the bottlenecks restricting the development of the world's mariculture industry. To carry out research on the pathogenic mechanism, transmission pathway and interaction between pathogenic organisms (e.g. bacteria, viruses, etc.) and their hosts is the basis for the development of effective prevention and control technology; at the same time, to carry out research on the molecular immunology and immunogenetics of marine aquaculture organisms and clarify the immune mechanism of marine fish, shrimp and shellfish is important for the cultivation of disease-resistant aquaculture varieties, and the effective prevention and control of aquaculture diseases. Therefore, pathogenic biology and immunity has become one of the key research areas of marine biotechnology, focusing on pathogenic microorganisms, disease-related genes, marine organisms, disease-related gene screening, cloning, the establishment of marine invertebrate cell lines, marine organisms, the exploration of immune mechanisms, DNA vaccine development.
2.4 Biological activity and its products of marine biological activity of the separation and utilization of substances is today's marine biotechnology is another research hot spot. Current research shows that a variety of marine organisms are widely present in the unique compounds used to protect themselves to survive in the ocean. Active substances from different marine organisms show great potential for application in biomedicine and disease control, such as sponges are an important resource for the isolation of natural drugs. In addition, there are some marine microorganisms with high or low temperature, high pressure, high salt and low nutritional functions, research and development of the use of these special functions of the marine extremophiles may be able to obtain the land can not get the new natural products, therefore, the extremophile research has also become the focus of marine biotechnology research in recent years. The focus of research in this field includes anti-tumor drugs, industrial enzymes and other special-purpose enzymes, screening of specific functional genes in extreme microorganisms, anti-microbial active substances, anti-reproductive drugs, immune-enhancing substances, antioxidants and industrial production.
2.5 Marine environmental biotechnology research in this field focuses on the development and application of marine bioremediation technology. Bioremediation technology is more extensive than the meaning of biodegradation, but also focus on biodegradation of the marine environment biotechnology. Its methods include the use of living organisms, or the production of products to degrade pollutants, reduce toxicity or transformed into non-toxic products, enrichment and immobilization of toxic substances (including heavy metals, etc.), large-scale bioremediation also includes ecological regulation in the ecosystem and so on. Application areas include large-scale aquaculture and factory farming, oil pollution, heavy metal pollution, urban sewage and other waste (water) treatment in the ocean. At present, the kinetic mechanism of microbial response to the environment, the biochemical mechanism of the degradation process, biosensors, the *** biotic relationship and mutual benefit mechanism between marine microorganisms and with other organisms, and the isolation and purification of anti-attachment substances are important research contents in this field.
3.Recent Research Progress in Frontier Areas
3.1 Developmental and Reproductive Regulation Applying the technique of regulating the maturation and reproduction of crustaceans by hormones such as GIH (gonadal inhibitory hormone) and GSH (gonadal stimulating hormone)[1], we investigated the regulation of thyroid hormones in the growth and development of the Jinshao, and found that the mRNA level of thyroid hormone receptor was highest in the brain, in the muscle and the lowest in liver, kidney and gill, indicating that thyroid hormone receptor plays an important role in the adult medaka brain [1], the homologous box (Homeobox) genes of sea squirts were characterized and 30 homologous box genes were isolated [1], the homologous box (Homeobox) genes of medaka were established [1], medaka embryonic stem cell lines were established and cell transplantation was performed to obtained chimeric medaka [1], established primary germ cell cultures of rainbow trout and isolated the Vasa gene [2], performed the isolation and characterization of reproductive inhibitory hormones in the spotted shrimp, Penaeus vannamei [2], applied receptor-mediated screening of GnRH analogues for fish reproduction [2], established sponge cell culture technology for drug screening [2], established the use of sea urchin embryos as a research gene expression model system [2], carried out research on sea urchin embryo engineering by gene transfer [2], studied the expression of human glucosyltransferase and rat hash glucokinase cDNAs in rainbow trout embryos [3], established a method for determining the rate of cell proliferation in marine fish fry by cell cycle protein-dependent kinase activity [3], investigated the expression of gibberellinase genes during moulting of Spotted Shrimp [4], isolated homologous frame genes from Sea Cucumber Homologous frame genes were isolated and sequenced[4].
3.2 Functional gene cloning established sequence signatures for the expression of mRN A in the liver and spleen of tooth flounder, a manipulator of pressure regulation was isolated from a pressure-tolerant bacterium in the deep sea, estrogen receptor and thyroxine receptor genes from Atlantic salmon, and gonadal inhibitory hormone genes from Norway prawn [1]; DNA microarray technology was applied on sponge cell culture, and the construction of the Banjo shrimp genetic linkage map, established marine red algae EST, isolated the catalytic subunit of mature proteasome from starfish oocytes, and preliminarily demonstrated that IGF-I proto E monopeptide of hardcore fish has anti-tumor effects [2]; constructed a plasmid vector for the marine yeast De-baryomyces hansenii, isolated and purified the carps serum from the carp protease inhibitor, an antimicrobial peptide-like substance from blood cells of the orchid crab, an actin promoter from the red abalone, and found that cell-cycle-dependent kinase activity can be used as a marker for cell proliferation in marine fish fry, cloned and sequenced eel cytochrome P4501A cD-NA, and analyzed the promoter region of the eel cytochrome P450IAI gene by a gene transfer method, and Isolated and cloned the eel cytochrome P450IAI gene, established polymorphic EST markers suitable for genetic mapping in Gulsho, constructed an EST database for yellow cap flounder and identified some new genes, established some tissue-specific EST markers for penaeid shrimp, and isolated 596 ESTs from lymphocytes of tooth flounder infected with Hirame Rhabdovirus virus from a cDNA clones [3]; PCR was used to clone an autotrophic hermaphrodite fish ? an actin gene, a peptide elongation factor EF-2 CDNA clone was isolated from a gilthead seabream cDNA library, and a TC1-like transposon element was found in the genome of lake trout [4]; genes identified and cloned included the following: antimicrobial peptide gene of the South American white shrimp (Litopenaeus vannamei), oyster allergen (allergen) gene, Atlantic eel and Atlantic salmon antibody genes, rainbow trout Vasa gene, medaka P53 genome gene, dinoflagellate eukaryotic initiation factor 5A gene, striped bass GtH (gonadotropin) receptor cDNA, abalone actin gene, cyanobacterial pyruvate kinase gene, carp optic violet gene regulatory series, and flounder lysozyme gene, etc [1-4].
3.3 Gene transfer The salmon IGF gene and its promoter were isolated and cloned, and the salmon IGF (insulin-like growth factor) gene expression vector was constructed[1]. The integration rate of exogenous gene transfer into zebrafish eggs was improved by nuclear localization of signaling factors [1], and a fast-growing transgenic tilapia line was established and evaluated for safety; triploidy induction was carried out in transgenic tilapia, and it was found that triploid transgenic tilapia, although not as fast-growing as the transgenic diploid, was superior to the non-transformed diploid fish, and at the same time the transgenic triploid females were totally sterile,. Thus, it has the value of popularization [2]; studied the technical method of ultrasonic treatment to promote the binding of exogenous DNA to the spermatozoa of the golden snapper, and used GFP as an indicator of transgene expression in cells and organisms; showed that the transgenic ditch catfish grows 33% faster than the control group, and that the transgenic fish is less able to escape from the enemy, and thus it can be released into the natural world without causing any major ecological hazards [3]; applied GFP as a genetic marker to study the optimization of conditions and expression efficiency of zebrafish transgenics[3]; in the breeding of disease resistance genetic engineering, the construction of marine organisms antimicrobial peptide and lysozyme gene expression vectors and gene transfer experiments[2]; in the species of transgenic research, it has now been gradually expanded from the economy of aquaculture fish to aquaculture of shrimp, shellfish, and certain ornamental fish[2.3]. Stable expression of exogenous genes was obtained in rainbow trout muscle by the gene gun method[4].
3.4 Molecular Marker Technology and Genetic DiversityThe feasibility of using fish gene introns as indicators of genetic diversity was investigated, and the genetic diversity of several marine organisms in the Atlantic Ocean and the Mediterranean Sea was studied by applying SSCP and sequencing methods[1]. The polymorphism of digestive enzyme genes in South American white shrimp was investigated[1]; the spacer region sequences of genomic DNA of parasitic protozoa and toxic methanogens were used as markers to detect the degree of contamination of these pathogenic organisms in environmental waters, and the sequence of the first internal spacer region (ITC-1) between the 18S and 5.8 S ribosomal RNA genes was applied as a marker to carry out crustacean inter- and intraspecific genetic diversity [2]; mitochondrial DNA polymorphisms in three populations of Penaeus vannamei were investigated, and species-specificity of Gobioid fry from Hawaii was identified by PCR. Intraspecific genetic diversity of South American white shrimp was revealed by determining intron sequences, genetic variation in different populations of brown trout was evaluated by using isozymes, microsatellite DNA and RAPD markers, 12 microsatellite DNAs were identified and isolated in flatfish, and highly variable microsatellite DNAs were found in California squid [3]; clarification of mitochondrial DNA in a deep-water fish (Gonostoma gracile) mitochondrial genome structure and discovered the first example of tRNA gene recombination in a scleractinian fish, determined satellite DNA sequences of commercially important marine rotifers, screened for microsatellite repetitive fragments in pangolins and soles using RAPD, isolated highly polymorphic microsatellite DNA from polychaete annelids, and investigated the eastern Thailand mud crab's The genetic diversity of the eastern Thai mud crab was investigated by RAPD[3]; the contribution of maternal genetic material in the genome of the female nucleus-developing striped bass was analyzed by AFLP[4].
3.5 DNA vaccine and disease control We constructed a DNA vaccine against fish necrosis virus [1]; we carried out a study on the construction of a DNA vaccine against rainbow trout IHNV and the prevention of disease, which showed that immunization of rainbow trout with a DNA vaccine encoding the IHNV glycoprotein gene induced a non-specific immune protective response, proving the feasibility of the DNA immunization pathway in fish, and we identified a DNA vaccine inducible by interferon in a cell line of rainbow trout [2]. A protein kinase inducible by interferon was identified [2]; an ELISA kit for viral pathogen detection in cultured shrimp was established, and viral pathogens in shrimp were identified by PCR and other molecular biology techniques, non-specific immune indicators in fish were used for monitoring the marine environment, and the feasibility of transferring disease resistance genes to improve the disease resistance of seabreams was studied, and antimicrobial defense reflections of salivary acid agglutinins in clams were investigated [2]; The antiviral activity of a marine biopolysaccharide and its derivatives was investigated[3]; a PCR-ELISA method for the determination of oyster pathogens was established[3]; and the immunolocalization of Latrunculin B toxin in red sponges was studied[4].
3.6 Bioactive substances New antioxidants were isolated from seaweeds [1], cell and tissue culture techniques of seaweeds for mass production of bioactive compounds were established, and a method for the preparation of antitumor compounds by in vitro culture of sponge cells was established [1]; anti-microbial peptides and their genes were identified and isolated from different organisms (e.g., shrimp and bacteria), and active peptides and their genes from fish hydrolysate that can be used as substrates for the growth of microorganisms were isolated [2]. used as substrates for microbial growth, the presence of anti-adhesion activators in marine organisms, the use of angiogenesis inhibitors as anti-conception agents, the extraction of immune activators from crabs and shrimps, the purification of photobacterial lethal compounds from marine algae and cyanobacteria, starfish extracts exhibiting batch spermatogonial cell formation in mice, the isolation of a non-toxic anti-adhesion activator compound from marine plant Zostera marina, and the isolation of a non-toxic anti-adhesion activator compound from sponges and cyanobacteria. attachment activity compound, isolation of anti-tumor compounds from sponge and sea squirt extracts, development of a natural inducer of coral metamorphosis, isolation of a new antioxidant drug from sea urchin, and identification of a long carbon chain highly unsaturated fatty acid (C28) in marine dinoflagellate phytoplankton, suggesting that marine fungi are an ideal source of isolation of biologically active compounds, such as anti-microbial peptides [2]; and discovery that the sulfate of Pseudomonas marina polysaccharide and its derivatives have antiviral activity, glutathione S-transferase was isolated from hard-shell clams, serine protease inhibitors were isolated from carp serum, aminoglycoproteinase was isolated from sponges, substances with DNA enzyme-like activity were isolated from a type of coral, and open sponge aquaculture systems have been established, which provide sufficient sponge raw materials for the large-scale preparation of biologically active compounds [3]; antioxidant fatty acids (C28) were isolated from the hydrolysis product of shrimp muscle, which indicates that shrimp muscle is an ideal source for the isolation of biologically active compounds such as antimicrobial peptides [4]. hydrolysis products were isolated to antioxidant peptide substances [4];
3.7 Bioremediation, extremophile microorganisms and anti-attachment studied the adsorption capacity of algae with trans-heavy metal sulfur protein genes for heavy metals in seawater environments, which was shown to be significantly greater than that of wild algae [1], and the therapeutic and application potentials of petroleum-degrading microorganisms for remediation of petroleum-polluted seawater environments were investigated [1]; and the marine magnetobacteria were investigated application potential in removing and recycling heavy metals in seawater environments [1]; Bacillus was used to remove nitrogen from fish farm effluent, and molecular techniques were used to screen microalgae used as bait for mariculture, and the potential of hexavalent chromium for bioremediation was developed, and cold-tolerant decane-degrading bacteria were isolated, and microbial degradation techniques for polyaromatized hydrocarbons in marine environments were investigated [2]; osmolality was isolated from salt-eating bacteria regulator gene from salt-eating bacteria and produced recombinant Ectoine (osmotic pressure regulator), isolated a heat-resistant bacterium from 2650 m deep sea, which can be used to isolate heat-resistant and heat-stable enzymes, discovered D-type amino acids and anaerobic ammonia acid dehydrogenase in the heat-resistant archaea, determined the genomic DNA sequences of three species of marine fireball bacteria, and carried out specific functional DNA sequence analysis by the help of CROSS／BLAST analysis. BLAST analysis for the screening of specific functional genes, collected more than 1,000 species of cold-phagic bacteria from seafloor sediments, seawater, and the Arctic Ocean, and isolated a variety of cold-adapted enzymes from these bacteria [2]; established a simple method for the determination of barnacle-attachment-inducing substances, investigated morphological interactions necessary for adhesion between Chlorophyta and ****-biotic bacteria, and investigated anti-attachment and anesthetic effects of coral anti-attachment substance (dterpene) analogues [3]; analyzed the initiation process of fouling in coastal environments and examined the effects of sedimentation and attachment [4].
4. Prospects and Suggestions
The above research analysis shows that marine biotechnology, as a brand new discipline, has become an important field of marine research and development in the 21st century, and is rapidly developing along three application directions. First, aquaculture, its goal is very clear is to enhance the traditional industry, prompting the aquaculture industry in the cultivation of fine varieties, disease prevention and control, large-scale production and many other aspects of leapfrog development; Second, marine natural product development, its goal is to explore the development of high value-added marine new resources, to promote the development of new marine pharmaceuticals, macromolecular materials and special function of the marine bio-active substances industrialization; Third, marine environmental protection, the goal is to ensure the sustainable use of the marine environment and the sustainable development of the industry. It is gratifying that this application development trend is consistent with the development needs of China's marine industry, especially with the high technology needs for the sustainable development and utilization of China's marine biological resources[5] . In fact, in the past five years, China's marine biotechnology research and application has made great progress, achieved a number of research results with world advanced level, and played an important role in promoting the development of marine industry. In the 21st century, it is not only of practical significance but also of strategic value to increase the support for Marine 863 to further promote the momentum of the rapid development of marine biotechnology in China. In addition, in the face of the challenges of scientific and technological globalization, it is also very important to strengthen international cooperation and exchanges through multiple channels to promote China's marine biotechnology innovation and industrialization to a higher level.
From the point of view of technology application, marine biotechnology is mainly to utilize the characteristics of marine environmental specificity and biodiversity, to develop and utilize the resources of marine biological groups from the molecular and cellular level, i.e., from the level of high technology on a multidimensional basis. Genetic resources and natural product resources, then the basic research related to this is very important. In fact, this is also an international research development trend. In order to make up for the shortcomings in this area, there is a need for multifaceted support and cooperation in the development of marine biotechnology in China, which not only needs to communicate and converge with the National Key Basic Research Development Plan, the National Natural Science Foundation of China and other relevant programs, but also needs to strengthen the basic construction. It is necessary to strengthen both the construction of pilot bases and industrialization bases, as well as infrastructure construction, such as strengthening the construction of open laboratories, research bases, biodiversity resource libraries, seed libraries, and information databases. These measures are of far-reaching significance to the development of China's marine biotechnology to a higher level.
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