Review and prospect the application of plant biotechnology in bamboo research, molecular genetics, tissue culture and cell engineering of bamboo disease resistance and insect resistance, bamboo flower in vitro, mutants, genetic transformation and genetic engineering Other aspects were analyzed and discussed, and pointed out that some problems existed in the introduction of biotechnology into the field of bamboo research. Since Watson and Crick discovered the double helix structure of DNA in the 1950s and the subsequent determination of the central law of genetic information transmission, the establishment of recombinant DNA technology divided the concepts and technologies of biology, namely genetics, into all areas of life science. . Over the past decade, plant biotechnology has developed rapidly on the basis of molecular biology and genetics. Significant progress has been made in cell and tissue culture, genetic transformation, cell engineering, and genetic engineering. Haploids Breeding, rapid propagation of test-tube seedlings, artificial seeds, etc. are also applied in production, resulting in better benefits. In recent years, due to the special nature of bamboo itself and its huge economic benefits, its research has received more and more attention, but it should be seen that the overall level of bamboo research is still relatively low, such as most Research is still focused on cultivation and macrobiological characteristics, and many new techniques and methods have not been applied and embodied in bamboo research. In recent years, some scientific researchers have begun to try to introduce biological high-tech into bamboo research, which has injected new vitality into the research in this field and has achieved certain results. For example, the Chinese Academy of Forestry Zhang Guangchu et al. Rapid propagation, genetic improvement of bamboo and breeding of elite hybrid clones have made breakthroughs. In the future, bamboo research will introduce more recent achievements in molecular biology and genetics and make its research truly reach the molecular level. 1. Molecular genetics of bamboo disease resistance and insect resistance Every year, bamboo suffers major economic losses due to pests and diseases. Since the 1990s, with the development of molecular biology, genetics, and plant diseases, the pest-resistant research of higher plants has developed rapidly. Scientists are studying the natural pest-resistant mechanisms of plants from the molecular level. In particular, several plant pest resistance genes have been isolated, transformed and expressed in the past ten years, and new varieties of pest resistant plants have been promoted, laying the foundation for the study of plant disease resistance mechanisms. Plant disease resistance depends on the dominant resistance gene (R) of the plant itself and the corresponding non-pathogenic gene (Avir) of the pathogen. This mechanism of genetic resistance to plant-pathogen interactions is ubiquitous in plants. In recent years, bamboo stem rot, bamboo shoot rot, bamboo stem rust, bamboo leaf rust, bamboo plexus disease, bamboo black worm disease, etc. have caused no small harm, causing damage to the bamboo industry. Pathogenic pathogens include fungi, bacteria, mycoplasma, and the like. Studies have shown that plants have similar resistance mechanisms against different pathogenic bacteria. After the pathogens recognize and interact with the receptors encoded by host resistance genes controlled directly or indirectly by the Avir gene, a series of Signal transduction and response, systematically through a series of processes such as the opening of plasma membrane ion channels, depolarization of membranes, activation of signal transduction molecules and redistribution in cells, activation of defence genes, and production of phytoalexins. Disease resistance. In the management of bamboo forests, insect pests cause various degrees of damage each year, and bamboo rafts, bamboo raft moths, bamboo oval eggs, bamboo wasps, bamboo shoots, and other glands fly often cause extensive damage. Plant insect resistance gene engineering Since the introduction of the Bt (Bacillus thurigiensis) toxin protein gene into tobacco and tomato for resistance to insects, work in this area has been extensively carried out, and insect-resistant transgenic maize, potato, cabbage, Cotton, poplar, etc. In bamboo research, the Bt toxin protein gene can be introduced into bamboo. However, because of the different strains of Bacillus thuringiensis, the insecticidal spectrum is quite different. It is possible to clone different Bt toxin protein bases or introduce several different ones at the same time. Toxin protein genes, in addition, can also be used to change the base of codons to make them beneficial to the resistance of bamboo to insects, thereby increasing their expression levels in plants to achieve resistance to insects, especially against Coleoptera and Diptera. . 2 . Cell Differentiation and Development Cell differentiation refers to the process in which plant cells form different structures that cause changes in the function or potential developmental patterns. Cell differentiation is a very complicated process in higher plants. In the process of plant growth and development, due to the active division of the apical meristem, a series of complex morphogenesis processes form different organs and tissues, finally flowering and fruiting to complete its life history. Cell differentiation has therefore become a central issue in developmental biology. Due to the "totality" of plant cells, plant tissue culture and cell culture developed on this basis have become very useful techniques for studying the differentiation and development of plant cells. Bamboo is a special higher plant, but bamboo cells also have "totality." According to the view of modern biology, the development process is characterized by the fact that different genes in the DNA strand are selectively activated or repressed in a certain temporal and spatial order, and the cell selectively expresses genetic information in what way, becoming a basic problem of cell differentiation. Problems such as "decision", differentiation, dedifferentiation, and redifferentiation during development have not yet been completely solved. In addition, interactions between cells and tissues, influences of various internal and external factors, and intracellular metabolism are different. All of these have made the study of cell differentiation more difficult. The research on the differentiation and development of bamboo cells needs to be further studied. This problem needs to be solved from different perspectives such as morphology, cytology, plant biology, biochemistry and molecular biology. research. 3. Tissue culture and cell engineering In the late 40's and early 50's, the regeneration of roots and buds was successfully induced by the cultivation of tobacco pulp tissue. Thereafter, the tissue culture technology was rapidly developed. The versatility and morphological plasticity of plant cells in tissue culture and cell culture can provide a very useful experimental system for the study of plant morphogenesis. Cultured plant tissues or cells can be induced by the callus pathway to form various organs (such as roots, shoots or branches, flowers, etc.) as whole plants, or to regenerate whole plants by means of somatic embryogenesis, which is the plant's The foundations of genetic improvement and plant genetic engineering have been laid. Tissue culture has a single source of research materials, consistent genetic background of clones (clones), economical convenience, high efficiency, controllable conditions, small errors, rapid growth, short cycle, and strong reproducibility; it can perform annual tests or production. Tissue culture and cell culture systems have become a good vehicle for the study of plant morphology changes, physiological and biochemical reactions, and genetic improvement, and will increasingly become an indispensable research tool. In recent years, there have been many reports on the in vitro culture of bamboo. The explants used include lateral buds, apical buds, nodes, leaves, and mature embryos. These explants induce callus and callus The formation of adventitious buds or somatic embryogenesis regenerates intact plants. Bamboo tissue culture can be used for micropropagation and cultivation of virus-free seedlings, and can be used for a series of studies such as the flowering of test tubes. At present, the propagation of test tube seedlings in bamboo tissue culture is mainly carried out in southern China and Southeast Asia. For example, Zhang Guihe of the Chinese Academy of Tropical Agricultural Sciences used tissue culture and rapid propagation of Dendrocalamus latiflorus embryos. Guangdong Academy of Forestry Zhang Guangchu et al. performed tissue culture on bamboo, Dendrocalamos oldhamii, Dendrocalamus brandisii, and Bambusa pervariabili X Dendrocalamus latiflorus No. 7. The problems still to be solved in the tissue culture of bamboo are mainly the rapid propagation of scattered bamboo tissue culture, the interaction factors affecting physiological and biochemical changes in bamboo tissue culture, and the mechanism of action. Cell engineering includes cell mutagenesis, protoplast fusion, cell reorganization, and genetic material transfer. It can purposely change the heredity of cells, and it can be applied to bamboo breeding, germplasm preservation, rapid propagation of clones, and production of useful materials. 4 . Flowering in vitro As the flowering phenomenon of bamboo is relatively rare, flowering often causes serious economic losses to the bamboo forest. Therefore, the research on bamboo flowering has only certain theoretical and practical significance. In the past, studies on the flowering of bamboo mainly focused on a small amount of historical data and observation records, and lacked strict systematic experiments. The mechanism of bamboo flowering, control and prevention of flowering, and sexual cross breeding are all problems to be solved. However, due to the long and unstable flowering cycle of bamboo seedlings, the research in this area has caused difficulties. The use of in vitro culture systems for bamboo flowering research is an effective way and a good idea to solve this problem. After half a century of development, the in vitro flowering research has achieved a great deal of results and is moving toward the molecular level. However, these research results and technical methods have been introduced into the field of bamboo research only in recent years. Domestic and foreign reports on this aspect are rare. For example, Zhang Guangchu et al. have reported the phenomenon of 5 months old tissue culture seedlings blossoming in vitro; Nadauda RS et al. have reported the flowering phenomenon of bamboo seedlings of Bambusa arundinacea. . Studies have shown that the laws of flower formation in vitro are basically similar to the laws of overall flower formation, and the formation of flowers in tissue culture tubes has the advantages of high flowering rate, good repeatability, and stable technology, and therefore it is widely used for flowering research. However, there are only a few plants that have actually studied in depth, only Arabidopsis, snapdragon and cucumber. Although bamboo flowering in vitro has been successful in clump bamboo, there are problems such as low flowering rate, poor system stability, and unclear mechanism. The solution of these problems will contribute to the improvement of bamboo genetic improvement. At present, there are still many problems in the study of flower formation in vitro. For example, due to the complexity of the factors involved in the flowering process itself, flower initiation, flower primordia, and floral organ primordia, it is difficult to conduct comprehensive monitoring of the flowering process. Only a certain stage of test results can be used as a measure, which makes it more difficult to explore the flowering mechanism. On the other hand, molecular genetic manipulations are still rarely developed in isolated flowering systems and need to be further strengthened. In terms of the research on the flower formation of bamboo in vitro, we should first establish a stable tissue culture test tube flowering experimental system, and then proceed to further study on this basis. 5. Mutant mutant screening and isolation is one of the major issues in plant cell engineering. A gene can contain many mutation units in one mutation. Gene mutations caused by physical or chemical methods are the ultimate source of all genetic variability. Mutants have become good materials for genetics, biochemistry, plant physiology and dominant breeding studies. The reason why artificial selection of lung screening mutations is of great significance is due to the fact that natural selection mutations have few chances of being fixed. On the other hand, it can promote population variation and the fixation of mutants that are more resilient than wild-type in populations. . In the past ten years, due to the advancement of molecular genetics, biochemistry and plant physiology, rapid development of mutants has been achieved. The "mutation breeding research" characterized by the isolation of mutants to produce new germplasm has become a plant biotechnology application. In addition to detoxification and rapid propagation, and the separation of genes to produce transgenic plants characterized by "molecular biology research" in addition to another important way. Currently. Studies of mutants are rarely involved in bamboo studies. This aspect is due to the particularity of bamboo as a higher plant, which makes it difficult to study it in the conventional way; on the other hand, it is also due to insufficient understanding of its importance. With the increasing status of bamboo in the national economy, the dominant breeding of bamboo will be more and more important. In the case of other technical methods can not meet the needs, mutant screening and separation may be an effective shortcut. 6 . Genetic transformation and genetic transformation of genetically engineered plant cells refers to the use of explant cultured plant tissues, cells and protoplasts. Exogenous genes are introduced into plant cells by certain techniques and approaches. Techniques for obtaining transgenic plants capable of stably expressing foreign genes. Foreign genes can be obtained from recombinant DNA constructed from plasmid vectors and foreign genes. The purpose of genetic transformation of plant cells is to transfer a specific target gene, such as disease-resistance, insect-resistance, and herbicide-resistance genes, to the defects or traits of a good cultivar of a plant, so that the transgenic plant can retain its original Various good qualities, while adding a good trait controlled by the new gene of interest. It can be said that genetic transformation has brought about broad prospects for plant variety improvement. There are a large number of plant species, such as soybeans, cereals, etc., which have been improved through genetic transformation. The transformation methods used were mainly Agrobacterium-mediated, electroporation, PEG-mediated, gene gun, pollen tube passage, and microinjection. The introduction of genetic transformation methods into bamboo research can increase effective ways to solve bamboo diseases, pest control and trait improvement. Work in this area is almost blank. The current problems in genetic transformation mainly include: how to improve the efficiency of the regenerated plants and the transformation system; how to rationally design the plasmid and optimize the screening conditions to reduce the copy number of the transgene and the fragmentation of the transgene.
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