The difficulties of defining the term "GM".
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This review introduced the recent research progress on transgenic methods and system of receptor in soybean. The major obstacles of genetic transformation in soybean and possible approach for solving the problem were also discussed. Cotyledon node via Agrobacterium tumefaciens-mediated and immature cotyledon via particle bombardment were thought to be the efficient systems of genetic transformation. Three problems existed in genetic transformation of soybean. The first one was that the system of tissue culture needs to be further improved. The second was that efficiency of genetic transformation was still low and difficult to be repeated. The last one was that restricted genotypes of soybean have been transformed successfully as a receptor. The path of solving these problems need to set a new and high efficient system for tissue culture in soybean. Also the number of target gene to be transformed will be increased from single gene to several genes at same time.
Chloroplast genetic transformation is a new way of plant genetic engineering. This paper reviews the construction methods of expression vector used in chloroplast genetic transformation. It contains the homologous recombinant fragments, the chloroplast specific promoter and terminater, the selectable marker genes and the interest genes whose expression in chloroplast have been achieved.
CpTI (Cowpea Trypsin Inhibitor) is a widely used insect resistance gene in the plant genetic engineering for its high insecticidal activity and the minimal ability of the insects to evolve resistance to it. To facilitate the safety assessment of genetically modified foods (GMFs) with CpTI protein, we need to produce gram quantities of this protein in microbes. With the pGEX fusion expression system, we expressed the GST-CpTI protein in E. coli BL21, which accounted for approximately 40% of germ proteins. By Glutathione Sephrose 4B affinity chromatography, GST-CpTI was obtained with the purity up to 90%. Overnight incubate the fusion proteins with Thrombin protease, we got the CpTI proteins cleavage of GST tag. Both of the GST-CpTI and CpTI proteins showed notable trypsin inhibitor activity. Immunization of rabbits with purified fusion protein generated high titer antibodies (> 20000), measuring by ELISA. Western Blotting also showed specific Ag-Ab binding band between the antiserum and the CpTI proteins no matter in the whole supersonic germ proteins or purified from the column. All these made a good ground for the further safety assessment of CpTI protein.
Study of beta-1,3-glucanase is one of the hot points in plant genetic engineering of disease resistance,big progress has been made in the past few years. This paper briefly reviewed the main biological characters and mechanism involved in the disease resistance,the classification,structure,function and the transformation of beta-1,3-glucanase genes.
A wide spectrum of strategies to genetically engineer potato plants resistant to potato tuber moth, Phthorimaea operculella (Zeller), have been investigated. The potato cv Iwa was transformed with a range of genes under the transcriptional control of the CaMV 35S promoter using Agrobacterium-mediated gene transfer. The transferred genes encode protease inhibitors (spleen inhibitor and alpha1-antitrypsin inhibitor), biotin-binding proteins (avidin and streptavidin) and Cry proteins (crylAc9, cry1Ba1, crylCa5 and cry9Aa2). Of these three transgenic approaches, cry genes have proved the most useful. In order to control the expression of the cry genes in foliage and not in the tubers a light-inducible Lhca3 promoter from potato was also used. The interaction of different cry genes was investigated using an experimental approach to simulate gene pyramiding in potato. Potato plants transgenic for both the crylAc9 and cryAa2 genes were developed and evaluated to help provide a more durable resistance to potato tuber moth.
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Indole alkaloids in Catharanthus roseus have been in focus because of their medicinal value. These alkaloids consist of an indole moiety provided by tryptamine and a terpenoid portion provided by the secologanin. The most important catharanthus alkaloids are vinblastine (VLB), vincristine (VCR) and ajmalicine. VLB and VCR are clinically useful anticancer agents whereas ajmalicine is used for the treatment of circulatory diseases. VCR and VLB are the most expensive because of their low abundance in the plant, and are formed by the coupling of monomeric indole alkaloids vindoline and catharanthine, catalysed by peroxidases. The pathway that lead to monomeric indole alkaloids involves more than 20 enzymes of which 16 enzymes have been isolated and characterized biochemically, and only three at the molecular level. The present state of knowledge on enzymes and genes involved in indole alkaloid biosynthesis and various aspects of their regulation has been discussed.
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L-Ascorbic acid (vitamin C) in fruits and vegetables is an essential component of human nutrition. Surprisingly, only limited information is available about the pathway(s) leading to its biosynthesis in plants. Here, we report the isolation and characterization of GalUR, a gene from strawberry that encodes an NADPH-dependent D-galacturonate reductase. We provide evidence that the biosynthesis of L-ascorbic acid in strawberry fruit occurs through D-galacturonic acid, a principal component of cell wall pectins. Expression of GalUR correlated with changing ascorbic acid content in strawberry fruit during ripening and with variations in ascorbic acid content in fruit of different species of the genus Fragaria. Reduced pectin solubilization in cell walls of transgenic strawberry fruit with decreased expression of an endogenous pectate lyase gene resulted in lower ascorbic acid content. Overexpression of GalUR in Arabidopsis thaliana enhanced vitamin C content two- to threefold, demonstrating the feasibility of engineering increased vitamin C levels in plants using this gene.
Recent studies in molecular biology and gene engineering were conducive to accumulation of numerous facts which help better understand the processes of plant genome functioning and follow up the effects of recombination processes in purposefully modified genome of a transformed plant on its expression. This review analyzes the major approaches to studies and results attained in plant gene engineering and plant genome imprinting.
The gene transfer into plants using the genetic engineering methods gives us the possibility to obtain transgeneric plants having acquired the new traits. Some bacterial genes can be used for this purpose. Obtaining of a transgeneric plant harbouring the cytokinin synthesis gene ipt (gene 4) from the T-DNA of Agrobacterium tumefaciens Ti-plasmid seems to be useful. The expression of tumor agrobacterial ipt gene in transformed plant cells interferes with the normal growth and regulation of the whole plant. The successful transfer of the cloned ipt gene from the recombinant plasmid pGV0319 into the tobacco plant using Agrobacterium vectors and succeeding regeneration of phenotypically normal transgenic plants are reported in the present paper.
The metabolism of one-carbon (C1) units is vital to plants. It involves unique enzymes and takes place in four subcellular compartments. Plant C1 biochemistry has remained relatively unexplored, partly because of the low abundance or the lability of many of its enzymes and intermediates. Fortunately, DNA sequence databases now make it easier to characterize known C1 enzymes and to discover new ones, to identify pathways that might carry high C1 fluxes, and to use engineering to redirect C1 fluxes and to understand their control better.