1.Establishment of a plant phosphorus utilization and weed control system based on phosphite and its dehydrogenase.
Guizhen YU ; Hang YUAN ; Zhu LUO ; Yanjuan LIU ; Xian LIU ; Yanxiu GAO ; Ming GONG ; Zhurong ZOU
Chinese Journal of Biotechnology 2019;35(2):327-336
Nowadays, available phosphorus (P) deficiency in soil and weed resistance to herbicides have emerged as two severe limiting factors for sustainable agriculture. Therefore, it is of urgent needs to improve plant absorption/utilization ability of the soil P, seek phosphate (Pi)-alternative P fertilizers, and develop new forms of weed control systems. Phosphite (Phi), as a P resource of relatively high amount only less than Pi in Earth, can be converted to utilizable Pi uniquely in some bacterial species by oxidization via its specific dehydrogenase (PTDH), but inhibits plant growth and development. This implies that Phi might rather become a suitable P fertilizer for plants if introducing a PTDH detoxifier from bacteria. Herein, we created the transgenic tobaccos harboring a Pseudomonas PTDH gene (PsPtx) amplified from the soil metagenome previously. RT-PCR showed that the exotic PsPtx gene could express similarly in root, stem and leaf tissues of all transgenic lines. PsPtx transgenic tobaccos could utilize Phi by oxidization as the sole Pi supply, and also outperformed wild-type tobacco with a remarkably dominant growth under Phi stress conditions. Moreover, the PsPtx gene was preliminarily evaluated with a notable quality as a potential candidate of the selection marker in plant genetic transformation. Conclusively, PsPtx and its encoded phosphite dehydrogenase might be applicable for developing a dual system of plant phosphorus utilization and weed control using Phi as P fertilizer and herbicide, and provide an effectual solution to some obstacles in the current crop transgenic studies.
Oxidoreductases
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Phosphites
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Phosphorus
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Plants, Genetically Modified
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Weed Control
2.Agricultural biotechnology safety assessment.
Scott MCCLAIN ; Wendelyn JONES ; Xiaoyun HE ; Gregory LADICS ; Andrew BARTHOLOMAEUS ; Alan RAYBOULD ; Petra LUTTER ; Haibin XU ; Xue WANG
Chinese Journal of Preventive Medicine 2015;49(1):78-87
Genetically modified (GM) crops were first introduced to farmers in 1995 with the intent to provide better crop yield and meet the increasing demand for food and feed. GM crops have evolved to include a thorough safety evaluation for their use in human food and animal feed. Safety considerations begin at the level of DNA whereby the inserted GM DNA is evaluated for its content, position and stability once placed into the crop genome. The safety of the proteins coded by the inserted DNA and potential effects on the crop are considered, and the purpose is to ensure that the transgenic novel proteins are safe from a toxicity, allergy, and environmental perspective. In addition, the grain that provides the processed food or animal feed is also tested to evaluate its nutritional content and identify unintended effects to the plant composition when warranted. To provide a platform for the safety assessment, the GM crop is compared to non-GM comparators in what is typically referred to as composition equivalence testing. New technologies, such as mass spectrometry and well-designed antibody-based methods, allow better analytical measurements of crop composition, including endogenous allergens. Many of the analytical methods and their intended uses are based on regulatory guidance documents, some of which are outlined in globally recognized documents such as Codex Alimentarius. In certain cases, animal models are recommended by some regulatory agencies in specific countries, but there is typically no hypothesis or justification of their use in testing the safety of GM crops. The quality and standardization of testing methods can be supported, in some cases, by employing good laboratory practices (GLP) and is recognized in China as important to ensure quality data. Although the number of recommended, in some cases, required methods for safety testing are increasing in some regulatory agencies, it should be noted that GM crops registered to date have been shown to be comparable to their nontransgenic counterparts and safe . The crops upon which GM development are based are generally considered safe.
Agriculture
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Animal Feed
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Animals
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Biotechnology
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China
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Consumer Product Safety
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Food, Genetically Modified
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Humans
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Models, Animal
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Plants, Genetically Modified
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Safety
3.Agroinfiltration, a useful technique in plant molecular biology research.
Zhao-Ming LIU ; Zong-Zhi LIU ; Qing-Wu BAI ; Rong-Xiang FANG
Chinese Journal of Biotechnology 2002;18(4):411-414
Agroinfiltration is a newly developed plant transient gene expression technique, which is simple, rapid and reproducible. It has been widely used in analyses of foreign gene expression, hypersensitive reaction, gene silencing, promoter activity and identification of new disease-resistance genes. In this paper, we describe the principle and the operation procedure of Agroinfiltration and its application in diverse aspects of plant molecular biology research. Our experiences in modification of the Agroinfiltration technique are also provided.
Agrobacterium tumefaciens
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genetics
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Genetic Vectors
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genetics
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Plants
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genetics
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Plants, Genetically Modified
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Research Design
4.Comparison of commercialization of transgenic crops in China and world-wide.
Chinese Journal of Biotechnology 2008;24(4):541-546
Currently, transgenic crops create huge economic, social and ecological benefits with the development of its commercial production. For China, the speed of development and commercialization of transgenic crops is a strategic issue for the sustainable agriculture development and the international competitiveness of our agricultural products. In this paper, we compared and analyzed the status of commercialization of transgenic crops in China and world-wide.
Agriculture
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methods
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trends
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China
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Crops, Agricultural
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genetics
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Gene Transfer Techniques
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trends
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Plants, Genetically Modified
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genetics
5.Bt transgenic crops for insect-resistance and modification of Bt protein and utilization of stacking strategy.
Chinese Journal of Biotechnology 2015;31(1):53-64
Insecticidal protein genes from Bacillus thuringiensis are currently the most widely used insect-resistant genes. They have been transferred to many crops for breeding and production. Among them, cotton, maize, potato and other insect-resistant crops are commercialized, creating considerable economic benefit. In this review, we summarized advances in identifying functional genes and transgenic crops for insect resistance, compared different strategies for enhancing vigor of insecticidal protein and utilizing gene stacking as well as listing valuable groups of stacked genes. In addition, the methods for multiple gene transformation was discussed.
Animals
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Bacterial Proteins
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genetics
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Crops, Agricultural
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genetics
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Endotoxins
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genetics
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Hemolysin Proteins
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genetics
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Insecta
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Plants, Genetically Modified
6.Advances in the research of genetic engineering of heavy metal resistance and accumulation in plants.
Ming-Lin LANG ; Yu-Xiu ZHANG ; Tuan-Yao CHAI
Chinese Journal of Biotechnology 2004;20(2):157-164
Using plants to remove or inactivate heavy metal pollutants from soils and surface waters provide a cheap and sustainable approach of Phytoremediation. However, field trials suggested that the efficiency of contaminant removal using natural hyperaccumulators is insufficient, due to that many of these species are slow growing and produce little shoot biomass. These factors severely constrain their potential for large-scale decontamination of polluted soils. Moreover, both the micronutrient and toxic metal content accumulated in crops determine the quality and safety of our food-chain. By a transgenic approach, the introduction of novel genes responsible for hyperaccumulating phenotype into high biomass plants and/or stable crops uptaking minerals as food is a promising strategy for the development of effective techniques of phytoremediation and improvement of nutritional value of stable food through a viable commercialization. Recently, the progress at molecular level for heavy metal uptaking, detoxification and hyperaccumulation in plants, and also the clarification of some functional genes in bacteria, yeasts, plants and animals, have advanced the research on genetic engineering plants of heavy metal resistance and accumulation, and on the functional genes (e . g. gsh1, MerA and ArsC) and their genetic transformated plants. These studies demonstrated commercialization potentials of phytoremediation. In this paper, the molecular approach, effects and problems in gene transformation were discussed in details, and also the strategy and emphases were probed into the future research.
Biodegradation, Environmental
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Genetic Engineering
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methods
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Metals, Heavy
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metabolism
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Plants, Genetically Modified
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genetics
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metabolism
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Soil Pollutants
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metabolism
8.HRCA and application in detection of genetically modified plant.
Zhen TAO ; Xing-Feng CAI ; Zhi-Qiang YAN ; Xiao-Bo HU ; Sheng-Li YANG ; Yi GONG
Chinese Journal of Biotechnology 2003;19(3):294-300
In this article primary studies of the application of hyperbranched rolling cycle amplification (HRCA) in exogenous genes detection of transgenic plants were done. Four padlock probes were designed according to the sequences of four genes/DNA fragments that are used widely in transgenic plants; part of the sequence of pKK233 was chosen as the linking part of padlock probes and a pair of HRCA primers was designed according to the sequence of linking part. Study of the specificity of ligation in HRCA with isotope labeled padlock probes indicated padlock probes could be ringed effectively only when corresponding target DNA exited in the same reaction system and could not be ringed when there was no corresponding target DNA exited. Ligation time is very different according to the characteristic of target DNA being used. 5 min to 10 min is enough if the target DNA is plasmid; 30 min to 60 min is needed if the target is genome DNA of plant because it's sequence is more complex than that of plasmid's. HRCA time was analyzed which indicated longer reaction time can obviously increase the amount of products. Quantity of enzyme in HRCA was also analyzed. Different amount of enzyme (from 0.5 unit to 4 units) can give similar result when other conditions are not changed. On the basis of the research, transgenic tobacco was detected with these four padlock probes and the results were just as prospective. In order to increase the efficiency of detection, multiplex HRCA (MHRCA)was used. In MHRCA more than one padlock probes are used at the same time in the same reaction system to detect more than one targets. Because the amplification products of MHRCA will be complex and it is almost impossible to analyze with electrophoresis, so reverse-blot is used. Detection results of transgenic tobacco with this method are the same with anticipation. Compare to MPCR method we established before MHRCA is more convenient to operate and more effective in detecting exogenous genes in transgenic plants.
Nucleic Acid Amplification Techniques
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methods
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Plants, Genetically Modified
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genetics
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Plasmids
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Tobacco
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genetics
9.Chloroplast genetic engineering: a new approach in plant biotechnology.
Tao SU ; Ya-Guang ZHAN ; Mei HAN ; Ai-Ping HAO
Chinese Journal of Biotechnology 2005;21(4):674-680
Chloroplast genetic engineering, offers several advantages over nuclear transformation, including high level of gene expression, increased biosafety, remedying some limitations associated with nuclear genetic transformation, such as gene silencing and the stability of transformed genes. It is now regarded as an attractive new transgenic technique and further development of biotechnology in agriculture. In this article we reviewed the characteristics, applications of chloroplast genetic engineering and its promising prospects were discussed.
Biotechnology
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methods
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Chloroplasts
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genetics
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Genetic Engineering
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methods
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Plants, Genetically Modified
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genetics
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Transformation, Genetic
10.Advances of resveratrol synthase gene in the application of genetic engineering and biofunctional investigation.
Shigang ZHENG ; Zhen LI ; Shancang ZHAO ; Qingguo WANG ; Wei LIU
Chinese Journal of Biotechnology 2014;30(3):341-354
Resveratrol synthase (RS) plays a key role in resveratrol (Res) biosynthesis. RS gene has been formerly reported to be transformed into many plant species and microorganisms, and to play certain roles in metabolic and regulation processes. In this paper, the transformations of RS gene in plants, and the related changes of biological properties, such as metabolites, anti-pathogen activities, anti-radical properties, and developmental characters in transgenic plants, as well as the production of resveratrol in microbes by utilizing RS gene were summarized. Moreover, the application prospects of RS gene in bioengineering were also addressed.
Acyltransferases
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genetics
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Genetic Engineering
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Plants, Genetically Modified
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enzymology
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genetics
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Stilbenes
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metabolism