1.Screening for UV-C irradiation-enhanced transcription factors that regulate the metabolism of phenolic compounds in tomato fruit.
Wenzhuo HAO ; Huanhuan ZHENG ; Changhong LIU ; Lei ZHENG
Chinese Journal of Biotechnology 2021;37(7):2483-2494
Solanum lycopersicum phenylalanine ammonia-lyase 5 (SlPAL5) gene regulates the metabolism of phenolic compounds. The study of transcription factors that regulate the expression of SlPAL5 gene is of great significance to elucidate the regulatory mechanism underlying the biosynthesis of phenolic compounds in tomato fruit induced by UV-C irradiation. Here, yeast one-hybrid library of tomato fruit was constructed, and the yeast one-hybrid technology was used to screen the transcription factors that regulate the expression of SlPAL5, the key gene related to the synthesis of phenolic compounds in tomato fruit. As a result, a transcription factor, SlERF7, was obtained and sequenced, followed by the blast homology analysis. Further experiments confirmed that SlERF7 interacted with the promoter of SlPAL5 gene. In addition, UV-C irradiation significantly increased the expression level of SlERF7. These results indicate that SlERF7, which is regulated by UV-C irradiation, might be involved in regulating the transcription of SlPAL5, which provided foundations for further studying the regulation mechanism of the biosynthesis of phenolic compounds in tomato fruit induced by UV-C irradiation.
Fruit
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Gene Expression Regulation, Plant
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Lycopersicon esculentum/metabolism*
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Phenols
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Plant Proteins/metabolism*
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Transcription Factors/metabolism*
2.Research progress in regulation model in different types of plant trichome.
Xiao MA ; Kui LI ; Zhimin WANG ; Dayong WEI ; Qinglin TANG
Chinese Journal of Biotechnology 2020;36(10):2051-2065
Plant trichomes are special structures that originate from epidermal outgrowths. Trichomes play an important role in plant defense against pests and diseases, and possess economic and medicinal values. Study on molecular mechanism of plant trichomes will contribute to the molecular design breeding and genetic improvement of crops. In recent years, the regulation mechanism of trichome development has been basically clarified in the model plant Arabidopsis thaliana, while great progresses are also found in other plant species. In this review, we focus on the developmental regulation of trichome formation from gene and phytohormones levels in Arabidopsis and cotton (with unicellular trichomes), as well as in tomato and Artemisia annua (with multicellular trichomes). The research progress associated with trichomes is also introduced in other typical monocotyledons and dicotyledons. Finally, the research and application of plant trichomes are prospected.
Arabidopsis/genetics*
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Gene Expression Regulation, Plant
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Gossypium/genetics*
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Lycopersicon esculentum
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Plant Growth Regulators/metabolism*
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Trichomes/genetics*
3.Transient expression of organophosphorus hydrolase to enhance the degrading activity of tomato fruit on coumaphos.
Journal of Zhejiang University. Science. B 2009;10(2):142-146
We constructed an expression cassette of the organophosphorus pesticide degrading (opd) gene under the control of the E8 promoter. Then opd was transformed into tomato fruit using an agroinfiltration transient expression system. beta-Glucuronidase (GUS) staining, reverse transcription-polymerase chain reaction (RT-PCR), wavelength scanning, and fluorescent reaction were performed to examine the expression of the opd gene and the hydrolysis activity on coumaphos of organophosphorus hydrolase (OPH) in tomato fruit. The results show that the agroinfiltrated tomato fruit-expressed OPH had the maximum hydrolysis activity of about 11.59 U/mg total soluble protein. These results will allow us to focus on breeding transgenic plants that could not only enhance the degrading capability of fruit and but also hold no negative effects on pest control when spraying organophosphorus pesticides onto the seedlings in fields.
Aryldialkylphosphatase
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genetics
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physiology
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Coumaphos
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pharmacology
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Insecticides
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pharmacology
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Lycopersicon esculentum
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genetics
;
metabolism
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Plants, Genetically Modified
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Promoter Regions, Genetic
4.Characteristics of fruit ripening in tomato mutant epi.
Zhong-feng WANG ; Tie-jin YING ; Bi-li BAO ; Xiao-dan HUANG
Journal of Zhejiang University. Science. B 2005;6(6):502-507
The characteristics of fruit ripening and expression of ripening-related genes were investigated in epi, an ethylene overproduction mutant of tomato (Lycopersicon esculentum Mill.). The epi produces apparently more ethylene than its wild type VFN8 at every stage of vegetative and fruit growth and ripening; compared to VFN8, the epi fruit showed higher CO2 evolution, faster descending of chlorophyll, slightly quicker increase of carotenoid and lycopene, and faster reduction in pericarp firmness during maturation and ripening; and the mRNAs of three ripening-related genes including E8, pTOM5 and pTOM6 were at higher levels in epi. The ripening-related characteristics changing of the fruit are consistent with the increase of ethylene production and ripening-related genes expression. These results suggest that epi mutation possibly did not affect the ethylene perception and signaling during fruit ripening, and that the modified characteristics of fruit ripening possibly resulted from the ethylene overproduction and increased expression of ripening-related genes.
Ethylenes
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metabolism
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Fruit
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growth & development
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metabolism
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Gene Expression Regulation, Plant
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physiology
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Hardness
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Lycopersicon esculentum
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growth & development
;
metabolism
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Mutation
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Plant Proteins
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metabolism
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Signal Transduction
;
physiology
5.Characteristics of transgenic tomatoes antisensed for the ethylene receptor genes LeETR1 corrected and LeETR2 corrected.
Zhong-feng WANG ; Tie-jin YING ; Ying ZHANG ; Bi-li BAO ; Xiao-dan HUANG
Journal of Zhejiang University. Science. B 2006;7(7):591-595
Two stable transformed lines containing antisense LeETR1 [corrected] or LeETR2 [corrected] sequences and their hybridized line were investigated to determine the effect of LeETR1 [corrected] and LeETR2 [corrected] specificity in the ethylene receptor family in tomato (Lycopersicon esculentum Mill.) on ethylene signaling. The transgenic line ale1 containing antisense LeETR1 [corrected] displayed shorter length of seedling grown in the dark and adult plant in the light, severe epinastic petiole, and accelerated abscission of petiole explant and senescence of flower explant, compared with its wild type B1. The transgenic line ale2 containing antisense LeETR2 [corrected] also exhibited shorter hypocotyls and slightly accelerated abscission. The phenotypes of cross line dale of LeETR1 [corrected] and LeETR2 [corrected] were close to ale1 in many aspects. These results suggested that LeETR1 [corrected] probably plays a relatively important role in ethylene signaling of tomato growth and development.
Ethylenes
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metabolism
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Gene Silencing
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physiology
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Lycopersicon esculentum
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physiology
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Plant Proteins
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genetics
;
metabolism
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Plants, Genetically Modified
;
physiology
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RNA, Antisense
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physiology
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Receptors, Cell Surface
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genetics
;
metabolism
6.Construction of transgenic tobacco expressing tomato GGPS2 gene and analysis of its low light tolerance.
Cuiping LI ; Weihua DONG ; Xingguo ZHANG
Chinese Journal of Biotechnology 2015;31(5):692-701
To explore the influence of low light on the synthesis of carotenoids, chlorophyll and the adaptability of transgenic plants with tomato Solanum lycopersicon L. GGPS2 gene, we constructed a vector containing a GGPS2 gene with green fluorescent protein (GFP) as report gene under the control of a cauliflower mosaic virus 35S promoter and introduced it into tobacco Nicotiana tabacum L. cv. Wisconsin 38 by Agrobacterium tumefaciens-mediated transformation. PCR analysis of the DNA from kanamycin resistant tobacco indicated that the transgenic tobacco containing the nptII gene, SlaGGPS2 gene and without contamination of Agrobacterium. We also detected the root tip of kanamycin resistant tobacco showing characteristic fluorescence. The contents of carotenoid, chlorophyll and photosynthesis of transgenic tobacco increased in comparison with wild tobacco after low light treatment. In addition, leaf mass per unit area, total dry weight, ratio of root to shoot in transgenic tobacco were all higher than that of the wild tobacco, which proved that the transgenic tobacco could increase the accumulation of biomass and promote it transport to root. The transgenic tobacco with SlaGGPS2 gene can increase the contents of carotenoid, chlorophyll, enhance the photosynthetic rate, promote the biomass accumulation and its distribution to root. Hence, the transgenic tobacco with SlaGGPS2 gene had increased low light tolerance and the SlaGGPS2 gene maybe can be used in other crops.
Agrobacterium tumefaciens
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Carotenoids
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analysis
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Chlorophyll
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analysis
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Gene Expression Regulation, Plant
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Genetic Vectors
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Light
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Lycopersicon esculentum
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genetics
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Photosynthesis
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Plants, Genetically Modified
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metabolism
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radiation effects
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Tobacco
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metabolism
;
radiation effects
7.Molecular cloning of tomato LeHsp110/ClpB gene and its effect on the thermotolerance in plant.
Jin-Ying YANG ; Ying SUN ; Ai-Qing SUN ; Shu-Ying YI ; Jian LIU
Chinese Journal of Biotechnology 2006;22(1):52-57
The heat shock protein ClpB is a member of the Clp family and functions as molecular chaperones. ClpB is related to the acquired thermotolerance in organisms. A cDNA of 3144 bp was screened out of a tomato cDNA library. The polypeptide deduced from the longest ORF contains 980 amino acid residues, and was classified into HSP100/ClpB family based on the result of molecular phylogenesis analysis. Thus it was named as LeHSP110/ClpB according to its calculated molecular weight. LeHSP110/ClpB was characteristic of heat-inducibility but no constitutive expression, and was demonstrated to locate in chloroplastic stroma. An antisense cDNA fragment of LeHsp110/ClpB under the control of CaMV 35S promoter was introduced into tomato by Agrobacterium tumefactions-mediated method. At high temperature, the mRNA levels of LeHsp110/ClpB in antisense transgenic plants were lower than those in control plants. The PS II of transgenic plants is more sensitive to high temperature than that of control plants according to data of Fv/Fm. These results clearly showed that HSP110/ClpB plays an important role in thermotolerance of high plants.
Adaptation, Physiological
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genetics
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Chloroplasts
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metabolism
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Cloning, Molecular
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Genes, Plant
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genetics
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HSP110 Heat-Shock Proteins
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genetics
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metabolism
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Hot Temperature
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Lycopersicon esculentum
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genetics
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physiology
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Photosystem II Protein Complex
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metabolism
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Plant Proteins
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genetics
;
metabolism
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Plants, Genetically Modified
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genetics
;
physiology
8.High expression of thymosin alpha 1 by injecting recombinant PVX vector into the tomato fruit.
Yanbing NI ; Zhengwen SHI ; Defu WANG ; Min YAO ; Mu QIAO ; Pingyi GUO
Chinese Journal of Biotechnology 2009;25(4):537-541
For expression of foreign genes in plant, plant virus vector provides many advantages, such as high expression level, short expression period and wider plant hosts. In the present study, we report the expression of thymosin alpha 1 (Talpha1) in tomato fruits by potato virus X (PVX) vector. Talpha1 gene fragment from plasmid pGEM-T containing Talpha1 gene was cloned into plant virus vector pGR107 and the resulting pGR107-Talpha1 plasmid was confirmed by digestion with Sal I and Cla I. To express the Talpha1 protein, Agrobacterium tumefaciens GV3101 transformed with pGR107-Talpha1 was directly injected into tomato fruits through the fruit stylar apex at different developmental stages. The ELISA results showed that Talpha1 protein was expressed successfully in fruits, and the highest expression level was obtained from 2.5-3 week-old tomato fruits inoculated by bacterium at 1.0 OD600 density.
Agrobacterium tumefaciens
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genetics
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Fruit
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genetics
;
metabolism
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Gene Expression Regulation, Plant
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Genetic Vectors
;
genetics
;
Lycopersicon esculentum
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genetics
;
metabolism
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Plants, Genetically Modified
;
metabolism
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Potexvirus
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genetics
;
metabolism
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Recombination, Genetic
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Thymosin
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analogs & derivatives
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biosynthesis
;
genetics
9.Effect of cadmium on growth and photosynthesis of tomato seedlings.
Jing DONG ; Fei-Bo WU ; Guo-Ping ZHANG
Journal of Zhejiang University. Science. B 2005;6(10):974-980
A hydroponic experiment carried out to study the effect of five Cd levels on growth and photosynthesis of two tomato cultivars showed that the addition of 0.1 micromol/L Cd induced a slight increase in plant height of Hezuo 903 and the SPAD (the Soil-Plant Analyses Development) value of the 2 cultivars. However, at higher Cd levels, i.e., 1 and 10 micromol/L, root length and volume, plant height, and SPAD value were all significantly reduced. On an average of the 2 cultivars, exposure to 1 and 10 micromol/L Cd for 33 d reduced plant height by 18.9% and 46.4% and SPAD value by 11.2% and 31.6%, compared with control, respectively. Similarly, root length was reduced by 41.1% and 25.8% and root volume by 45.2% and 63.7%, respectively. The addition of Cd in the growth medium also had significant deleterious effect on net photosynthetic rate (Pn) and intracellular CO(2) concentration (Ci), with Pn being reduced by 27.2% and 62.1% at 1 micromol/L and 10 micromol/L Cd treatments compared to the control, respectively, while Ci increased correspondingly by 28.4% and 39.3%.
Cadmium
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toxicity
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Lycopersicon esculentum
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drug effects
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growth & development
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metabolism
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Photosynthesis
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drug effects
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Plant Leaves
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drug effects
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growth & development
;
metabolism
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Plant Roots
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drug effects
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growth & development
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Seedlings
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drug effects
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growth & development
;
metabolism
10.Functional analysis of specific promoter using vecotors harboring GFP/RFP double fluorescent marker genes.
Tao YIN ; Qiaoping QIN ; Shanglong ZHANG ; Jingmei LIU ; Daming CHEN
Chinese Journal of Biotechnology 2008;24(12):2106-2110
Most studies related to determining the expression profile of genes and specific promoters used histochemical localization of the reporter gene, gusA. While the histochemical method for visualizing gusA expression suffers from several limitations in the determination of gene expression and location, especially in the tissues with high background acitivty. To solve this problem, a transient expession vector pBI221-RFP/GFP, was constructed using GFP and RFP as double fluorescent marker genes. This vector used CaMV 35S promoter to drive GFP and determine the transforming efficiency. It analyzed expression profile of the target gene and promoter through the RFP activities of the tranformed tissues. Through the specific promoter AGPL1 from watermelon and E8 promoter from tomato, it is resistible to use this vector to study the expression patterns of promoters. Results indicated that the pBI221-RFP/GFP is a very efficient transient expression vector that can be verify the functions of the genes and promoters.
Citrullus
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genetics
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Gene Expression Regulation, Plant
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Genes, Plant
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Genes, Reporter
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Genetic Vectors
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genetics
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Green Fluorescent Proteins
;
genetics
;
metabolism
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Luminescent Agents
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metabolism
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Luminescent Proteins
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genetics
;
metabolism
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Lycopersicon esculentum
;
genetics
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Promoter Regions, Genetic