scholarly journals Construction of CRISPR/Cas9 expression vectors habouring gRNA targeted on SLIAA9 gene of tomato

2020 ◽  
Vol 18 (1) ◽  
pp. 147-156
Author(s):  
Bui Manh Minh ◽  
Ha Hong Hanh ◽  
Le Thi Thu Hien ◽  
Huynh Thi Thu Hue
Keyword(s):  
Transgenic Plants ◽  
Genome Editing ◽  
Tomato Fruit ◽  
Expression System ◽  
Secondary Compounds ◽  
Expression Vectors ◽  
Tomato Plants ◽  
Normal Morphology ◽  
Transgenic Tomato Plants ◽  
Pcr Test

Tomato (Solanum lycopersicum) is a nutritious fruit containing many secondary compounds with health benefits. The formation of tomato fruit through fertilization is controlled by auxin through Aux/IAA9 and ARF8 proteins. The mutated SlIAA9 gene leads to the parthenocarpic development of fruit or seedless tomato fruit. Nowadays, the CRISPR/Cas9 genome editing system is becoming increasingly popular in modifying desired genes on plant objects. In this study, gRNAs which target on tomato SlIAA9 gene were designed and inserted into CRISPR/Cas9 vectors. In addition, two strains of A. tumefaciens harboring pRGEB31-IAA9G2 and pRGEB32-IAA9G2 vectors carrying CRISPR/Cas9 expression system towards SlIAA9 gene in tomato were successfully created. The strain of A. tumefaciens harboring pRGEB31- IAA9G2 plasmid was used to develop transgenic tomato plants from Micro-Tom variety. PCR test showed that 5/14 plants had the presence of Cas9 gene in T0 plants. The transgenic plants have a normal morphology in comparation with the controls. The evaluation of mutant efficiency, type, and stability of mutations on the SlIAA9 will be conducted on next-generation plants when the mutations are stable and segregated into descendents.

MODIFIED COMPOSITION OF MICROBIAL COMMUNITY IN THE RHIZOSPHERE OF TRANSGENIC TOMATO PLANTS WITH A GLYCINE BETAINE SYNTESIS GENE

2020 ◽  
pp. 18-29
Author(s):  
A.A. ANTONOV ◽  
◽  
E.N. BARANOVA ◽  
A.A. GULEVICH ◽  
L.V. KURENINA ◽  
...  
Keyword(s):  
Microbial Community ◽  
Transgenic Plants ◽  
Root Zone ◽  
Microbial Community Composition ◽  
Transgenic Tomato ◽  
Choline Oxidase ◽  
Soil Culture ◽  
Oxidase Gene ◽  
Tomato Plants ◽  
Transgenic Tomato Plants

The change in the composition of soil microbiota as a result of the cultivation of various crops in agrocenoses is currently of great interest. The authors studied the effect of the root system of transgenic tomato plants (Solanum lycopersicum L.) grown in soil culture on the microbial community in the rhizosphere. The results showed that as a result of the cultivation of transgenic plants with the choline oxidase gene, the microbial community composition in the rhizosphere has changed significantly. A significant increase in the proportion (73%) and species diversity (Shannon index 2.25) of actinobacteria in the soil root zone of tomato transgenic codA plants as compared with control plants (10% and 0.95, respectively) has been revealed. The content of pseudomonads and micromycetes is significantly reduced (25 and 12% in transgenic plants; 70 and 81% in control plants, respectively). Thus, genetically modified plants are able to influence the microbial community structure in the rhizosphere.

scholarly journals Transgenic Expression of Pear PGIP in Tomato Limits Fungal Colonization

2000 ◽  
Vol 13 (9) ◽  
pp. 942-950 ◽  
Author(s):  
Ann L. T. Powell ◽  
Jan van Kan ◽  
Arjen ten Have ◽  
Jaap Visser ◽  
L. Carl Greve ◽  
...  
Keyword(s):  
Fungal Pathogen ◽  
Heterologous Protein ◽  
Tomato Fruit ◽  
Fungal Growth ◽  
Fungal Colonization ◽  
Pear Fruit ◽  
Tomato Plants ◽  
Transgenic Expression ◽  
Transgenic Tomato Plants ◽  
Tissue Maceration

Transgenic tomato plants expressing the pear fruit polygalacturonase inhibitor protein (pPGIP) were used to demonstrate that this inhibitor of fungal pathogen endopolygalacturonases (endo-PGs) influences disease development. Transgenic expression of pPGIP resulted in abundant accumulation of the heterologous protein in all tissues and did not alter the expression of an endogenous tomato fruit PGIP (tPGIP). The pPGIP protein was detected, as expected, in the cell wall protein fraction in all transgenic tissues. Despite differential glycosylation in vegetative and fruit tissues, the expressed pPGIP was active in both tissues as an inhibitor of endo-PGs from Botrytis cinerea. The growth of B. cinerea on ripe tomato fruit expressing pPGIP was reduced, and tissue breakdown was diminished by as much as 15%, compared with nontransgenic fruit. In transgenic leaves, the expression of pPGIP reduced lesions of macerated tissue approximately 25%, a reduction of symptoms of fungal growth similar to that observed with a B. cinerea strain in which a single endo-PG gene, Bcpg1, had been deleted (A. ten Have, W. Mulder, J. Visser, and J. A. L. van Kan, Mol. Plant-Microbe Interact. 11:1009–1016, 1998). Heterologous expression of pPGIP has demonstrated that PGIP inhibition of fungal PGs slows the expansion of disease lesions and the associated tissue maceration.

scholarly journals Transgenic Plants Expressing Geminivirus Movement Proteins: Abnormal Phenotypes and Delayed Infection by Tomato mottle virus in Transgenic Tomatoes Expressing the Bean dwarf mosaic virus BV1 or BC1 Proteins

2000 ◽  
Vol 13 (3) ◽  
pp. 297-308 ◽  
Author(s):  
Yu-Ming Hou ◽  
Rick Sanders ◽  
Virgina M. Ursin ◽  
Robert L. Gilbertson
Keyword(s):  
Transgenic Plants ◽  
Mosaic Virus ◽  
Viral Disease ◽  
Transgenic Tomato ◽  
Wild Type ◽  
Significant Delay ◽  
Tomato Plants ◽  
Movement Proteins ◽  
Transgenic Tomato Plants ◽  
Bean Dwarf Mosaic Virus

Transgenic tomato plants expressing wild-type or mutated BV1 or BC1 movement proteins from Bean dwarf mosaic virus (BDMV) were generated and examined for phenotypic effects and resistance to Tomato mottle virus (ToMoV). Fewer transgenic plants were recovered with the wild-type or mutated BC1 genes, compared with the wild-type or mutated BV1 genes. Transgenic tomato plants expressing the wild-type or mutated BV1 proteins appeared normal. Interestingly, although BDMV induces only a symptomless infection in tomato (i.e., BDMV is not well adapted to tomato), transgenic tomato plants expressing the BDMV BC1 protein showed a viral disease-like phenotype (i.e., stunted growth, and leaf mottling, curling, and distortion). This suggests that the symptomless phenotype of BDMV in tomato is not due to a host-specific defect in the BC1 protein. One transgenic line expressing the BC1 gene did not show the viral disease-like phenotype. This was associated with a deletion in the 3′ region of the gene, which resulted in expression of a truncated BC1 protein. Several R0 plants, expressing either wild-type or mutated BV1 or BC1 proteins, showed a significant delay in ToMoV infection, compared with non-transformed plants. R1 progeny plants also showed a significant delay in ToMoV infection, but this delay was less than that in the R0 parents. These results also demonstrate that expression of viral movement proteins, in transgenic plants, can have deleterious effects on various aspects of plant development.

scholarly journals Transgenic Tomato Plants Overexpressing Tyramine N-Hydroxycinnamoyltransferase Exhibit Elevated Hydroxycinnamic Acid Amide Levels and Enhanced Resistance to Pseudomonas syringae

2014 ◽  
Vol 27 (10) ◽  
pp. 1159-1169 ◽  
Author(s):  
Laura Campos ◽  
Purificación Lisón ◽  
María Pilar López-Gresa ◽  
Ismael Rodrigo ◽  
Laura Zacarés ◽  
...  
Keyword(s):  
Transgenic Plants ◽  
Pseudomonas Syringae ◽  
Acid Amide ◽  
Hydroxycinnamic Acid ◽  
Transgenic Tomato ◽  
Tomato Plants ◽  
Antibacterial And Antifungal Activities ◽  
Enhanced Resistance ◽  
Transgenic Tomato Plants ◽  
Key Enzyme

Hydroxycinnamic acid amides (HCAA) are secondary metabolites involved in plant development and defense that have been widely reported throughout the plant kingdom. These phenolics show antioxidant, antiviral, antibacterial, and antifungal activities. Hydroxycinnamoyl-CoA:tyramine N-hydroxycinnamoyl transferase (THT) is the key enzyme in HCAA synthesis and is induced in response to pathogen infection, wounding, or elicitor treatments, preceding HCAA accumulation. We have engineered transgenic tomato plants overexpressing tomato THT. These plants displayed an enhanced THT gene expression in leaves as compared with wild type (WT) plants. Consequently, leaves of THT-overexpressing plants showed a higher constitutive accumulation of the amide coumaroyltyramine (CT). Similar results were found in flowers and fruits. Moreover, feruloyltyramine (FT) also accumulated in these tissues, being present at higher levels in transgenic plants. Accumulation of CT, FT and octopamine, and noradrenaline HCAA in response to Pseudomonas syringae pv. tomato infection was higher in transgenic plants than in the WT plants. Transgenic plants showed an enhanced resistance to the bacterial infection. In addition, this HCAA accumulation was accompanied by an increase in salicylic acid levels and pathogenesis-related gene induction. Taken together, these results suggest that HCAA may play an important role in the defense of tomato plants against P. syringae infection.

scholarly journals Expression of erythropoietin in Indian tetraploid potato variety

F1000Research ◽  
2012 ◽  
Vol 1 ◽  
pp. 26
Author(s):  
Priti N Desai ◽  
Harish Padh
Keyword(s):  
Transgenic Plants ◽  
Mammalian Cells ◽  
Capital Investment ◽  
Transgene Expression ◽  
Present Report ◽  
Expression System ◽  
Expression Vectors ◽  
Pcr Analysis ◽  
Tissue Specific ◽  
Tissue Specific Promoter

With the advent of protein-based biotech drugs in the market, the quest for the “perfect” protein expression system, which is both economical and effective, has come into focus. Currently bacteria, yeast, insect cells, mammalian cells, transgenic animal and transgenic plants are widely used for the expression of therapeutic proteins. Among these, transgenic plants provide advantages in terms of low production cost, lower capital investment in infrastructure, and suitable post-translational modifications. The major limitation of plants as an expression host is the low level of transgene expression. To increase the expression of heterologous proteins in plants, a number of approaches have been used. One of the approaches is to increase the transgene expression by using tissue-specific promoter(s) which can concentrate the protein of interest in targeted tissues and, thus, prove advantageous in downstream purification. In the present report, a protocol for expression of heterologous protein erythropoietin in potato tuber using patatin, the tuber-tissue-specific promoter, was standardized. Expression vectors for production of the erythropoietin gene under tissue-specific promoter were successfully constructed. For production of a transgenic plant, tissue culture techniques for regeneration of the whole plant from single explants were standardized. Polymerase chain reaction (PCR) analysis was performed to confirm the stable integration of the erythropoietin gene in the potato plant by using sequence-specific primers.

scholarly journals Morphological Features of the Anther Development in Tomato Plants with Non-Specific Male Sterility

Biology ◽  
2020 ◽  
Vol 9 (2) ◽  
pp. 32 ◽  
Author(s):  
Inna A. Chaban ◽  
Neonila V. Kononenko ◽  
Alexander A. Gulevich ◽  
Liliya R. Bogoutdinova ◽  
Marat R. Khaliluev ◽  
...  
Keyword(s):  
Transgenic Plants ◽  
Male Sterility ◽  
Molecular Mechanisms ◽  
Asymmetric Cell Division ◽  
Male Gametophyte ◽  
Embryo Sac ◽  
Pollen Grain ◽  
Parthenocarpic Fruit ◽  
Tomato Plants ◽  
Transgenic Tomato Plants

The study was devoted to morphological and cytoembryological analysis of disorders in the anther and pollen development of transgenic tomato plants with a normal and abnormal phenotype, which is characterized by the impaired development of generative organs. Various abnormalities in the structural organization of anthers and microspores were revealed. Such abnormalities in microspores lead to the blocking of asymmetric cell division and, accordingly, the male gametophyte formation. Some of the non-degenerated microspores accumulate a large number of storage inclusions, forming sterile mononuclear pseudo-pollen, which is similar in size and appearance to fertile pollen grain (looks like pollen grain). It was discussed that the growth of tapetal cells in abnormal anthers by increasing the size and ploidy level of nuclei contributes to this process. It has been shown that in transgenic plants with a normal phenotype, individual disturbances are also observed in the development of both male and female gametophytes. The reason for the developmental arrest of some ovules was the death of endosperm at different stages of the globular embryo. At the same time, noticeable hypertrophy of endothelial cells performing a secretory function was observed. In the ovules of transgenic plants with abnormalities, the endothelium forms a pseudo-embryo instead of the embryo sac, stimulating the development of parthenocarpic fruits. The data obtained in this study can be useful for a better understanding of the genetic and molecular mechanisms of cytoplasmic male sterility and parthenocarpic fruit development in tomatoes.

Identification of α-Tomatine 23-Hydroxylase Involved in the Detoxification of a Bitter Glycoalkaloid

2019 ◽  
Vol 61 (1) ◽  
pp. 21-28 ◽  
Author(s):  
Masaru Nakayasu ◽  
Ryota Akiyama ◽  
Midori Kobayashi ◽  
Hyoung Jae Lee ◽  
Takashi Kawasaki ◽  
...  
Keyword(s):  
Fruit Ripening ◽  
Tomato Fruit ◽  
Transgenic Tomato ◽  
Metabolic Enzyme ◽  
Gene Encoding ◽  
Tomato Plants ◽  
Tomato Fruit Ripening ◽  
Transgenic Tomato Plants ◽  
Key Enzyme ◽  
Esculeoside A

Abstract Tomato plants (Solanum lycopersicum) contain steroidal glycoalkaloid α-tomatine, which functions as a chemical barrier to pathogens and predators. α-Tomatine accumulates in all tissues and at particularly high levels in leaves and immature green fruits. The compound is toxic and causes a bitter taste, but its presence decreases through metabolic conversion to nontoxic esculeoside A during fruit ripening. This study identifies the gene encoding a 23-hydroxylase of α-tomatine, which is a key to this process. Some 2-oxoglutarate-dependent dioxygenases were selected as candidates for the metabolic enzyme, and Solyc02g062460, designated Sl23DOX, was found to encode α-tomatine 23-hydroxylase. Biochemical analysis of the recombinant Sl23DOX protein demonstrated that it catalyzes the 23-hydroxylation of α-tomatine and the product spontaneously isomerizes to neorickiioside B, which is an intermediate in α-tomatine metabolism that appears during ripening. Leaves of transgenic tomato plants overexpressing Sl23DOX accumulated not only neorickiioside B but also another intermediate, lycoperoside C (23-O-acetylated neorickiioside B). Furthermore, the ripe fruits of Sl23DOX-silenced transgenic tomato plants contained lower levels of esculeoside A but substantially accumulated α-tomatine. Thus, Sl23DOX functions as α-tomatine 23-hydroxylase during the metabolic processing of toxic α-tomatine in tomato fruit ripening and is a key enzyme in the domestication of cultivated tomatoes.

scholarly journals Transgenic Tomato Plants Expressing Satellite RNA Are Tolerant to Some Strains of Cucumber Mosaic Virus

1994 ◽  
Vol 119 (3) ◽  
pp. 642-647 ◽  
Author(s):  
P.B. McGarvey ◽  
M.S. Montasser ◽  
J.M. Kaper
Keyword(s):  
Transgenic Plants ◽  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Unit Length ◽  
Transgenic Tomato ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Satellite Rna ◽  
Tomato Plants ◽  
Transgenic Tomato Plants

Transgenic tomato plants (Lycopersicon esculentum Mill.) expressing cucumber mosaic virus (CMV) satellite RNA fused to a gene for β-glucuronidase were produced using Agrobacterium-mediated transformation. The R1 progeny of self-crossed R0 plants were challenge-inoculated with virion or RNA preparations of CMV or tomato aspermy virus (TAV). The transgenic plants challenged with CMV-1 showed mild disease symptoms in the first 2 weeks postchallenge followed by a decrease in symptoms, resulting in little difference between the transgenic and uninfected control group by the fourth week. Enzyme-linked immunosorbent assay results showed about a 10-fold decrease in virus accumulation in the transgenic plants compared to controls. Tolerance was evident only in plants that contained the recombinant insert and produced mature unit-length satellite RNA after CMV infection. Plants challenged with TAV showed no significant tolerance to virus-induced symptoms.

scholarly journals Production of vaccines for treatment of infectious diseases by transgenic plants

2016 ◽  
Vol 107 (1) ◽  
pp. 191 ◽  
Author(s):  
Kristina LEDL ◽  
Zlata LUTHAR
Keyword(s):  
Transgenic Plants ◽  
Infectious Diseases ◽  
Disease Virus ◽  
Hepatitis B Surface Antigen ◽  
Expression System ◽  
Expression Vectors ◽  
Norwalk Virus ◽  
Mouth Disease Virus ◽  
Avian Influenza Virus H5n1 ◽  
Transformation Methods

<p>Since the first pathogen antigen was expressed in transgenic plants with the aim of producing edible vaccine in<em> </em>early 1990s, transgenic plants have become a well-established expression system for production of alternative vaccines against various human and animal infectious diseases. The main focus of plant expression systems in the last five years has been on improving expression of well-studied antigens such as porcine reproductive and respiratory syndrome (PRRSV), bovine viral diarrhea disease virus (BVDV), footh and mouth disease virus (FMDV), hepatitis B surface antigen (HBsAg), rabies G protein, rotavirus, Newcastle disease virus (NDV), Norwalk virus capsid protein (NVCP), avian influenza virus H5N1, <em>Escherichia coli</em> heat-labile enterotoxin subunit B (LT-B), cholera toxin B (CT-B), human immunodeficiency virus (HIV), artherosclerosis, ebola and anthrax. Significant increases in expression have been obtained using improved expression vectors, different plant species and transformation methods.</p>

scholarly journals Expression of the Human Hepatitis B Virus Large Surface Antigen Gene in Transgenic Tomato Plants

2007 ◽  
Vol 14 (4) ◽  
pp. 464-469 ◽  
Author(s):  
Xiao-Ming Lou ◽  
Quan-Hong Yao ◽  
Zhen Zhang ◽  
Ri-He Peng ◽  
Ai-Sheng Xiong ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Surface Antigen ◽  
Tomato Fruit ◽  
Transgenic Tomato ◽  
Tomato Plants ◽  
Antigen Gene ◽  
Transgenic Tomato Plants ◽  
Pathogenesis Related Protein ◽  
B Virus

ABSTRACT The original hepatitis B virus (HBV) large surface antigen gene was synthesized. In order to optimize the expression of this gene in tomato plants, the tobacco pathogenesis-related protein S signal peptide was fused to the 5′ end of the modified gene and the sequence encoding amino acids S, E, K, D, E, and L was placed at the 3′ end. The gene encoding the modified HBV large surface antigen under the control of a fruit-specific promoter was constructed and expressed in transgenic tomato plants. The expression of the antigen from transgenic plants was confirmed by PCR and reverse transcriptase PCR. Enzyme-linked immunoassays using a monoclonal antibody directed against human serum-derived HBsAg revealed that the maximal level of HBsAg was about 0.02% of the soluble protein in transgenic tomato fruit. The amount of HBsAg in mature fruits was found to be 65- to 171-fold larger than in small or medium fruits and leaf tissues. Examination of transgenic plant samples by transmission electron microscopy proved that HBsAg had been expressed and had accumulated. The HBsAg protein was capable of assembling into capsomers and virus-like particles. To our knowledge, this is the first time the HBV large surface antigen has been expressed in plants. This work suggests the possibility of producing a new alternative vaccine for human HBV.

Sign in / Sign up

Export Citation Format

Share Document