Production of santalenes and bergamotene in Nicotiana tabacum plants

Trap Crop ◽

Chloroplast Targeting ◽

Tobacco Plants ◽

Aphid Infestation

AbstractTerpenes play an important role in plant–insect relationships, and these relationships can potentially be modified by altering the profile of terpenes emitted from plants using metabolic engineering methods. Transgenic plants generated by employing such methods offer the prospect of low-cost sustainable pest management; in this regard, we used chloroplast targeting and cytosolic mevalonic acid pathway enhancement in this study to investigate the interaction of santalenes and bergamotene with insects. The santalene- and bergamotene-emitting transgenic tobacco plants thus generated were utilized to study host preference in the green peach aphid Myzus persicae (Sulzer). The results showed that co-expression of either 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) or truncated HMGR with santalene synthase led to the production of higher amounts of santalenes and bergamotene in transgenic tobacco plants, and that these santalene- and bergamotene-emitting plants were attractive to green peach aphids. We accordingly propose that such transgenic plants may have potential application in pest management as a trap crop to prevent green peach aphid infestation of wild-type tobacco plants.

Overexpression of the pitaya phosphoethanolamine N-methyltransferase gene (HpPEAMT) enhanced simulated drought stress in tobacco

Plant Cell Tissue and Organ Culture (PCTOC) ◽

10.1007/s11240-021-02040-3 ◽

2021 ◽

Author(s):

Ai-Hua Wang ◽

Lan Yang ◽

Xin-Zhuan Yao ◽

Xiao-Peng Wen

Keyword(s):

Drought Stress ◽

Stress Response ◽

Transgenic Plants ◽

Drought Tolerance ◽

Transgenic Tobacco ◽

Sequence Similarity ◽

Amino Acid Sequence Similarity ◽

Wild Type ◽

AbstractPhosphoethanolamine N-methyltransferase (PEAMTase) catalyzes the methylation of phosphoethanolamine to produce phosphocholine and plays an important role in the abiotic stress response. Although the PEAMT genes has been isolated from many species other than pitaya, its role in the drought stress response has not yet been fully elucidated. In the present study, we isolated a 1485 bp cDNA fragment of HpPEAMT from pitaya (Hylocereus polyrhizus). Phylogenetic analysis showed that, during its evolution, HpPEAMT has shown a high degree of amino acid sequence similarity with the orthologous genes in Chenopodiaceae species. To further investigate the function of HpPEAMT, we generated transgenic tobacco plants overexpressing HpPEAMT, and the transgenic plants accumulated significantly more glycine betaine (GB) than did the wild type (WT). Drought tolerance trials indicated that, compared with those of the wild-type (WT) plants, the roots of the transgenic plants showed higher drought tolerance ability and exhibited improved drought tolerance. Further analysis revealed that overexpression of HpPEAM in Nicotiana tabacum resulted in upregulation of transcript levels of GB biosynthesis-related genes (NiBADH, NiCMO and NiSDC) in the leaves. Furthermore, compared with the wild-type plants, the transgenic tobacco plants displayed a significantly lower malondialdehyde (MDA) accumulation and higher activities of the superoxide dismutase (SOD) and peroxidase (POD) antioxidant enzymes under drought stress. Taken together, our results suggested that HpPEAMT enhanced the drought tolerance of transgenic tobacco.

Allium Sativum Leaf Agglutinin (ASAL) Endowed Enhanced Resistance Against Myzus Persicae under Both Constitutive and Phloem Specific Promoters

10.21203/rs.3.rs-1042830/v1 ◽

2021 ◽

Author(s):

Noroza Umer ◽

Rubab Zahra Naqvi ◽

Imran Rauf ◽

Naveed Anjum ◽

Hamid Anees Siddiqui ◽

...

Keyword(s):

Transgenic Tobacco ◽

Myzus Persicae ◽

Plant Viruses ◽

Specific Primers ◽

Tobacco Plants ◽

Aphid Infestation ◽

Variable Expression ◽

Enhanced Resistance ◽

Plant Transformation Vector

Abstract Globally, aphid, Myzus persicae is an economically significant, polyphagous crop pest that feeds on more than 400 plant species and transmits more than 100 plant viruses. Aphid infestation is mostly managed by insecticides that cause heavy environmental contamination and insect resistance. Cloning of plant derived insecticidal genes to develop transgenic plants under suitable promoter is a promising technology. In the present study, ASAL (MN820725) was isolated from native garlic and cloned in plant transformation vector, pGA482 through Agrobacterium mediated tobacco transformation. PCR of genomic DNA of transgenic tobacco plants using gene specific primers confirmed the presence of asal gene of 546 bp. To detect the integration of gene Southern blot analysis was conducted that revealed stable integration of asal gene while, gene expression was analyzed through qRT-PCR that showed variable expression of asal gene in transgenic tobacco plants. Efficacy of asal gene was evaluated through aphid bioassay. Aphid bioassay revealed that transgenic tobacco lines LS-17, LS-20, LR-1, and LR-7 exhibited 100% aphid mortality and significantly reduced the aphid population. These findings suggested the potential of ASAL against aphids that can be further used against other notorious sap sucking pests.

Transgenic tobacco expressing an Arisaema heterophyllum agglutinin gene displays enhanced resistance to aphids

Canadian Journal of Plant Science ◽

10.4141/p03-036 ◽

2004 ◽

Vol 84 (3) ◽

pp. 785-790 ◽

Cited By ~ 6

Author(s):

Jianhong Yao, Xiuyun Zhao ◽

Huaxiong Qi, Bingliang Wan ◽

Fei Chen, Xiaofen Sun ◽

Shanqian Yu ◽

Kexuan Tang

Keyword(s):

Transgenic Plants ◽

Transgenic Tobacco ◽

Myzus Persicae ◽

Insect Bioassay ◽

Tobacco Plants ◽

Potato Aphid ◽

Enhanced Resistance ◽

Multiple Copies ◽

Peach Potato Aphid

Tobacco leaf discs were transformed with a plasmid, pBIAHA, containing the selectable marker neomycin phosphotransferase gene (nptII) and an Arisaema heterophyllum agglutinin gene (aha) via Agrobacterium tumefaciens-mediated transformation. Thirty-two independent transgenic tobacco plants were regenerated. PCR and Southern blot analyses confirmed that multiple copies of the aha gene had integrated into the plant genome. Northern blot analysis revealed that the aha gene was expressed at various levels in the transgenic plants. Insect bioassay test showed that transgenic plants expressing multiple copies of the aha gene reduced the rate of population increase of the peach potato aphid (Myzus persicae Sulzer). This is the first report that transgenic tobacco plants expressing the aha gene display enhanced resistance to aphids. Key words: Insect bioassay, Arisaema heterophyllum agglutinin, transformation, transgenic tobacco, peach potato aphid (Myzus persicae Sulzer)

The Arabidopsis expansin gene (AtEXPA18) is capable to ameliorate drought stress tolerance in transgenic tobacco plants

10.21203/rs.3.rs-204710/v1 ◽

2021 ◽

Author(s):

Alireza Abbasi ◽

Meysam Malekpour ◽

Sajjad Sobhanverdi

Keyword(s):

Drought Stress ◽

Transgenic Plants ◽

Transgenic Tobacco ◽

Dry Weight ◽

Conventional Polymerase Chain Reaction ◽

Specific Gene ◽

Severe Drought ◽

Tobacco Plants ◽

Reverse Transcription Pcr

Abstract Expansins are cell wall proteins that, due to changes in pH, causing the expansion of the cell walls. In this study, a previously gene construct designed based on a root-specific gene, AtEXPA18, was utilized to assess its potential roles on different morphological, physiological, and cellular levels of generated transgenic tobacco plants in response to moderate and severe drought stress. AtEXPA18 gene was successfully transferred to the tobacco plants through an agrobacterium-mediate transformation system. Upon obtaining the second generation, tobacco transgenic plants were confirmed by conventional polymerase chain reaction (PCR) technique alongside reverse transcription PCR (RT-PCR) using specific primers. Under drought stress, the transgenic lines showed remarkable growth and significantly improved based on morphological traits such as height and stem diameter, leaf area, leaf number, root dry weight, and Abscisic acid (ABA) levels of leaves compared control plants. As a result, the Cytokinin content of transgenic plants has increased under severe stress levels. Notably, the area's expansion for abaxial epidermal cells under the microscope confirmed in transgene cells compared with the -transgene cells. These results, altogether, could support the AtEXPA18 gene implication in cell expansion and improving tolerance capacity of transgenic crops under drought stress.

Overexpression of Grapevine VvIAA18 Gene Enhanced Salt Tolerance in Tobacco

International Journal of Molecular Sciences ◽

10.3390/ijms21041323 ◽

2020 ◽

Vol 21 (4) ◽

pp. 1323 ◽

Cited By ~ 1

Author(s):

Wei Li ◽

Changxi Dang ◽

Yuxiu Ye ◽

Zunxin Wang ◽

Laibao Hu ◽

...

Keyword(s):

Salt Stress ◽

Salt Tolerance ◽

Transgenic Plants ◽

Transgenic Tobacco ◽

Transcriptional Activation ◽

Late Embryogenesis Abundant Protein ◽

Pcr Analysis ◽

Vitis Vinifera L ◽

In plants, auxin/indoleacetic acid (Aux/IAA) proteins are transcriptional regulators that regulate developmental process and responses to phytohormones and stress treatments. However, the regulatory functions of the Vitis vinifera L. (grapevine) Aux/IAA transcription factor gene VvIAA18 have not been reported. In this study, the VvIAA18 gene was successfully cloned from grapevine. Subcellular localization analysis in onion epidermal cells indicated that VvIAA18 was localized to the nucleus. Expression analysis in yeast showed that the full length of VvIAA18 exhibited transcriptional activation. Salt tolerance in transgenic tobacco plants and Escherichia. coli was significantly enhanced by VvIAA18 overexpression. Real-time quantitative PCR analysis showed that overexpression of VvIAA18 up-regulated the salt stress-responsive genes, including pyrroline-5-carboxylate synthase (NtP5CS), late embryogenesis abundant protein (NtLEA5), superoxide dismutase (NtSOD), and peroxidase (NtPOD) genes, under salt stress. Enzymatic analyses found that the transgenic plants had higher SOD and POD activities under salt stress. Meanwhile, component analysis showed that the content of proline in transgenic plants increased significantly, while the content of hydrogen peroxide (H2O2) and malondialdehyde (MDA) decreased significantly. Based on the above results, the VvIAA18 gene is related to improving the salt tolerance of transgenic tobacco plants. The VvIAA18 gene has the potential to be applied to enhance plant tolerance to abiotic stress.

Expression and Chloroplast Targeting of Cholesterol Oxidase in Transgenic Tobacco Plants

PLANT PHYSIOLOGY ◽

10.1104/pp.126.3.1116 ◽

2001 ◽

Vol 126 (3) ◽

pp. 1116-1128 ◽

Cited By ~ 50

Author(s):

David R. Corbin ◽

Robert J. Grebenok ◽

Thomas E. Ohnmeiss ◽

John T. Greenplate ◽

John P. Purcell

Keyword(s):

Transgenic Tobacco ◽

Cholesterol Oxidase ◽

Chloroplast Targeting ◽

Effects of salt stress on proline content, expression of delta-1-pyrroline-5-carboxylate synthetase, and activities of catalase and ascorbate peroxidase in transgenic tobacco plants

Biological Letters ◽

10.2478/v10120-009-0002-4 ◽

2009 ◽

Vol 46 (2) ◽

pp. 63-75 ◽

Cited By ~ 12

Author(s):

Roya Razavizadeh ◽

Ali Ehsanpour

Keyword(s):

Salt Stress ◽

Transgenic Plants ◽

Transgenic Tobacco ◽

Expression Pattern ◽

Ascorbate Peroxidase ◽

Proline Content ◽

35S Promoter ◽

Tobacco Plants ◽

Moth Bean

Effects of salt stress on proline content, expression of delta-1-pyrroline-5-carboxylate synthetase, and activities of catalase and ascorbate peroxidase in transgenic tobacco plantsIn arid and semiarid regions, soil salinity limits crop production. Proline accumulation in transgenic plants results in increased stress tolerance, but the underlying mechanism was unclear. To elucidate it, effects of salt stress on the expression pattern of Δ1-pyrroline-5-carboxylate synthetase (P5CS), proline content, catalase (CAT), and ascorbate peroxidase (APX) activities were analyzed in transgenic tobacco (Nicotiana tabacumcv. Wisconsin). Transgenic tobacco plants containing CaMV 35S promoter and theP5CSgene from moth bean (Vigna aconitifolia), linked to theNPTIIgene, were culturedin vitrowith or without 300 mM NaCl. The expression pattern ofP5CSwas evaluated using semiquantitative RT-PCR (reverse transcription-polymerase chain reaction). Time-course experiments showed an increase in proline content after 4 h of the treatment. The level ofP5CStranscripts was increased significantly in leaves and roots of transgenic plants after 24 and 48 h of treatment. This rise in transcripts was concomitant with the highest increase in proline content. In addition, CAT and APX activities increased under salt stress, and their highest activities were observed after 24 and 48 h of NaCl treatment. These results suggest thatP5CSis an inducible gene regulating the activities of CAT and APX and the accumulation of proline in plants subjected to salt stress.

Transformation of tobacco and potato with cDNA encoding the full-length genome of Potato leafroll virus: evidence for a novel virus distribution and host effects on virus multiplication

Journal of General Virology ◽

10.1099/0022-1317-80-11-2813 ◽

1999 ◽

Vol 80 (11) ◽

pp. 2813-2822 ◽

Cited By ~ 34

Author(s):

Liliana F. Franco-Lara ◽

Kara D. McGeachy ◽

Uli Commandeur ◽

Robert R. Martin ◽

Mike A. Mayo ◽

...

Keyword(s):

Transgenic Plants ◽

Transgenic Tobacco ◽

Transgenic Potato ◽

Mesophyll Cells ◽

Tobacco Plants ◽

Potato Plants ◽

The Mean ◽

Leafroll Virus ◽

Tissue Prints

A full-length cDNA copy of the genome of Potato leafroll virus (PLRV) was introduced into the genome of tobacco and potato plants by Agrobacterium tumefaciens-mediated transformation. Transgenic lines were obtained in which the transgene was readily detected by PCR with DNA extracted from T1 tobacco seedlings and clonally multiplied potato plants. PLRV-specific genomic and sub- genomic RNAs, coat protein antigen and virus particles were detected in transgenic plants. Aphids fed on the transgenic tobacco plants readily transmitted PLRV to test plants. Infected transgenic tobacco plants, like non-transgenic (WT) PLRV-infected plants, displayed no symptoms of the infection but transgenic plants of potato were severely stunted. In parallel tests, the mean PLRV titres in WT tobacco plants and transgenic tobacco plants were 600 and 630 ng virus/g leaf, respectively, although differences in PLRV titres among transgenic plants were much greater than those among infected WT plants. In similar tests with potato, the mean PLRV titre of WT plants was 50 ng virus/g leaf whereas higher concentrations (up to 3400 ng virus/g leaf) accumulated in transgenic potato plants. In tissue prints of stems, PLRV was detected in similar proportions of phloem cells in transgenic and infected WT plants. In transgenic tobacco and potato plants, but not in infected WT plants, a few stem epidermal cells also contained virus. From tissue prints of transgenic tobacco leaves, it was estimated that about one in 40000 mesophyll cells contained virus, but in transgenic potato, a greater proportion of mesophyll cells was infected.

Functional expression of a biologically active fragment of soluble gp130 as an ELP-fusion protein in transgenic plants: purification via inverse transition cycling

Biochemical Journal ◽

10.1042/bj20060544 ◽

2006 ◽

Vol 398 (3) ◽

pp. 577-583 ◽

Cited By ~ 33

Author(s):

Meng Lin ◽

Stefan Rose-John ◽

Joachim Grötzinger ◽

Udo Conrad ◽

Jürgen Scheller

Keyword(s):

Transgenic Plants ◽

Fusion Protein ◽

Transgenic Tobacco ◽

Molecular Farming ◽

Biologically Active ◽

Tobacco Plants ◽

Inverse Transition Cycling ◽

Inverse Transition ◽

Soluble Gp130

In murine models of Crohn's disease, rheumatoid arthritis and colon cancer, IL-6 (interleukin-6) signalling via the sIL-6R (soluble IL-6 receptor; termed IL-6 trans-signalling) has been shown to promote the pathology associated with these conditions. These detrimental activities can, however, be selectively blocked by soluble forms of the gp130 (glycoprotein 130) receptor. Although sgp130 (soluble gp130) therefore represents a viable therapeutic modality for the treatment of these conditions, the mass manufacture of such biologics is often expensive. The advent of molecular farming has, however, provided an extremely cost-effective strategy for the engineering of recombinant proteins. Here, we describe the expression and production of a biologically active sgp130 variant that is expressed in transgenic tobacco plants as an ELP (elastin-like peptide)-fusion protein (mini-gp130–ELP). Mini-gp130–ELP consists of the first three domains of gp130 (Ig-like domain and cytokine binding module) fused to 100 repeats of ELP. Expression of mini-gp130–ELP did not affect the growth rate or morphology of the transgenic plants, and purification was achieved using inverse transition cycling. This approach led to an overall yield of 141 μg of purified protein per g of fresh leaf weight. The purified mini-gp130–ELP specifically inhibited sIL-6R-mediated trans-signalling as measured by binding to the IL-6–sIL-6R complex and through its ability to block sIL-6R-mediated activation of STAT3 (signal transducer and activator of transcription 3) phosphorylation and proliferation in human hepatoma cells and murine pre-B-cells. Consequently, the present study validates the potential application of molecular farming in transgenic tobacco plants as a strategy for the expression and purification of therapeutically advantageous biologics such as sgp130.

Productivity and stress-tolerance of transgenic tobacco plants with constitutive expression of rapeseed glutathione synthetase gene BnGSH

Ecological genetics ◽

10.17816/ecogen15112-19 ◽

2017 ◽

Vol 15 (1) ◽

pp. 12

Author(s):

Bulat R Kuluev ◽

Zoya A Berezhneva ◽

Elena V Mikhaylova ◽

Bogdan N Postrigan ◽

Aleksey V Knyazev

Keyword(s):

Transgenic Plants ◽

Transgenic Tobacco ◽

Metal Tolerance ◽

Growth Parameters ◽

Heavy Metal Tolerance ◽

Constitutive Expression ◽

Glutathione Synthetase ◽

35S Promoter ◽

Glutathione is the most important part of plant antioxidant defense system. Biosynthesis of glutathione in the cells is performed by two enzymes: glutamylcysteine ligase and glutathione synthetase, the latter catalyzing the attachment of glycine to a dipeptide glutamylcysteine. In literature there is information on the improvement of heavy metal-tolerance of transgenic plants due to the increase in the expression level of glutathione synthetase genes. However there is not enough data on the tolerance of these plants to other types of abiotic stress. Therefore the aim of our research was to make transgenic tobacco plants with constitutive expression of rapeseed glutathione synthetase gene BnGSH and to estimate their growth parameters in normal conditions and under salt, drough and cold stress. Using agrobacterial transformation method, we generated 17 lines of transgenic plants containing rapeseed BnGSH gene under control of 35S promoter. The presence of transgenes was confirmed by PCR method and histochemical analysis of the activity of GUS reporter gene. 12 lines with the highest expression of BnGSH gene were chosen on the basis of the results of RT-PCR. We performed morphological analysis, including measurements of stem hight, leaf area, flower length, fresh and dry weight of shoots and root length. Some transgenic plants demonstrated increased productivity in normal conditions as well as under NaCl stress. However, no change in drought and cold tolerance was observed in transgenic plants.