scholarly journals Overexpression of Medicago sativa glutamate-semialdehyde aminotransferase (GSA) gene in tobacco increased photosynthesis efficiency

2019 ◽  
Author(s):  
Maryam Ghasemzadeh ◽  
Mahdi Khozeai ◽  
Hamzeh Amiri
Keyword(s):  
Aminolevulinic Acid ◽  
Photosynthetic Capacity ◽  
Cauliflower Mosaic Virus ◽  
35S Promoter ◽  
Total Biomass ◽  
Tobacco Plants ◽  
Over Expression ◽  
Photosynthesis Efficiency ◽  
Individual Enzyme

AbstractTo investigate the effect of increased glutamate-semialdehyde aminotransferase (GSA) on photosynthetic capacity and growth, tobacco (Nicoliana tabacum L. Xanti) plants with increased levels of glutamate-semialdehyde aminotransferase protein were produced. This was achieved using a cassette composed of a full-length Medicago sative cDNA under the control of the cauliflower mosaic virus 35S promoter. The results revealed distinct impacts of GSA activity on photosynthesis rate and growth in GSA over expression tobacco plants. In transgenic plants with increased GSA activity, an increase in soluble and insoluble sugars accumulation was evident. Total biomass, leaf area, plant height and internode 3-4 were increased in GSA sense plants, compared with equivalent wild-type tobacco plants. Moreover, transgenic tobacco plants with increased GSA activity exhibit higher levels of 5-aminolevulinic acid (ALA) accumulation and increased in content of chlorophyll and carotenoids pigments. Collectively, our data suggest that higher level of GSA activity gives an advantage to photosynthesis, growth in tobacco plants. This work also provides a case study that an individual enzyme in the biosynthesis of chlorophyll pathway may serve as a useful target for genetic engineering to improve photosynthesis and growth in plants.HighlightOverexpression of glutamate-semialdehyde aminotransferase (GSA) increase photosynthetic capacity, growth in tobacco.

scholarly journals Expression of soybean nodulin 26 in transgenic tobacco. Targeting to the vacuolar membrane and effects on floral and seed development.

1995 ◽  
Vol 6 (1) ◽  
pp. 109-117 ◽  
Author(s):  
Y Zhang ◽  
D M Roberts
Keyword(s):  
Transgenic Tobacco ◽  
Plasma Membranes ◽  
Pollen Viability ◽  
Physiological Effect ◽  
Cauliflower Mosaic Virus ◽  
35S Promoter ◽  
Subcellular Targeting ◽  
Western Blots ◽  
Tobacco Plants ◽  
Capsule Size

Nodulin 26 is an integral membrane protein of the symbiosome membrane of nitrogen-fixing soybean nodules. We expressed a nodulin 26 cDNA in transgenic tobacco (TN26 tobacco) under the control of the cauliflower mosaic virus 35S promoter to study subcellular targeting and the physiological effect(s) of its expression. Based on Northern and Western blots, the expression of nodulin 26 mRNA and protein in transgenic plants is high in apical shoot sections, flowers, and stems, low in mature leaves, and absent in roots. Western blot analysis revealed high levels of transgenic nodulin 26 protein in tonoplast membranes. In contrast, nodulin 26 protein was not found in isolated plasma membranes, the soluble fraction, nor in chloroplast and mitochondria-enriched membrane fractions. About 50-60% of the flowers and pods from TN26 tobacco plants abscised prematurely. Seed capsule size and seed fill per capsule from the remainder of surviving flowers were about 50% of that of control plants. Pollen viability was found to be normal, but flowers from TN26 tobacco plants showed shorter anther filaments compared with control plants. Normal seed production and capsule size was restored by manually crossing the stigmas from TN26 plants with isolated pollen from either transgenic or control plants. Thus, the aberrant filament growth could have resulted in the reproductive defects associated with the plants.

scholarly journals SpPKE1, a Multiple Stress-Responsive Gene Confers Salt Tolerance in Tomato and Tobacco

2019 ◽  
Vol 20 (10) ◽  
pp. 2478 ◽  
Author(s):  
Jinhua Li ◽  
Chunrui Chen ◽  
Juanjuan Wei ◽  
Yu Pan ◽  
Chenggang Su ◽  
...  
Keyword(s):  
Salt Stress ◽  
Salt Tolerance ◽  
Wild Species ◽  
Amino Acid Sequences ◽  
Cauliflower Mosaic Virus ◽  
Stress Factors ◽  
35S Promoter ◽  
Over Expression ◽  
Abiotic Tolerance ◽  
Cultivated Species

Understanding the mechanism of abiotic-tolerance and producing germplasm of abiotic tolerance are important in plant research. Wild species often show more tolerance of environmental stress factors than their cultivated counterparts. Genes from wild species show potential abilities to improve abiotic resistance in cultivated species. Here, a tomato proline-, lysine-, and glutamic-rich type gene SpPKE1 was isolated from abiotic-resistant species (Solanum pennellii LA0716) for over-expression in tomato and tobacco for salt tolerance. The protein encoded by SpPKE1 was predominantly localized in the cytoplasm in tobacco. SpPKE1 and SlPKE1 (from cultivated species S. lycopersicum cv. M82) shared 89.7% similarity in amino acid sequences and their transcripts abundance in flowers and fruits was reduced by the imposition of drought or oxidative stress and the exogenous supply of abscisic acid. The DNA of the PKE1 promoter was highly methylated in fruit and leaf, and the methylation of the coding sequence in leaf was significantly higher than that in fruit at different development stages. The over-expression of SpPKE1 under the control of a CaMV (Cauliflower Mosaic Virus) 35S promoter in transgenic tomato and tobacco plants enhanced their tolerance to salt stress. PKE1 was downregulated by abiotic stresses but enhanced the plant’s salt stress tolerance. Therefore, this gene may be involved in post-transcriptional regulation and may be an important candidate for molecular breeding of salt-tolerant plants.

scholarly journals Polyubiquitin Promoter-Based Binary Vectors for Overexpression and Gene Silencing in Lotus japonicus

2008 ◽  
Vol 21 (4) ◽  
pp. 375-382 ◽  
Author(s):  
Takaki Maekawa ◽  
Mitsumasa Kusakabe ◽  
Yoshikazu Shimoda ◽  
Shusei Sato ◽  
Satoshi Tabata ◽  
...  
Keyword(s):  
Rna Interference ◽  
Transgenic Plants ◽  
Lotus Japonicus ◽  
Cauliflower Mosaic Virus ◽  
35S Promoter ◽  
Binary Vectors ◽  
Gus Reporter Gene ◽  
Over Expression ◽  
Gus Reporter ◽  
Alternative Choice

In this study, we compared the transcriptional activities between Cauliflower mosaic virus (CaMV)35S promoter and polyubiquitin (Ljubq1) promoter from Lotus japonicus using β-glucuronidase (gus) reporter gene in transgenic plants of L. japonicus. The promoter analysis demonstrated that the Ljubq1 promoter possessed higher activity than the CaMV35S promoter in leaves, stems, roots, nodules, and pollen. Finally, we created GATEWAY conversion technology-compatible binary vectors for over-expression and RNA interference under the Ljubq1 promoter. These materials could provide alternative choice for studies in L. japonicus.

scholarly journals Complementation of Coat Protein Mutants of Pepper Huasteco Geminivirus in Transgenic Tobacco Plants

1999 ◽  
Vol 89 (7) ◽  
pp. 540-545 ◽  
Author(s):  
R. G. Guevara-González ◽  
P. L. Ramos ◽  
R. F. Rivera-Bustamante
Keyword(s):  
Coat Protein ◽  
Transgenic Tobacco ◽  
Cauliflower Mosaic Virus ◽  
Wild Type Virus ◽  
35S Promoter ◽  
Wild Type ◽  
Tobacco Plants ◽  
Protein Mutants ◽  
Infectivity Analysis

The role of the pepper huasteco virus (PHV) coat protein (CP) gene during the infection was investigated in three different hosts by using mutations that produced truncated proteins and by complementation assays in transgenic plants. The infectivity analysis revealed that mutants that express truncated CP (CP7 and CP191) behave like the wild-type virus when inoculated onto pepper and Nicotiana benthamiana plants in terms of symptom expression and viral DNA movement. On the contrary, the CP7 mutant was unable to systemically infect tobacco plants, whereas only 10% of the plants inoculated with the CP191 mutant became infected. The CP7 mutant was complemented by coinoculating it with another geminivirus (taino tomato mottle virus). No complementation was observed in plants from nine transgenic tobacco lines expressing CP under the control of the cauliflower mosaic virus (CaMV) 35S promoter. However, 3 out of 10 lines expressing CP under the control of its own promoter (693 nucleotides) were able to complement the CP7 mutant. Interestingly, upon infection, the levels of CP mRNA in 693CP plants increased dramatically, probably due to transactivation of the CP promoter by the viral protein AC2.

scholarly journals Overexpression of the Golden SNP-Carrying Orange Gene Enhances Carotenoid Accumulation and Heat Stress Tolerance in Sweetpotato Plants

Antioxidants ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 51
Author(s):  
So-Eun Kim ◽  
Chan-Ju Lee ◽  
Sul-U Park ◽  
Ye-Hoon Lim ◽  
Woo Sung Park ◽  
...  
Keyword(s):  
Heat Stress ◽  
Stress Tolerance ◽  
Ipomoea Batatas ◽  
Radical Scavenging Activity ◽  
Radical Scavenging ◽  
Cauliflower Mosaic Virus ◽  
Carotenoid Content ◽  
35S Promoter ◽  
Storage Roots ◽  
Heat Stress Tolerance

Carotenoids function as photosynthetic accessory pigments, antioxidants, and vitamin A precursors. We recently showed that transgenic sweetpotato calli overexpressing the mutant sweetpotato (Ipomoea batatas [L.] Lam) Orange gene (IbOr-R96H), which carries a single nucleotide polymorphism responsible for Arg to His substitution at amino acid position 96, exhibited dramatically higher carotenoid content and abiotic stress tolerance than calli overexpressing the wild-type IbOr gene (IbOr-WT). In this study, we generated transgenic sweetpotato plants overexpressing IbOr-R96H under the control of the cauliflower mosaic virus (CaMV) 35S promoter via Agrobacterium-mediated transformation. The total carotenoid contents of IbOr-R96H storage roots (light-orange flesh) and IbOr-WT storage roots (light-yellow flesh) were 5.4–19.6 and 3.2-fold higher, respectively, than those of non-transgenic (NT) storage roots (white flesh). The β-carotene content of IbOr-R96H storage roots was up to 186.2-fold higher than that of NT storage roots. In addition, IbOr-R96H plants showed greater tolerance to heat stress (47 °C) than NT and IbOr-WT plants, possibly because of higher DPPH radical scavenging activity and ABA contents. These results indicate that IbOr-R96H is a promising strategy for developing new sweetpotato cultivars with improved carotenoid contents and heat stress tolerance.

Aberrant promoter methylation occurred from multicopy transgene and SU(VAR)3 - 9 homolog 9 (SUVH9) gene in transgenic Nicotiana benthamiana

2012 ◽  
Vol 39 (9) ◽  
pp. 764 ◽  
Author(s):  
Gi-Ho Lee ◽  
Seong-Han Sohn ◽  
Eun-Young Park ◽  
Young-Doo Park
Keyword(s):  
Gene Silencing ◽  
Nicotiana Benthamiana ◽  
Fluorescent Protein ◽  
Cauliflower Mosaic Virus ◽  
Camv 35S Promoter ◽  
35S Promoter ◽  
Camv 35S ◽  
Histone Deacetylase 1 ◽  
Transgenic Lines ◽  
Green Fluorescent

The chemical modification of DNA by methylation is a heritable trait and can be subsequently reversed without altering the original DNA sequence. Methylation can reduce or silence gene expression and is a component of a host’s defence response to foreign nucleic acids. In our study, we employed a plant transformation strategy using Nicotiana benthamiana Domin to study the heritable stability of the introduced transgenes. Through the introduction of the cauliflower mosaic virus (CaMV) 35S promoter and the green fluorescent protein (GFP) reporter gene, we demonstrated that this introduced promoter often triggers a homology-dependent gene-silencing (HDGS) response. These spontaneous transgene-silencing phenomena are due to methylation of the CaMV 35S promoter CAAT box during transgenic plant growth. This process is catalysed by SU(VAR)3–9 homologue 9 (SUVH9), histone deacetylase 1 (HDA1) and domains rearranged methylase 2 (DRM2). In particular, we showed from our data that SUVH9 is the key regulator of methylation activity in epigenetically silenced GFP transgenic lines; therefore, our findings demonstrate that an introduced viral promoter and transgene can be subject to a homology-dependent gene-silencing mechanism that can downregulate its expression and negatively influence the heritable stability of the transgene.

Chloroplast-dependent and light-independent expression of the tobacco rbcS promoter–GUS chimeric gene in black spruce

1996 ◽  
Vol 26 (6) ◽  
pp. 909-917
Author(s):  
Madoka Gray-Mitsumune ◽  
Bong Y. Yoo ◽  
Pierre J. Charest
Keyword(s):  
Fusion Gene ◽  
Black Spruce ◽  
Gus Activity ◽  
Chimeric Gene ◽  
Cauliflower Mosaic Virus ◽  
35S Promoter ◽  
Zygotic Embryos ◽  
Ribulose Bisphosphate Carboxylase ◽  
Bisphosphate Carboxylase ◽  
Rbcs Promoter

The tobacco rbcS (ribulose bisphosphate carboxylase small subunit) promoter, fused to the β-glucuronidase (GUS) reporter gene, was delivered to black spruce (Piceamariana (Mill.) BSP) tissues via microprojectile DNA bombardment, and its regulation was studied. The expression of the tobacco rbcS promoter–GUS chimeric gene was dependent on the presence of chloroplasts in black spruce tissues, as demonstrated in two ways: (i) there was no GUS activity expressed in zygotic embryos where no chloroplasts were observed, whereas it was expressed in light- and dark-grown seedlings that contained mature or immature chloroplasts; (ii) a herbicide, Norflurazon, destroyed chloroplast structure in seedlings and inhibited the expression of the tobacco rbcS promoter–GUS chimeric gene. A control chimeric gene, the cauliflower mosaic virus (CaMV) 35S promoter–GUS fusion gene was not inhibited by Norflurazon. Unlike in angiosperms, light had no effect on the expression of tobacco rbcS promoter–GUS chimeric gene. Both light- and dark-grown seedlings showed GUS activity, and expression in dark-grown seedlings was not enhanced by light. These results suggest that the tissue-specific regulation of the rbcS promoter may be conserved between angiosperms and conifers, but that the light regulation of this promoter may not be conserved.

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