scholarly journals Agrobacterium-mediated transformation of tobacco (Nicotiana tabacum L.) with human lactoferrin gene and analysis of transgenic lines

2020 ◽  
Vol 26 ◽  
pp. 164-168
Author(s):  
A. Yu. Buziashvili ◽  
A. I. Yemets
Keyword(s):  
Nicotiana Tabacum ◽  
Transgenic Tobacco ◽  
Cauliflower Mosaic Virus ◽  
Human Lactoferrin ◽  
35S Promoter ◽  
Root Cells ◽  
Agrobacterium Mediated Transformation ◽  
Nicotiana Tabacum L ◽  
Lactoferrin Gene ◽  
Transgenic Lines

Aim. Obtaining of tobacco lines (Nicotiana tabacum L.) stably transformed with human lactoferrin gene (hLf), analysis of transgenic lines. Methods. Agrobacterium-mediated transformation of tobacco was carried out with the use of pBin35LF plasmid containing human lactoferrin gene under control of 35S promoter of cauliflower mosaic virus (CaMV 35S) and selective npt II gene encoding neomicyne phosphotransferase II providing the resistance to kanamycin. The selection of transgenic lines was carried out for 3 months on MS medium supplemented with 100 mg/l of kanamycin. Integration of lactoferrin gene was confirmed with the use of PCR with primers specific to hLf gene. The investigation of karyotype of transgenic lines was carried out after the staining of the root cells with 1 % solution of acetoorseine. Results. After selection, 2 transgenic tobacco lines with confirmed integration of hLf gene were obtained. The transformation efficiency was 6.4 %. The chromosome number in transgenic and control lines was 2n=48. Conclusions. The obtained transgenic tobacco lines have the similar morphology and karyotype to control lines, so they could be considered to use as plant systems for the expression of recombinant human lactoferrin. Keywords: human lactoferrin gene hLf, Nicotiana tabacum L., Agrobacterium-mediated transformation, PCR, chromosomes, transgenic plants.

scholarly journals Genetic transformation of Nicotiana tabacum with the yeast TPS1 and TPS2 genes involved in trehalose synthesis

2020 ◽  
Vol 17 (2) ◽  
pp. 150-158 ◽  
Author(s):  
A. Yu. Kvasko ◽  
S. V. Isayenkov ◽  
E. E. Krasnoperova ◽  
K. V. Dmytruk ◽  
A. I. Yemets
Keyword(s):  
Nicotiana Tabacum ◽  
Genetic Transformation ◽  
Target Genes ◽  
Leaf Explants ◽  
Cauliflower Mosaic Virus ◽  
Selected Lines ◽  
Agrobacterium Mediated Transformation ◽  
Transgenic Lines ◽  
Yeast Genes ◽  
Trehalose Synthesis

Aim. The aim of the study was the creation of vector constructions with yeast genes of trehalose synthesis TPS1 and TPS2 and their using for Agrobacterium-mediated transformation of N. tabacum. Methods. Strain of Agrobacterium tumefaciens GV3101 carrying vector constructions — pGWВ2-TPS1 and pGWВ2-TPS2 with TPS1 and TPS2 target genes respectively under 35S promoter of cauliflower mosaic virus and selectable hpt gene of hygromycin phosphotransferase has been used for plants transformation. N. tabacum leaf explants were used for Agrobacterium-mediated transformation. The medium with the addition of hygromycin was applied to transgenic lines selection. Results. The created vector constructions pGWВ2-TPS1 and pGWВ2-TPS2 has been used in genetic N. tabacum transformation. Target genes TPS1 and TPS2 were integrated applied of Agrobacterium-mediated transformation and transgenic lines of N. tabacum were selected with addition at 25 mg/L hygromycin into the medium. Molecular analysis confirmed the transgenic nature of selected lines. Conclusions. Sensitivity of selected lines for sugars content into the medium was established for shoots and roots formation of tobacco plants. Conditions for increase of transformation frequency and rooting of transgenic lines of plant after integrated of target TPS1 and TPS2 genes were investigated. Keywords: trehalose, yeast genes TPS1, TPS2, genetic transformation, Nicotiana tabacum.

Co-expression of AGPS and AGPL genes encoded for AGPase from cassava to enhance starch accumulation in transgenic tobacco

2016 ◽  
Vol 14 (2) ◽  
pp. 287-293
Author(s):  
Nguyễn Văn Đoài ◽  
Nguyễn Minh Hồng ◽  
Lê Thu Ngọc ◽  
Nguyễn Thị Thơm ◽  
Nguyễn Đình Trọng ◽  
...  
Keyword(s):  
Transgenic Tobacco ◽  
Higher Plants ◽  
Starch Content ◽  
Genetically Modified Crops ◽  
Starch Accumulation ◽  
35S Promoter ◽  
Effective Strategies ◽  
Plant Expression Vector ◽  
Transgenic Lines ◽  
Cassava Varieties

The AGPase (ADP-Glucose pyrophosphorylase) is one of the ubiquitous enzymes catalyzing the first step in starch biosynthesis. It plays an important role in regulation and adjusts the speed of the entire cycle of glycogen biosynthesis in bacteria and starch in plants. In higher plants, it is a heterotetramer and tetrameric enzyme consisting two large subunits (AGPL) and two small subunits (AGPS) and encoded by two genes. In this paper, both AGPS and AGPL genes were sucessfully isolated from cassava varieties KM140 and deposited in Genbank with accession numbers KU243124 (AGPS) and KU243122 (AGPL), these two genes were fused with P2a and inserted into plant expression vector pBI121 under the control of 35S promoter. The efficient of this construct was tested in transgenic N. tabacum. The presence and expression of AGPS and AGPL in transgenic plants were confirmed by PCR and Western hybridization. The starch content was quantified by the Anthrone method. Transgenic plant analysis indicated that that two targeted genes were expressed simultaneously in several transgenic tobacco lines under the control of CaMV 35S promoter.  The starch contents in 4 analyzed tobacco transgenic lines displays the increase 13-116%  compared to WT plants. These results indicated that the co-expression of AGPS and AGPL is one of effective strategies for enhanced starch production in plant. These results can provide a foundation for developing other genetically modified crops to increase starch accumulation capacity.

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.

scholarly journals Lead Phytoremediation Potential of Wild Type and Transgenic Tobacco Plants

2021 ◽  
Vol 5 (1) ◽  
pp. 168-182
Author(s):  
Hatice DAGHAN ◽  
Veli UYGUR ◽  
Abdullah EREN
Keyword(s):  
Nicotiana Tabacum ◽  
Transgenic Tobacco ◽  
Transgenic Tobacco Plants ◽  
Wild Type ◽  
Tobacco Plants ◽  
Nicotiana Tabacum L ◽  
Lead Phytoremediation

Genetiği değiştirilmiş bitkiler, kurşunun (Pb) kökten yer üstü kısımlarına translokasyonunu geliştirmek için büyük bir potansiyele sahip olabilir. Transgenik olmayan ( Nicotiana tabacum L. cv. Petit Havana SR1) ve transgenik (p-cV-ChMTII GFP) tütün bitkileri tarafından Pb alımının sağlanması araştırmak için Çin hamsteri metalotiyonin II gezen bir kap deneyi yapıldı . Transgenik ve transgenik olmayan tütün bitkileri, 0, 1000, 2500, 5000 mg Pb kg- 1 ile Pb (NO 3 ) 2 olarak işlenmiş topraklarda yetiştirildi. Kelimede bir büyüme bölümünde 6 hafta boyunca çiçeklenme aşamasına kadar.Bitkilerin büyümesi, klorofil içeriği, mineral besin elementleri ve düşük glutatyon (GSH) bezleri, bitkilerin Pb alım potansiyeli ile birlikte incelenmiştir. Hem transgenik hem de transgenik olmayan bitkiler için Pb uygulamasındaki artışa bağlı olarak yer üstü biyokütle çevrildi aşamalı bir düşüş gözlendi. Yaprak besinlerinin bulaştığı, aşırı Pb işlemlerinden olumsuz etkilenmiştir, bunlardan en büyük düşüşü. Sürgün Pb yüksek derecesi 76.0 mg kg kadar ulaşan -1 transgenik ve 70.9 mg kg -1 transgenik olmayan bitkilerde. Pb alımı, p-cV-ChMTII GFP'nin tütün bitkisine aktarılmasıyla iyileştirildi; ancak, Pb fitoremediasyonunda yeterli değildi. 

scholarly journals Expression of thaumatin, a new type of alternative sweetener in rice

2020 ◽  
Vol 48 (3) ◽  
pp. 1276-1291
Author(s):  
Shahina AKTER ◽  
Md. Amdadul HUQ ◽  
Yu-Jin JUNG ◽  
Kwon-Kyoo KANG
Keyword(s):  
Transgenic Rice ◽  
Oryza Sativa L ◽  
Plasmid Vector ◽  
Single Copy ◽  
Cauliflower Mosaic Virus ◽  
Pcr Analysis ◽  
35S Promoter ◽  
Alternative Sweetener ◽  
Sweet Proteins ◽  
Transgenic Lines

  Sweet proteins are the natural alternative to the artificial sweeteners as well as flavor enhancers. Among other sweet protein, thaumatin protein was isolated from Thaumatococcus daniellii Benth plant fruit. In this study, pinII Ti plasmid vector was constructed with thaumatin gene, where thaumatin was placed under the control of the duel cauliflower mosaic virus 35S promoter into rice (Oryza sativa L. var. japonica cv. ‘Dongjinbyeo’) by Agrobacterium-mediated transformation to generate transgenic plants. Thirteen plant lines were regenerated and the transgenic rice lines were confirmed by different molecular analysis. The genomic PCR result revealed that all of the plant lines were transgenic. The single copy and intergenic plant lines were selected by Taqman PCR analysis and FST analysis, respectively. Expression of thaumatin gene in transgenic rice resulted in the accumulation of thaumatin protein in the leave. Thaumatin protein was also accumulated in leave of T1 generation. Sensory analysis result suggested that the thaumatin protein expressing transgenic lines exerted sweet tasting activity. These results demonstrated that thaumatin was expressed in transgenic rice plants.

A detoxification gene in transgenicNicotiana tabacumconfers 2,4-D tolerance

Weed Science ◽  
1999 ◽  
Vol 47 (4) ◽  
pp. 401-404 ◽  
Author(s):  
David I. Last ◽  
Danny J. Llewellyn
Keyword(s):  
Arabidopsis Thaliana ◽  
Nicotiana Tabacum ◽  
Pisum Sativum ◽  
Histone Gene ◽  
Cauliflower Mosaic Virus ◽  
35S Promoter ◽  
Spray Drift ◽  
Degree Of Protection ◽  
Dioxygenase Gene ◽  
Detoxification Gene

TransgenicNicotiana tabacumwith tolerance to 2,4-D has previously been produced using a bacterial 2,4-D-dioxygenase gene (tfdA) driven by the 35S promoter of cauliflower mosaic virus. Using promoters from thePisum sativumplastocyanin gene (petE) and anArabidopsis thalianahistone gene (H4A), we demonstrate that similar protection from 2,4-D can be obtained in transgenicN. tabacumby targeting expression oftfdAto either meristematic tissues or chloroplast-containing tissues. As with the 35S promoter constructs, the plants are tolerant but not completely resistant; very young seedlings in particular are only slightly protected. However, the levels of tolerance observed could offer a useful degree of protection from accidental spray drift.

Carbon metabolism enzymes and photosynthesis in transgenic tobacco (Nicotiana tabacum L.) having excess phytochrome

Planta ◽  
1991 ◽  
Vol 185 (3) ◽  
Author(s):  
ThomasD. Sharkey ◽  
TerryL. Vassey ◽  
PeterJ. Vanderveer ◽  
RichardD. Vierstra
Keyword(s):  
Nicotiana Tabacum ◽  
Transgenic Tobacco ◽  
Carbon Metabolism ◽  
Nicotiana Tabacum L ◽  
Metabolism Enzymes
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