scholarly journals Agrobacterium-mediated Genetic Transformation of Lentil (Lens culinaris Medik.) with Chitinase Gene followed by In vitro Flower and Pod Formation

2019 ◽  
Vol 29 (1) ◽  
pp. 99-109
Author(s):  
Subroto K Das ◽  
Kishwar Jahan Shethi ◽  
MI Hoque ◽  
RH Sarker
Keyword(s):  
Genetic Transformation ◽  
Lens Culinaris ◽  
Target Gene ◽  
Gene Selection ◽  
Chitinase Gene ◽  
Pcr Analysis ◽  
Agrobacterium Strain ◽  
Half Strength ◽  
Selection Of

To investigate the integration of chitinase gene in lentil (Lens culinaris Medik.) namely, BARI masur-4 (BM-4), BARI masur-5 (BM-5) and BARI masur-6 (BM-6) through Agrobacterium-mediated genetic transformation was performed using Agrobacterium strain EHA 105 harboring bar (resistant to phosphinotrycin) and chitinase (gene of interest) gene. Selection of transformed shoots was carried out by gradually increasing the concentration of phosphinotrycin (PPT) up to 2.0 mg/l. Transgenic lentil shoots were produced with an overall frequency of 0.36 in case of BM-4 and BM-6 and 0.34 in case of BM-5, respectively. Most of the selected shoots developed in vitro flowers and pods following their sub-culture on half strength of MS supplemented with 20 mg/l IBA, 0.5 mg/l NAA with 50 mg/l ticarcillin. Seedlings germinated from the seeds were successfully transferred to soil for the development of further progeny. Stable integration of target gene was confirmed through PCR analysis. Plant Tissue Cult. & Biotech. 29(1): 99-109, 2019 (June)

scholarly journals Agrobacterium-mediated Genetic Transformation in Lentil (Lens culinaris Medik.) followed by In vitro Flowering and Seed Formation

2012 ◽  
Vol 22 (1) ◽  
pp. 13-26 ◽  
Author(s):  
Subroto K. Das ◽  
Kishwar Jahan Shethi ◽  
M. I. Hoque ◽  
R. H. Sarker
Keyword(s):  
Genetic Transformation ◽  
Lens Culinaris ◽  
Multiple Shoots ◽  
Cotyledonary Node ◽  
In Vitro Flowering ◽  
Pcr Analysis ◽  
Seed Formation ◽  
Half Strength ◽  
Genetic Transformation System

Genetic transformation system was developed for two microsperma varieties of lentil (Lens culinaris Medik.), namely Bari Masur-4 (BM-4) and Bari Masur-5 (BM-5) using Agrobacterium tumefaciens strain LBA4404 harbouring binary plasmid pBI121, containing GUS and nptII genes. Three different types of embryo explants, namely cotyledonary node (CN), decapitated embryo (DE) and cotyledone attached decapitated embryo (CADE) were used. Highest GUS positive expression was found in DE followed by CADE as detected by transient assays. Following Agrobacterium infection CADE showed better response in developing multiple shoots on MS supplemented with 2.22 µM BAP, 2.32 µM Kn, 0.29 µM GA3 and 30.35 µM tyrosine. Selection of the transformed shoots was carried out by gradually increasing the concentration of kanamycin up to 200 mg/l. Transgenic lentil shoots were produced with an overall frequency of 1.009%. In vitro rooting appeared to have a limitation in obtaining complete plantlets in lentil, therefore in vitro flowering and seed formation were induced in transformed shoots of lentil with a view to recovering of the transgenic progenies.  GUS positive shoots were found to produce in vitro flowers and pods on half-strength MS containing 98.4 µM IBA and 2.69 µM NAA. Expression of gene was detected in various tissues of the transformed shoots. Stable integration of GUS gene was also confirmed through PCR analysis. Plant Tissue Cult. & Biotech. 22(1): 13-26, 2012 (June) DOI: http://dx.doi.org/10.3329/ptcb.v22i1.11243 

scholarly journals In vitro Regeneration and Agrobacterium-mediated Genetic Transformation of Local Varieties of Mungbean (Vigna radiata (L). Wilczek)

2019 ◽  
Vol 29 (1) ◽  
pp. 81-97
Author(s):  
Sujay Kumar Bhajan ◽  
Setara Begum ◽  
Mohammad Nurul Islam ◽  
M Imdadul Hoque ◽  
Rakha Hari Sarker
Keyword(s):  
Genetic Transformation ◽  
Vigna Radiata ◽  
Zygotic Embryo ◽  
In Vitro Regeneration ◽  
Multiple Shoots ◽  
Direct Organogenesis ◽  
Pcr Analysis ◽  
Root Induction ◽  
Half Strength

An efficient Agrobacterium-mediated transformation compatible in vitro regeneration protocol was developed for two important varieties of mungbean (Vigna radiata (L.) Wilczek) cultivated in Bangladesh, namely Binamoog-5 and BARI Mung-6. Two different zygotic embryo derived explants, such as cotyledonary node (CN) and cotyledon attached decapitated embryo (CADE) were used for direct organogenesis of shoot. MS supplemented with 4.0 μM BAP was found to be the best for the development of highest number of multiple shoots from CADE in both the varieties of mungbean. While in case CN the best shoot formation was achieved on MS containing 4.0 μM BAP and 0.5 μM NAA in both varieties. Half strength of MS with 2.0 μM IBA was found to be most effective for producing healthy root from regenerated shoots. Following root induction, the in vitro raised plantlets were successfully transplanted to soil for their establishment. Considering overall responses, genetic transformation efficiency was found to be better with CADE explant using Agrobacterium tumefaciens strain LBA4404 harboring the binary plasmid pBI121 conferring GUS and nptII genes. Different factors influencing transformation was optimized during this study. Selection of transformed shoots was carried out by gradually increasing the concentration of kanamycin and such transformed shoots were eventually selected using 200 mg/l kanamycin. Stable expression of the GUS gene was detected in various parts of regenerated transformed plantlets. Transformed shoots were rooted on half strength MS containing 2.0 μM IBA and 100 mg/l ticarcillin. Rooted transformed plantlets were successfully transferred to soil. Stable integration of GUS and nptII genes in the putative transformed shoots was confirmed through PCR analysis. Plant Tissue Cult. & Biotech. 29(1): 81-97, 2019 (June)

scholarly journals Agrobacterium-mediated Genetic Transformation for Local Cultivars of Potato (Solanum tuberosum L.) Using Marker Genes

1970 ◽  
Vol 20 (2) ◽  
pp. 145-155 ◽  
Author(s):  
Rita Sarah Borna ◽  
M. I. Hoque ◽  
R. H. Sarker
Keyword(s):  
Solanum Tuberosum ◽  
Genetic Transformation ◽  
Solanum Tuberosum L ◽  
In Vitro Regeneration ◽  
Direct Regeneration ◽  
Pcr Analysis ◽  
Marker Genes ◽  
Transformation Rate ◽  
Best Responses

Genetic transformation using nodal and internodal segments from three economically important potato (Solanum tuberosum L.) varieties namely, Diamant, Cardinal and Granola was conducted using an Agrobacterium tumefaciens strain LBA4404 harbouring binary plasmid pBI12 containing the GUS and nptII genes. Node and internodal segments were used for direct regeneration as well as regeneration with the intervention of callus. best responses were  obtained for direct regeneration of shoots when the explants were cultured on MS supplemented with 4.0 mg/l BAP +1.0 mg/l IAA, 1.5 mg/l BAP  + 0.5 mg/l IAA and 5.0 mg/l BAP +1.0 mg/l IAA in Diamant, Cardinal  and  Granola, respectively. In Diamant spontaneous in vitro microtuberization was obtained from these proliferated shoots. Further culturing of these in vitro grown green microtubers regenerated a large number of shoots on MS containing 4.0 mg/l BAP +1.0 mg/l IAA. By combining the best treatments, this protocol yielded an average transformation rate of 87% of treared explants. Stable expression of GUS gene was visualized in the various parts of transformed shoots through histochemical assay. Genomic DNA was isolated from transformed shoots and stable integration of the GUS and nptII genes was confirmed by PCR analysis.   Key words:  Potato, in vitro regeneration, transformation   D.O.I. 10.3329/ptcb.v20i2.6894   Plant Tissue Cult. & Biotech. 20(2): 145-155, 2010 (December)

Copper affects the binding of HIF-1α to the critical motifs of its target genes

Metallomics ◽  
2019 ◽  
Vol 11 (2) ◽  
pp. 429-438 ◽  
Author(s):  
Zhijuan Wu ◽  
Wenjing Zhang ◽  
Y. James Kang
Keyword(s):  
Dna Binding ◽  
Target Gene ◽  
Target Genes ◽  
Gene Selection ◽  
Hypoxic Conditions ◽  
Selection Of

Copper regulates the target gene selection of HIF-1α under hypoxic conditions by affecting HIF-1α-DNA binding patterns across the genome.

scholarly journals Mutant ACTB mRNA 3′UTR Promotes Hepatocellular Carcinoma Development by Regulating miR-1 and miR-29a

2019 ◽  
Author(s):  
Yong Li ◽  
Hong-Bin Ma ◽  
Chang-Ying Shi ◽  
Fei-Ling Feng ◽  
Liang Yang
Keyword(s):  
Hepatocellular Carcinoma ◽  
Target Gene ◽  
Molecular Mechanisms ◽  
Luciferase Reporter ◽  
Migration And Invasion ◽  
Pcr Analysis ◽  
Limited Data ◽  
Cancer Types

AbstractIn recent years, mounting studies have shown that ACTB is closely related to various tumors. Although ACTB is dysregulated in numerous cancer types, limited data are available on the potential function and mechanism of ACTB in hepatocellular carcinoma (HCC). This study evaluated the expression and biological roles of mutant ACTB mRNA 3′UTR in HCC. Transcriptome sequence and qRT-PCR analysis determined that mutant ACTB mRNA 3′UTR was high expression in HCC tissues. Luciferase reporter assay showed that the ACTB mRNA 3′UTR mutations made it easier to interact with miR-1 and miR-29a. Moreover, mutant ACTB mRNA 3′UTR regulated miR-1 and miR-29a degradation via AGO2. Furthermore, mutant ACTB mRNA 3′UTR promoted hepatocellular carcinoma cells migration and invasionin vitroandin vivoby up-regulating miR-1 target gene MET and miR-29a target gene MCL1. In a word, our study demonstrates that 3′UTR of ACTB plays a key role in the tumor growth of hepatocellular carcinoma (HCC) and highlights the molecular mechanisms of ACTB-involved cancer growth and development.

scholarly journals Glyphosate selection of maize transgenic callus lines among genotypes of Ukrainian plant breeding

1970 ◽  
pp. 237-242
Author(s):  
I. O. Nitovska ◽  
I. K. Komarnytskyi ◽  
B. V. Morgun
Keyword(s):  
Plant Breeding ◽  
Transgenic Maize ◽  
Pcr Analysis ◽  
Agrobacterium Mediated Transformation ◽  
Maize Germplasm ◽  
Maize Transformation ◽  
Callus Lines ◽  
Selection Of ◽  
Maize Callus

Aim. Glyphosate selection has a number of advantages over other commonly used selectable markers for maize. There is some natural variability within maize germplasm for degree of sensitivity to glyphosate. We investigated the selective effect of glyphosate for production transgenic maize callus after Agrobacterium-mediated transformation among geno-types of Ukrainian plant breeding. Methods. Agrobacterium-mediated transformation, glyphosate selection in vitro, and PCR analysis were used to obtain transgenic maize callus and to confirm its status. Results. An efficient selectable marker system for production transgenic maize callus lines tolerant to herbicide glyphosate was proposed. Calluses of four maze genotypes of Ukrainian plant breeding and pCB135 vector containing CP4epsps gene were used in Agrobacterium-mediated transformation experiments. Three callus maize lines of DK267×PLS61 genotype containing CP4epsps gene were obtained. Conclusions. The use of glyphosate as a selective agent after Agrobacterium-mediated transformation proved to be effective for transgenic maize callus lines production containing the gene CP4epsps. The success of Agrobacterium-mediated transformation of maize callus strongly depended on the genotype of source ma-terial. Keywords: Agrobacterium-mediated maize transformation, CP4epsps gene, glyphosate selection, PCR analysis.

scholarly journals Global transcriptional analysis identifies a novel role for SOX4 in tumor-induced angiogenesis

eLife ◽  
2018 ◽  
Vol 7 ◽  
Author(s):  
Stephin J Vervoort ◽  
Olivier G de Jong ◽  
M Guy Roukens ◽  
Cynthia L Frederiks ◽  
Jeroen F Vermeulen ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Transcription Factor ◽  
Poor Prognosis ◽  
Target Gene ◽  
Gene Selection ◽  
Mammary Tumorigenesis ◽  
Breast Tumors ◽  
Transcriptional Analysis

The expression of the transcription factor SOX4 is increased in many human cancers, however, the pro-oncogenic capacity of SOX4 can vary greatly depending on the type of tumor. Both the contextual nature and the mechanisms underlying the pro-oncogenic SOX4 response remain unexplored. Here, we demonstrate that in mammary tumorigenesis, the SOX4 transcriptional network is dictated by the epigenome and is enriched for pro-angiogenic processes. We show that SOX4 directly regulates endothelin-1 (ET-1) expression and can thereby promote tumor-induced angiogenesis both in vitro and in vivo. Furthermore, in breast tumors, SOX4 expression correlates with blood vessel density and size, and predicts poor-prognosis in patients with breast cancer. Our data provide novel mechanistic insights into context-dependent SOX4 target gene selection, and uncover a novel pro-oncogenic role for this transcription factor in promoting tumor-induced angiogenesis. These findings establish a key role for SOX4 in promoting metastasis through exploiting diverse pro-tumorigenic pathways.

scholarly journals Production of hairy root cultures of lettuce (Lactuca sativa L.)

2014 ◽  
Vol 9 (12) ◽  
pp. 1196-1205 ◽  
Author(s):  
Tadić Vojin ◽  
Milošević Snežana ◽  
Cingel Aleksandar ◽  
Petrić Marija ◽  
Trifunović Milana ◽  
...  
Keyword(s):  
Hairy Root ◽  
Lactuca Sativa ◽  
Target Genes ◽  
Pcr Analysis ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Lactuca Sativa L ◽  
Half Strength ◽  
Independent Transformation

AbstractHairy root cultures of lettuce (Lactuca sativa L.) were obtained by inoculation of cotyledonary leaves of in vitro lettuce seedlings (cvs. Nansen and Ljubljanska ledenka) with Agrobacterium rhizogenes A4M70GUS. Approximately in 96.7% cvs. Nansen and in 91.2% Ljubljanska ledenka inoculated explants produced hairy root when they were incubated on Murashige and Skoog (MS) half-strength medium without plant growth regulators. A total of 54% of all hairy root cultures expressed GUS activity. Every hairy root represented an independent transformation event. Line Ljubljanska ledenka 18 showed the highest biomass (5.5 times the biomass of control root). A PCR analysis of the genomic DNA confirmed the presence of marker and target genes in 15 hairy roots examined.

scholarly journals Genetic transformation of new perspective winter wheat genotypes in vitro

2020 ◽  
Vol 26 ◽  
pp. 270-275
Author(s):  
L. V. Slivka ◽  
O. V. Dubrovna
Keyword(s):  
Transgenic Plants ◽  
Genetic Transformation ◽  
Winter Wheat ◽  
Target Gene ◽  
Callus Cultures ◽  
Genetic Construct ◽  
Wheat Genotypes ◽  
Agrobacterium Mediated Transformation ◽  
New Perspective

Aim. Optimization of conditions for genetic transformation of new perspective winter wheat genotypes and production of transgenic plants. Methods. Agrobacterium-mediated transformation in vitro culture using callus cultures. Results. The influence of the optical density of cells of agrobacterial suspension, the concentration of the antibiotic cefotaxime, the duration of coculture on the frequency of obtaining kanamycin-resistant regenerants of new winter wheat genotypes by genetic transformation of callus cultures were investigated. By Agrobacterium-mediated transformation of morphogenic calluses of new perspective winter wheat genotypes were obtained plant-regenerants in the genome of which revealed the complete incorporation of a genetic construct containing oat and nptII transgenes. Conclusions. Agrobacterium-mediated transformation of callus cultures of new perspective winter wheat genotypes was optimized, and transgenic plants with the target gene of ornithine-δ-aminotransferase were obtained. Keywords: Triticum aestivum, Agrobacterium-mediated transformation, callus cultures, ornithine-δ-aminotransferase gene.

scholarly journals Agrobacterium-mediated Genetic Transformation of Two Varieties of Jute (Corchorus capsularis L.)

1970 ◽  
Vol 18 (1) ◽  
pp. 7-16 ◽  
Author(s):  
Rakha Hari Sarker ◽  
G.M. Al-Amin ◽  
Fathi Hassan ◽  
M.I. Hoque
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Genetic Transformation ◽  
Plant Tissue ◽  
Genomic Dna ◽  
Mature Embryo ◽  
Gus Expression ◽  
Pcr Analysis ◽  
Gus Gene ◽  
Corchorus Capsularis ◽  
Selection Of

Transformation experiments were carried out using different explants of two varieties of white jute (Corchorus capsularis L.), namely, CVL-1 and CVE-3 with Agrobacterium tumefaciens strain (LBA4404/pBI121) containing the GUS and nptII genes. Maximum transformation ability was obtained from petiole-attached cotyledons and mature embryo explants. Kanamycin at a concentration of 200 mg/l was found optimum for selection of transformed shoots developed frommature embryos. Histochemical assay revealed the stable expression of the GUS gene within the various tissues of transformed plantlets. Stable integration of GUS and nptII genes were confirmed by PCR analysis of genomic DNA isolated from these transformed shoots. Key words: Jute, Transformation, GUS expression, PCR analysis D.O.I. 10.3329/ptcb.v18i1.3245 Plant Tissue Cult. & Biotech. 18(1): 7-16, 2008 (June)

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