scholarly journals Gamma Radiation Effect on Agrobacterium tumefaciens-Mediated Gene Transfer in Potato (Solanum tuberosum L.)

2021 ◽  
Author(s):  
Murat Aycan ◽  
Muhammet Cagri Oguz ◽  
Yasin Ozgen ◽  
Burak Onol ◽  
Mustafa Yildiz
Keyword(s):  
Solanum Tuberosum ◽  
Agrobacterium Tumefaciens ◽  
Transgenic Plants ◽  
Gamma Radiation ◽  
Solanum Tuberosum L ◽  
Pcr Analysis ◽  
Axillary Meristems ◽  
Technological Research Council ◽  
Selection Medium ◽  
Transformation Methods

Potato (Solanum tuberosum L.) is one of the major crops of the world. Significant improvements can be achieved in terms of yield and quality by the determination of efficient transformation methods. On the other hand, low transformation frequency seriously limits the application of molecular techniques in obtaining transgenic crops. In the present study, the effect of gamma radiation on Agrobacterium tumefaciens-mediated transformation to the potato was firstly investigated. Sterile seedlings of potato cv. ‘Marabel’, which was grown on Gamborg’s B5 medium in Magenta vessels, were irradiated with different gamma radiation doses (0-control, 40, 80, 120 Gy 60Co). Stem parts having axillary meristems were excised from irradiated seedlings and inoculated by A. tumefaciens (GV2260), which harbors the binary plasmid p35S GUS-INT contains and GUS (β-glucuronidase) gene controlled by 35S promoter (CaMV) and nptII (neomycin phosphotransferase II) gene driven by NOS (nopaline synthase) promoter). Inoculated stem parts having axillary meristems explants were then directly transported to a selection medium containing duocid (500 mg l−1), and kanamycin (100 mg l−1), 4 mg l−1 gibberellic acid, 1 mg l−1 BAP and 0.1 mg l−1 NAA. The adult transgenic plants were detected by polymerase chain reaction (PCR) analysis. According to the number of transgenic plants determined by PCR analysis, results obtained from explants treated with 40 Gy gamma gave the best results compared to the control (0 Gy) application. The doses over 40 Gy were also found statistically significant compared to the control (0 Gy). It is expected that the protocol described in this study make the transformation studies easier by skipping the stages of ‘co-cultivation’, ‘culturing explants on selection medium’ and ‘recovery of transgenic shoots on selection medium’ not only for potato but also for other crop plants. This study was supported by a grant from the Scientific and Technological Research Council of Turkey (TUBİTAK) (Grant number 113O280 to Prof. Dr. Mustafa YILDIZ).

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)

scholarly journals In vitro Regeneration and Agrobacterium-mediated Transformation of Potato (Solanum tuberosum L.) Cultivars Grown in Mexico

2013 ◽  
Vol 22 (2) ◽  
pp. 93-105 ◽  
Author(s):  
Rose Onamu ◽  
Juan P Legaria ◽  
Jaime C Sahagún ◽  
José L Rodríguez ◽  
Joel N Pérez
Keyword(s):  
Solanum Tuberosum ◽  
Solanum Tuberosum L ◽  
In Vitro Regeneration ◽  
Potato Cultivars ◽  
Pcr Analysis ◽  
Nptii Gene ◽  
Histochemical Assay ◽  
Gus Histochemical Assay ◽  
Binary Plasmid

Prior to Agrobacterium-mediated genetic transformation in vitro regeneration protocol was established for three potato cultivars (Alfa, Cambray Rosa Morelos and Atlantic) grown in Mexico using leaf, node and internodal explants. Regeneration protocol was developed with or without the intervention of callus. Two potato cultivars, namely, Cambray Rosa Morelos and Alpha were transformed using Agrobacterium tumefaciens strain LBA4404 harboring binary plasmid pBI121 containing the GUS and nptII genes. GUS histochemical assay and PCR analysis were conducted on rooted shoots grown in media without hormones but supplemented with antibiotics. Transformed shoots tested positive through GUS histochemical assay and integration of nptII gene was confirmed by PCR analysis DOI: http://dx.doi.org/10.3329/ptcb.v22i2.14193 Plant Tissue Cult. & Biotech. 22(2): 93-105, 2012 (December)

scholarly journals Transformasi Genetik Tanaman Kentang (Solanum tuberosum L.) dengan Gen acvB Menggunakan Vektor Agrobacterium tumefaciens

2021 ◽  
Vol 11 (1) ◽  
pp. 63
Author(s):  
DARWIN SILALAHI ◽  
I GEDE PUTU WIRAWAN ◽  
MADE SRITAMIN
Keyword(s):  
Solanum Tuberosum ◽  
Agrobacterium Tumefaciens ◽  
Treatment Group ◽  
Genetic Transformation ◽  
Solanum Tuberosum L ◽  
Control Group ◽  
Shoot Height ◽  
Specific Pcr ◽  
Convenient Technique

Agrobacterium tumefaciens Mediated Genetic Transformation of acvB Gene in Potato (Solanum tuberosum L.). Genetic transformations are now routinely applied to plant mediated by Agrobacterium tumefaciens as the most convenient technique. This study aimed to prove the success of A. tumefaciens mediated genetic transformation in potato. A. tumefaciens LBA (pBI 121) and explant of potato shoot were used in this study. Explants were grown in vitro on Murashige and Skoog media. Transformation was implemented using smear technique by smearing A. tumefaciens to injured explant. Experimental groups consisted of two groups: control group which did not receive transformation treatment and treatment group receiving transformation treatment. Explant growth was observed through the presence of shoots, branches and the shoot height. Explants in the treatment group resulted in a higher number of shoots, branches, and shoot heights compared to control. Phenol compounds appear in explant epidermal tissue, indicating the wounds produced by A. tumefaciens infection, thus the gene predicted to be transformed. Identification by PCR is needed to prove the existence of the acvB gene in potato plants genome, using acvB specific PCR primer as the marker, such as (5?-CCCT CTAG AGAC CCGC GCCA AGGCG-3?) and (5?CGCG TCGA CCTT GTCG GAAAG -3?) with 540-bp in base pair size produced.

scholarly journals EXPRESSION OF REPORTER GENES IN TRANSFORMED INDICA RICE THROUGH Agrobacterium MEDIATED METHOD

2007 ◽  
Vol 20 (1) ◽  
pp. 01-08
Author(s):  
Shamsul H. Prodhan ◽  
A. Komamine
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Oryza Sativa L ◽  
Indica Rice ◽  
Kanamycin Resistance ◽  
Pcr Analysis ◽  
Gus Gene ◽  
Immature Inflorescence ◽  
Seed Fertility ◽  
Selection Medium ◽  
Mature Seeds

Genetic transformation of rice (Oryza sativa L.) mediated by Agrobacterium tumefaciens has been confirmed for japonica varieties and extended to include more recalcitrant indica varieties. Scutellum-derived calli from mature seeds of Kasalath and BR-5 were used. The Agrobacterium tumefaciens strain, EHA101, harboring the binary vector pIG121Hm/Km/GUS was used for transformation. The vector contains b-glucuronidase (GUS) gene as a reporter gene and hygromycin resistance (HPT) as well as kanamycin resistance gene (NPTII) as selection genes in the T-DNA region. After co-cultivation with the bacteria, calli were inoculated on selection medium in which hygromycin concentration was 50 mg/l for Kasalath and 20 mg/l for BR-5. Carbenicillin (500 mg/l) was used for removal of Agrobacterium after co-cultivation. Inclusion of acetosyringone 50–100 mM in the Agrobacterium suspension in co-culture medium increased the frequency of transformation. Frequency of transformed calli (hygromycin resistant cells) was 82% in Kasalath and 6% in BR-5. Regeneration efficiency from transformed calli in Kasalath was about 63% and in BR-5 was about 34%. Most of the transgenic plants were morphologically normal but seed fertility was lower than the control. In transformed calli, roots and immature inflorescence showed positive response in GUS assay. Presence of GUS, HPT and NPTII genes was confirmed by PCR analysis and PCR Southern blot analysis. Expression of GUS gene was 100% in T1 progeny of Kasalath, whereas that of HPT gene was 51%. BR-5 could not be tested because of low seed fertility of T0 plants. In T1 plants, seed fertility of transformed Kasalath was 79% which is lower than that of the respective non-transformants.DOI: http://dx.doi.org/10.3329/bjpbg.v20i1.10633

scholarly journals CLAVATA3/EMBRYO SURROUNDING REGION (CLE) Gene Family in Potato (Solanum tuberosum L.): Identification and Expression Analysis

Agronomy ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 984
Author(s):  
Maria Gancheva ◽  
Irina Dodueva ◽  
Maria Lebedeva ◽  
Ludmila Lutova
Keyword(s):  
Solanum Tuberosum ◽  
Gene Family ◽  
Solanum Tuberosum L ◽  
Expression Patterns ◽  
Pcr Analysis ◽  
Regulatory Sequences ◽  
Functional Studies ◽  
Cle Peptides ◽  
Using Data ◽  
Potato Growth

CLE genes encode a group of small secretory peptides, which regulate cell proliferation and differentiation in plants. CLE genes have been studied in many plants; however, little is known about this gene family in potato. In this study, we characterized members of the CLE gene family in potato Solanum tuberosum (StCLE) and comprehensively analyzed their phylogenetic relationships, structure, and expression patterns. Using available transcriptomic data, we found a relative high expression level of StCLE8, StCLE12, and StCLE13 in stolons and tubers. Real-time PCR analysis showed that the StCLE23 gene was upregulated by water deficiency, whereas the expression of StCLE4 and StCLE10 was induced by nitrogen supply. Besides that, using data from transcriptomic studies obtained previously for plants with the induction the StBEL5 gene, a positive regulator of tuber development, we found that StCLE4 was among genes upregulated in response to StBEL5 induction, suggesting that StCLE4 could be a target of StBEL5 transcription factor. However, we did not reveal a direct binding of StBEL5 to the regulatory sequences of StCLE4 using yeast one-hybrid assay. Taken together, our data provide basic information for future functional studies of CLE peptides in potato growth and tuberization and in response to various environmental stimuli.

Gamma radiation degradation of chitosan for application in growth promotion and induction of stress tolerance in potato (Solanum tuberosum L.)

2019 ◽  
Vol 210 ◽  
pp. 289-301 ◽  
Author(s):  
Abhijeet B. Muley ◽  
Prashant R. Shingote ◽  
Ajit P. Patil ◽  
Sunil G. Dalvi ◽  
Penna Suprasanna
Keyword(s):  
Solanum Tuberosum ◽  
Gamma Radiation ◽  
Stress Tolerance ◽  
Growth Promotion ◽  
Solanum Tuberosum L ◽  
Radiation Degradation

scholarly journals Pengaruh Jenis Bakteri Pelarut terhadap Pertumbuhan dan Hasil Benih G3 Kentang (Solanum tuberosum L.) Varietas Medians.

2020 ◽  
Vol 8 (2) ◽  
pp. 131
Author(s):  
Indriana Ulfah ◽  
Linlin Parlinah ◽  
Noertjahyani Noertjahyani ◽  
Rohana Abdullah
Keyword(s):  
Solanum Tuberosum ◽  
Agrobacterium Tumefaciens ◽  
Solanum Tuberosum L ◽  
Potato Crop ◽  
Growth And Yield ◽  
Phosphate Solubilizing Bacteria ◽  
Group Design ◽  
Number Of Leaves ◽  
Phosphate Solubilizing ◽  
Experimental Garden

This article aims to examine the effect of phosphate solubilizing bacteria (PSB) on the growth and yield of G3 seeds of potato (solanum tuberosum L.) varieties of medians. The experiment was conducted in the experimental garden of The Faculty of Agriculture of Winaya Mukti University Tanjungsari, Sumedang with a height of 850 meters above sea level with the order of land Andisol. Time trials are conducted from May to August 2019. The design used is a simple RandomIzed Group Design consisting of five types of phosphate Solubilizing bacteria, namely without PSB, Pseudacidovorax intermedius, Agrobacterium fabrum, Agrobacterium tumefaciens, Serratia nematodiphila and repeated five times each. The results of the experiment showed that the administration of this type of PSB was able to increase the height of the plant and the number of leaves at the age of 28 DAP and 42 DAP and give the result of the number of potatoes and the weight of the perpetak tubers on the potato crop of the medians varieties better.

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