Comparison of transformation efficiency of five Agrobacterium tumefaciens strains in Nicotiana Tabacum L.

Bakhsh, Allah; Anayol, Emine; Özcan, Sancar Fatih

doi:10.9755/ejfa.v26i3.16437

Cited by 41 publications

(27 citation statements)

References 18 publications

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“…Thus, the transformation efficiency of MNH-786 and VH-289 was 3.75 % and 2.88 %, respectively (Table 3). Bakhsh et al [48] reported higher transformation efficiency (20 %) of Agrobacterium strain (LBA4404) in tobacco plants [48], but cotton is harder to transform, and our results were similar to the work of Majeed et al [49] in which transformation efficiency in cotton was 5.17 % [49, 50]. …”

Section: Discussionsupporting

confidence: 89%

Amplicon-Based RNA Interference Targeting V2 Gene of Cotton Leaf Curl Kokhran Virus-Burewala Strain Can Provide Resistance in Transgenic Cotton Plants

et al. 2016

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The conserved coat or V2 gene of begomoviruses is responsible for viral movement in the plant cells. RNAi technology was used to silence V2 gene for resistance against these viruses in transgenic plants. The transformation of the RNAi-based gene construct targeting V2 gene of CLCuKoV-Bur, cloned under 35S promoter, was done in two elite cotton varieties MNH-786 and VH-289 using shoot apex cut method of gene transformation. The transformation efficiency was found to be 3.75 and 2.88 % in MNH-786 and VH-289, respectively. Confirmation of successful transformation was done through PCR in T 0, T 1, and T 2 generations using gene-specific primers. Transgenic cotton plants were categorized on the basis of the virus disease index in T 1 generation. Copy number and transgene location were observed using FISH and karyotyping in T 2 generation which confirmed random integration of V2 RNAi amplicon at chromosome 6 and 16. Real-time quantitative PCR analyses of promising transgenic lines showed low virus titer compared to wild-type control plants upon challenging them with viruliferous whiteflies in a contained environment. From the results, it was concluded that amplicon V2 RNAi construct was able to limit virus replication and can be used to control CLCuV in the field.

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Section: Discussionsupporting

confidence: 89%

Amplicon-Based RNA Interference Targeting V2 Gene of Cotton Leaf Curl Kokhran Virus-Burewala Strain Can Provide Resistance in Transgenic Cotton Plants

et al. 2016

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“…The Agrobacterium-mediated binary vector system is an efficient system of natural exchange of genetic material. Based on our earlier experiments, we used the LBA4404 agrobacterium strain harboring pRD400 plasmid to infect apical meristems in genetic transformation experiments [39]. The transformation efficiency achieved in the present study has been low compared to earlier studies [40][41][42]25].…”

Section: Discussionmentioning

confidence: 89%

Development of insect-resistant cotton lines with targeted expression of insecticidal gene

Bakhsh

Anayol

Khabbazi

et al. 2016

ARCH BIOL SCI BELGRA

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In order to address biosafety concerns regarding the constitutive expression of foreign genes in crops, we applied a strategy aimed at confining foreign gene expression in insect wounding sites of cotton. For this purpose, a plant expression construct was designed by cloning the AoPR1 promoter (pathogenesis-related protein gene isolated from Asparagus officinalis) upstream from the insecticidal gene cry1Ac. The Turkish cotton cultivar cv. STN-468 was transformed using the Agrobacterium tumefaciens strain LBA4404 containing the recombinant binary vector pRD400 harboring cry1Ac under a wound-inducible promoter. The neomycin phosphotransferase (nptII) gene was used as a selectable marker at a concentration of 100 mg/L. The primary transformants were analyzed for T-DNA integration and expression using standard molecular approaches. The efficacy of insecticidal gene control of the AoPR1 promoter was investigated using leaf bioassays with 2 nd instar larvae of Helicoverpa armigera and Spodoptera littoralis. Positive primary transformants from T 0 progeny were further raised under greenhouse conditions to obtain progeny (T 1 ). The introduced gene was properly inherited and expressed in T 1 progeny. The mechanical wounding of plants resulted in increased cry1Ac protein levels during 0-48 h of the wounding period. The transgenic lines exhibited appreciable levels of resistance against targeted insect pests in the leaf bioassays. The use of a wound-inducible promoter to drive insecticidal gene expression is a valuable insect resistant management strategy as gene expression will remain limited to the insect biting sites of plant and crop, food and environmental concerns can be minimized.

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“…The suitability of C58C1 strain harbouring various plasmids for the transformation of many economic plants has already been reported (Bakhsh et al 2014). This study used shoot tip explants taken from in vitro grown stock material multiplied on MS medium containing 1.0 mg/l BA for 6 weeks following Karataş and Aasim (2015).…”

Section: Resultsmentioning

confidence: 99%

“…43 out of 78 plants (55.13%) plants showing regeneration on MS medium supplemented with Kanamycin were speculated a escapes that were further confirmed by GUS histochemical analysis. Although Kanamycin is believed to be a good selection agent for putative plants, still escapes have been documented in various genetic transformation studies (Bakhsh et al, 2014;McCormick et al, 1986).…”

Section: Resultsmentioning

confidence: 99%

First Report of Agrobacterium tumefaciens mediated genetic transformation of aquatic Rice paddy herb (Limnophila aromatica)

Bakhsh

Aasım

Zia

et al. 2016

Turkish JAF Sci.Tech.

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The study presents first report of Agrobacterium mediated genetic transformation in Rice paddy herb (Linmophila aromatica). A. tumefaciens strain C58C1 harboring pBin19 Plasmid containing β-glucuronidase (GUS) and neomycin phosphotransferase II (nptII) gene, under the control of 35S promoter and NOS terminator was used. Shoot tip explants were inoculated for 30 min followed by co-cultivation for 72 h and selected on agar semi solidified MS medium containing 100 mg/l Kanamycin and 1.0 mg/l BA; whereas total number of 78 putative transgenic shoots were obtained. The shoots were rooted on MS medium containing 1.0 mg/l IBA and 100 mg/l Kanamycin where 43 plants survived and rooted. Expression of GUS gene in the putative transgenics was confirmed by histochemical GUS assay. Visible localised gus expression was noted in a few cells and callus tissues of 4 plantlets that were photographed using compound light microscope.

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Comparison of transformation efficiency of five Agrobacterium tumefaciens strains in Nicotiana Tabacum L.

Cited by 41 publications

References 18 publications

Amplicon-Based RNA Interference Targeting V2 Gene of Cotton Leaf Curl Kokhran Virus-Burewala Strain Can Provide Resistance in Transgenic Cotton Plants

Amplicon-Based RNA Interference Targeting V2 Gene of Cotton Leaf Curl Kokhran Virus-Burewala Strain Can Provide Resistance in Transgenic Cotton Plants

Development of insect-resistant cotton lines with targeted expression of insecticidal gene

First Report of Agrobacterium tumefaciens mediated genetic transformation of aquatic Rice paddy herb (Limnophila aromatica)

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