IMPROVEMENT OF GENETIC TRANSFORMATION AND PLANT REGENERATION VIA SUSPENSION CULTURES IN Cucurbitaceae FAMILY

Khidr, Y. A.; Nasr, M. I.

doi:10.21608/ejgc.2012.10550

Cited by 4 publications

(5 citation statements)

References 35 publications

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“…Leaves were detached from10 days old of in vitro grown seedling and incubated with Agrobacteium as described by Khidr and Nasr (2012). Briefly, 50 leaf discs of in vitro grown of 10 Nicotiana bentamiana plants were cocultivated with Agrobacterium for two days at 25 °C on MS medium (Murashige and Skoog 1962) with 0.1 g/L NAA and 1mg/L BAP.…”

Section: Plant Transformation and Regenerationmentioning

confidence: 99%

Determination of Integrated Sequences of Non-T-Dna in Genome of Transgenic Nicotiana Benthamiana Plants

Khidr¹

2018

Journal of Agricultural Chemistry and Biotechnology

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Genetic transformation of plants by useful genes is widely made by Agrobacterium tumifacienes harboring manipulated T-DNA sequences of the binary plasmid. Determining of the integrated T-DNA in the final step of the transformation is critical for understanding the mechanism of this process. The integration of non-T-DNA sequences in transgenic Nicotiana benthamiana plants obtained through Agrobacterium inoculation was investigated. Genomic DNAs from ten plants were subjected to PCR amplification on the sequences located outside the left or right T-DNA borders. Three plants out of ten were found to have 30 base pair of non-T-DNA sequences including the entire left T-DNA border. The same plants were also contained 107 base pair of the non-T-DNA sequences with the entire right border sequence. The percentage of integrated non-T-DNA sequences represented 30 % whether it belongs to sequences followed the right or left T-DNA side. Integrated T-DNA sequences were confirmed by southern blot analysis.

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Section: Plant Transformation and Regenerationmentioning

confidence: 99%

Determination of Integrated Sequences of Non-T-Dna in Genome of Transgenic Nicotiana Benthamiana Plants

Khidr¹

2018

Journal of Agricultural Chemistry and Biotechnology

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“…Plant transformation was performed as described by Khidr and Nasr (2012). In brief, Leaf discs of in vitro grown Nicotiana tabacum cv.…”

Section: Plant Materials and Generation Of Transgenic Plantsmentioning

confidence: 99%

Functional Analysis of Introns for Their Effect on Transgene Expression in Plant

Khidr

Nasr

2018

Journal of Productivity and Development

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Most plant genes contain intervening sequences known as introns that are found in nearly all plant genes and they are transcribed into pre-mRNA, the later will be removed by splicing machinery. Proteins are encoded by exons which separated by intron segments while the noncoding exons are conserved in the mature transcript. In this study, level of spliced and non-spliced introns was measured in transgenic Nicotiana tabacum lines expressing gusint-gfpint fusion gene controlled by the CaMV 35S promoter. Transcription analysis of the transgenes was performed by (RT)-PCR. All transcripts of transgenic lines generated two different length of fragments for gfp and gus genes. The results indicated that the gene expression of transcripts (without intron) in which their intron was spliced out, was higher than those of non-spliced transcript (with intron) in which the intron was maintained i.e. the both introns inside gus and gfp were partially and not completely spliced out of the transcripts. Furthermore, the relative quantifications of band intensity of gfp transcripts revealed that the percentage of non-spliced transcripts compared to the housekeeping gene, elongation factor 1-alpha (ef1α) using different amplification cycles (

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“…Regeneration via organogenesis and somatic embryogenesis has been described in this species, and commonly used explants include cotyledons [26][27][28][29], cotyledonary nodes [26,[30][31][32], hypocotyls [27,[33][34][35], shoot tips [36][37][38][39], and leaves [27,37,40,41]. Regeneration of cucumber has also been reported from protoplasts [42][43][44] and suspension cultures [45][46][47]. Cucumber regeneration is largely dependent on genotype, explant type, and culture medium [24,26,34].…”

Section: Introductionmentioning

confidence: 99%

Influence of High Concentrations of Copper Sulfate on In Vitro Adventitious Organogenesis of Cucumis sativus L.

Miguel

2023

IJPB

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Copper (Cu) is an essential plant micronutrient. This report is the first to assess the effects of high copper sulfate (CuSO4) levels on in vitro callus and shoot formation of cucumber. Four-day-old cotyledon explants from the inbred line Wisconsin 2843 and the commercial cultivars Marketer and Negrito were used. Murashige and Skoog (MS)-derived callus and shoot induction medium containing 0.5 mg L−1 indole-3-acetic acid (IAA) and 2.5 mg L−1 6-benzylaminopurine (BAP) was supplemented with CuSO4 (0.2–5 mg L−1). The response on callus-derived shoots showed that the optimal concentration of CuSO4 was 8- to 200-fold greater than in standard MS medium. Shoot frequency (SF) and shoot number (SN) were assessed, and Marketer > Negrito > Wisconsin 2843, where 1, 0.2 and 5 mg L−1 CuSO4 produced the highest results, respectively. SF and SN increased 6- and 10-fold in Wisconsin 2843 and twice in the other cultivars. All explants formed calluses, and in two of the three cultivars, callus extension was significantly affected by CuSO4 application. SN showed a strong relationship with CuSO4 levels and no association with callus extension. The results show that specific CuSO4 concentrations higher than in standard MS medium increase adventitious cucumber shoot organogenesis.

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IMPROVEMENT OF GENETIC TRANSFORMATION AND PLANT REGENERATION VIA SUSPENSION CULTURES IN Cucurbitaceae FAMILY

Cited by 4 publications

References 35 publications

Determination of Integrated Sequences of Non-T-Dna in Genome of Transgenic Nicotiana Benthamiana Plants

Determination of Integrated Sequences of Non-T-Dna in Genome of Transgenic Nicotiana Benthamiana Plants

Functional Analysis of Introns for Their Effect on Transgene Expression in Plant

Influence of High Concentrations of Copper Sulfate on In Vitro Adventitious Organogenesis of Cucumis sativus L.

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