Evaluation of the uniformity and stability of T-DNA integration and gene expression in transgenic apple plants

Flachowsky, Henryk; Riedel, Marko; Reim, Stefanie; Hanke, Magda‐Viola

doi:10.2225/vol11-issue1-fulltext-10

Cited by 32 publications

(26 citation statements)

References 22 publications

(29 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Domínguez et al (2004) juga menemukan bahwa tingkat terbentuknya kimera pada proses pembentukan jeruk transgenik dengan bantuan A. tumefaciens sangat tinggi, yaitu 12% dari total tanaman yang dihasilkan. Pembentukan kimera ini juga teramati pada kentang (Rakosy-Tican et al, 2007), apel (Flachowsky et al, 2008), dan anggur (Costa et al, 2009). Fenomena tersebut juga terlihat dalam penelitian ini, walaupun pada pengujian anakan tidak ditemukan perbedaan jumlah salinan T-DNA.…”

Section: Analisis Jumlah Salinan Transgen Beberapa Rumpun Padi Nipponunclassified

Keragaman Jumlah Salinan Transgen Galur T0 Padi Kultivar Nipponbare Berdasarkan Analisis qPCR dengan Penanda Gen hptII

Polosoro¹,

Enggarini²

2018

J. AgroBiogen

View full text Add to dashboard Cite

The development of transgenic crop using Agrobacterium tumefaciens produces different inserted transgenes, whether copy numbers or location in the plant genome. The research was performed to detect chimeric phenomena based on the transgene quantity analysis in tillers of a clump and of some clumps which were derived from the same calli. CsNitr1-L gene and hptII gene as a marker on a binary plasmid pCAMBIA1300 was transformed into the Nipponbare rice genome using A. tumefaciens strain LBA 4404. Molecular analysis was carried out on three tillers of each four clumps of Nipponbare transgenic T 0 generation (events number 1, 2, 3, and 4) and four groups of T 0 clump derived from one callus. Three T 0 clump samples were collected from each of the four groups of T 0 clumps. The results of qPCR analysis showed that the transgene copy numbers of tillers which were derived from one T 0 clump were the same. qPCR analysis also discovered that not all plants from one callus demonstrated the same transgene copy numbers. This implies that each T 0 rice clump which grows from the transformed calli was needed to be split in the acclimatization step so that the uniform T 1 seeds would be obtained. Keywords:Copy number, qPCR, Agrobacterium tumefaciens, Nipponbare. ABSTRAKPerakitan tanaman transgenik dengan bantuan Agrobacterium tumefaciens menghasilkan penyisipan transgen yang berbeda, baik dalam jumlah salinan maupun letak transgen dalam genom tanaman. Penelitian ini bertujuan mendeteksi keberadaan kimera berdasarkan analisis jumlah salinan transgen pada tiap anakan dalam satu rumpun dan beberapa rumpun padi Nipponbare transforman T 0 yang berasal dari kalus yang sama. Gen CsNitr1-L dan hptII sebagai penanda pada plasmid biner pCAMBIA1300 ditranformasikan ke dalam genom tanaman padi kultivar Nipponbare dengan bantuan A. tumefaciens strain LBA 4404. Analisis molekuler dilakukan terhadap tiga anakan masing-masing dari empat rumpun tanaman Nipponbare transgenik generasi T 0 (event 1, 2, 3, dan 4) dan empat kelompok rumpun tanaman T 0 yang berasal dari kalus yang sama. Dari setiap kelompok tersebut diambil tiga rumpun T 0 . Hasil analisis qPCR menunjukkan bahwa anakan yang berasal dari rumpun tanaman T 0 yang sama memiliki jumlah salinan transgen yang seragam. Selain itu, hasil analisis qPCR juga menunjukkan bahwa tidak semua tanaman yang berasal dari kalus yang sama memiliki jumlah salinan transgen yang seragam. Implikasi hasil penelitian ini adalah setiap rumpun padi T 0 yang tumbuh dari kalus hasil transformasi perlu dipisahkan saat aklimatisasi agar benih T 1 yang dihasilkan seragam.

show abstract

Section: Analisis Jumlah Salinan Transgen Beberapa Rumpun Padi Nipponunclassified

Keragaman Jumlah Salinan Transgen Galur T0 Padi Kultivar Nipponbare Berdasarkan Analisis qPCR dengan Penanda Gen hptII

Polosoro¹,

Enggarini²

2018

J. AgroBiogen

View full text Add to dashboard Cite

show abstract

“…In the rest, four plants did not express the antibiotic marker gene, one had lost its promoter and in other three, the promoter was silenced (not functional). In a second experiment, 26 lines carrying the attacin E gene from Hyalophora cecropia, the -glucuronidase (gus) gene and the nptII gene were propagated vegetatively in vitro without selective agents for 4 years (50 generations) and then analysed [87]. Neither expression nor integration remained stable in some lines, differences were found between plants of a single line and several plants were chimaeras of expressing and non-expressing cell clones.…”

Section: Transgene Instability and The Illegality Of Gmosmentioning

confidence: 99%

The New Genetics and Natural versus Artificial Genetic Modification

Ho¹

2013

Entropy

View full text Add to dashboard Cite

Abstract:The original rationale and impetus for artificial genetic modification was the "central dogma" of molecular biology that assumed DNA carries all the instructions for making an organism, which are transmitted via RNA to protein to biological function in linear causal chains. This is contrary to the reality of the "fluid genome" that has emerged since the mid-1970s. In order to survive, the organism needs to engage in natural genetic modification in real time, an exquisitely precise molecular dance of life with RNA and DNA responding to and participating in "downstream" biological functions. Artificial genetic modification, in contrast, is crude, imprecise, and interferes with the natural process. It drives natural systems towards maximum biosemiotic entropy as the perturbations are propagated and amplified through the complex cascades of interactions between subsystems that are essential for health and longevity.

show abstract

“…The routine generation of transgenic plants involves analysis of transgene integration into the host genome as well as the evaluation of the transgene expression using standard molecular techniques such as PCR, RT-PCR among others (Flachowsky et al, 2008). The presence of the transgene in selected putative transgenic lines is presented in Figure 3, and the agarose gel electrophoresis of RT-PCR to show the expression of the transgene (AtAOX1a) at the transcript level is presented in Figure 4.…”

Section: Screening Of Transgenic Linesmentioning

confidence: 99%

Transgenic cassava lines carrying heterologous alternative oxidase (AtAOX1a) showed impaired quantitative and qualitative response to embryogenesis

Afuape¹,

S²,

Sayre³

et al. 2013

Afr. J. Biotechnol.

View full text Add to dashboard Cite

In our approach to control reactive oxygen species produced as a result of oxidative stress experienced by cassava roots during harvesting, a phenomena which causes postharvest physiological deterioration in the roots, we transformed cassava variety TMS 60444 with the AtAOX1a gene, driven by the 35S promoter, using agrobacterium-mediated approach. Extracted genomic DNAs of putative transgenic lines were screened using polymerase chain reaction technique (PCR). Messenger RNA was extracted from selected PCR-positive lines for reverse transcription-PCR analysis for gene expression. To screen positive lines for gene function, leaf lobes from two transgenic lines with a line carrying an empty vector and the wild type were subjected to somatic embryogenesis (SE), a known oxidative stress process. The results show that the wild type, at 16 days after initiation (DAI) of the leaf lobes on callus initiation medium, had the highest (100%) number of leaf lobes that produced at least one observed organised embryogenic structure (OES). This was followed by PEV-3, the empty vector plant with 50% OES production, while PB-3 had the least percent (20%) of leaf lobes with OES. PB-3 line also had no OES at all in five out of the seven periods of data collection. During the period, the wild type recorded the highest attainable OES quality score of 2.0 (on a scale of 1-5 where 1=bad and 5=excellent) at the first initiation cycle. Both the transgenic lines and the empty-vector plantlet recorded quality score of 1.0. It seems AtAOX1a only hinders OES development, but exerts little effect on the quality, if OES does not degenerate after development.

show abstract

Evaluation of the uniformity and stability of T-DNA integration and gene expression in transgenic apple plants

Cited by 32 publications

References 22 publications

Keragaman Jumlah Salinan Transgen Galur T0 Padi Kultivar Nipponbare Berdasarkan Analisis qPCR dengan Penanda Gen hptII

Keragaman Jumlah Salinan Transgen Galur T0 Padi Kultivar Nipponbare Berdasarkan Analisis qPCR dengan Penanda Gen hptII

The New Genetics and Natural versus Artificial Genetic Modification

Transgenic cassava lines carrying heterologous alternative oxidase (AtAOX1a) showed impaired quantitative and qualitative response to embryogenesis

Contact Info

Product

Resources

About