Integration and Expression Stability of Transgenes in Hybriding Transmission of Transgenic Rice Plants Produced by Particle Bombardment

Zhao, Yan; Guo, Longbiao; Wang, Huizhong; Danian, Huang

doi:10.5376/mpb.2011.02.0008

Cited by 6 publications

(6 citation statements)

References 20 publications

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“…Independent studies have also assessed genetic stability of GMO events not only with different approaches (e.g. cytogenetics and Southern Blot) [10, 11] but also with more rigorous criterions, such as seasonal and tissue-specific ELISA analysis [12] and RNA expression using Northern blot technique [11]. In addition, studies have detected transgene rearrangements, such as sequence deletions [13–15] and nucleotide addition of undesired fragments into the transgene sequence [16], and also, alterations in mRNA expression levels were also observed [17].…”

Section: Introductionmentioning

confidence: 99%

Levels of DNA methylation and transcript accumulation in leaves of transgenic maize varieties

et al. 2016

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BackgroundPrior to their release in the environment, transgenic crops are examined for their health and environmental safety. In addition, transgene expression needs to be consistent in order to express the introduced trait (e.g. insecticidal and/or herbicide tolerance). Moreover, data on expression levels for GM events are usually required for approval, but these are rarely disclosed or they are considered insufficient. On the other hand, biosafety regulators do not consider epigenetic regulation (e.g. DNA methylation, ncRNAs and histone modifications), which are broadly known to affect gene expression, within their risk assessment analyses. Here we report the results of a DNA methylation (bisulfite sequencing) and transgene transcript accumulation (RT-qPCR) analysis of four Bt-expressing single transgenic maize hybrids, under different genetic backgrounds, and a stacked transgenic hybrid expressing both insecticidal and herbicide tolerance traits.ResultsOur results showed differences in cytosine methylation levels in the FMV promoter and cry2Ab2 transgene of the four Bt-expressing hybrid varieties. The comparison between single and stacked hybrids under the same genetic background showed differences in the 35S promoter sequence. The results of transgene transcript accumulation levels showed differences in both cry1A.105 and cry2Ab2 transgenes among the four Bt-expressing hybrid varieties. The comparison between single and stacked hybrids showed difference for the cry2Ab2 transgene only.ConclusionsOverall, our results show differences in DNA methylation patterns in all varieties, as well as in transgene transcript accumulation levels. Although the detection of changes in DNA methylation and transgenic accumulation levels does not present a safety issue per se, it demonstrates the need for additional studies that focus on detecting possible safety implications of such changes.Electronic supplementary materialThe online version of this article (doi:10.1186/s12302-016-0097-2) contains supplementary material, which is available to authorized users.

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Section: Introductionmentioning

confidence: 99%

Levels of DNA methylation and transcript accumulation in leaves of transgenic maize varieties

et al. 2016

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“…On the effect of selection agent on oil palm transformation, minimal inhibitory concentration studies showed that herbicides Basta and hygromycin are the most effective selectable agents as they could kill the untransformed cells at a very low concentration (Parveez et al, 1996;. Basta and hygromycin were also shown to be the effective selection agents for other monocots such as rice (Zhao et al, 2011;Htwe et al, 2014), turfgrass (Song et al, 2013) and finger millet (Ignacimuthu and Ceasar, 2012). Initial regeneration of transgenic oil palm was obtained after transformed oil palm calli were selected on herbicide Basta.…”

Section: Introductionmentioning

confidence: 99%

Regeneration of Transgenic Oil Palm Carrying GFP Gene Used as a Visual Selectable Marker

Majid¹,

Parveez²

2016

JOPR

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“…Various ways to deliver foreign deoxyribonucleic acid (DNA) into the oil palm genome have been evaluated such as bombardment (Parveez et al, 2000;2015a), Agrobacterium-mediated transfer (Izawati et al, 2015;, Polyethylene glycol (PEG)mediated transfection and DNA microinjection (Masani et al, 2014). DNA delivery by bombardment has been widely used and proven in many plants such as soyabean (Khalafalla et al, 2005), rice (Tian et al, 2015;Zhao et al, 2011;Higuchi et al, 2001;Datta et al, 2000), wheat (Melchiorre et al, 2002) and banana (Mahdavi et al, 2014;Sreeraman et al, 2005). However, the transformation efficiency of oil palm by bombardment was low at about 1%-1.5% (Parveez, 2000).…”

Section: Introductionmentioning

confidence: 99%

Short Communication: Optimisation of the Bombardment Parameters for Transformation and Regeneration of Stable Oil Palm Transformants

Rahman¹

2020

JOPR

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Genetic transformation approaches by using particle bombardment have been an integral part in efforts to improve palm oil quality. The bombardment parameters were previously evaluated to efficiently deliver deoxyribonucleic acid (DNA) during bombardment. However, the evaluation was carried out at transient stage and therefore the results may not be very precise. In order to improve the DNA delivery and regeneration of the oil palm carrying transgenes, a study at stable stage was performed and discussed in this paper. Oil palm embryogenic calli were bombarded at several parameters using pAHC25 plasmid. Selection on media containing 1 mg litre-1 of bialaphos was initiated after one month of bombardment. Then, the concentration of bialaphos was increased to 2 mg litre-1 and subsequently to 3 mg litre-1 to effectively eliminate the regeneration of untransformed cells. Generated shoots were subjected to polymerase chain reaction (PCR) analysis. The best bombardment condition was determined based on the highest number of shoots carrying the transgene. The bombardment at 1100 psi helium pressure with 7.5 cm distance between stopping plate to tissue and one time bombardment using 2.0 µg DNA per bombardment was obtained as the best condition for DNA delivery and also minimised tissue damage. Hence, the results of this study will be useful as a guideline to generate transformed oil palm.

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Integration and Expression Stability of Transgenes in Hybriding Transmission of Transgenic Rice Plants Produced by Particle Bombardment

Cited by 6 publications

References 20 publications

Levels of DNA methylation and transcript accumulation in leaves of transgenic maize varieties

Levels of DNA methylation and transcript accumulation in leaves of transgenic maize varieties

Regeneration of Transgenic Oil Palm Carrying GFP Gene Used as a Visual Selectable Marker

Short Communication: Optimisation of the Bombardment Parameters for Transformation and Regeneration of Stable Oil Palm Transformants

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