Molecular characterization and efficacy evaluation of a transgenic corn event for insect resistance and glyphosate tolerance

Liu, Miaomiao; Zhang, Xiaojing; Gao, Yan; Shen, Zhicheng; Lin, Chaoyang

doi:10.1631/jzus.b1700345

Cited by 14 publications

(8 citation statements)

References 26 publications

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“…Comparing the relationship between Cry1C protein content and C. suppressalis mortality rate, the lowest thresholds given in the study can only be used to evaluate the resistance of rice materials to C. suppressalis during the tillering and heading stages observed in this experiment. For example, comprehensive evaluation of the resistance of Bt transgenic rice to C. suppressalis is required, and it is necessary to test the borer resistance throughout the whole growth period of each strain (Ferré & Van Rie, 2002; Liu et al., 2018). When transgenic rice is planted together with nontransgenic rice, the insect resistance of the nontransgenic rice will be improved.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Chilo suppressalis resistance and analysis of CRY1C expression in transgenic rice

et al. 2022

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The insect resistance of Bacillus thuringiensis (Bt) transgenic rice (Oryza sativa L.) is mainly determined by the transcription of the CRY1C gene and the translation process of the Cry1C protein. With this in mind, we analyzed CRY1C expression and Cry1C protein content in a transgenic Bt insect‐resistant restorer line and its F1 hybrids, evaluated the resistance to Chilo suppressalis of various lines of rice, and determined the 50% lethal concentration (LC50) of Cry1C for C. suppressalis. In four transgenic rice lines, the relative expression of CRY1C was highest at the heading stage in most tissues. Among different tissues from the same developmental stage, CRY1C expression was highest in leaves, followed by stems and panicles. The relative expression of CRY1C was higher in the parent restorer line than in the F1 hybrids. The LC50 of the Cry1C protein for C. suppressalis was 4.016 μg g−1, and the Cry1C protein expression level of transgenic insect‐resistant rice exceeded this threshold at the heading stages and in stems. We next analyzed the lethality of each strain toward C. suppressalis. After 48 h, the mortality rate of second‐instar C. suppressalis larvae feeding on transgenic stem tissue was higher at the heading stage than at the tiller stage and varied from 66.7 to 91.7%. By documenting the temporal and spatial expression of CRY1C and evaluating C. suppressalis resistance to Bt transgenic rice, the risk of pests and diseases during the rice production process can be greatly reduced.

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Section: Discussionmentioning

confidence: 99%

Evaluation of Chilo suppressalis resistance and analysis of CRY1C expression in transgenic rice

et al. 2022

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“…Finally, gene editing will provide precision breeding technology to improve desirable characteristics (Shan et al, 2014;Svitashev et al, 2016;Liang et al, 2017). Multiplex editing may be possible soon using the similar technology developed for GMO crop (Liu et al, 2018). This will shorten the breeding cycle by incorporating speed breeding and tissue-culture-free techniques (Maher et al, 2020).…”

Section: Perspectivesmentioning

confidence: 99%

Breeding crops by design for future agriculture

2020

J. Zhejiang Univ. Sci. B

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“…For example, Southern blotting is used to determine the copy number of exogenous genes and to confirm the presence/absence of vector backbone sequences; PCR‐based methods coupled with Sanger sequencing are used to identify exogenous DNA integrated sites and flanking sequences, examples of which include thermal asymmetric interlaced PCR (TAIL‐PCR), ligation‐mediated PCR (LM‐PCR) and inverse PCR (iPCR). To date, molecular characterization of most commercialized GM crop events has been revealed through combinations of these techniques (Liu et al ., 2018 ; Safaei et al ., 2019 ). However, these approaches are time‐consuming and the obtained results are often incomplete (Liang et al ., 2014 ).…”

Section: Introductionmentioning

confidence: 99%

LIFE‐Seq: a universal Large Integrated DNA Fragment Enrichment Sequencing strategy for deciphering the transgene integration of genetically modified organisms

Zhang

Guo

et al. 2022

Plant Biotechnology Journal

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Summary Molecular characterization of genetically modified organisms (GMOs) yields basic information on exogenous DNA integration, including integration sites, entire inserted sequences and structures, flanking sequences and copy number, providing key data for biosafety assessment. However, there are few effective methods for deciphering transgene integration, especially for large DNA fragment integration with complex rearrangement, inversion and tandem repeats. Herein, we developed a universal L arge I ntegrated DNA F ragments E nrichment strategy combined with PacBio Seq uencing (LIFE‐Seq) for deciphering transgene integration in GMOs. Universal tilling DNA probes targeting transgenic elements and exogenous genes facilitate specific enrichment of large inserted DNA fragments associated with transgenes from plant genomes, followed by PacBio sequencing. LIFE‐Seq were evaluated using six GM events and four crop species. Target DNA fragments averaging ~6275 bp were enriched and sequenced, generating ~26 352 high fidelity reads for each sample. Transgene integration structures were determined with high repeatability and sensitivity. Compared with next‐generation whole‐genome sequencing, LIFE‐Seq achieved better data integrity and accuracy, greater universality and lower cost, especially for transgenic crops with complex inserted DNA structures. LIFE‐Seq could be applied in molecular characterization of transgenic crops and animals, and complex DNA structure analysis in genetics research.

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Molecular characterization and efficacy evaluation of a transgenic corn event for insect resistance and glyphosate tolerance

Cited by 14 publications

References 26 publications

Evaluation of Chilo suppressalis resistance and analysis of CRY1C expression in transgenic rice

Evaluation of Chilo suppressalis resistance and analysis of CRY1C expression in transgenic rice

Breeding crops by design for future agriculture

LIFE‐Seq: a universal Large Integrated DNA Fragment Enrichment Sequencing strategy for deciphering the transgene integration of genetically modified organisms

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