Genetic transformation and expression of transgenic lines of Populus x euramericana with insect-resistance and salt-tolerance genes

Yang, Ruochen; Wang, A X; Zhang, J; Dong, Yan; Yang, Minsheng; Wang, J M

doi:10.4238/gmr.15028635

Cited by 17 publications

(20 citation statements)

References 29 publications

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“…A comparison of the insect resistance conferred by different multigene vector structures and transgenic lines is presented in Table 5 . Neither the transgenic strains of Yang et al. (2016) nor those of Liu et al.…”

Section: Discussionmentioning

confidence: 99%

Expression of Multiple Exogenous Insect Resistance and Salt Tolerance Genes in Populus nigra L.

et al. 2020

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Four exogenous genes, Cry3A , Cry1Ac, mtlD, and BADH , were inserted into the p1870 vector to obtain multigenic transgenic Populus nigra L. with improved insect resistance and salt tolerance. During vector construction, different promoters were used for each gene, the AtADH 5′-UTR enhancer was added between the Cry1Ac promoter and the target gene, and the matrix attachment region (MAR, GenBank: U67919.1) structure was added at both ends of the vector. It was then successfully transferred into the genome of European black poplar by Agrobacterium-mediated leaf disk transformation, and a total of 28 transgenic lines were obtained by kanamycin screening. Five events with the highest insect resistance were selected based on preliminary tests: nos. 1, 7, 9, 12, and 17. PCR, real-time PCR, and enzyme-linked immunosorbent assays (ELISA) were used to detect the expression of exogenous genes and to analyze the Bt protein toxin levels in transgenic lines from June to October. PCR results showed that all four genes were successfully introduced into the five selected lines. Fluorescence quantitative PCR showed no significant differences in the transcript abundance of the four exogenous genes between different lines. A Bt protein toxin assay showed that the Cry3A protein toxin content was significantly higher than the Cry1Ac protein toxin content by approximately three orders of magnitude. Levels of the two toxins were negatively correlated. Over the course of the growing season, Cry1Ac content raised and varied between 0.46 and 18.41 ng·g −1 . Cry3A content decreased over the same time period and varied between 2642.75 and 15775.22 ng·g −1 . Indoor insect feeding assay showed that the transgenic lines had high insect resistance, with mortality rates of 1–2-year-old Hyphantria cunea larvae reaching more than 80%, and those of Plagiodera versicolora larvae and nymphs reaching 100%. No. 17 and no. 12 lines had better insect resistance to Lepidoptera and Coleoptera pests. There was no clear improvement in salt tolerance of the transgenic lines, but comprehensive evaluation of 11 salt tolerance indicators showed that lines no. 17 and no. 7 had certain degrees of salt tolerance.

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Section: Discussionmentioning

confidence: 99%

Expression of Multiple Exogenous Insect Resistance and Salt Tolerance Genes in Populus nigra L.

et al. 2020

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“…Although expression of modified Bt genes in plants has improved greatly, with the advent of large-scale planting of transgenic plants the evolution of insect resistance cannot be ignored. To slow this process, previous researchers have used a multi-gene strategy (Yang et al, 2003, 2016; Klocko et al, 2014; Zhang et al, 2016; Ding et al, 2017). Additionally, an H .…”

Section: Discussionmentioning

confidence: 99%

Optimization of the cry1Ah1 Sequence Enhances the Hyper-Resistance of Transgenic Poplars to Hyphantria cunea

Wei

Wang

et al. 2019

Front. Plant Sci.

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Increased expression of the insect control protein genes of Bacillus thuringiensis in Populus has been critical to the development of genetically improved plants with agronomically acceptable levels of insect resistance. Bacillus thuringiensis ( Cry1Ah1 ) proteins with highly specific toxicity against Hyphantria cunea were screened using an indoor bioactivity assay to obtain hyper-resistant transgenic poplars. Then, the Cry1Ah1 sequence was optimized and transformed according to the optimal codon in poplar using software of our own design ( http://120.79.60.226:8080/u/chen/w/codonpoplar ). A vector was constructed to transform poplar NL895. The Cry1Ah1 gene was transformed to poplar NL895 and six transgenic lines were obtained. The expression and insecticidal effect of the Cry1Ah1 gene in transgenic poplar were evaluated by PCR and ELISA, and the specific indoor activity and field insecticidal activity against H . cunea were compared with a control. We concluded that the insecticidal activity of the transgenic NL895 was significantly better against lower instar larvae of H. cunea than against higher instar larvae. The mortality and pupation rates clearly differed among the various instar larvae and between transgenic and non-transgenic poplar. We obtained poplar seedlings with hyper-resistance to H . cunea by screening Bt genes and optimizing their genetic sequence.

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“…The protein encoded by the BADH catalyzes the synthesis of glycinebetaine, an important quaternary ammonium compound produced in response to salt and other osmotic stressors by many organisms (Jia et al, 2002). Multigene plant transformation vectors carrying Cry1Ac + BADH , Cry1Ac + NTHK1 , Cry1Ac + Cry3A + NTHK1 (Du et al, 2014; Liu et al, 2014), and Cry1Ac + Cry3A + BADH (Yang et al, 2012, 2016; Liu et al, 2016) were created at the Hebei Key Laboratory for Tree Genetic Resources and Forest Protection, Hebei province. The genes were successfully transformed into tobacco and various poplars; regenerated plants exhibited both pest-resistance and an increase in salt tolerance compared with controls (Ren et al, 2015).…”

Section: Multigene Transformationmentioning

confidence: 99%

The Current Status and Development of Insect-Resistant Genetically Engineered Poplar in China

et al. 2018

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Poplar is one of the main afforestation tree species in China, and the use of a single, or only a few, clones with low genetic diversity in poplar plantations has led to increasing problems with insect pests. The use of genetic engineering to cultivate insect-resistant poplar varieties has become a hot topic. Over the past 20 years, there have been remarkable achievements in this area. To date, nearly 22 insect-resistant poplar varieties have been created and approved for small-scale field testing, environmental release, or pilot-scale production. Here, we comprehensively review the development of insect-resistant genetically modified (GM) poplars in China. This review mostly addresses issues surrounding the regulation and commercialization of Bt poplar in China, the various insecticidal genes used, the effects of transgenic poplars on insects, toxic protein expression, multigene transformation, the stability of insect resistance, and biosafety. The efficacy of GM poplars for pest control differed among different transgenic poplar clones, larval instars, and insect species. The Bt protein analysis revealed that the expression level of Cry3A was significantly higher than that of Cry1Ac. Temporal and spatial studies of Bt protein showed that its expression varied with the developmental stage and tissue. The inheritance and expression of the exogenous gene were reviewed in transgenic hybrid poplar progeny lines and grafted sections. Biosafety issues, in terms of transgene stability and the effects on soil microorganisms, natural enemies of insects, and arthropod communities are also discussed.

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Genetic transformation and expression of transgenic lines of Populus x euramericana with insect-resistance and salt-tolerance genes

Cited by 17 publications

References 29 publications

Expression of Multiple Exogenous Insect Resistance and Salt Tolerance Genes in Populus nigra L.

Expression of Multiple Exogenous Insect Resistance and Salt Tolerance Genes in Populus nigra L.

Optimization of the cry1Ah1 Sequence Enhances the Hyper-Resistance of Transgenic Poplars to Hyphantria cunea

The Current Status and Development of Insect-Resistant Genetically Engineered Poplar in China

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