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

Wang, Guiying; Dong, Yan; Liu, Xiaojie; Yao, Guosheng; Yu, Xiaoyue; Yang, Minsheng

doi:10.3389/fpls.2018.01408

Cited by 36 publications

(27 citation statements)

References 144 publications

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“…The low strength of the promoters used for the salt-tolerance genes help to explain the poor salt tolerance of the transgenic lines. In the process of translation, the insect-resistant genes and the salt-tolerant genes compete with each other for photosynthate ( Winicov and Seemann, 1990 ), and the salt-tolerance genes may be inhibited due to their inferior position, resulting in poor salt tolerance ( Jiang et al., 2003 ; Si et al., 2007 ; Wang et al., 2018 ). Under salt stress, poplar undergoes a series of physiological and biochemical responses that affect photosynthesis and respiration ( Zhou et al., 2004 ; Zhou, 2006 ; Luo et al., 2017 ).…”

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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“…Transgenes encoding the spider knottin Hv1a (or its close otholog ω‐hexatoxin‐Ar1a) have been engineered into cotton, tobacco (alone or as a fusion to thioredoxin, onion leaf lectin, or Cry1Ac), poplar (fused to a C‐terminal fragment of CryIAb), and Arabidopsis (as a fusion to GNA or luteovirus coat protein). Transgenic plants expressing Hv1a/Ar1a are more resistant to a diverse range of insect pests, including bollworms ( Helicoverpa armigera ) and leafworms ( Spodoptera littoralis ) on cotton, peach‐potato aphids ( Myzus persicae ) on Arabidopsis , gypsy moth ( Lymantria dispar ) on poplar, and peach‐potato aphids, bollworms, leafworms, and whiteflies ( Bemisia tabaci ) on tobacco.…”

Section: Crops With Fangs: Engineering Plants To Express Spider Knottmentioning

confidence: 99%

Tying pest insects in knots: the deployment of spider‐venom‐derived knottins as bioinsecticides

King

2019

Pest Management Science

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Spider venoms are complex chemical arsenals that contain a rich variety of insecticidal toxins. However, the major toxin class in many spider venoms is disulfide‐rich peptides known as knottins. The knotted three‐dimensional fold of these mini‐proteins provides them with exceptional chemical and thermal stability as well as resistance to proteases. In contrast with other bioinsecticides, which are often slow‐acting, spider knottins are fast‐acting neurotoxins. In addition to being potently insecticidal, some knottins have exceptional taxonomic selectivity, being lethal to key agricultural pests but innocuous to vertebrates and beneficial insects such as bees. The intrinsic oral activity of these peptides, combined with the ability of aerosolized knottins to penetrate insect spiracles, has enabled them to be developed commercially as eco‐friendly bioinsecticides. Moreover, it has been demonstrated that spider‐knottin transgenes can be used to engineer faster‐acting entomopathogens and insect‐resistant crops. © 2019 Society of Chemical Industry

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“…Pb29 is the main transgenic poplar 741 line planted commercially. It theoretically carries two insect resistance genes (BtCry1AC and API) that confer high levels of resistance to lepidopteran pests, such as Hyphantria cunea and Clostera anachoreta [2,36,37]. The leaves of poplar 741 and Pb29 were collected, immediately placed into liquid nitrogen, and preserved at 80° for subsequent DNA extraction.…”

Section: Plant Materialsmentioning

confidence: 99%

“…It is one of the important industrial timber species that is widely used in the paper-making industry and panel processing. However, with the continuous increase of poplar planting area, the ensuing insect attack has become more and more serious, which has brought huge losses to forestry production [2]. In order to reduce the economic losses caused by insect pests, decrease the need for chemical pesticides, and protect the ecological environment, the cultivation of insect-resistant transgenic varieties is particularly important [3].…”

Section: Introductionmentioning

confidence: 99%

Whole-Genome Resequencing using Next-Generation and Nanopore Sequencing for Molecular Characterization of T-DNA Integration in Transgenic Poplar 741

Chen

Dong

Huang

et al. 2020

Preprint

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BackgroundWith the rapid development of transgenic technology, transgenic plants have been planted all over the world, and transgenic forest trees have also been commercialized. At the same time, the potential threat of transgenic plants to human health and the natural environment has aroused widespread concern. Therefore, safety assessments before field release and commercial planting of transgenic plants are necessary. By determining the copy number and integration sites of T-DNA in transgenic plants, the safety of transgenic plants at the genomic level can be assessed.ResultsIn this study, we performed genome resequencing of Pb29, a transgenic high-resistance poplar 741 line that has been commercialized, using next-generation and Nanopore sequencing. The results revealed that there are two T-DNA insertion sites, located at 9,283,905–9,283,937 bp on chromosome 3 (Chr03) and 10,868,777–10,868,803 bp on Chr10. The accuracy of the T-DNA insertion locations and directions was verified using polymerase chain reaction amplification. Through sequence alignment, different degrees of base deletions were detected on the T-DNA left and right border sequences, and in the flanking sequences of the insertion sites. An unknown fragment was inserted between the Chr03 insertion site and the right flanking sequence, but the Pb29 genome did not undergo chromosomal rearrangement. It is worth noting that we did not detect the API gene in the Pb29 genome, indicating that Pb29 is a transgenic line containing only the BtCry1AC gene. On Chr03, the insertion of T-DNA disrupted a gene encoding TAF12 protein, but the transcriptional abundance of this gene did not change significantly in the leaves of Pb29. Additionally, except for the gene located closest to the T-DNA integration site, the expression levels of four other neighboring genes did not change significantly in the leaves of Pb29. ConclusionsThis study provides important molecular information for safety assessments and management of transgenic poplar 741. Our findings also provide a theoretical basis for safety assessments of other transgenic poplar.

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

Cited by 36 publications

References 144 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.

Tying pest insects in knots: the deployment of spider‐venom‐derived knottins as bioinsecticides

Whole-Genome Resequencing using Next-Generation and Nanopore Sequencing for Molecular Characterization of T-DNA Integration in Transgenic Poplar 741

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