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

ABSTRACT: 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.

SUBMITTER: Xu C

PROVIDER: S-EPMC6443852 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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

Xu Chen C Wei Hui H Wang Like L Yin Tongming T Zhuge Qiang Q

Frontiers in plant science 20190326

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 t ...[more]

PMID: 30972085

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Project description:BackgroundBaculoviruses act as effective biological control agents against the invasive pest Hyphantria cunea Drury. In this study, two Chinese Hyphantria cunea nucleopolyhedrovirus (HycuNPV) isolates, HycuNPV-BJ and HycuNPV-HB, were deep sequenced and compared with the Japanese isolate, HycuNPV-N9, to determine whole-genome level diversity and evolutionary history.ResultsThe divergence of the phylogenetic tree and the K2P distances based on 38 core-gene concatenated alignment revealed that two Chinese HycuNPV isolates were a novel species of Alphabaculovirus that infected Hyphantria cunea in China. The gene contents indicated significant differences in the HycuNPV genomes between the Chinese and Japanese isolates. The differences included gene deletions, acquisitions and structural transversions, but the main difference was the high number of single nucleotide polymorphisms (SNPs). In total, 10,393 SNPs, corresponding to approximately 8% of the entire HycuNPV-N9 genome sequence, were detected in the aligned reads. By analyzing non-synonymous variants, we found that hotspot mutation-containing genes had mainly unknown functions and most were early expressing genes. We found that the hycu78 gene which had early and late promoter was under positive selection. Biological activity assays revealed that the infectivity of HycuNPV-HB was greater than that of HycuNPV-BJ, and the killing speed of HycuNPV-HB was faster than that of HycuNPV-BJ. A comparison of molecular genetic characteristics indicated that the virulence differences between the two isolates were affected by SNP and structural variants, especially the homologous repeat regions.ConclusionsThe genomes of the two Chinese HycuNPV isolates were characterized, they belonged to a novel species of Alphabaculovirus that infected Hyphantria cunea in China. We inferred that the loss or gain of genetic material in the HycuNPV-HB and HycuNPV-BJ genomes resulted in new important adaptive capabilities to the H. cunea host. These results extend the current understanding of the genetic diversity of HycuNPV and will be useful for improving the applicability of this virus as a biological control agent.

Dataset Information

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

Publications

Optimization of the <i>cry1Ah1</i> Sequence Enhances the Hyper-Resistance of Transgenic Poplars to <i>Hyphantria cunea</i>.

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